ANNUAL REPORT EDUCATION ACTIVITIES 339 STAFF 341

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2 PREFACE ANNUAL REPORT MARIO NEGRI INSTITUTE, MILAN DEPARTMENTS Department of Oncology.. 7 Department of Environmental Health Sciences Department of Neuroscience.. 77 Department of Cardiovascular Research Department of Molecular Biochemistry and Pharmacology Department of Epidemiology Department of Public Health LABORATORIES AND CENTERS Laboratory of Regulatory Policies Centre of Computer Science Engineering. 247 Italian Cochrane Center The Catullo and Daniela Borgomainerio Center 259 Library NEGRI BERGAMO LABORATORIES DEPARTMENTS Department of Molecular Medicine Department of Biomedical Engineering. 285 Laboratory of Biology and Therapy of Metastasis ALDO and CELE DACCO CENTER DEPARTMENT Department of Renal Medicine. 305 LABORATORIES AND CENTERS Laboratory of Coordination of Diagnosis and Information on Rare Diseases The Transplant Research Center. 337 EDUCATION ACTIVITIES 339 STAFF 341 All the staff of the Institute is listed on its website PUBLICATIONS A comprehensive list of the Institute s publications is available on the website Section Publications 1

3 Edited by Isabella Bordogna printed April

4 PREFACE This booklet provides a brief description of the research and training work done at the Mario Negri Institutes in Milan and Bergamo. It is divided by departments, and in some cases by single laboratories. Details of the results are provided in the text itself, so here we shall just draw a few general remarks. This is our first year in the new Milan Headquarters: during this time we devoted most of our efforts improving the new available technologies. Particularly, we have carried out translational research following the mouse clinic patterns. Nuclear magnetic resonance, micro cat scanner, ultrasound, Doppler, photon microscope are some of the methodologies used to do research on human diseases in mice, following the techniques utilized in medical clinic. This will allow to transfer data in more effective ways, to study more in depth and to use a lower number of animals in experimental research. Imaging will play an important role, thanks to the installation of electronic microscopy, of contrast, time- lapse microscopy and atomic force microscopy. New important technologies will also allow us to develop our proteomics research. The tendency nowadays is towards widespread use of molecular biology techniques, especially for studying the mechanism of action of drugs. In vitro studies are an essential basis for thorough investigations although a significant number of in vivo experiments are still needed as this is the only mean we have of validating in vitro findings and establishing models that resemble human diseases as closely as possible. This has led to a substantial increase in the use of transgenic animals. The Institute is still concentrating on its traditional research lines: oncology, neurosciences, cardiovascular and renal diseases, organ transplants with strong cell biology and molecular biochemistry connotations. Significant work has been carried out on the environment and health as a whole. Experimental, clinical and epidemiological research on rare diseases and orphan drugs is growing all the time. The Mario Negri Institute strives to develop a multifaceted approach to all these research themes, ranging from basic research, to pharmacokinetics, pharmacology, controlled clinical trials, epidemiological analysis and whenever possible the epidemiology of services. So far we have published more than articles on peer reviewed scientific journals. If research is to continue young scientists must be continually trained. Working in the laboratory not only gives them an outlet for their ideas, but enables them to obtain worthwhile qualifications by taking part in the Institute s training schemes, which are recognised by the Lombardy Region in Italy, or by working for a Ph.D. awarded by the Open University, UK. Training courses are also available on biomedical statistics, for general practitioners, family pediatricians, and clinical trial nurses. A vital part of the Institute s work involves providing information, at all levels. This is done, in particular, through the Rare Diseases Information Center, the Center for Information on Medicinal Drugs and the Website ( We also provide information to medical doctors, nurses, patients associations and lay people, through the media and through a recently developed website: Because research is going through a very difficult time it is vital to be supported by the Government, by private and public bodies as well as by foundations and private citizens. Silvio Garattini Director 3

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6 Mario Negri INSTITUTE FOR PHARMACOLOGICAL RESEARCH Milan ANNUAL REPORT 2008 departments and laboratories 5

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8 DEPARTMENT OF ONCOLOGY STAFF Chief Maurizio D INCALCI, M.D. Oncological Studies Office and Documentation Scientific Documentalist Stefania FILIPPESCHI, Chemist Laboratory of Cancer Pharmacology Head Biophysics Unit Head Flow Citometry Unit Head Cancer Clinical Pharmacology Unit Head Maurizio D INCALCI, M.D. Paolo UBEZIO, Phys.D. Eugenio ERBA, Biochem.D Massimo ZUCCHETTI, Chem.Pharm.D. Laboratory of Molecular Pharmacology Head DNA Repair Unit Head Massimo BROGGINI, Ph.D. Giovanna DAMIA, M.D. Laboratory of Biology and Therapy of Metastasis Head Tumor Angiogenesis Unit Head Unit located in Bergamo Molecular Cancer Therapeutics Unit Head Raffaella GIAVAZZI, Biol.Sci.D., Ph.D. Giulia TARABOLETTI, Biol.Sci.D. Maria Rosa BANI, Biol.Sci.D., Ph.D. 7

9 Laboratory for the development of new pharmacological strategies Head Valter TORRI, M.D. Laboratory of Clinical Trials Head Irene FLORIANI, Dr.Sci.Biol., Dr.Stat., Ph.D. Laboratory of Translational and Outcome Research in Oncology Head Gynecology Oncology Unit Head Giovanni APOLONE, M.D. Roldano FOSSATI, M.D. CERP: Center for the Evaluation and Research on Pain Head Giovanni APOLONE, M.D. Oscar CORLI, M.D. Laboratory of Medical Research and Consumer Involvement Head Paola MOSCONI, Biol.Sci.D. 8

10 CURRICULA VITAE Maurizio D'Incalci obtained his Medical Degree cum Laude from the University of Milan in After specializing in Pharmacology at the Mario Negri Institute of Milan and in Oncology at the University of Genoa, he worked in the Laboratory of Molecular Pharmacology of the National Cancer Institute in Bethesda, MD, USA. Since 1986 he has been chief of the Laboratory of Cancer Chemotherapy at the Mario Negri Institute and since 1996 he has become chief of the Department of Oncology at the Mario Negri Institute. He has been President of the Pharmacology and Molecular Mechanisms Group of the European Organization for Research and Treatment of Cancer (EORTC). From 1994 to 1997 he was Chairman of the New Drug Development Coordinating Committee and from 1997 to 2000 he was chairman of the Research Division of the EORTC. He has been member of the Board of the EORTC from April 2000 to From 1997 he is the Preclinical Coordinator of the Southern Europe New Drug Organization (SENDO) and since 2006 the Chairman of the New Agents Committee (NAC). He is on the editorial board of many international cancer-related scientific journals and since September 2000 he is Editor for Experimental Oncology of the European Journal of Cancer. Dr D'Incalci is author of more than 440 papers on cancer chemotherapy published in peer reviewed international journals, and of several chapters in books on cancer chemotherapy. Selected publications Grosso F., Jones R.L. Demetri G.D., Judson I.R., Blay J.Y., Le Cesne A., Sanfilippo R., Casieri P., Collini P., Dileo P., Spreafico C., Stacchiotti S., Tamborini E., Tercero J.C., Jimeno J., D Incalci M., Gronchi A., Fletcher J.A., Pilotti S., Casali P.G. Efficacy of Trabectedin (ET-743) in advanced pre-treated myxoid liposarcomas. Lancet Oncology, 2007; 8: Salvati E., Leonetti C., Rizzo A., Scarsella M., Mottolese M., Galati R., Sperduti I., Stevens M., D Incalci M., Blasco M., Chiorino G., Horard B., Gilson E., Zupi G., Biroccio A. Telomere damage promotes antitumoral activity of the G- quadruplex ligand RHPS4. J. Clin. Invest., 2007; 117: Zongaro S., de Stanchina E., Colombo T., D Incalci M., Giulotto E., Mondello C. Stepwise neoplastic transformation of a telomerase immortalized fibroblast cell line. Cancer Research, 2005; 65:(24): Allavena P., Signorelli M., Chieppa M., Erba E., Bianchi G., Marchesi F., Garbi A., Lissoni A., de Braud F., Jimeno J. and D Incalci M. Anti-inflammatory properties of the novel antitumor agent Yondelis (Trabectedin): inhibition of macrophage differentiation and cytokine production. Cancer Res., 2005; 65(7): Tavecchio M., Natoli C., Ubezio P., Erba E., D Incalci M. Dynamics of cell cycle phase perturbations by Trabectedin (ET-743) in nucleotide excision repair (NER)-deficient and NER-proficient cells, unravelled by a novel mathematical simulation approach. Cell proliferation, 2007; 40: Marchini S, Mariani P, Chiorino G, Marrazzo E, Bonomi R, Fruscio R, Clivio L, Garbi A, Torri V, Cinquini M, Dell'Anna T, Apolone G, Broggini M, D'Incalci M Analysis of gene expression in early-stage ovarian cancer Clin Cancer Res : Giovanni Apolone, got his Medical degree in 1982 (Pavia, Italy) and his post-doctoral specializations in Internal Medicine in 1987 (Pavia, Italy) and Pharmacological Research (1992). He is Head of the Laboratory of Translational and Outcome Research. He is also Vice-President of the Ethics Committee of the European Institute of Oncology in Milan (Italy) and listed as National Expert at the European Agency for the Evaluation of Medicinal products (EMEA) in London (UK). His main fields of interest are: Methodological, ethical and regulatory aspects of clinical research, with special emphasis on oncology and the cancer pain. Health care evaluation with special emphasis on oncology; Development and validation of case-mix and patient-reported outcome measures; Education and health promotion research and programs. He has authored or co-authored over 200 publications. Selected publications Trotta F, Apolone G, Garattini S, Tafuri G Stopping a trial early in oncology: for patients or for industry?ann Oncol :

11 Apolone G, Corli O, Greco M T, Zagonel V, CPOR SG Investigators Factors influencing the decision to take or reject opioids for cancer pain: are we on target? Ann Oncol : Deandrea S, Montanari M, Moja L, Apolone G Prevalence of undertreatment in cancer pain. A review of published literature Ann Oncol : Apolone G, Tafuri G, Trotta F, Garattini S A new anti-cancer drug in the market: good news for investors or for patients? Eur J Cancer : Apolone G, Patarnello F The value of a drug: From innovation to the payment via Karl MarxJ Ambul Care Manage : Massimo Broggini followed the faculty of Science of the University of Milan, got the specialization in Biochemistry at Mario Negri Institute, and the PhD degree at the Open University, London,UK. He worked for a period in the laboratory of Molecular Pharmacology of the National Cancer Institute of Bethesda, Md, in From 1991 is the head of the Molecular Pharmacology Unit of the Mario Negri Institute and from 1999 of the Laboratory of Molecular Pharmacology of the same Institute. His main fields of interest are the study of the mechanism of action of new anticancer agents, the search of proteins and genes altered in human cancer and the study of oncosuppressor genes. He is member of the "Pharmacology and Molecular Mechanisms Group" of the European Organisation for the Research and Treatment of Cancer (EORTC) and of the American Association for Cancer Research. He is in the Editorial board of the European Journal of Cancer. He is author of more than 100 articles published in international journals. Selected publications Zangrossi S, Marabese M, Broggini M, Giordano R, D'Erasmo M, Montelatici E, Intini D, Neri A, Pesce M, Rebulla P, Lazzari L. Oct-4 expression in adult human differentiated cells challenges its role as a pure stem cell marker. Stem Cells. 2007;25(7): Marrazzo E, Marchini S, Previdi S, Broggini M. Questioning the oncogenic role of DeltaNp73alpha in different cell lines expressing p53 or not. Cancer Biol Ther. 2006;5(7): Polato F, Codegoni A, Fruscio R, Perego P, Mangioni C, Saha S, Bardelli A, Broggini M. PRL-3 phosphatase is implicated in ovarian cancer growth. Clin Cancer Res : Maffucci T, Piccolo E, Cumashi A, Iezzi M, Riley AM, Saiardi A, Godage HY,Rossi C, Broggini M, Iacobelli S, Potter BV, Innocenti P, Falasca M. Inhibition of the phosphatidylinositol 3-kinase/Akt pathway by inositol pentakisphosphate results in antiangiogenic and antitumor effects. Cancer Res. 2005;65: Marabese M, Marchini S, Sabatino MA, Polato F, Vikhanskaya F, Marrazzo E, Riccardi E, Scanziani E, Broggini M. Effects of inducible overexpression of DNp73alpha on cancer cell growth and response to treatment in vitro and in vivo. Cell Death Differ. 2005;12: Sabatino MA, Colombo T, Geroni C, Marchini S, Broggini M. Enhancement of in vivo antitumor activity of classical anticancer agents by combination with the new, glutathione-interacting DNA minor groove-binder, brostallicin. Clin Cancer Res. 2003;9: Irene Floriani got her degree in Biological Sciences at the University of Milan in 1988, her degree in Biostatistics and Experimental Statistics at the University of Milan in 2003 and her phd in Life Sciences at Open University of London (UK) in After ten-year experience in pharmaceutical industry, in 2002 she became Head of the Biometry and Data Management Unit of the Laboratory of Clinical Research in Oncology and since 2006 she is Head of Laboratory of Clinical Trials. She is also member as bio-statistician of three Italian Ethics Committees. Her main fields of interest are: statistical aspects of methodology of clinical research with focus on Controlled Clinical Trials in Oncology; Systematic Overview of the medical literature and Methodological aspects of diagnostic test evaluation. Selected publications Graziano F, Ruzzo A, Loupakis F, Canestrari E, Santini D, Catalano V, Bisonni R, Torresi U, Floriani I, Schiavon G, Andreuzzi B, Maltese P, Rulli E, Humar B, Falcone A, Giustini L, Tonini G, Fontana A, Masi Gianluca, Magnani M Pharmacogenetic profiling for cetuximab/irinotecan therapy in patients with refractory advanced colorectal cancer. J Clin Oncol 2008 ; 26 : Ludovini V, Gori S, Colozza M, Pistola L, Rulli E, Floriani I, Pacifico E, Tofanetti F R, Sidoni A, Basurto C, Rulli A, Crino' L. Evaluation of serum HER2 extracellular domain in early breast cancer patients: correlation with clinicopathological parameters and survival. Ann Oncol 2008 ; 19 : Di Costanzo F, Gasperoni S, Manzione L, Bisagni G, Labianca R, Bravi S, Cortesi E, Carlini P, Bracci R, Tomao S, Messerini L, Arcangeli F, Torri V, Bilancia D, Floriani I, Tonato M, Italian Oncology Group for Cancer Research. Adjuvant chemotherapy in completely resected gastric cancer: a randomized phase III trial conducted by GOIRC. J Natl Cancer Inst 2008 ; 100 : Cascinu S, Berardi R, Labianca R, Siena S, Falcone A, Aitini E, Barni S, Di Costanzo F, Dapretto E, Tonini G, Pierantoni C, Artale S, Rota S, Floriani I, Scartozzi M, Zaniboni A, GISCAD. Cetuximab plus gemcitabine and cisplatin 10

12 compared with gemcitabine and cisplatin alone in patients with advanced pancreatic cancer: a randomised, multicentre, phase II trial. Lancet Oncol 2008 ; 9 : Santini D, Loupakis F, Vincenzi B, Floriani I, Stasi I, Canestrari E, Rulli E, Maltese P, Andreoni F, Masi Gianluca, Graziano F, Baldi G G, Salvatore L, Russo A, Perrone G, Tommasino M R, Magnani M, Falcone A, Tonini G, Ruzzo A. High concordance of KRAS status between primary colorectal tumors and related metastatic sites: implications for clinical practice. Oncologist 2008 ; 13 : Cascinu S, Labianca R, Barone C, Santoro A, Carnaghi C, Cassano A, Beretta GD, Catalano V, Bertetto O, Barni S, Frontini L, Aitini E, Rota S, Torri V, Floriani I. Adjuvant treatment of high-risk, radically resected gastric cancer patients with 5-fluorouracil, leucovorin, cisplatin, and epidoxorubicin in a randomized controlled trial. J Natl Cancer Institute (2007); 99: Raffaella Giavazzi got her Biological Sciences degree (1979) at the University of Milan, and her Ph.D. in Pharmacology at the Mario Negri Institute of Milan (1984), followed by a specialization in pharmacology (1994) at the University of Milan. From 1981 to 1983 she was a Fellow in the Cancer Metastasis and Treatment Laboratory, NCI-FCRDC, Frederick, MD., and from 1983 to 1985 Assistant Professor at the Department of Cell Biology of M.D. Anderson Hospital and Tumour Institute, University of Texas System Cancer Centre in Houston (TX). Raffaella Giavazzi s research interests are in the field of tumour biology and pharmacology. Specifically, she is studying aspects related to the metastatic process and angiogenesis. She is involved in the pre-clinical evaluation of new therapeutic strategies against cancer focusing on the angiogenesis inhibitors and combination therapies. From 1986 to 1993 she was Head of the Cancer Metastasis Treatment Unit and since 1993 she has been the Head of the Laboratory of Biology and Treatment of Metastasis at Mario Negri Institute for Pharmacological Research. She is also adjutant Professor in Oncology, Medical School-University of Brescia, member of the Teaching Committee for the PhD course in Physiology-Pharmacology-Molecular and Cellular Tossicology-University of Siena, member of the Executive Committee at SENDO (South Europe New Drug Development Organization), member of the Pezcoller Foundation Scientific Committee and member of the Executive Committee of the European Association for Cancer Research (EACR). She was consulting scientist for the NCI-Drug Therapeutics Program, USA ( ); She is a member of the American Association for Cancer Research (AACR), International Metastases Research Society, EORTC-Screening and Pharmacology Group, European Association for Cancer Research (EACR), Italian Cancer Society (SIC) of which she was President ( ). She is on the Editorial Board of the European Journal of Cancer, Journal of Clinical Experimental Metastasis, Journal Exp. Therapeutic & Oncology, The International Journal of Biological Markers, Current Cancer Therapy Reviews and Journal of Chemotherapy. She is on the Editorial Board of several international scientific journals. She has published approximately 200 articles on peer reviewed scientific journals. Selected publications Belotti D., Calcagno C., Garofalo A., Caronia D., Riccardi E., Giavazzi R., Taraboletti G. Vascular Endothelial Growth Factor stimulates organ-specific host matrix metalloproteinase-9 expression and ovarian cancer invasion. Molecular Cancer Research, 6(4):525-34, Valentini G., D Andrea C., Ferrari F., Pifferi A., Cobeddu R., Martinelli M., Natoli C., Ubezio P., Giavazzi R. In-vivo measurement of vascular modulation in experimental tumors using a fluorescent contrast agent. Photochemistry and Photobiology, 84(5): , Ghilardi C., Chiorino G., Dossi R., Nagy Z., Giavazzi R., Bani M.R. Identification of novel vascular markers through gene expression profiling of tumor-derived endothelium. BMC Genomics, 30(9), 201, Martinelli M., Bonezzi K., Riccardi E., Kuhn E., Frapolli R., Zucchetti M., Ryan A.J., Taraboletti G., Giavazzi R.: Sequence dependent antitumour efficacy of the vascular disrupting agent ZD6126 in combination with paclitaxel. British Journal of Cancer 97:888-94, Giavazzi R., Bani M.R.,Taraboletti G.: Tumor host interaction in the optimization of paclitaxel-based combination therapies with vascular targeting compounds. Cancer Metastasis Rev. 26:481 88, 2007 Naumova E., Ubezio P., Garofalo A., Borsotti P., Cassis L., Riccardi E., Scanziani E., Eccles S.A., Bani M.R., Giavazzi R.: The vascular targeting property of paclitaxel is enhanced by SU6668, a receptor tyrosine kinase inhibitor, causing apoptosis of endothelial cells and inhibition of angiogenesis. Clin. Cancer Research 12(6): , Paola Mosconi got his Biological Science degree (Milan 1982) and the specialisation in Pharmacological Research (Milan 1984). Her main areas of interest are: development of strategies to involve patients-consumer associations in health debate, and research projects; assessment of quality of life, translation and cultural adaptation of questionnaires for quality of life; 11

13 studies to evaluate the type of information on diseases and treatments received by patients, mainly in cancer patients; set-up of websites targeted on consumers/patients Mattioli.it ; studies to evaluate the consumers level of satisfaction with the health services and cure. Paola Mosconi has participated as a teacher, or coordinator, to the realization of training course on Methodological aspects of clinical research or Evaluation of quality of life for health care professionals and representatives of voluntary associations. Selected publications Lucca U, Tettamanti M, Mosconi P, Apolone G, Gandini F, Nobili A, Tallone MV, Detoma P, Giacomin A, Clerico M, Tempia P, Guala A, Fasolo G, Riva E. Association of mild anemia with cognitive, functional, mood and quality of life outcomes in the elderly: the health and anemia study. PlosONE April 2008, Vol 3, Issue 4, e1920: 1-8 Mosconi P, Apolone G, Mingardi G. Quality of life assessment and instruments in end-stage renal disease. Journal of Nephrology 2008; 21(suppl 13): S107-S112 Mosconi P, Colombo C, Guella F, Pierotti B, Vimercati F. Are Italian medical societies bridging the distance from citizen and patients associations? Result of a survey. Journal of Preventive Medicine and Hygiene 2008: 3 Mosconi P, Colombo C, Satolli R, Liberati A. PartecipaSalute, an Italian project to involve lay people, patients associations and scientific-medical representatives on the health debate. Health Expectations 10: , Mosconi P, Satolli R, Colombo Cinzia, Liberati A, Donati S, Mele A. Hormone replacement therapy and information: in Italy a Consensus Conference to help woman decision. British Medical Journal, Valter Torri got his Medical degree in 1985 and the specialization in medical Oncology in 1989 at the University of Milano. Education: 1985: MD Degree with full honors cum Laude, University of Milano; 1988 Post-Doctoral Degree in Pharmacological Research, Mario Negri Institute, Milano; 1989 Post-Doctoral Degree in Medical Oncology, University of Milano; Research Fellow at the Biometric Research Branch of Cancer Treatment Evaluation Program, NCI, Bethesda, MD (USA) Areas of Interest: Statistical aspects of clinical research methodology with focus on Controlled Clinical Trials in Oncology; Systematic Overview of the medical literature; Methodological aspects of diagnostic test evaluation. Present Position: Head of Laboratory of Clinical Research In Oncology, Oncology Department, Mario Negri Institute, Milano. Chronology of Professional Appointments: : Clinical research Fellow in Internal Medicine at the University Hospital, University of Milan; : Research assistant at the Clinical Trial Unit of the Laboratory of Clinical Epidemiology, Mario Negri Institute for Pharmacological Research, Milano; : Research fellow at the Biometric Research Branch of Cancer Treatment Evaluation Program, NCI, Bethesda, MD (USA); 1994: Head of Biometric Unit of the Laboratory of Cancer Clinical Epidemiology, Oncology Department, Mario Negri Institute for Pharmacological Research, Milano, Italy; 1995 Vice Director of the Italian Cochrane Center; 2001: Head of Laboratory of Clinical Research In Oncology, Oncology Department, Mario Negri Institute, Milano. 2006: Head of Laboratory for the development of new pharmacological strategies, Oncology Department, Mario Negri Institute, Milano.. Selected publications Marchini S, Mariani P, Chiorino G, Marrazzo E, Bonomi R, Fruscio R, Clivio L, Garbi A, Torri V, Cinquini M, Dell'Anna T, Apolone G, Broggini M, D'Incalci M. Analysis of gene expression in early-stage ovarian cancer. Clin Cancer Res Dec 1;14(23): Benedetti Panici P, Basile S, Maneschi F, Alberto Lissoni A, Signorelli M,Scambia G, Angioli R, Tateo S, Mangili G, Katsaros D, Garozzo G, Campagnutta E,Donadello N, Greggi S, Melpignano M, Raspagliesi F, Ragni N, Cormio G, Grassi R, Franchi M, Giannarelli D, Fossati R, Torri V, Amoroso M, Crocè C, Mangioni C. Systematic pelvic lymphadenectomy vs. no lymphadenectomy in early-stage endometrial carcinoma: randomized clinical trial. J Natl Cancer Inst Dec 3;100(23): Mitry E, Fields AL, Bleiberg H, Labianca R, Portier G, Tu D, Nitti D, Torri V,Elias D, O'Callaghan C, Langer B, Martignoni G, Bouché O, Lazorthes F, Van Cutsem E, Bedenne L, Moore MJ, Rougier P. Adjuvant chemotherapy after potentially curative resection of metastases from colorectal cancer: a pooled analysis of two randomized trials. J Clin Oncol Oct 20;26(30): Cascinu S, Labianca R, Barone C, Santoro A, Carnaghi C, Cassano A, Beretta GD, Catalano V, Bertetto O, Barni S, Frontini L, Aitini E, Rota S, Torri V, FlorianiI; Italian Group for the Study of Digestive Tract Cancer, Adjuvant treatment of high-risk, radically resected gastric cancer patients with 5-fluorouracil, leucovorin, cisplatin, and epidoxorubicin in a randomised controlled trial. J Natl Cancer Inst. 2007;99(8):

14 Graziano F, Kawakami K, Ruzzo A, Watanabe G, Santini D, Pizzagalli F, Bisonni R, Mari D, Floriani I, Catalano V, Silva R, Tonini G, Torri V, Giustini L, Magnani M Methylenetetrahydrofolate reductase 677C/T gene polymorphism, gastric cancer susceptibility and genomic DNA hypomethylation in an at-risk Italian population. Int J Cancer : Torri V: Clinical trials and data management In: Oxford textbook of oncology, 2nd. ed. Vol. 1. Oxford Univ. Press, Oxford; 2002 : Maria Rosa Bani got her Biological Sciences degree at the University of Milan in 1998 attaining the Italian Government Qualification to practice as Biologist in She obtained the specialization in Pharmacological Research from the Department of Education of the Regional Government of Lombardia in 1991 and the specialization in Biomedical Research from the Department of Education of the Regional Government of Abruzzo in In 2005 she was awarded the degree of Doctor of Philosophy (PhD), Discipline of Life Sciences of the Open University Research School (UK). From 1991 to 1995 she was a Post Doctoral Fellow in the Cancer Research Division, Sunnybrook Health Science Centre, University of Toronto (Canada); from 2000 to 2001 she was Guest Scientist at the Advance Technology Centre, National Cancer Institute, National Institute of Health (USA). At the MarioNegri Institute for Pharmacological Research she was a Fellow Research Scientist in the Laboratory of Biology and Treatment of Metastasis, Bergamo from 1996 to 2002 and she became a Staff Research Scientist in In April 2004 she became Head of the Molecular Cancer Therapeutics Unit. She is the Scientific Manager of STROMA(since 2004) and ADAMANT(since2008), two Integrated Projects within the 6 th and 7 th Framework Programs of the European Commission. She is a member of the American Association for Cancer Research (AACR), the European Association for Cancer Research (EACR) and the Italian Cancer Society (SIC). Maria Rosa Bani research interests are in the field of cancer biology and molecular therapeutics. She is co-author of 34 peer reviewed publications, 2 book chapters and 65 proceedings of which 15 selected for oral presentation at international meetings. Selected publications Ghilardi C., Chiorino G., Dossi R., Nagy Z., Giavazzi R., Bani M.R. Identification of novel vascular markers through gene expression profiling of tumor-derived endothelium. BMC Genomics, 30(9), 201, Giavazzi R., Bani M.R.,Taraboletti G.: Tumor host interaction in the optimization of paclitaxel-based combination therapies with vascular targeting compounds. Cancer Metastasis Rev. 26:481 88, Naumova E., Ubezio P., Garofalo A., Borsotti P., Cassis L., Riccardi E., Scanziani E., Eccles S.A., Bani M.R. & Giavazzi R. The vascular targeting property of paclitaxel is enhanced by SU6668, a receptor tyrosine kinase inhibitor, causing apoptosis of endothelial cells and inhibition of angiogenesis. Clinical Cancer Research 12: , Bani M.R., Nicoletti M.I., Alkharouf N.W., Ghilardi C., Petersen D., Erba E., Sausville E.A., Liu E.T. and Giavazzi R. Gene expression correlating with response to paclitaxel in ovarian carcinoma xenografts. Molecular Cancer Therapeutics 3: , Vikhanskaya F.*, Bani M.R.*., Borsotti P., Ghilardi C., Ceruti R., Ghisleni G., Marabese M., Giavazzi R., Broggini M. & Taraboletti G. p73 overexpression increases VEGF and reduces thrombospondin-1 production: implication for tumor angiogenesis. Oncogene 20 : , Taraboletti G., Sonzogni L., Vergani V., Hosseini G., Ceruti R., Ghilardi C., Bastone A., Toschi E., Borsotti P., Scanziani E., Giavazzi R., Pepper M.S., Stetler-Stevenson W.G. & Bani M.R. Post-transcriptional stimulation of endothelial cell matrix metalloproteinases 2 and 1 by endothelioma cells. Experimental Cell Research 258 : , Giovanna Damia obtained her Medical Degree cum Laude from the University of Milan in After specializing in Pharmacology at the Mario Negri Institute of Milan and in Oncology at the University of Milan, she worked as a post-doctoral fellow in the Laboratory of Experimental Immunology of the National Cancer Institute, Frederick, USA. She worked as a research fellow in the Laboratory of Cancer Chemotherapy at the Mario Negri Institute and since April 2003 she has become chief of the DNA Repair Unit at the Mario Negri Institute. From 1992 to1995 she has been consultant of the General Secretariat of the Progetto Finalizzato CNR "Applicazioni Cliniche della Ricerca Oncologica". Since September 2005 she is Deputy Editor for Experimental Oncology of the European Journal of Cancer. Her main fields of interest are: mechanism of action of anticancer drugs, cell cycle checkpoints and natural compounds. Selected publications Simone M, Erba E, Damia G, Vikhanskaya F, Di Francesco A M, Riccardi R, Baldeyrou B, Bailly C, Cuevas C, Sousa- Faro J M F, D'Incalci M. Variolin B and its derivate deoxy-variolin B: New marine natural compounds with cyclindependent kinase inhibitor activity. Eur J Cancer 2005; 41:

15 Carrassa L, Broggini M, Erba E, Damia G. Chk1, but not Chk2, is involved in the cellular response to DNA damaging agents. Differential activity in cells expressing or not p53. Cell Cycle 2004; 3: Damia G, Broggini M. Improving the selectivity of cancer treatment by interfering with cell response pathways. Eur J Cancer 2004; 40: Damia G, Broggini M. Cell cycle checkpoint proteins and cellular response to treatment by anticancer agents. Cell Cycle 2004; 3: Carrassa L, Broggini M, Vikhanskaya F, Damia G. Characterization of the 5' flanking region of the human chk1 gene. Identification of E2F1 functional sites. Cell Cycle 2003; 2: Damia G, Sanchez Y, Erba E, Broggini M. DNA damage induces p53-dependent down-regulation of hchk1. J Biol Chem 2001; 276: Eugenio Erba has obtained his Biological and Biochemmistry Analysis Degree at the University of Urbino. He worked as a research fellow in the Laboratory of Cancer Chemotherapy at the Mario Negri Institute and since 1984 he is head of the Flow Cytometry Unit in the Department of Oncology at the Mario Negri Institute of Milan. He has worked as a visiting fellow in the Department of Istochemistry and Cytochemistry of the University of Leiden, The Netherlands in Since 1997 he is Teacher of Post-Graduate Studies in Cytometry at the University of Milan and Co-ordinator and Teacher of Post- Graduate Studies in Cytometry for the Italian Cytometry Group. He has been President of the Italian Cytometry Group from 1999 to Since 2001 he is member of the Executive Board of the Italian Cytometry Group. Scientific areas of interest: studies on the mechanism of action of different compounds with provided antitumoral activity evaluating the mechanism of cell death and cell cycle phase perturbations induced on different human cancer cell lines by using flow cytometry. Co-ordinator of working-group in a quality control study on flow cytometric DNA content analysis in human tumors. Selected publications C. Valli, G. Paroni, A. Di Francesco, R. Riccardi, M. Tavecchio, E. Erba, A. Boldetti, M. Giannì, M. Fratelli, C. Pisano, L. Merlini, A. Antoccia, C. Cenciarelli, M. Terao, E. Garattini. Atypical retinoids ST1926 and CD437 are S-phase specific agents causing DNA double-strand breaks: significance for the cytotoxic and antiproliferative activity. Mol. Ca. Ther. 7(9), , 2008 L. Roncoroni, L. Elli, E. Delfini, E. Erba, E. Dogliotti, C. Terrai, L. Doneda, MG. Grimoldi, MT. Bardella. Resveratrol inhibits cell growth in a human cholangiocarcinoma cell line. Liver Int. 28(10), , 2008 M.Tavecchio, M. Simone, E.Erba, I. Chiolo, G. Liberi, M. Foiani, M. D Incalci, G. Damia. Role of homologous recombination in trabectedin-induced DNA damage. Eur. J. Ca 44: (2008) Paulis M., Bensi M., Orioli D., Mondello C., Mazzini G., D Incalci M., Falcioni C., Radaelli E., Erba E., Raimondi E., De Carli L. Transfer of a Human Chromosomal Vector from a Hamster Cell Line to a Mouse Embryonic Stem Cell Line. Stem Cell, 25: (2007) Tavecchio M., Natoli C., Ubezio P., Erba E., D Incalci M. Dynamics of cell cycle perturbations by trabectedin (ET-743) in nucleotide excision repair (NER) deficient and NER-proficient cells, unravelled by a novel mathematical simulation approach. Cell Prolif., 40: (2007) Tognon G., Bernasconi S., Celli N., Faircloth G.T. Cuevas C., Jimeno J., Erba E., D Incalci M. Induction of resistance to Aplidin in a human ovarian cancer cell line related to MDR expression. Cancer Biology and Therapy, 4(12): (2005). Roldano Fossati got his Medical Degree cum Laude from the University of Milan in 1980, his Post- Doctoral Degree in Endocrinolgy cum Laude from the University of Verona in 1983 and his Post- Doctoral Degree in Medical Statistics from the University of Milan in He has been consultant at the Mario Negri Institute since 1983 and, at present, he is head of the Gynecology and Oncology Unit of the Laboratory of Translational and Outcome Research. Areas of Interest: Statistical and methodologic aspects of clinical research with focus on Controlled Clinical Trials in Oncology; Systematic Overview of the medical literature. Selected publications Labianca R., Fossati R., Zaniboni A., Torri V., Marsoni S., Nitti D., Boffi L., Scatizzi M., Tardio B., Mastrodonato N., Banducci S., Consani G., Pancera G. on behalf of ACOI/GIVIO/GISCAD investigators. Randomized Trial of Intraportal and/or Systemic Adjuvant Chemotherapy in Patients with Colon Carcinoma. J Natl Cancer Inst 96:750-8;2004 P. Benedetti Panici, A. Maggioni, N. Hacker, F. Landoni, S. Ackermann, E. Campagnutta, K. Tamussino, R. Winter, A. Pellegrino, S. Greggi, R. Angioli, N. Manci, G. Scambia, T. Dell'Anna, R. Fossati, I. Floriani, R.S. Rossi, R. Grassi, G. Favalli, F. Raspagliesi, D. Giannarelli, L. Martella, C. Mangioni. Systematic Aortic and Pelvic Lymphadenectomy versus Resection of Bulky Nodes Only in Optimally Debulked Advanced Ovarian Cancer: A Randomized Clinical Trial J Natl Cancer Inst 97:1-6;2005 Buda A, Fossati R, Colombo N, Fei F, Floriani I, Gueli Alletti D, Katsaros D, Landoni F, Lissoni A, Calzoni C, Sartori E, Scollo P, Torri V, Zola P, Mangioni C. Randomized trial of neoadjuvant chemotherapy comparing paclitaxel, ifosfamide, 14

16 and cisplatin with ifosfamide and cisplatin followed by radical surgery in patients with locally advanced squamous cell cervical carcinoma: The SNAP01 (Studio Neo-Adjuvante Por. J Clin Oncol 2005; 23: Maggioni A, Benedetti Panici P, Dell'anna T, Landoni F, Lissoni A, Pellegrino A, Rossi RS, Chiari S, Campagnutta E, Greggi S, Angioli R, Manci N, Calcagno M, Scambia G, Fossati R, Floriani I, Torri V, Grassi R, Mangioni C.Randomised study of systematic lymphadenectomy in patients with epithelial ovarian cancer macroscopically confined to the pelvis. Br J Cancer Sep 18;95(6): Maggi R, Lissoni A, Spina F, Melpignano M, Zola P, Favalli G, Colombo A, Fossati R. Adjuvant chemotherapy vs radiotherapy in high-risk endometrial carcinoma: results of a randomised trial. Br J Cancer Aug 7;95(3): Fruscio R, Colombo N, Lissoni AA, Garbi A, Fossati R, Ieda' N, Torri V, Mangioni C.A phase II randomised clinical trial comparing cisplatin, paclitaxel and ifosfamide with cisplatin, paclitaxel and epirubicin in newly diagnosed advanced epithelial ovarian cancer: long-term survival analysis. Br J Cancer Feb 5. Giulia Taraboletti got her degree cum laude in Biological Sciences at the University of Pavia (Pavia, Italy) in 1983, and the specialization in Pharmacological Research at the Mario Negri Institute, Milano, Italy in From 1986 to 1988 she was a post-doctoral fellow at the Laboratory of Pathology, NCI, NIH, Bethesda, MD, and from research scientist at Mario Negri Institute in Bergamo, Italy. Since 1995 she is Head of the Unit of Tumor Angiogenesis, at Mario Negri Institute, in Bergamo. Research interests include tumor angiogenesis, endogenous inhibitors of angiogenesis (thrombospondin- 1) and preclinical studies of antiangiogenic and vascular disrupting compounds, including tubulintargeting agents. She is member of Metatasis Research Society (MRS), American Association for Cancer Research (AACR), European Association for Cancer Research (EACR), and the Italian Society of Oncology (SIC). She is on the editorial board of European Journal of Cancer. Selected publications Margosio B, Rusnati M, Bonezzi K, Cordes B-lA, Annis DS, Urbinati C, Giavazzi R, Presta M, Ribatti D, Mosher DF, and Taraboletti G. Fibroblast growth factor-2 binding to the thrombospondin-1 type III repeats, a novel antiangiogenic domain. Int J Biochem Cell Biol 40: , Giavazzi R., Bani M.R.,Taraboletti G. Tumor host interaction in the optimization of paclitaxel-based combination therapies with vascular targeting compounds. Cancer Metastasis Rev, 26:481 88, Martinelli M., Bonezzi K., Riccardi E., Kuhn E., Frapolli R., Zucchetti M., Ryan A.J., Taraboletti G., Giavazzi R. Sequence dependent antitumour efficacy of the vascular disrupting agent ZD6126 in combination with paclitaxel. Br J Cancer 97:888-94, Margosio B., Marchetti D., Vergani V., Giavazzi R., Rusnati M., Presta M., and Taraboletti G. Thrombospondin-1 as a scavenger for matrix-associated fibroblast growth factor-2. Blood 102: , Taraboletti G. Micheletti G, Rieppi M, Poli M, Turatto M, Rossi C, Borsotti P, Roccabianca P, Scanziani E, Nicoletti MI, Bombardelli E, Morazzoni P, Riva A, and Giavazzi R. Antiangiogenic and antitumor activity of IDN 5390, a new taxane derivative. Clin Cancer Res. 8: , 2002 Taraboletti G., Morbidelli L., Donnini S., Parenti A., Granger H.J., Giavazzi R., and Ziche M.The heparin binding 25 kda fragment of thrombospondin-1 promotes angiogenesis and modulates gelatinase and TIMP-2 production in endothelial cells. FASEB J., 14: , Paolo Ubezio got his B.Sc. degree in Physics at the University of Milan, in 1982, and the specialisation in Pharmacological Research Specialist" at the Mario Negri Institute for Pharmacological Research in Main activities are: i) Study of cell-cycle mathematical models; ii) Development of flow cytometric methods; iii) Optimization of anticancer drug scheduling. Since 1991 is Head of the Unit of Biophysics at the Mario Negri Institute Selected publications Basse, B., Ubezio, P. (2007) A generalised age and phase structured model of human tumour cell populations both umperturbed and exposed to a range of cancer therapies. Bull. Math. Biol. 69: Lupi, M., Matera, G., Natoli, C., Colombo, V., Ubezio, P. (2007) The Contribution of p53 in the Dynamics of Cell Cycle Response to DNA Damage Interpreted by a Mathematical Model. Cell Cycle 6: Lupi, M., Matera, G., Branduardi, D., D'Incalci M. and Ubezio, P. (2004) Cytostatic and cytotoxic effects of topotecan decoded by a novel mathematical simulation approach. Cancer Res. 64: Matera, G., Lupi, M.,and Ubezio, P. (2004) Heterogeneous cell response to topotecan in a CFSE-based proliferation test. Cytometry 62A: Ubezio, P. (2004) Unraveling the complexity of cell cycle effects of anticancer drugs in cell populations.discrete and Continuous Dynamical Systems-Series B 4: Montalenti, F., Sena, G., Cappella, P., and Ubezio, P. (1998) Simulating cancer-cell kinetics after drug treatment: Application to cisplatin on ovarian carcinoma. Phys. Rev. E, 57: Massimo Zucchetti obtained his Chem. Pharm. Degree from the University of Milan in After specializing in Pharmacology at the Mario Negri Institute of Milan (1988), he worked in the Laboratory 15

17 of Clinical Pharmacology of Department of Oncology at San Giovanni Hospital, Bellinzona, Switzerland ( ). Since 1996 he has been chief of the Cancer Clinical Pharmacology Unit at the Mario Negri Institute. He is member of the Pharmacology and Molecular Mechanisms Group of the European Organization for Research and Treatment of Cancer (EORTC) from 1988 up to date. His main field of interest are: - Clinical pharmacology, phase I and Phase II studies - Analysis of drugs, pharmacokinetic and pharmacodynamic studies in humans in GCP and GLP conditions - Pharmacokinetic and metabolic studies in animals - Pharmacokinetic drug interaction Dr Zucchetti is author of more than 80 papers on pre-clinical and clinical cancer chemotherapy published in peer reviewed international journals. Selected publications Gambacorti-Passerini CB, Tornaghi L, Marangon E, Franceschino A, Pogliani EM, D'Incalci M, Zucchetti M.. Imatinib concentrations in human milk Blood Feb 15;109(4):1790. Marangon E, Sala F, Caffo O, Galligioni E, D'Incalci M, Zucchetti M. Simultaneous determination of gemcitabine and its main metabolite, dfdu, in plasma of patients with advanced non-small-cell lung cancer by high-performance liquid chromatography-tandem mass spectrometry. J Mass Spectrom Feb;43(2): Frapolli R., Marangon E., Zaffaroni M., Colombo T., Falcioni C., Bagnati R., Simone M., D Incalci M., Manzotti C., Fontana G., Morazzoni P., Zucchetti M. Pharmacokinetics and metabolism in mice of IDN 5390 (13-(N-Boc-3-ibutylisoserinoyl)-C-7,8-seco-10-deacetylbaccatin III), a new oral C-seco-taxane derivative with antiangiogenic property effective on paclitaxel-resistant tumors. Drug Metabolism and Disposition, 34(12): (2006). Rizzari C., Citterio M., Zucchetti M., Conter V., Chiesa R., Colombini A., Malguzzi S., D Incalci M. Pharmacological study on pegylated asparaginase used in front-line treatment of children with acute lymphoblastic leukemia. Hematologica, 91: (2006). Fruscio R., Lissoni A.A., Frapolli R., Corso S., Mangioni C., D Incalci M., Zucchetti M. Clindamycin-Paclitaxel pharmacokinetic interaction in ovarian cancer patients. Cancer Chemother. Pharmacol., 58(3): (2006). Gambacorti Passerini C, Zucchetti M, Russo D, Frapolli R, Verga M, Bungaro S, Tornaghi L, Rossi F, Pioltelli P, Pogliani E, Alberti D, Corneo G, D'Incalci M Alpha 1 acid glycoprotein binds to imatinib (STI571) and substantially alters its pharmacokinetics in chronic myeloid leukemia patients Clin Cancer Res 2003; 9:

18 INTRODUCTION TO THE DEPARTMENT'S ACTIVITIES The Oncology Department comprises three preclinical experimental laboratories (Laboratory of Cancer Pharmacology, Laboratory of Molecular Pharmacology and Laboratory of Biology and Therapy of Metastasis) and four laboratories dealing with clinical research and clinical trials (Laboratory for the Development of New Pharmacological Strategies, Laboratory of Clinical Trials, Laboratory of Translational and Outcome Research in Oncology and Laboratory for Medical Research and Consumer Involvement). The Oncology department hosts the coordination center of two networks of hospitals that carry on clinical research in gynecologic cancer (MaNGO: Mario Negri Gynecologic Oncology) and in cancer pain (CPOR-SG: Cancer Pain Outcome Research Study Group) and a center for cancer pain assessment and research (CERP:Center for the Evaluation and Research on Pain). In some cases research projects are carried out by single laboratories or research units, in other cases by collaborations between different laboratories of the Oncology Department or other departments, or other groups outside the Institute (see National and International Collaborations). Preclinical laboratories focus on the discovery and development of new antitumor and antimetastatic drugs and their new combinations; on tumor biology, not only to acquire new scientific knowledge, but particularly as a base for more selective therapeutic approaches and to identify and evaluate experimental models for discovering and studying new drugs or treatments. Clinical new drug development involves close participation in the activity of SENDO (South Europe New Drug Development Organization) and studies driven by the Laboratory of Cancer Pharmacology, the Laboratory of Molecular Pharmacology and the Laboratory of Biology and Therapy of Metastasis. The Laboratory for the Development of New Pharmacological Strategies, the Laboratory of Clinical Trials, the Laboratory of Translational and Outcome Research in Oncology and the Laboratory for Medical Research and Consumer Involvement are involved in the evaluation of the effects of new therapeutic modalities in phase I/II and in phase III comparative and effectiveness outcome studies. Outcome Research implies organizing trials to clarify the results of certain health care practices and interventions in clinical practice. Observational (surveys) and outcome research (effectiveness) studies are carried out, in collaboration with regional and national health authorities and other scientific associations. At the preclinical and clinical level there are studies of various human tumors, with particular emphasis on ovarian tumors and more recently on soft tissue sarcomas. FINDINGS/MAIN RESULTS At nanomolar concentrations, ET-743 (Yondelis, Trabectedin) affects the regulatory mechanisms of the transcription. Nucleotide excision repair deficient cells that are hypersensitive to UV rays and to other DNA damaging drugs are resistant to Trabectedin. The selective activity of ET-743 against human myxoid liposarcoma appears related to the drug ability to modulate the transcription of genes involved in adipocytic differentiation. Use of mathematical models of tumor growth and anticancer treatment to interpret experimental data and to manage the complexity of underlying biological phenomena. 17

19 The theoretical relationship between proliferation, quiescence and cell loss leading to growth control of tumor cell populations was found. Gene profiling analysis shows specific molecular signatures according to the histotype and prognosis of stage I ovarian carcinoma. The expression of a truncated form of p63 (DNp63) increases with the increased malignancy of ovarian cancer. Patients expressing high levels of DNp63 have a worst prognosis. DNp63 represents therefore a new potential target for selective therapies in this malignancy. CHK1 downregulation by specific inhibitors or sirna, increases the antitumor activity of 5- fluorouracil in vivo. This effect is particularly evident in p53 deficient tumors indicating that this combination increases the selectivity of this anticancer agent. An anthracycline derivative, Nemorubicin, has a peculiar mechanism of action and is active against tumors resistant to drugs such as cisplatin. A mechanism of resistance against this drug has been identified, which involves the abrogation of the expression of a nucleotide excision repair gene. Overexpression of a truncated form of p73 (DNp73beta) in human cancer cells lead to growth arrest and formation of tetraploid cells, suggesting a role of this isoform in mitosis. A new mechanism of p73 activation mediated by the protease HtrA2 has been identified. After apoptotic stimuli this protease cleaves p73 in the C-terminal region increasing its apoptotic potential. Inositol pentaphosphate analogues interfere with the PI3-kinase-induced phosphorylation of akt and possess antitumor activity in vitro and in vivo, particularly when combined with other anticancer agents. Human umbilical cord-derived stem cells express checkpoints proteins only in specific differentiation stages. It is likely that this is related to the different susceptibility of the cells. Inhibition of PLC gamma, through sirna technology, reduces the in vivo growth of tumors and reduces the formation of metastasis. Identification of genes preferentially expressed by tumor associated endothelial cells. VEGF released by cancer cells modulatesgene expression in the tumor microenvironment (stroma). VEGF produced by ovarian tumor cells stimulates host MMP9 expression; the anti-vegf antibody bevacizumab (Avastin ) inhibited MMP9 expression and abolished ovarian tumor invasion. Soluble human VEGFC levels in serum and ascites correlate with tumor progression and metastatic capability of ovarian carcinoma models. A new antiangiogenic domain of thrombospondin-1 (an endogenous inhibitor of angiogenesis) that binds the angiogenic factor FGF-2 has been identified and characterized. New antineoplastic compounds directed against the tumor vasculature (vascular disrupting agents) have been identified. 18

20 The histone deacetylase inhibitor SAHA potentiates the cytotoxic effect of paclitaxel in human ovarian carcinoma cells resistant to paclitaxel. The effect is mediated by the acetylation of tubulin. The expression of protease-activated receptor-1 (PAR-1) correlates with the malignant phenotype of human melanomas and is accountable for their motility and invasive features ICON4, a, randomised trial of second-line chemotherapy in advanced ovarian cancer, coordinated by the Mario Negri Institute and by MRC, showed for the first time a reduction in mortality in favour of platinum and paclitaxel chemotherapy. The response to chemotherapy was a good surrogate endpoint of survival in patients with locally advanced cervix carcinoma. Adjuvant chemotherapy with the regimen vindesin, mitomycin C and cisplatin (MVP) did not improve survival of non small cell lung cancer (NSCLC) patients compared with surgery alone. The website of the project PartecipaSalute ( has a very innovative character in comparison with the other health Italian sites because introduces and develops with ad-hoc instruments the information transfer in an active way. The project PartecipaSalute ( has an innovative character of the website panorama in Italy. This web - compared to other Italian health websites - introduces and develops ad hoc methods to transfer in an active way information on health. A randomized phase III trial has shown that pelvic systematic lymphadenectomy in early endometrial cancer does not improve overall survival. As pelvic systematic lymphadenectomy is not devoid of side effects, this trial will spare patients with early endometrial cancer important early and late surgical morbidities. A twin trial in ovarian carcinoma, published in 2005, came to similar conclusions. Bupropion more than doubled the odds of continuous abstinence from smoking from week 4 to 7 and from week 4 to 12 months in a way similar to that observed in academic studies. The adherence of GPs and participants to the protocol was excellent, making our findings robust and easy to generalize to the context of primary care. A randomized phase II trial has shown that the efficacy and tolerability of a chemotherapic regimen containing paclitaxel and cisplatin (TP) in the neoadjuvant treatment of locally advanced squamous cell cervical cancer is inferior to the triplet containing also iphosphamide (TIP). Adding up to previous similar findings, the TIP regimen (TP plus ifosfamide) has been proving to be one the best chemotherapic regimen in this setting of patients as it can adequately reduce the cancer volume allowing the surgical ablation with curative intent in most of the patients. An observational longitudinal study carried out in 110 Italian centers and involving about 1800 patients with metastatic cancer and pain have documented that in Italy most patients at the time of study inclusion were still on active anti-cancer treatment (>50%), were not aware of their prognosis (about 70%). In addition, up to 45% did not received an appropriate analgesic treatment (under-treatment), especially in some sub-groups. In terms of analgesics effectiveness, although the observational design, results suggest that each drugs prescribed by investigators (morphine, fentanyl, buprenorphine and oxycodone) were able to reduce the intensity of pain of about 2 points on a 11-eleven point numerical rating scale 19

21 (p<0.001). The application of specific pre-planned algorithm identified about 30% cases who were classified as non-responders. Preliminary analyses documented some differences between drugs in terms of size of the analgesic effect, dosages required and side effects reported. A systematic review of the oncologic literature (last 11 years) have shown that the utilization of interim analysis to stop early RCT for apparent benefit is frequent (35% of the trials stopped), is increasing in last years, especially in RCT carried out for regulatory purposes (78%). The implications of such phenomenon are twofold: results of trials stopped earlier should be interpreted with caution because the stop for apparent benefit does statistically exaggerate the true benefit; unresolved clinical questions generate controversies and ethical problems either for patients included in the study, in physicians and society. Most of the drugs recently approved by the European Agency (EMEA) for hematologic malignancies had some information debt about their true risk-benefit profiles and, notwithstanding they are prescribed to thousand of patients. A survey of all the assessments carried out by EMEA over the last decade gave evidence that in as many as 2/3 of new drugs were approved by EMEA without a real therapeutic benefit over the old ones A randomized trial of patients with high risk (stage IcG3, IIG3 with myometrial invasion >50%, and III) endometrial carcinoma showed the substantial equivalence between radiotherapy or chemotherapy as an adjuvant therapy after surgery. Although both radiotherapic and chemotherapic approaches are still unsatisfactory, since the risk of progression or death remains high, this encouraging evidence of clinical activity suggest a possible use of their concurrent or sequential use in an adjuvant setting. The estimates of the prevalence and impact of cancer pain in a large and representative sample of cancer patients (1800) recruited by several Italian centers (more than 120), with the evaluation of the actual proportion of cases that received a substantial analgesic under-treatment (about 25%), mainly attributable to a sub-optimal utilization of opiods. An evaluation of the activity of the EMEA over the last 10 years, has documented that most of the new anti-cancers drugs has actually received an approval on the basis of very preliminary findings: in 48% of case the approval was based on studies using surrogate endpoints, and in 40% of cases the design of the study was non-comparative and non-randomized. The activity of training and information organized with the associations of citizens & patients in the framework of the PartecipaSalute project has been finalized to the organization of the Parita task Participate to the research project with the associations. Parita is organised to discuss with the scientific community the grey areas of the medical assistance and clinical research identified from the patients and their associations, and to develop ad hoc protocols for future research programs. An ad hoc data collection with the cooperation of patient s associations has highlighted the missmacht between patients and researchers agenda. Development and validation of a new short questionnaire, the PGWBI-short version available to be used in large sample of citizens or patients. 20

22 NATIONAL COLLABORATIONS ASR, Agenzia Sanitaria Regionale, Bologna AIFA, Agenzia Italiana del Farmaco (Roma) Azienda Sanitaria Locale, Rimini Assessorato Sanità, Regione Emilia Romagna Azienda Sanitaria Unica Regionale, Regione Marche Casa Sollievo della Sofferenza, San Giovanni Rotondo (IRCCS) CNPDS, Centro Nazionale per la prevenzione e Difesa Sociale, Milano CNR IGBE, Pavia CNR, Istituto di Chimica del Riconoscimento Molecolare, Milano Cochrane Collaboration Fondazione IRCCS Istituto Nazionale dei Tumori (INT), Milano Fondazione LUVI, Milano Fondazione Nerina e Mario Mattioli Onlus, Milano Fondazione Salvatore Maugeri, Pavia Fondazione SmithKline (FSK), Milano Fondo Edo Tempia, Laboratorio di Farmacogenomica, Biella I.A.S.I., Roma Istituto Clinico Humanitas, Rozzano MI Istituto Dermopatico dell'immacolata, Roma Istituto FIRC per l Oncologia Molecolare (IFOM) Istituto Ortopedico Galeazzi, Milano Istituti Ortopedici Rizzoli, Bologna Istituto Europeo di Oncologia (IEO), Milano Istituto di Fisica, Politecnico di Milano Istituto di Genetica Molecolare CNR, Sezione di Istochimica e Citometria, Pavia Istituto Nazionale per la Ricerca sul Cancro (IST), Genova Istituto Nazionale Tumori Fondazione G. Pascale, Napoli Istituto Regina Elena, Roma Istituto Superiore di Sanità Laboratorio Cell factory, Policlinico di Milano Ospedale San Gerardo, Monza, Milano Ospedale San Matteo, Pavia Rete Oncologica Lombarda (ROL), Milano Unità di Tossicologia e Scienze Biomediche, ENEA Centro Ricerche, Roma Università Cattolica del Sacro Cuore, Roma Università di Bari Università di Brescia Università di Chieti Università di L Aquila Università di Milano Università di Milano Bicocca Università di Modena e Reggio Emilia Università di Monza Università di Catania Università di Padova Università di Pisa Università di Siena 21

23 Università La Sapienza, Roma Zadig, Agenzia di Giornalismo scientifico INTERNATIONAL COLLABORATIONS ADAMANT Consortium, IP 7th FP, EC ARCAGY (Association de Recherche sur les Cancers Gynécologiques), Francia Breakthrough Breast Cancer Center, Instutite of Cancer Reasearch, Londra, Gran Bretagna Cancer Biomarkers and Prevention Group, University of Leicester, Gran Bretagna Cancer Research UK, Londra, Gran Bretagna Cancerdegradome Consortium, IP 6th FP, EC EORTC, Bruxelles, Belgio EUROPA DONNA European Agency for the Evaluation of Medicinal Products (EMEA), Londra, Gran Bretagna European Association for Palliative Care (EAPC) Eusoma (European Society of Breast Cancer Specialist) Firenze, Italy Executive Board of GCIG (Gynecologic Cancer Intergroup) Frontier science & technology Research Foundation Southern Europe (FSE) Genome Institute of Singapore (GIS), Singapore German Cancer Research Center, Division of Toxicology and Cancer Risk Factors, Heidelberg, Germania Goteborg University, Lundberg Laboratory for Cancer Research, Goteborg, Svezia Helios Klinikum Erfurt GmbH, Institute of Pathology, Germania Istituto Oncologico della Svizzera Italiana Johns Hopkins University, USA Ludwig Institute for Cancer Research, Londra, Gran Bretagna National Cancer Center, Singapore Stony Brook University, NY USA Massachusetts General Hospital and Harvard Medical School, USA MD Anderson Cancer Center, Houston, Texas, USA MRC, Londra, Gran Bretagna National Cancer Institute (NCI), Bethesda and Frederick, MD, USA Ospedale San Giovanni, Bellinzona, Svizzera Paterson Institute for Cancer Research, Manchester, Gran Bretagna Southern Europe New Drug Organization (SENDO), Milano, Italia Stroma Consortium, IP 6th FP, EC Swiss Federal Institute of Technology, Zurigo, Svizzera The Sackler Institute, University College Londra, Gran Bretagna Tumor Biology and Metastasis Institute of Cancer Research, Sutton, Gran Bretagna University College, London Medical School, Londra, Gran Bretagna University of Birmingham, Gran Bretagna University of Cincinnati, USA University of Crete Medical School, Greece University of Newcastle, Gran Bretagna University of Pau, Francia University of Wisconsin, Madison, WI, USA Kyoto University, Giappone 22

24 Weizmann Institute of Science, Israele EDITORIAL BOARD MEMBERSHIP Attualità in Senologia (Paola Mosconi) British Journal of Cancer (Maurizio D Incalci) Chemotherapy (Maurizio D Incalci) Clinical Experimental Metastasis (Raffaella Giavazzi) Current Opinion in Oncologic, Endocrine and Metabolic Drugs (Maurizio D Incalci) Current Cancer Therapy Reviews (Raffaella Giavazzi) European Journal of Cancer (Maurizio D Incalci, Giovanna Damia, Raffaella Giavazzi, Massimo Broggini e Giulia Taraboletti) Health and Quality of Life Outcomes (Giovanni Apolone, Paola Mosconi) International Journal of Biological Markers (Raffaella Giavazzi) International Journal for Quality in Health Care (Giovanni Apolone) Journal of Ambulatory Care and Management (Giovanni Apolone) Journal of B.U.ON. (Maurizio D Incalci) Journal of Cancer Microenvironment (Raffaella Giavazzi) Journal of Chemotherapy (Raffaella Giavazzi) Journal of Experimental Therapeutics and Oncology (Raffaella Giavazzi) Journal of Medicine and the Person (Giovanni Apolone) Journal of Preventive Medicine anf Hygiene (Giovanni Apolone) Molecular Cancer Therapeutics (Maurizio D Incalci) Oncology Research (Maurizio D Incalci) Tumori (Maurizio D Incalci, Raffaella Giavazzi) (Paola Mosconi) (Paola Mosconi) PEER REVIEW ACTIVITIES American Journal of Pathology, Annals of Oncology, Anti-cancer Drugs, Biochemical Pharmacology, BioMed Central Editorial, British Journal of Cancer, British Journal of Pharmacology, British Medical Journal, Cancer Chemotherapy and Pharmacology, Cancer Detection and Prevention, Cancer Letters, Cancer Research, Carcinogenesis, Chemico- Biological Interactions, Clinical & Experimental Metastasis, Clinical Cancer Research, Cytometry, European Journal of Cancer, European Journal of Immunology, Faseb Journal, Gynecologic Oncology, Health and Quality of Life Outcomes, Health Expectations, European Journal of Neurology, Intensive Care Medicine, International Journal of Biological Markers, International Journal of Cancer, International Journal for Quality in Health Care, Journal of Ambulatory Care and Management, Journal of Biological Chemistry, Journal of Biological Markers, Journal of Cell Biochemistry, Journal of Clinical Oncology, Journal of Experimental Therapeutics and Oncology, Journal of Medicine and the Person, Journal of the National Cancer Institute, Journal of Neurology, Journal of Preventive Medicine and Hygiene, Journal of the National Cancer Institute, Leukemia, Molecular Cancer Therapeutics, Nature, Nature 23

25 Biotechnology, Nature Reviews, Oncology Research, PharmacoEconomics, Quality of Life Research, Science, Tumori, ZEG Centre for Epidemiology & Health Research. NATIONAL AND INTERNATIONAL COMMITTEE MEMBERSHIP Ethical Committee, Centro di Riferimento Oncologico, Aviano PN, Italy Ethical Committee, Ente Ospedaliero San Paolo, Milan, Italy Ethical Committee, Istituto Europeo di Oncologia, Milan, Italy Ethical Committee, Istituto Neurologico Carlo Besta, Milan, Italy Ethical Committee, Istituto Scientifico Eugenio Medea, Bosisio Parini, Lecco, Italy Ethical Committee, Ospedale San Gerardo, Monza, Milan, Italy Ethical Committee, Ospedale Sant Anna, Como, Italy Ethical Committee, Ospedale della Valtellina e Valchiavenna, Sondrio, Italy Ethical Committee, IRCCS MultiMedica, Sesto San Giovanni, Milan, Italy Ethical Committee, Azienda USL di Bologna, Italy Executive Board of GCIG (Gynecologic Cancer Intergroup) Scientific Committee, Associazione Italiana Ematologia e Oncologia Pediatrica, Monza, Milan, Italy Scientific Committee, Pezcoller Foundation, Trento, Italy Technical-Scientific Commitee, Associazione Italiana per la Ricerca sul Cancro, Milan, Italy Board of Directors, Fondazione Nerina e Mario Mattioli Onlus, Milan, Italy Board of Directors, Società Italiana di Cancerologia (SIC) Board of Directors, Società Italiana di Citometria (GIC) Directional Council Areas-CCI National Advisory Board 8th World Congress of Psycho-Oncology Developmental Therapeutics Program, National Cancer Institute (NCI) Decision Network and Executive Committee, South Europe New Drug Organization (SENDO) Executive Board, Europa Donna Executive Committee, European Asociation for Cancer Research (EACR) External Scientific Committee, Angiotargeting Consortium EU Sixth Framework Programme, University of Bergen, Norvegia NHS R&D National Coordinating Centre for Health Technology Assessment, UK Scientific Committee, Swiss Cancer League University Medical School of Siena, Italy EVENT ORGANIZATION Investigator Meeting Studio ITACAS Istituto di Ricerche Farmacologiche Mario Negri, Milan, February 18, Investigator Meeting Studio Head & Neck Congresso AIOM, Verona, October 12, Investigator Meeting Studio Tailor Congresso AIOM, Verona, October 12, I Corso di Epidemiologia Clinica e Metodologia della Ricerca, Rimini Progetto Il dolore nel paziente con cancro, Rimini, October 27 and 28,

26 Simposio Satellite Progetto Il dolore nel paziente con cancro XV Congresso Nazionale Società Italiana di Cure Palliative, Giardini Naxos, November 3/6, Workshop Progetto Fragilità ed Equità in Sanità, Milan, November 7, 2008 Conferenza di consenso Quale informazione per la donna in menopausa sulla terapia ormonale sostitutiva?, Torino, May 16-17, International Clinical Trials Day. Donne & ricerca clinica, Milan, May 21, I corso di formazione per componenti laici di Comitati Etici 2008, Milan, September January III edizione percorso di formazione Orientarsi in salute & sanità per fare scelte consapevoli,. Milan, September-December PARTICIPATION IN EVENTS IN WHICH THE DEPARTMENT WAS INVOLVED Meeting: 29th Winter Meeting of the Pharmacology and Molecular Mechanisms Group. "Selective Modulation of Transcription Regulation by Natural Products with Antitumor Properties". Palermo (Italy), January 30 February 2, Meeting: Cytosquelette microtubulaire & cancer. Microtubule-affecting agents for vascular targeting therapy. Marseil (France), February 26, Conference: Gordon Research Conference on Molecular Mechanisms in Lymphatic Function and Disease. Ovarian carcinoma xenografts as a model to study lymphangiogenesis. Ventura CA, USA, March 2-7, Conference: 1 st International Ovarian Cancer Action. Angiogenesis: basic aspects. Angiogenesis-preclinical development. Londra (UK), March 7-8, Meeting: AACR Annual Meeting. The effect of vandetanib on tumor vasculature remodeling and paclitaxel uptake in a xenograft model of ovarian carcinoma. San Diego (USA), April 12-16, Meeting: AACR Annual Meeting. Characterization of predictive markers and target genes of ProLindac, a novel DACH-platinum compound, compared to oxaliplatin and cisplatin in human cancer cell lines. San Diego (USA), April 12-16, Meeting: AACR Annual Meeting. Anti-inflammatory properties of the novel anti-tumor agent trabectedin: Reduction of ccl2, il-6 and pentraxin-3 by monocytes/macrophages and tumor cells. San Diego (USA), April 12-16, Congress: XXIV International Congress ISAC: Cytometry in the Age of Systems Biology. Budapest Sportarena, Budapest, May 17-21,

27 Conference: CancerSim 2008 Euroconference on Modeling and Simulation of Cancer Growth and Therapy. Targeting the tumor vasculature: pharmacological end-points for the design of combination treatments. Torino, May 19-21, Meeting: Workshop SIICA Angiogenesi: basi molecolari ed implicazioni terapeutiche II. Certosa di Pontignano, Siena, May 21-23, Pharmacological end-points to develop combination therapies with vascular targeting agents. Lymphangiogenesis in ovarian carcinoma xenografts. Meeting: 5th Research Forum of the European Association for Palliative Care (EAPC) Trondheim, Norway, May 29-31, Pain in cancer. An Outcome Research Project to evaluate the epidemiology, the quality and the effects of pain treatment in cancer patients. Meeting: 9 th International Conference, Angiogenesis: Basic Science and Clinical Applications. Pharmacological end-points to develop combination therapies with inhibitors of angiogenesis. Patras (Grecia), June 22-26, Meeting: 20 th Meeting of the European Association for Cancer Research. Large scale comparative proteomic study of accessible vascular proteins in mouse liver metastases and normal liver. Lione (Francia), July 5-8, Conference: Joint Metastasis Research Society-AACR Conference on Metastasis Tubulintargeting agents as inhibitors of cell motility Vancouver, BC, Canada, August 3-7, Congress: IASP - 12 th World Congress on Pain, Glasgow (Scozia), August 17-22, Pain in cancer. An Outcome Research Project to evaluate the epidemiology, the quality and the effects of pain treatment in cancer patients. Course: XXV Incontro di Aggiornamento e Formazione Scuola Nazionale di Citometria La Citometria nella Clinica e nella Biologia: metologie di base e protocolli applicativi, Campus Scientifico Università di Urbino, October 1-4, Congress: 50 th Annual Meeting of the Italian Cancer Society. Napoli, October 6-9, Soluble vegfr-1 expression in human melanoma cell lines established from primary or metastatic lesions. Novel markers of tumor vasculature identified through gene expression analysis of endothelial cells. Meeting: Cancer Degradome Symposium. Bevacizumab inhibits organ-specific host MMP9 expression and ovarian tumor invasion. London, UK, October 8-9, Simposium: 20th EORTC-NCI-AACR symposium on Molecular Targets and Cancer Therapeutics. Ginevra (Svizzera), October 21-24, The effects of treatment sequencing on the antitumor activity of vandetanib and paclitaxel in a xenograft model of human ovarian carcinoma. Simposium: 20th EORTC-NCI-AACR symposium on Molecular Targets and Cancer Therapeutics. Ginevra (Svizzera), October 21-24, Tubulin acetylation and the antimotility effects of tubulin-targeting agents. 26

28 Simposium: 20th EORTC-NCI-AACR symposium on Molecular Targets and Cancer Therapeutics. Ginevra (Svizzera), October 21-24, The complexity of cell cycle dynamic of anticancer drugs unraveled by the use of mathematical models suitable for a quantitative assessment of G1, S and G2M checkpoint activities. Simposium: 20th EORTC-NCI-AACR symposium on Molecular Targets and Cancer Therapeutics. Ginevra (Svizzera), October 21-24, The novel taxane derivative, IDN6140, crosses the Blood Brain Barrier and has a promising activity in CNS tumors. Simposium: 20th EORTC-NCI-AACR symposium on Molecular Targets and Cancer Therapeutics. Ginevra (Svizzera), October 21-24, Expression of genes involved in DNA damage response pathways in ovarian cancers. Workshop: 2 nd Workshop CM 0602 Cost Action: Inhibitors of Angiogenesis Design, synthesis and biological exploitation. Angiogenesis inhibitors: strategies for combination therapies in cancer treatment. Favignana (Trapani), October 24-26, Congress: XV Congresso Nazionale Società Italiana di Cure Palliative, Giardini Naxos November 3-6, Prevalenza dell'undertreatment nel trattamento del dolore da cancro. Risultati di una revisione sistematica e di uno studio osservazionale. Conference: Conferenza ECTA2008 : 3 rd European Conference on Tumor Angiogenesis and Antiangiogenic Therapy. Abano Terme (PD), November 6-8, Molecular profile of the stroma of human ovarian carcinoma xenografts equipped with different angiogenic phenotypes. Preclinical studies on combinations with vascular-targeting and anti-angiogenic agents and chemotherapy. Meeting: British Gynaecologic Cancer Society, Liverpool (UK), November 13-14, Radical (type II) vs Extrafascial Hysterectomy (type I) for Clinical Stage I Endometrial Carcinoma: Final Results of an Italian Randomized Clinical Trial. GRANTS AND CONTRACTS Arcispedale Santa Maria Nuova di Reggio-Emilia Azienda Sanitaria Locale - Rimini Azienda Sanitaria Unica Regionale - MARCHE CNPDS, Centro Nazionale per la Prevenzione e Difesa Sociale (CNPDS), Milano Ministero della Salute (Sesto Progetto Integrato Oncologia), Roma Grunenthal Italia, Milano AIFA Agenzia Italiana del Farmaco AIL Associazione Italiana contro le Leucemie, Padova 27

29 Amgem SpA, Milano AIRC Associazione Italiana per la Ricerca sul Cancro ASL Padova ASL Provincia di Lodi Astra Zeneca SpA Astra Zeneca UK AVAPO (Associazione Volontari Assistenza Pazienti Oncologici) Aventis Pharma Azienda Ospedaliera Fatebenefratelli e Oftalmico- Milano Azienda Sanitaria Locale, Rimini Azienda Sanitaria Unica Regionale, Marche Azienda Ospedaliera Spedali Civili di Brescia Bracco Imaging SpA, Milan Centro Cochrane Italiano Chiesi Farmaceutici SpA CIPOMO (Collegio Italiano dei Primari Oncologi Medici Ospedalieri) CNPDS, Centro Nazionale per la prevenzione e Difesa Sociale, Milano CNR Consiglio Nazionale delle Ricerche CNR-MIUR Ministero Istruzione Università e Ricerca Compagnia di San Paolo CTI Cell Therapeutics, Inc. Cyclacel Ltd. Dompé Eli Lilly Italia SpA Elsevier Science Ltd. EORTC-European Organization for Research and Treatment of Cancer EOS SpA European Commission - 7th Framework Programme (ADAMANT) FIRB-MIUR Fondo per gli Investimenti della Ricerca di Base-Ministero Istruzione Università e Ricerca FIRC Fondazione Italiana per la Ricerca sul Cancro Fondazione Cassa di Risparmio delle Province Lombarde Fondazione Lu.V.I. Fondazione Nerina e Mario Mattioli Onlus Fondo Edo Tempia FSE GISCAD(Gruppo Italiano Studi di Carcinomi Apparato Digerente) GlaxoSmithKline, Verona Grunenthal, Milano Indena SpA Institut de Recherche Pierre Fabre Istituto Clinico Humanitas Rozzano Istituto Superiore di Sanità Italfarmaco Komen Italia Onlus Lottomatica Madaus Srl Medac Merck Sharp & Dome Ministero della Salute NCI SAIC Frederick 28

30 Nerviano Medical Science S.r.l. Novartis Novartis Farma SpA Oncoethix Optigenex Inc. Pfizer Global Research and Development Pfizer Italia Pharma Mar, SA Pharmatex Pharminox Ltd, UK Policlinico di Padova / C.O.R. PTC Pharma AG Regione Emilia Romagna Regione Lombardia Regione Veneto Sanofi-Aventis Pharma Sara Bet, Roma SENDO-Tech Srl Sigma-Tau SpA Università degli Studi di Padova Università Federico II Napoli (Dipartimento di Endocrinologia ed Oncologia molecolare e clinica) 29

31 SELECTION OF SCIENTIFIC PUBLICATIONS FROM 2008 Corica F, Corsonello A, Apolone G, Mannucci E, Lucchetti M, Bonfiglio C, Melchionda N, Marchesini G, QUOVADIS Study Group Metabolic syndrome, psychological status and quality of life in obesity: The QUOVADIS study Int J Obes : Marchini S, Marabese M, Marrazzo E, Mariani P, Cattaneo D, Fossati R, Compagnoni A, Fruscio R, Lissoni A A, Broggini M ΔNp63 expression associates with poor survival in ovarian cancer Cancer Res : Graziano F, Ruzzo A, Loupakis F, Canestrari E, Santini D, Catalano V, Bisonni R, Torresi U, Floriani I, Schiavon G, Andreuzzi B, Maltese P, Rulli E, Humar B, Falcone A, Giustini L, Tonini G, Fontana A, Masi Gianluca, Magnani M Pharmacogenetic profiling for cetuximab/irinotecan therapy in patients with refractory advanced colorectal cancer J Clin Oncol : Mosconi P, Apolone G, Mingardi G Quality of life assessment and instruments in end-stage renal disease J Nephrol Suppl. 13 : S107-S112 Marangon E, Sala F, Caffo O, Galligioni E, D'Incalci M, Zucchetti M Simultaneous determination of gemcitabine and its main metabolite, dfdu, in plasma of patients with advanced nonsmall-cell lung cancer by high-performance liquid chromatography-tandem mass spectrometry J Mass Spectrom : Cooper W N, Dickinson R E, Dallol A, Grigorieva E V, Pavlova T V, Hesson L B, Bieche I, Broggini M, Maher E R, Zabarovsky E R, Clark G J, Latif F Epigenetic regulation of the ras effector/tumour suppressor RASSF2 in breast and lung cancer Oncogene : Tavecchio M, Simone M, Erba E, Chiolo I, Liberi G, Foiani M, D'Incalci M, Damia G Role of homologous recombination in trabectedin-induced DNA damage Eur J Cancer : Marabese M, Marchini S, Marrazzo E, Mariani P, Cattaneo D, Fossati R, Compagnoni A, Signorelli M, Moll U M, Codegoni A M, Broggini M Expression levels of p53 and p73 isoforms in stage I and stage III ovarian cancer Eur J Cancer : Marabese M, Mazzoletti M, Vikhanskaya F, Broggini M HtrA2 enhances the apoptotic functions of p73 on bax Cell Death Differ : Marchini S, Mariani P, Chiorino G, Marrazzo E, Bonomi R, Fruscio R, Clivio L, Garbi A, Torri V, Cinquini M, Dell'Anna T, Apolone G, Broggini M, D'Incalci M Analysis of gene expression in early-stage ovarian cancer Clin Cancer Res : Belotti D, Calcagno C, Garofalo A, Caronia D, Riccardi E, Giavazzi R, Taraboletti G Vascular endothelial growth factor stimulates organ-specific host matrix metalloproteinase-9 expression and ovarian cancer invasion Mol Cancer Res : Apolone G, Patarnello F The value of a drug: From innovation to the payment via Karl Marx J Ambul Care Manage : Ferketich A K, Gallus S, Colombo P, Fossati R, Apolone G, Zuccaro P, La Vecchia C Physician-delivered advice to quit smoking among italian smokers Am J Prev Med :

32 Corsonello A, Lucchetti M, Corica F, Apolone G, Marchesini G Metabolic syndrome and health-related quality of life: does psychological well-being matter? Ann Epidemiol : Ludovini V, Pistola L, Gregorc V, Floriani I, Rulli E, Di Carlo L, Semeraro A, Daddi G, Darwish S, Stocchi L, Tofanetti F R, Bellezza G, Sidoni A, Tognellini R, Crino' L, Tonato M Biological markers and DNA flow cytometric analysis in radically resected patients with non-small cell lung cancer. A study of the Perugia Multidisciplinary Team for Thoracic Tumors Tumori : Ludovini V, Gori S, Colozza M, Pistola L, Rulli E, Floriani I, Pacifico E, Tofanetti F R, Sidoni A, Basurto C, Rulli A, Crino' L Evaluation of serum HER2 extracellular domain in early breast cancer patients: correlation with clinicopathological parameters and survival Ann Oncol : Ludovini V, Pistola L, Gregorc V, Floriani I, Rulli E, Piattoni S, Di Carlo L, Semeraro A, Darwish S, Toffanetti F R, Stocchi L, Mihaylova Z, Bellezza G, Del Sordo R, Daddi G, Crino' L, Tonato M Plasma DNA, microsatellite alterations, and p53 tumor mutations are associated with disease-free survival in radically resected non-small cell lung cancer patients. A study of the Perugia Multidisciplinary Team for Thoracic Oncology J Thorac Oncol : Trotta F, Apolone G, Garattini S, Tafuri G Stopping a trial early in oncology: for patients or for industry? Ann Oncol : Ganzinelli M, Carrassa L, Crippa F, Tavecchio M, Broggini M, Damia G Checkpoint kinase 1 down-regulation by an inducible small interfering RNA expression system sensitized in vivo tumors to treatment with 5-fluorouracil Clin Cancer Res : Cecchinato V, Erba E, Basile A, Scarpati B, Fazi C, Brando B, Comi P, Chiaramonte R Hexamethylene bisacetamide inhibits malignant phenotype in T-ALL cell lines Leuk Res : Tavecchio M, Simone M, Bernasconi S, Tognon G, Mazzini G, Erba E Multi-parametric flow cytometric cell cycle analysis using TO-PRO-3 iodide (TP3): Detailed protocols Acta Histochem : Apolone G, Patarnello F Finance fibrillation and research rhythm: do we need a pace-maker? Reply to J Ambul Care Manage : Fruscio R, Colombo N, Lissoni A A, Garbi A, Fossati R, Ieda N, Torri V, Mangioni C A phase II randomised clinical trial comparing cisplatin, paclitaxel and ifosfamide with cisplatin, paclitaxel and epirubicin in newly diagnosed advanced ovarian cancer: long-term survival analysis Br J Cancer : Giusti I, D'Ascenzo S, Millimaggi D, Taraboletti G, Carta G, Franceschini N, Pavan A, Dolo V Cathepsin B mediates the ph-dependent proinvasive activity of tumor-shed microvesicles Neoplasia : Apolone G, Corli O, Greco M T, Zagonel V, CPOR SG Investigators Factors influencing the decision to take or reject opioids for cancer pain: are we on target? Ann Oncol : Porcu L, Poli D, Torri V, Rulli E, Cropalato di Tullio M, Cinquini M, Bajetta E, Labianca R, Di Costanzo F, Nitti D, Floriani I Impact of recent legislative bills regarding clinical research on Italian ethics committee activity J Med Ethics : Benedetti Panici P, Basile S, Maneschi F, Lissoni A A, Signorelli M, Scambia G, Angioli R, Tateo S, Mangili G, 31

33 Katsaros D, Garozzo G, Campagnutta E, Donadello N, Greggi S, Melpignano M, Raspagliesi F, Ragni N, Cormio G, Grassi R, Franchi M, Giannarelli D, Fossati R, Torri V, Amoroso M, Crocè C, Mangioni C Systematic pelvic lymphadenectomy vs no lymphadenectomy in early-stage endometrial carcinoma: randomized clinical trial J Natl Cancer Inst : Deandrea S, Montanari M, Moja L, Apolone G Prevalence of undertreatment in cancer pain. A review of published literature Ann Oncol : Margosio B, Rusnati M, Bonezzi K, Cordes B A, Annis D S, Urbinati C, Giavazzi R, Presta M, Ribatti D, Mosher D F, Taraboletti G Fibroblast growth factor-2 binding to the thrombospondin-1 type III repeats, a novel antiangiogenic domain Int J Biochem Cell Biol : Lucca U, Tettamanti M, Mosconi P, Apolone G, Gandini F, Nobili A, Tallone M V, Detoma P, Giacomin A, Clerico M, Tempia P, Guala A, Fasolo G, Riva E Association of mild anemia with cognitive, functional, mood and quality of life outcomes in the elderly: The Health and Anemia" study PLoS One : e1920 Mosconi P, Colombo C, Guella F, Pierotti B, Vimercati F. Are Italian medical societies bridging the distance from citizen and patients associations? Result of a survey. Journal of Preventive Medicine and Hygiene : Di Costanzo F, Gasperoni S, Manzione L, Bisagni G, Labianca R, Bravi S, Cortesi E, Carlini P, Bracci R, Tomao S, Messerini L, Arcangeli F, Torri V, Bilancia D, Floriani I, Tonato M, Italian Oncology Group for Cancer Research Adjuvant chemotherapy in completely resected gastric cancer: a randomized phase III trial conducted by GOIRC J Natl Cancer Inst : Cascinu S, Berardi R, Labianca R, Siena S, Falcone A, Aitini E, Barni S, Di Costanzo F, Dapretto E, Tonini G, Pierantoni C, Artale S, Rota S, Floriani I, Scartozzi M, Zaniboni A, GISCAD Cetuximab plus gemcitabine and cisplatin compared with gemcitabine and cisplatin alone in patients with advanced pancreatic cancer: a randomised, multicentre, phase II trial Lancet Oncol : 39-4 Apolone G, Tafuri G, Trotta F, Garattini S A new anti-cancer drug in the market: good news for investors or for patients? Eur J Cancer : Rossi A, Torri V, Gridelli C Paclitaxel plus bevacizumab for metastatic breast cancer N Engl J Med : 1637 Labianca R, Garassino M, Torri V Predicting response of molecular targeted therapies: a still possible challenge? Ann Oncol : Santini D, Loupakis F, Vincenzi B, Floriani I, Stasi I, Canestrari E, Rulli E, Maltese P, Andreoni F, Masi Gianluca, Graziano F, Baldi G G, Salvatore L, Russo A, Perrone G, Tommasino M R, Magnani M, Falcone A, Tonini G, Ruzzo A High concordance of KRAS status between primary colorectal tumors and related metastatic sites: implications for clinical practice Oncologist : Valli C, Paroni G, Di Francesco A M, Riccardi R, Tavecchio M, Erba E, Boldetti A, Gianni M, Fratelli M, Pisano C, Merlini L, Antoccia A, Cenciarelli C, Terao M, Garattini E Atypical retinoids ST1926 and CD437 are S-phase-specific agents causing DNA double-strand breaks: significance for the cytotoxic and antiproliferative activity Mol Cancer Ther : Ferketich A K, Fossati R, Apolone G An evaluation of the Italian version of the Fagerstrom Test for Nicotine Dependence 32

34 Psychol Rep : Casciani E, Polettini E, Carmenini E, Floriani I, Masselli G, Bertini L, Gualdi G F Endorectal and dynamic contrast-enhanced MRI for detection of local recurrence after radical prostatectomy AJR Am J Roentgenol : Torri V, Floriani I, Poli D, Rulli E, Ocular Hypertension Treatment Study Group, European Glaucoma Prevention Study Group (EGPS) The accuracy and clinical application of predictive models for primary open-angle glaucoma in ocular hypertensive individuals Ophthalmology : Lecchi C, Ceciliani F, Bernasconi S, Franciosi F, Bronzo V, Sartorelli P Bovine alpha-1 acid glycoprotein can reduce the chemotaxis of bovine monocytes and modulate CD18 expression Vet Res : 50 Cazzaniga M E, Pronzato P, Mustacchi G, De Matteis A, Di Costanzo F, Rulli E, Floriani I The anthracyclines and the clinical practice: do all breast cancer patients benefit? Results from the NORA study Ann Oncol : Pignon J-P, Tribodet H, Scagliotti G V, Douillard J Y, Shepherd F A, Stephens R J, Dunant A, Torri V, Rosell R, Seymour L, Spiro S G, Rolland E, Fossati R, Aubert D, Ding K, Waller D, Le Chevalier T Lung adjuvant cisplatin evaluation: a pooled analysis by the LACE Collaborative Group J Clin Oncol : Ceresoli G L, Castagneto B, Zucali P A, Favaretto A, Mencoboni M, Grossi F, Cortinovis D, Del Conte G, Ceribelli A, Bearz A, Salamina S, De Vincenzo F, Cappuzzo F, Marangolo M, Torri V, Santoro A Pemetrexed plus carboplatin in elderly patients with malignant pleural mesothelioma: combined analysis of two phase II trials Br J Cancer 2008 ; 99 : Floriani I, Rotmensz N, Albertazzi E, Torri V, De Rosa M, Tomino C, de Braud F Approaches to interim analysis of cancer randomised clinical trials with time to event endpoints: a survey from the Italian National Monitoring Centre for Clinical Trials Trials : 46 Broggini M. Nemorubicin. In: Anthracycline Chemistry and Biology II, Mode of Action, Clinical Aspects and New Drugs. Topics in Current Chemistry, Ed. K. Krohn, Springer-Verlag Berlin Heidelberg : Valentini G., D Andrea C., Ferrari R., Pifferi A., Cubeddu R., Martinelli M., Natoli C., Ubezio P., Giavazzi R. In vivo measurement of vascular modulation in experimental tumors using a fluorescent contrast agent. Photochemistry and Photobiology : Ubezio P., Cameron D. Cell kiling and resistance in pre-operative breast cancer chemotherapy. BMC Cancer, : Sala G., Dituri F., Raimondi C., Previdi S., Maffucci T., Mazzoletti M., Rossi C., Iezzi M., Lattanzio R., Piantelli M., Iacobelli S., Broggini M., Falasca M. Phospholipase C gamma1 is required for metastasis development and progression. Cancer Res (24): Leonetti C., Scarsella M., Riggio G., Rizzo A., Salvati E., D Incalci M., Staszewsky L., Frapolli R., Stevens M.F., Stoppacciaro A., Mottolese M., Antoniani B., Gilson E., Zupi G., Biroccio A. The G-quadruplex ligand RHPS4 potentiates the antitumor activity of camptothecins in preclinical models of solid tumors. Clin. Cancer Res (22): Ghilardi C., Chiorino G., Dossi R., Nagy Z., Giavazzi R., Bani M.R. Identification of novel vascular markers through gene expression profiling of tumor-derived endothelium. BMC Genomics (9): 201. Mitry E, Fields AL, Bleiberg H, Labianca R, Portier G, Tu D, Nitti D, Torri V, Elias D, O'Callaghan C, Langer B, Martignoni G, Bouché O, Lazorthes F, Van Cutsem E, Bedenne L, Moore MJ, Rougier P. Adjuvant chemotherapy after potentially curative resection of metastases from colorectal cancer: a pooled analysis of two randomized trials. J Clin Oncol (30):

35 Giavazzi R, Bonezzi K, Taraboletti G Microtubule targeting agents and the tumor vasculature In: Cancer drug discovery and development: the role of microtubules in cell biology, neurobiology, and oncology Humana Press, Totowa, NJ, 2008; LAY PRESS SELECTION PUBLISHED IN 2008 Porcu L, Floriani I GISCAD DIDATTICA. Punta sul cavallo vincente.. Medical Oncology Progress & Perspectives 2008 ; 29 : Costato A, Braun C, D'Incalci M, Pasina L, Nobili A Malati e ricercatori alleati contro il tumore gastrointestinale Partecipasalute 2008 ; Klotchenko S A, Tsymbalenko N V, Solov'ev K V, Skvortsov A N, Zatulovskii E A, Babich P S, Platonova N A, Shavlovskii M M, Puchkova L V, Broggini M The effect of silver ions on copper metabolism and expression of genes encoding copper transport proteins in rat liver Dokl Biochem Biophys 2008 ; 418 : Mangano S, Negri Emanuele, Apolone G Guida all'uso del programma. ; Il dolore nel paziente con cancro; In :Il dolore nel paziente con cancro. Riflessioni e approfondimenti dalla valutazione alle terapie Selecta Medica, Pavia, 2008; Apolone G, Corli O Prefazione In :Il dolore nel paziente con cancro. Riflessioni e approfondimenti dalla valutazione alle terapie Selecta Medica, Pavia, 2008; 5-10 Apolone G, Negri Emanuele, Mangano S, Greco M T, Montanari M Dolore nel paziente con cancro: un progetto multidisciplinare per valutare e migliorare la qualità del trattamento In :Il dolore nel paziente con cancro. Riflessioni e approfondimenti dalla valutazione alle terapie Selecta Medica, Pavia, 2008; Zagonel V, Apolone G Cancro, curare humanum est Sole 24 Ore Sanita : 37 Apolone G, Mosconi P Le politiche che regolano il mercato dei farmaci: il ruolo delle agenzie regolatorie In :La dispensa di Partecipasalute: orientarsi in salute e sanità per fare scelte consapevoli IRFMN, Milano, 2008; Apolone G, Corli O, Mosconi P Oncologi: Cure migliori se c'è la comunicazione" Sole 24 Ore Sanità 2008 n.42 : Mosconi P La Commissione oncologica nazionale e la rappresentanza dei pazienti: occasione persa? Partecipasalute 2008 Mosconi P, Liberati A, Satolli R Il progetto Partecipasalute In :La dispensa di Partecipasalute: orientarsi per fare scelte consapevoli IRFMN, Milano, 2008; IX-X Braun C, Mosconi P DHEA: l'integratore da consigliare al posto della terapia ormonale sostitutiva? Partecipasalute 2008 Pasina L, Colombo Cinzia, Mosconi P, Nobili A, Perego L, Taddei G C 34

36 Fare chiarezza sui farmaci equivalenti Ricerca & Pratica 2008 n.141 : Braun C, Mosconi P E' tempo di vacanza: la profilassi antimalarica Partecipasalute 2008 Simi S, Mosconi P Così l'italia della salute si allea con i pazienti Sole 24 Ore Sanita : Colombo Cinzia, Mosconi P L'associazionismo sta cambiando: dall'assistenza alla nascita di un progetto In :La dispensa di Partecipasalute: orientarsi in salute e sanità per fare scelte consapevoli IRFMN, Milano, 2008; Mosconi P, Satolli R, Colombo Cinzia, Liberati A, Mele A L'informazione sulla terapia ormonale post menopausale Bollettino Informazione Farmaci : Mosconi P, Colombo Cinzia, Satolli R, Donati S, Mele A, Liberati A Quando usare la terapia ormonale sostitutiva Ricerca & Pratica 2008 n.143 : 209 Mosconi P L'advocacy, voce di chi non ha voce Janus : RESEARCH ACTIVITIES Laboratory of Cancer Pharmacology Mode of action of Ecteinascidins A project ongoing since several years is about the characterization of marine natural products possessing antitumor activity. In particular we carried on the studies on the effects of ET-743 in cells defective for some DNA repair mechanisms. Cells deficient for Homologous Recombination (HR) are very sensitive to the drug, while cells deficient for Non Homologous End-Joining (NHEJ) are only slightly more sensitive, but surpraisingly cell lines defective for Nucleotide Excision Repair (NER) are less sensitive to ET-743. Flow cytometric analysis coupled to a software of computer simulation, developed in our laboratory, has demonstrated that NER defective cells showed, after ET-743 treatment, cell cycle perturbations different than those occurring in NER proficient cells, probably for the activation of different and more efficient repair mechanisms. We study also a functional evaluation of the DNA repair mechanisms by the cell capacity to recognize and repair double helix breaks with a recently introduced test that is very sensitive to detect the phosphorylation of histone H2AX. An in vitro study is ongoing with flow cytometry and immunofluorescence techniques to evaluate in different tumor cell lines the phosphorylation level of histone H2AX in relation to the distribution of the cells in the different phases of the cell cycle and the cytotoxic effect induced after treatment with ET-743. Studies are in progress on the mechanism of action of new ET-743 derivates compounds that have shown antitumoral activity on cell lines with different DNA repair mechanisms. A new project is the study of the selective action of ET-743 on mixoid lyposarcoma, a pathology representing 10% of all soft tissue sarcomas, trying to understand if the significative 35

37 antitumor effect is due to a selective action of the compound on pathogenetic alterations characteristic of this pathology. In particular we are trying to evaluate how ET-743 interact with the transcriptional modifications of specific genes due to the translocation FUS-CHOP that characterizes mixoid sarcomas or those caused by the interaction host-tumor, modifying inflammatory and angiogenetic processes. Studies are in progress to obtain cell lines and xenografts of mixoid lyposarcomas exhibiting the same molecular features of the patients tumors. Combinations of natural products of marine origin with other anticancer drugs We have observed additive or synergistic activity of ET-743 combined with other anticancer drugs such as cisplatin, doxorubicin, campthotecin and inhibitors of telomerase. Flow Cytometric analysis of the DNA content in human ovarian cancer: clinical correlations Conflicting results have been published on the prognostic significance of DNA aneuploidy on advanced ovary carcinoma (stage III or IV). The Citometry unit has reported one of the largest studies of the scientific literature indicating that the aneuploidy in the advanced ovary carcinoma is not an independent prognostic factor. In a large number of cases of stage I and II ovary tumors DNA content and the percentage of cells in S phase of the cell cycle, has been measured with flow cytometry, demonstrating that in the early stages of the disease DNA content is a prognostic factor important for ovary tumor. Analysis of cell cycle data and interactions of different drugs The Biophysics Unit is engaged in theoretical and methodological studies aimed at a critical evaluation of current techniques of investigation of drug effects on heterogeneous cell populations. Several computing tools have been produced to simulate the cell proliferation at different levels (from molecular interactions to in vivo growth of solid tumours) and the process of measure. Collaborations are ongoing with other research groups for design and data analysis of drug combination studies in vitro. In this field, a number of computer programs have been developed, allowing comparative data analysis with the most common models of drug interaction. Evaluation of the complexity of the response of cell populations to treatment with anticancer drugs This project of the Biophysics Unit addresses the issue of establishing a connection between the intracellular drug interactions and the resulting cell cycle perturbations. It starts from the singlecell level of investigation to reach the cell-population level where the relevant end points of treatment efficacy are evaluated by flow cytometry and growth inhibition/cytotoxicity assays. The model adopted for data analysis and interpretation is the result of the merging of two mathematical models. One model describes the cell cycle, exploiting the results of the theory of age-structured cell population dynamics. The second model describes the response to the drug's challenge, using distinct parameters ("effect descriptors") measuring either the strength of cell cycle arrest, damage repair or cell death in every phase (G1, S and G2M). In this way, it is possible to reach a quantitative interpretation of the experimental results, overcoming the current qualitative and partial approaches to this problem, which are unable to resolve the overlapping of cytostatic and cytotoxic effects, and to establish a connection with phase-related events. Applying this procedure we demonstrated complex but biologically consistent patterns of time and dose-dependence for each cell cycle effect descriptor, following a short treatment with melphalan on a reference cell line of ovarian carcinoma. These results add to the previously 36

38 reported studies on topotecan, cisplatin and taxol. Eventually, this project will produce a database containing the values associated to the new effect descriptors, related to few compounds but rich of information about them, especially in the dose and time dependence of the effects. This database will be used to compare the treatment response of the most common drugs adopted in the ovarian carcinoma. Cell cycle dysregulation in erlotinib-based treatments decoded by flow cytometry and mathematical modeling Epidermal growth factor receptor (EGFR) inhibitors represent one of the most promising class of anticancer compounds, some of them, like erlotinib, are already used for clinical therapy. Nevertheless, so far, the research has focused on molecular interaction of these compounds, somewhat neglecting the study of the dynamics of cell cycle perturbations and underscoring the importance of this issue for the optimization of both single and multidrug therapies. In order to fill this gap, the Biophysics Unit will apply the multidisciplinary approach, exploiting cell cycle simulation tools, to the study of cell cycle perturbations induced by erlotinib as a representative EGFR inhibitor. Studies of the time- and dose-dependence of perturbations induced by treatments are ongoing for single erlotinib treatment or for erlotinib combined with gemcitabine, irinotecan and oxaliplatin. The appreciation and the quantification of these effects will provide an important contribution to the comprehension of the mode of action of erlotinib alone or in combination. Such perspectives are vital if the events are to be decoded and used in predictive models for the exploration of pharmacodynamic actions and in understanding the origins of treatment failure. Anticancer Drug Effects Decoded by Time-Lapse Imaging, Flow Cytometry and Modeling We aim to use flow cytometric (cell-population based analysis) and time-lapse imaging (single cell lineage based analysis) techniques to generate data that will be used to predict drug responses in term of the two major determinant of cytostatic/cytotoxic actions of anticancer drugs: specific cell cycle perturbations (detecting accumulation or depletion of cells in G1, S and G2M phases) and the commitment to cell death (apoptosis). The response to a treatment with the anticancer drugs will be investigated in a human osteosarcoma cell line U2OS. engineered in order to express fluorescent reporter (a fusion cyclin B1-GFP protein), so that it is now possible to follow the cells through G1, S and G2M using time-lapse microscopy. The working hypothesis is that quantitative analysis of time-lapse microscopy data could be integrated with the information provided by flow cytometry - allowing for the first time a joint interpretation of both kind of experiments through a common mathematical model that simulate the underlying phenomena. The final goal is to provide new levels of understanding and simulation tools to the cancer research community. Timing the changes of the cellular content of specific proteins inside G1, in exponentially growing cells We developed a method for measuring the content of immunocytochemically detected proteins in individual cells progressing through G1 phase. The feasibility was demonstrated in the analysis of cyclin E levels. The sequence of G1 events is tracked in unaltered cycling conditions, in a cell line in the phase of balanced growth in vitro, to avoid the pitfalls of synchronization. The method is based on i) a bromodeoxyuridine (BrdUrd) pulse-and-chase experimental plan; ii) triparametric flow cytometric detection of DNA, BrdUrd and cyclin E; iii) data analysis supported by the basic mathematical theory of asynchronous growing populations with variable cell cycle phase durations. 37

39 Establishing a CFSE-based method for a quantitative measure of cytostatic effects of drugs on tumor cell populations Carboxyfluorescein diacetate succinimidyl ester (CFSE) is currently used to investigate migration and proliferation of hematopoietic cells. Several technical problems had precluded until now its use in the studies of the antiproliferative activity of anticancer drugs. We analysed several critical steps of the procedure, providing the way to overcome potential pitfalls. The project was eventually successful and the previous limitations were overcome. The outcome was a new standardised procedure of cytometry and data-analysis allowing a measure of the dynamics of cell cycle blockades after drug treatment. The new method was applied in the study of the drug topotecan, measuring the variability of response to treatment in terms percentage of cells immediately blocked, divided once or more times and then blocked or unaffected by the treatment. Pharmacokinetic of new taxane derivates Preclinical studies have been done on new taxane derivatives, chemically and biologically different from the conventional ones. For two of these compounds we evaluated the bioavailability after oral administration and the kinetic and metabolic profile has been entirely characterized by applying analytical methods based on HPLC/MS/MS technique developed in the Cancer Clinical Pharmacology Unit. They showed biological activity even in tumors with low susceptibility to other taxanes (cerebral tumor), suggesting a potential clinical interest. According to the hypothesis that these drugs act through an antiangiogenic mechanism, put forward by the Laboratory of the Biology and Therapy of Metastasis, it would be important to investigate prolonged chronic treatments and therefore we are investigating the pharmacokinetic properties after oral administration and after prolonged daily treatment. An interesting pk profile was found for a third compound, the 14-β-hydroxy-10- deacetylbaccatin III derivative (IDN 6140). He showed good bioavailability and high distribution in brain and glioblastoma achieving high concentration in comparison to that of plasma, demonstrating high ability to cross the blood-brain-barrier. These data make this compound of great potential interest for the therapy of Central Nervous System tumors and metastasis. Pharmacokinetic of sanguinarine The pharmacokinetic of SA, a quaternary phenanthridine alkaloid with potent antiinflammatory, antibacterial and antitumor activity, was studied in mice after single and repeated administrations, given the drug both by oral and intravenous route. Firstly, we developed an analytical methods based on HPLC/MS/MS technique to measure SA and its main circulating metabolite DHSA in plasma of mice. The drug showed low absorption, being the bioavailability lower than 10% and achieved only after the intravenous treatment, concentrations potentially able to exert cytotoxic activity. No accumulation of drug was found in plasma after repeated treatments. DHSA was the main metabolite of SA, present in plasma only at concentrations less than 10% of those of the parent drug. Clinical pharmacokinetics of gimatecan we are conducting a study in collaboration with SENDO on the clinical pharmacokinetics and pharmacodynamic of a new proteosome inhibitor, CEP18770, in cancer patients during a phase I investigation. The study is still in progress and shows that this compound is highly available, has a long half life, with consequent long exposure to the drug. Pharmacokinetic parameters will be correlated to the pharmacological data, i.e. inhibition of the proteosome activity, to define the pharmacodinamy of this compound. 38

40 Other relevant clinical studies ST The clinical pharmacokinetic of ST1926 a new oral retinoid derivative was investigated in women with ovarian cancer. In particular we studied the pharmacokinetics, the bioavailability of the drug and its metabolism during a Phase I study, providing for the first time data on the main plasma metabolite of ST 1926 (the glucuronide conjugate). Our data show variable absorption of the drug and high conversion to the glucuronide. Antitumoral activity and pharmacokinetic properties of new drugs and combinations The antitumor activity, pharmacokinetic properties and toxicity of novel anticancer drugs with specific targets (e.g. different kinase inhibitors), conventional anticancer drugs (camptothecin) and combinations is being investigated using rodent tumors and human tumor xenografts. Development of a software framework for a rationalized process of Microarrays analysis It is currently active in the Institute a multidisciplinary group involved in the rationalization of the various microarray analysis aspects, with many external collaborations. The different possible analysis procedures have been discussed, compared and formalized in order to obtain a common work flow to be accepted from the scientific community. Thanks to this activity it is now possible to automate some aspects of the analysis making them faster end better reproducible for people managing the analysis itself. This activity has involved the development of the necessary software for data analysis and the implementation of a Beowulf computer cluster, using the old computers dismissed by the desktop users in order to obtain a sufficient power. Internal collaborations: Department of Biochemistry and Molecular Pharmacology (ref.: Maddalena Fratelli, Mario Salmona) Department of Oncology (ref.: Sergio Marchini, Mariarosa Bani, Maurizio D'Incalci) External collaborations: Fondo Edo Tempia (ref.: Giovanna Chiorino) Istituto Toscano Tumori (ref: Duccio Cavalieri) Istituto Weizmann (ref. Eytan Domany) Università di Birmingham (ref. Francesco Falciani) Università di Aberdeen (ref. Tony Travis) Microarray data analysis for the Oncology Department The data analysis activities concern the biology of sensible versus resistant to therapy human ovarian carcinoma and the study of the ET-743 working mechanism in human sarcoma, sensible and resistant. Related to the projects: Department of Oncology (ref. Sergio Marchini, Maurizio D'Incalci, Massimo Broggini, Mirko Marabese) Agilent platform Metastasis/Relapses Project (phase III ovarian) LPS2 Project (cell lines, response to ET-743) Department of Oncology (ref. Mariarosa Bani, Raffaella Giavazzi) Affymetrics platform Stroma Project Parenchyma Project External collaborations: Osp. S.Gerardo, Monza (ref. Robert Fruscio) 39

41 Development of the new database handling software for the ovarian tumors bio-bank, compliant with the new privacy related laws for data handling for clinical trials and with a new reporting engine for a better data extration Internal collaborations: Department of Oncology (ref.: Sergio Marchini, Mariarosa Bani, Raffaella Giavazzi, Maurizio D'Incalci, Massimo Broggini, Mirko Marabese) External collaborations; Ospedale S.Gerardo, Monza (ref. Robert Fruscio) Implementation of the data management engine for the Italian Registry Domiciliar Artificial Nutrition, in collaboration with the Scientific Society SINPE (ref. Loris Pironi, S.Orsola, Bologna) This activity involves all aspects of the data management, statistical analysis, help desk, informatic infrastructures and software for handling an observational clinical study. For this purpose a new ibrid online/offline database has been developed, compliant with the new privacy related laws about the data protection issues. External collaborations: Osp. S. Orsola, Bologna (ref. Loris Pironi) Società Scientifica SINPE Laboratory of Molecular Pharmacology G2 checkpoint and cell cycle A new system able to specifically inhibit CHK1 expression in vivo in nude mice transplanted with human tumors has been developed. The system has been proved to be able to reduce the expression only in cancer cells. This plasmid allows the expression of the sirna only after induction with tetracycline and is therefore a unique tool to determine the effect of CHK1 inhibition in human tumors growing in nude mice following treatment with anticancer agents. The use of these cells derived from the human colocarcinoma cell line HCT116 with an inducible expression of sirna directed against CHK1, allowed us to demonstrate that in vivo too we have a nice downregulation of chk1 only when tetracycline is added in the water. Chk1 downregulation does not induce a reduction in tumor growth but induce a strong sensitisazion of the tumors to treatment with the anticancer agent 5-fluorouracil. This effect is particularly evident in p53 deficient tumors, indicating that the combination is likely to result in increased selectivity. Characterization of new potential oncosuppressor genes DRAGO gene, identified and cloned in our laboratory is one of the most interesting projects of the group. The characterization of the response of KO mice for DRAGO to ionising radiation is similar to normal mice. The characterization of the transcriptional regulation of DRAGO indicated that the gene is not only a p53-responsive gene, but, inside the p53 family, p73 has a strong ability to induce the transcription. Drago therefore, represents a new p73 responsive gene. Molecular characterization of ovarian carcinoma The molecular characterization of stage I ovarian carcinomas has been further studied. The gene expression profile analysis has showed interesting results. 40

42 It is possible to identify genes able to discriminate in ovarian cancer the different histotypes, suggesting that specific biological and molecular characteristics are responsible for the differences in morphology and clinical behaviour. These studies can help in identifying new specific molecular targets for the different subclasses of ovarian cancer that have a different clinical outcome. It is possible to identify patients with higher probability of relapse because there are genes able to differentiate these two classes of patients. We have demonstrated that stage I borderline patients have a gene expression profile similar to that of grade 1 patients, while they are well distinguishable from grade 2 and grade 3 patients. This can have important clinical implications for the treatment of this particular subgroup of patients. We are currently evaluating the differential expression of genes, microrna and proteins in tumor biopsies of patients at first surgery and after relapse, with the idea of finding genes associated with response to treatment. Expression of gene involved in DNA repair in human ovarian cancer By Real Time PCR, the expression of genes involved in DNA repair has been evaluated in 90 stage I and 90 stage III ovarian cancer. The genes analysed include those belonging to the nucleotide excision repair, in the fanconi anemia repair, in the base excision repair. In addition, genes important for the cellular response to damage, such as chk1 and claspin have been studied. The analysis that is still ongoing, suggests that there are difference in the expresison of some of the genes between early (stage I) and late (stage III) ovarian tumors. Thes information will be useful for the determinatin of the mechanisms responsible for tumor progression. We are also attempting to identify correlation between gene expression and response to chemotherapy. These data will help in identifying patients more likely to respond to platinum based therapy, which represents so far the standard therapy for this type of malignancy. Inhibition of the signal mediated by PI3K/akt In ovarian carcinoma cells PI3K gene is often overexpressed or mutated and this kinase is then constitutively activated. The consequence is the presence of proteins involved in cell survival, like akt, constitutively phosphorylated and then active. This project evaluated the capacity of tetra and pentaphosphate inositols and of some analogues to inhibit akt recruitment to the membrane and its further phosphorylation. IP5 and some new synthetic molecules, were shown to induce a reduction of the phosphorylation of akt and therefore to reduce cell growth. The activity of these molecules is not restricted to ovarian cancer, but has been shown also for other tumors such as breast and prostate. The possibility to combine PI3K inhibitors and mtor ( a kinase downstream to PI3K and akt) inhibitors has been evaluated. It has in fact been shown that some mtor inhibitors induce an aberrant phosphorylation, and hence activation of akt, and the combination of these inhibitors can block these undesired effects. The in vitro results show that the combination has at least an additive effect and open the way to find out new treatment schedules to verify whether the sequence of combination can be important for the antiproliferative effects. Role of phospholipase C γ1 in the development of metastasis We have evidentiated an important role for phospholipase C γ1, in determining metastasis formation. By generating tumor model with inducible downregulation of phospholipase C γ1, we have demonstrated in vitro a reduced motility and migration of cancer cells and in vivo a reduced formation of metastasis. The results have been obtained in different cell lines of human and murine origin. Our result strongly suggest phospholipase C γ1 as a target for the development of new molecules with antimetastatic activity. 41

43 Oncosuppressors p53 and p73 The p53 analogue, p73 is present in different isoforms derived from alternative splicing of the C-terminal. Among these isoforms there is one called DNp73 in which the transactivation domain in the N-terminal of the protein is absent. This DN form of p73 is an antagonist of p53. The DNp73 isoforms can be further processed through alternative splicing, to generate a number of isoforms with a not yet clarified biological activity. We have previously shown that the alpha form of DNp73 does not modify either the growth in vitro and in vivo of cancer cells or the response to treatment with anticancer agents. We have therefore generated clones derived from the human lung cancer derived cell line H1299 which express DNp73 beta following induction. These clones show, upon induction, interesting effects on cell growth suggesting that the presence of high levels of this isoform can have unpredicted effects. Since the data generated on the expression of p73 isoforms in cancer patients indicate that the beta isoform of DNp73 is indeed present in human cancer, the results obtained have a particular relevance and will be further analysed to verify which are the effects linked to the overexpression of this isoform. Identification of a new proteolytic mechanism of activation of p73 Another member of the p53 family, p73, has been characterised in the laboratory for its ability to modify the growth and resposne to therapy of cancer cells. We have further studied this potential tumor suppressor by studying new possible mechanisms of activation. We have found an additional regulation mechanism for the p73-alpha form that occurs through proteolytic cleavage connected to the activity of the serine protease HtrA2. Following apoptotic stimuli, HtrA2 accumulates in the nucleus and cleaves p73alpha in the C-terminal portion, enabling the protein to increase its transactivation activity on the apoptotic gene bax but not on the cell-cycle regulator gene p21. In the presence of HtrA2, p73 is more prone to cause caspase activation and nuclei fragmentation: p73 needs HtrA2 to activate and enhance its apoptotic functions. This new relation between p73 and HtrA2 may help to understand the different behavior of the p73 protein in cell physiology and in the responses of cancer cells to chemotherapy. Mechanisms of action of new antitumor drugs The mechanism of action of a new anthracycline derivative, nemorubicin (methoxy morpholino doxorubicin) has been characterised. Nemorubicin presents a pattern of antitumor activity in vitro and in vivo different from that of doxorubicin. We have found that cell lines selected for resistance to Nemorubicin have a reduced DNA repair ability particularly they show defects in the nucleotide excision repair (NER). In particular, all the cell lines selected fro resistance do not have detectable levels of XPG protein and have a collateral sensitivity to UV light, while they are maintaining the same sensitivity to ionising radiations. Furthermore, the mechanism of resistance to nemorubicin seems similar to that observed for the molecule of marine origin ET-743 and in fact cells resistant to nemorubicin are also resistant to ET-743. Cells resistant to nemorubicin maintain their resistance also when transplanted in nude mice and the tumor growing in vivo has the same molecular characteristics of the cells growing in vitro. The combintin studies between the distamycin derivative brostallicin and the demethylating agent zebularin have been completed. The rational for this combination derives from previous data generated in the lab indicating that brostallicin activity depends on the presence of GST and GSH. In tumors like prostate cancer, the GST gene (and particularly the pi isoform) is methylated and hence the protein is not expressed. In these cells the pretreatment with compounds able to revert the methylation increases the in vitro activity of brostallicin. The study has been conducted in vivo using different treatment schemes with the demethylating agent zebularin and brostallicin. We have found that zebularin, which does not have antitumor activity per se, increases the activity of brostallicin. This effect is associated with an increased 42

44 GST activity. However, from a molecular point of view, we did not find re-expression of the pi isoform of GST, probably because in these cells the gene is heavily methylated. We have found that another isoform of GST, the GST-M, is also methylated (much less than the pi isoform) and hence the increased GST activity observed could be due to re-expression of this specific isoform. Generation of new cellular systems for in vivo imaging We have generated new cell clones derived from human cancer cells growing in vitro, which stably express fluorescent or luminescent probes which can allow us to follow in vivo the growth of primary tumors and metastasis in mice. These systems generated in human ovarian, breast and prostate cancer cell lines, can be implanted in nude mice and the growth and response to therapy followed by either optical and luminescent imaging or microtac analysis. We have in particular set up models derived from human breast cancer, which are able to metastasis to the bone which can be evidentiated by optical imaging and microtc techniques in laboratory animals. Utilising different reporter genes, we have generated fluorescent and luminiscent human cancer cell lines which can be transplanted in immunodeficient mice. These cells can then be visualised in organs such as peritoneum and lungs were these cells were previously be observed only after sacrifice of the animal. The cells generated to be fluorescent or bioluminiscent will also have specific gene defects which will be useful for understanding the mechanism of action of new molecules. These systems will be particularly useful to study the antimetastatic potential of new drugs. Characterization of response of stem cells to damage The therapeutic use of stem cells is continuously increasing. This project aims to investigate the ability of stem cells isolated from umbilical cord to respond to stress induction with particular emphasis on their ability to activate checkpoint proteins. Stem cells isolated and maintained in vitro with specific cytokine cocktails able to induce partial differentiation, have shown a peculiar expression of proteins controlling the cell cycle. In particular, there is a specific time point in the differentiation process in which cell cycle checkpoints proteins are present at high levels. This could imply that this represents the time point in which these cells are more susceptible and must be protected from external damages. The characterization of these important molecular aspects will be further studied. Identification of cancer stem cells from ovarian cancer This project is aimed at isolating and characterizing a possible cancer stem cell from ovarian cancers. There are increasing evidences supporting the idea that few important multipotent cancer cells, termed cancer stem cells, are among the most relevant cells to be killed in a tumor. Normally present as quiescent cells inside the tumors, they are able to rapidly generate dividing and growing cancer cells. The current hypothesis is that normally dividing cancer cells can be preferentially killed by chemotherapy while the cancer stem cells would be more difficult to kill and would be responsible for the relapse following treatment. The possibility to identify and characterize the cancer stem cell would theoretically open the way to the selection of new generation molecules able to preferentially kill these cells. Cancer stem cells have been already identified in different human tumors including breast and hematopoietic tumors. We will focus our attention on human ovarian cancer by the use of antibodies directed against surface marker proteins to identify such potentially relevant cancer sub populations. 43

45 Determination of the impact of EGFR mutations in the activity of tyrosine kinase inhibitors in patients with NSCLC We have started a multicenter, three year project aimed at identifying those patients who can have the chance to respond to tyrosine kinase inhibitors. The study will define whether patients without mutations in the EGFR have more chance to respond to conventional therapy rather than to treatment with TKI. Our laboratory is involved in the detection of mutations of EGFR in patients with NSCLC either in the tumor tissue or in the blood of patients with a secondary aim of defining whether is possible to detect mutations in the blood circulating DNA which are representative for the tumor. DNA sequencing and scorpion arms-based PCR methods are used for these studies. Laboratory of Biology and Therapy of Metastasis Physiologic regulation of angiogenesis Angiogenesis, the formation of blood vessels from existing ones is a fundamental process in tumor progression. A delicate balance between pro- and antiangiogenic factors finely tunes this process. In the last years we have been interested in endogenous angiogenesis-regulatory factors. During 2008 we have continued the study of trombospondin-1 (TSP-1), an endogenous inhibitor of angiogenesis. TSP-1 directly binds to angiogenic factors, in particular FGF-2 (Fibroblast Growth Factor-2), inhibiting their bioavailability and activity. In a structure/function relationship analysis of different active domains of TSP-1, we have identified the FGF-2 binding site of TSP-1. This sequence is now used as a model to design new antiangiogenic compounds based on the active sequence of TSP-1. We are studying the involvement of matrix metalloproteinases (MMPs) in angiogenesis and tumor progression. In 2008, we have shown the cross-talk between MMPs and Vascular Endothelial Growth Factor (VEGF), a factor that stimulates angiogenesis and vessel permeability, during ovarian carcinoma progression. In addition we have evaluated the possibility of using VEGF inhibitors to affect MMP-dependent ovarian tumor invasion. We are currently analyzing modifications in metastasis/invasion-related gene expression profile in stroma cells induced by tumor VEGF. Lymphangiogenesis in ovarian carcinoma Lymphatic spread in early ovarian cancer is a predictor of outcome with potential clinical value. With the aim to clarify the molecular mechanisms involved in the process of lymphangiogenesis in ovarian cancer, the expression of VEGFC, a factor that stimulates lymphangiogenesis, has been measured in serum and ascites of mice bearing human ovarian carcinoma xenografts and correlated with lymphonode infiltration by neoplastic cells. To better represent the clinical features of ovarian neoplasia, an orthotopic model of human ovarian carcinoma xenograft has been created by injecting ovarian tumor cells under the bursa of the ovary. Infection of ovarian carcinoma cells with lentivirus vectors carrying the coding sequence of VEGFC, the firefly luciferease gene and fluorescent probes are ongoing; the invasive and metastatic potential of tumor cells producing VEGFC will be evaluated using optical imaging techniques, in mice bearing tumor. How the microenvironment affects endothelial cell gene expression It is fundamental to understand qualitative and functional differences between tumor and normal tissue endothelial cells (EC) and the molecular mechanisms that drive the angiogenic process. This could lead to the identification of selective markers of the vascular endothelium associated to pathologies and/or of target molecules for the development of novel therapeutic strategies. To this purpose we analysed the gene expression profile of endothelial cells isolated from ovarian 44

46 carcinoma and adrenal glands exposed or not to an angiogenic/tumor environment reconstituted in vitro. We have found that: i) the angiogenic/tumor environment is able to modulate EC gene expression ii) few genes are preferentially expressed by tumor associated endothelial cells. Preliminary results suggest that those genes might be of interest as markers of tumor endothelium. Their expression is higher in endothelial cells from tumor specimens than from normal tissues and are not expressed by tumor cells. The putative new tumor endothelial markers have been further validated as anti-cancer / vascular targets by in situ hybridization analysis of normal and tumor tissues. The putative targets (identified as mrna transcript) are being produced as recombinant proteins, with the aim to produce antibodies, directed against the recombinant proteins, suitable for immunohistochemical analyses. Preclinical models: role of Vascular Endothelial Growth Factor (VEGF) on tumor growth, vascularization and response to therapy To study the role of VEGF induced angiogenesis in the response of cancer to chemotherapy we have generated a variant of the human ovarian carcinoma A2780/1A9 by stable transfection with the VEGF121 isoform (1A9-VS1) or the antisense (1A9-VAS3). In mice 1A9-VS1 xenografts i) show dilated blood vessels sc ii) produce ascites when implanted orthotopically in the peritoneal cavity, iii) release human VEGF in the plasma, iv) the level of circulating VEGF correlates with tumor burden. We have found that the response to chemotherapy (e.g. paclitaxel) differ in xenografts producing high levels of VEGF. The blockade of VEGF, by the administration of Avastin, greatly improved the antitumor activity by paclitaxel treatment of 1A9-VS1. Studies are ongoing to elucidate the mechanism, associated to modification of the tumor microenvironment, that are responsible of these differences. This model is therefore being used to study gene expression. Based on circulating VEGF levels and morphological analysis of the tumor vasculature, samples were chosen and tissue slices of the 1A9-VS1 and 1A9-VAS3 were microdissected (PALM Microlaser system, in collaboration with the Institute of Pathology at Helios Klinikum, Germany) in order to isolate the stroma compartment to be evaluated by mean of Affymetrix s GeneChip Arrays technology. The microarray hybridisation data were analyzed with the aid of specialized software (i.e. GeneSpring and Rosetta Resolver) and differentially expressed genes were identified. Specifically, we have discovered that in the stroma of tumors 294 genes were up- and 162 were down-regulated in consequence of the VEGF produced by the cancer cells. Gene Ontology (GO) enquiry (using Expression Analysis Systematic Explorer EASE), identified overrepresented categories of potentially biologically relevant genes in the stroma of 1A9VS1 (high VEGF) tumors. Structural molecule activity, cell organization and biogenesis, and basement membrane were among the up-regulated categories. Preclinical evaluation of inhibitors of angiogenesis and vascular targeting agents Antineoplastic therapies directed against the tumor vascular system may be designed with two different strategies. Antiangiogenic therapy prevents the formation of new vessels, while vascular disrupting agents (VDA) aim to selectively destroy the already formed tumor vessels. In 2008 we have investigated the activity of several inhibitors of angiogenesis and VDA. We have investigated the antiangiogenic activity and mechanism of action of new molecules peptides or non-peptidic small molecules which mimic endogenous inhibitors of angiogenesis, including thrombospondin (TSP-1). Among the VDA, we have studied the properties of novel tubulin binding agents (analogues of colchicines and combretastatins), which cause microtubules depolymerization, selective damage 45

47 to tumor blood vessels and tumor necrosis in experimental models in vivo. In collaboration with Prof. Bellina at the Department of Chemistry, University of Pisa (Prof. Bellina and Prof. Rossi), we have screened classes of compounds with such properties. The lead compound/s will be further characterized for pharmacological and vascular targeting/antineoplastic properties. The challenge of combination The optimization of biological therapies against selective targets in combinations with chemotherapy is one of the interest of this laboratory. We are investigating small molecule receptor tyrosine kinase inhibitors, that affects angiogenesis. In 2008 we studied vandetanib, an inhibitor of VEGFR, EGFR and Ret in combination with pacltaxel (PTX) on a model of human ovarian carcinoma xenograft. In a study designed to determine the relationship between the effects of vandetanib on tumor vascular morphological and functional changes and paclitaxel uptake i)we did not find significant change in vessel number, while a reduced number of large vessels, together with an increase in the percentage of mature vessels were particularly evident after 5 days of treatment; ii) pre-treatment with vandetanib resulted in decreased tumor levels of paclitaxel within one hour from its injection, though at 24 hours the levels were comparable to controls. The combination therapy, following two different sequences of PTX administration (PTX before and after vandetanib), was more efficacious compared to each of the single agent alone. However the greatest efficacy was obtained with PTX administered before vandetanib; this outcome was enforced by enhanced fibrotic tissue reorganization in the PTX vandetanib tumors. These findings imply that the analysis of vascular changes and paclitaxel uptake in vandetanib treated tumors may assist to guide the schedule of this combination. High PAR-1 expression is associated to a malignant phenotype Protease-activated receptor-1 (PAR-1) over-expression has been associated to a variety of human cancers, and increasing evidence implicates PAR-1 as a contributor to human melanoma malignancy. We investigated human melanoma cells, isolated from lesions representing various stages of disease progression, for the expression of PAR-1 (in collaboration with Prof. Naldini at the University of Siena) and evaluated their migratory and metastatic capabilities. Cells from advanced stage melanomas expressed higher levels of PAR-1 than those from early stages. The metastatic capability showed by the melanoma cells which overexpressed PAR-1 allowed these cells to colonize the lungs in % of the mice. Accordingly, melanoma cells overexpressing PAR-1 had higher migrated (chemotaxis assay) and invading (invasion assaymatrigel) cell counts than those expressing low PAR-1. Migration and invasion were decreased by silencing PAR-1 (sirna) and treating with SCH9797, a PAR-1 specific inhibitor. Laboratory for the development of new pharmacological strategies The laboratory was born out of the consideration that the advent of oncological drugs endowed with mechanisms of action different from those of traditional chemiotherapics, introduces new treatment opportunities. At the same time, new problems arise concerning the choice of the most appropriate and effective design for research into the clinical activity profile of these new treatments. The traditional paradigm where the choice of dose is based on the maximal tolerated toxicity, and the screening of therapeutic activity focus on tumor mass reduction, may not necessarily be suitable for the evaluation of new agents whose targets may include the extracellular compartment or specific molecular targets. 46

48 The clinical development of non toxic anti tumor molecules requires a critical review of the existing models as well as of all the aspects relative to the conduction of clinical trials including: dose selection criteria, methods for determination and confirmation of pharmacological activity, and the validation of new technologies and laboratory methods. This is where the need for a profound integration of the clinical screening and the preclinical research lies. It is a prerequisite for the construction of the pharmacological rationale for the identification of the most interesting molecules, the choice of dose, the hypotheses of combination with other drugs, and of the most appropriate indicators of clinical activity. The acquisition of know how and the development and application of new designs for clinical activity studies, including the use of randomization, the introduction of groups of patients treated with placebo, and new discontinuation designs, proceed in parallel to the above. Another fundamental issue in laboratory research is the recognition that the genomic characterization of any single tumor may now play a more relevant role in drug development and treatment identification. This notwithstanding, numerous uncertainties remain regarding the role of biomarkers in drug development and in the implementation of genomic technologies in clinical trials. It is therefore necessary to improve the methodology and more biomarkers evaluation already in the early stages of research, thus shifting translational research from a simple process of correlation search to one producing knowledge regarding the predictive role of the clinical activity of the investigational treatments. Therefore, the primary focus of the laboratory is the optimization of the methods for evaluating the activity of cytotoxic drugs, but mostly for those therapies aimed at specific molecular targets, as well as the identification of factors predictive of therapeutic response. In 2008 two phase II studies exploring the activity of sorafenib in patients with pancreatic cancer and colorectal cancer have been initiated. Laboratory of Clinical Trials The Laboratory of Clinical Trials is involved in the planning, coordination and analysis of randomized clinical trials in oncology, conducted in cooperation with a network of medical oncologists. Main covered research areas are gastric, colorectal, breast and lung cancer. Moreover the laboratory works on a single arm, prospective, experimental study in children with affected by primary glaucoma, refractory to surgical procedures. The study is sponsored by the Azienda Ospedaliera Spedali Civili Di Brescia and supported by the Agenzia Italiana del Farmaco (AIFA). Gastric cancer ITACAS Intergruppo Nazionale Adiuvante Gastrico study is a multicenter, randomized, controlled, open-label, superiority, phase III trial aimed at assessing the role of adjuvant chemotherapy in the treatment of gastric cancer. It compares the efficacy and safety of a sequential treatment (irinotecan plus flurouracil/leucovorin, followed by docetaxel and cisplatin) versus flurouracil/leucovorin regimen, used as standard reference in patients with radically resected adenocarcinoma of the stomach or of the gastro-esophageal junction. The study, sponsored by Mario Negri Institute, involves 11 Italian oncologic collaborative groups and is being conducted in more than 110 Italian experimental centers. From February 2005, more than 950 patients out of the 1100 planned have been enrolled and it is expected to conclude the recruitment in the first half of Lung cancer On November 2007 a a phase III, Italian, multicentre, open label, randomized trial was started. 47

49 Aim of the study is to assess whether it is possible to optimize second-line treatment in advanced non small cell lung cancer (NSCLC) patients using biological and clinical markers. The trial compares the efficacy in terms of overall survival of erlotinib vs. docetaxel, given as second line therapy in pts without EGFR mutations. In particular, the predictive value of K- RAS mutation, EGFR protein expression and EGFR gene amplification in determining the effect of erlotinib as compared to chemotherapy will be assessed. Even if erlotinib is registered in all patients affected by NSCLC in second and subsequent lines with a small benefit, recent evidence suggest that it should be possible to select patients according with clinical and biological features. Most of these proofs are drawn from case series or post hoc analyses and not from properly planned randomized clinical trials making their interpretation controversial. EGFR mutations, EGFR copy number and EGFR expression should positive select responders, while K-RAS mutations should predict negative outcome. All these data are suggesting that a "tailored therapy" based on individual molecular features may result in better responses and optimization of sources and costs. The study, sponsored by Azienda Ospedaliera Fatebenefratelli e Oftalmico of Milan and supported by AIFA, will enroll approximately 1500 patients in 3 years. Colon cancer 3 studies are being conducted in this area On June 2007 started the accrual of a randomised, controlled, with a factorial design, phase III clinical trial aimed at identifying the best therapeutic adjuvant strategy in radically resected colon cancer patients. The study, sponsored by Fondazione Giscad per la Cura dei Tumori and supported by AIFA, will address the following two questions: 1) Optimal duration of FOLFOX-4 regimen (3 vs 6 months), by means a non-inferiority trial between 3-month FOLFOX-4 vs. a 6-month FOLFOX-4, considered as the standard treatment. 2) benefit of the addition of bevacizumab to FOLFOX-4 regimen, only in high risk stage III patients. For both questions, primary efficacy endpoint will be recurrence free survival. On February 2009 more than 120 Italian centres have been involved with the enrollment of approximately 700 patients out of the targeted A multicenter, randomized, controlled, open-label, superiority, phase III study is now starting aimed at compare the efficacy of the addition of cetuximab to FOLFIRI vs. FOLFIRI alone given as first line therapy in patients K-RAS wild type with advanced colorectal cancer. In particular, the predictive value of PTEN mutation will be assessed in determining the effect of cetuximab+folfiri as compared to chemotherapy alone. The efficacy will be evaluated in terms of progression free survival. This study foresees the involvement of approximately 30 experimental centers and the enrollment of 300 K-RAS non mutated patients. Another open-label, randomized, parallel group, phase III, multicenter trial, sponsored by Fondazione Giscad per la Cura dei Tumori and supported by AIFA, started. Aim of this study is to compare the efficacy and safety of two different sequences of chemotherapeutic agents (Irinoteca/Cetuximab followed by FOLFOX-4 vs. FOLFOX-4 followed by Irinotecan/Cetuximab) in order to optimize the treatment of patients with metastatic colorectal cancer progressed to a first line chemotherapy with FOLFIRI and bevacizumab. Primary endpoint will be overall survival. The maximum estimated study duration is approximately 52 months and about 350 patients will be enrolled. Breast cancer The TOP (Trastuzumab Optimisation trial) study is aimed at increasing the knowledge on the efficacy of herceptin in the treatment of locally advanced or metastatic breast cancer patients. It includes two randomised, open label, phase III clinical trials. The first is aimed at assessing 48

50 whether a maintenance therapy with herceptin is superior in terms of progression free survival to no further herceptin treatment in patients with locally advanced and/or metastatic breast cancer over expressing HER2 and not progressed to a first line chemotherapy containing trastuzumab. The second evaluates whether a second line chemotherapy with trastuzumab after failure of a first line regimen of chemotherapy plus trastuzumab is superior in terms of overall survival to chemotherapy alone in patients with locally advanced and/or metastatic breast cancer over expressing HER2. The results of TOP study will allow to evaluate the cost/benefit ratio of herceptin treatment and to optimize the efficacy of such a drug, already approved in Italy, but used without clear data supporting evidence of benefit in the considered setting. This study is sponsored by Regione Lombardia and supported by AIFA. Head and neck On March 2008 started the accrual of a randomized, multicenter, open-label, with a factorial design, phase III trial evaluating the overall survival in patients with locally advanced squamous cell carcinoma of head and neck treated with loco-regional treatment (radiotherapy plus concomitant chemotherapy or cetuximab) with or without neo-adjuvant chemotherapy. Patients will be randomized to receive 3 cycles of neo-adjuvant chemotherapy (TPF - docetaxel, cisplatin e 5-flurouracil) followed by radiotherapy plus concomitant chemo or cetuximab, or radiotherapy plus concomitant chemo or cetuximab alone. The primary objectives of the study are to compare the overall survival between neo-adjuvant and no neo-adjuvant arm and to compare the toxicity between concomitant chemo-radiotherapy and radiotherapy plus cetuximab. This study, sponsored by AVAPO (Associazione Volontari Assistenza Pazienti Oncologici) Ricerche of the Ospedale Civile SS. Giovanni e Paolo, Venezia, will be conducted by a multidisciplinary team, composed by oncologists, radiotherapists and othorinolaryngologists and will enroll approximately 350 patients in Italian centers. Laboratory of Translational and Outcome Research in Oncology The Laboratory is mainly aimed at documenting, by using either Systematic Literature Review, Randomized or Outcome Research studies, the value of new diagnostic and therapeutic interventions in oncology, paying particular attention to two critical steps: the passage from early to late clinical research (from the activity to efficacy evaluation) and from phase III to clinical practice (from efficacy to effectiveness). The principal lines of research are three: cancer pain evaluation, clinical research on gynecologic cancers and evalution of the effectiveness of complex clinical programs in oncology care. In order to facilitate the research activities and optimize the outputs, the Laboratory hosts the Coordination Centers of two multidisciplinary Groups (MANGO: Mario Negri Gynecologic Oncology and the CP-OR: Cancer Pain Outcome Research Study Groups). As from 2007 on, all the activities of research and training in the field of chronic pain has been coordinated by a dedicated center (CERP:Center for the Evaluation and Research on Pain). CERP The objective of this newly set up center is in line with the Mario Negri Institute s purpose: CERP is aimed at advancing the scientific knowledge of chronic pain and particularly the cancer pain and at improving the quality of palliative care. Activities concerns three fields: preclinical and clinical research, education and information. Multidisciplinary and multiinstitutional approach are used with special care to pharmacologic therapy. Multidiscipinary teams have been created to value any contribution from either physicians or patients and with the involvement of scientific societies and patients associations. Several educational and clinical activities focusing on patients with cancer pain are currently on going and are being conducted with both private and public funds. Major activities the following: 49

51 - Analysis and dissemination of results from the Outcome Research study carried out between Design of a large 4-arm multicenter RCT comparing the efficacy of 4 strong opioids that will be launched in Implementation of several educational courses for members of the CPOR-SG addressing epidemiological statistical and methodological topics - Implementation of new experiments in vivo, in collaboration with the Laboratory of Experimental Psychology to test new approaches in animal models - Collaboration with the Laboratory of Medical Research and Consumers Involvement in the implementation and coordination of a population-based information and education project to change the knowledge, opinions, attitudes and utilization of opioid drugs in cancer patients that is at present ongoing in an Italian Region (Le Marche). - Activation of collaborations with European Research Networks As to the first point, in the context of a wide multidisciplinary project, a nationwide multicenter, prospective outcome research study was launched in Italy in 2006 to investigate the epidemiology of cancer pain, the pattern and quality of analgesic-drug therapy, and the evolution of health outcomes over time. In a large, prospective, cohort of advanced cancer patients reporting pain, investigators collected predictive and prognostic variables, information about type of care, as well as several patientreported-outcomes, such as pain, quality of life, satisfaction with analgesic care using standardized questionnaires and data collections forms. 110 centers recruited 1801 patients from February 2006 to March Preliminary results were presented to investigators during a meeting held at the Mario Negri Institute on December Final results, reported on 4 papers already published or forthcoming, document that most of patients did not receive exhaustive information about prognosis, the prevalence of undertreatment was quite high at the time of study inclusion (up to 45%), and that analgesic drugs prescribed by physicians to control cancer pain awere effective (reducing the average pain reported by patients of about 35%) but with some differences in terms of dosages and side effects. These results are the basis for the design of the next RCT that has the objective to confirm these preliminary findings. The study will involve 80 centers and about 1000 patients. The collaborative group in clinical gynecologic oncology named MaNGO The Mario Negri Gynecologic Oncology group (MaNGO) is a new name for a collaborative group that has been active in clinical gynecologic oncology for several years. Infact, this group consolidated its network and logistics while running the ICONs studies which were conducted in very close partnership with researchers at the Medical Research Council, Clinical Trial Unit, UK. MaNGO was formally set up in May 2006 and is mainly representative of the northern part of Italy, although there are important sites in the central and southern part of the country too. Participating centers are either general public and private hospitals or university clinics. One of MaNGO s main statutory objectives was to foster an active collaboration with the Gynecologic Cancer Intergroup (GCIG), a true International Forum that circulates the scientific proposals from fifteen collaborative groups through ten countries. In 2008, two randomized clinical trials in ovarian cancer completed the inclusion phase, one sponsored by a French group (CALYPSO study) and the other sponsored by EORTC (TARCEVA study). MaNGO launched the PORTEC 3 study in Italy: this is a randomized phase III trial in endometrial cancer promoted by the Dutch collaborative. In 2008 MaNGO was involved in finalizing the protocols of two randomized clinical trials in ovarian cancer with new antiangiogenic drugs. These trials should be internationally coordinated by the German onco-gynecologic group named AGO 50

52 During 2008, MaNGO s Technical-Scientific Committe met quarterly while MaNGO affiliates were conveyed in one General Assembly and new representatives of the TS Commitee were elected. Colo-rectal cancer The assessment of efficacy of screening for relapses of colorectal carcinoma has been debated for a long time, with controversial results. GILDA is an open label, international, randomised study comparing two different strategies of post surgical surveillance in colorectal cancer (Dukes B2-C stage): minimalist versus intensive. Primary endpoints of this trial are disease free survival (which is used to assess diagnostic anticipation of metastases), overall survival, health related quality of life, direct and indirect costs evaluation. At present, GILDA trial is the largest randomised study evaluating the efficacy of two follow-ups in colorectal carcinoma. The trial was closed to patient entry in September 2006 when a total of 1200 patients had been enrolled. At the end of 2008 a manuscript for an Italian journal has been prepared to present the progress of the trial and in 2009 the final analyses will be submitted to an international peer-review journal. Ovarian cancer: clinical and translational studies During 2008, the Unit of Gynecology-Oncology has coordinated the participation of a selected network of Italian hospitals to two international randomized clinical trials. The CALYPSO trial was sponsored by ARCAGY, France and compared two chemotherapy regimens (carboplatintaxol vs carboplatin- pegylated liposomal doxorubicin ) in patients with epithelial ovarian cancer in late relapse. The TARCEVA trial was sponsored by EORTC and evaluated the impact of adding erlotinib for two years in patients with no evidence of progression after first line, platinum-base chemotherapy for ovarian cancer. Both studies reached their target sample size well in advance to the anticipated date of recruitment closure, thanks to really succesful international collaboration. Results will be available in about two years. During 2008, the Gynecology-Oncology Unit has started the collection of retrospective data aimed to describe the therapeutic approaches used to manage the ovarian cancer recurrence that develop between 6 and 12 months from the end of a first line platinum-based chemotherapy. This study should give clues to optimize the choice of drug combination in this specific clinical setting. MaNGO will be a partner in the European network involved in two randomized clinical trials sponsored by the German onco-gynecologic group AGO. These two trial wills be exploring the effect of two angiogenic drugs, BIBF 1120 and pazopanib, in combination with the standard first line chemotherapic treatment (carboplatinum-taxol) of ovarian carcinoma. The Unit of Gynecology-Oncology has prepared an ancillary translational proposal to assess a panel of circulating proteins (VEGFC, VEGFB, VEGFA and their soluble receptors VEGFR2 and 3)that are possible critical signalling effectors of the angiogenic pathway. Endometrial cancer: clinical studies In 2008, the Gynecology Oncology Unit has started randomization for the PORTEC 3 protocol. This is an international randomized phase III sponsored by the Dutch Cooperative Gynecologic Oncology Group and it is aimed at comparing concurrent chemoradiation and adjuvant chemotherapy with pelvic radiation alone in high risk and advanced stage Endometrial Carcinoma. During 2008 a prospective observational study, assessing the value of trans-vaginal ultrasound imaging in predicting the myometrial infiltration of endometrial carcinoma, has been launched and recruitment is expected to last for 12 months. Should this diagnostic procedure show satisfactory concordance rate with final histopathological examination, clinicians could use it to improve surgical planning. In 2008 the results of a coordinate effort with the Sapienza University, Rome was published in the Journal of the National Cancer Institute. It was a randomized clinical trial that demonstrated that the systematic pelvic lymphadenectomy does 51

53 not impact on disease free survival and overall survival of patients with early endometrial cancer. In the end of 2008, A draft of a manuscript of a prospective pooling of the data with a trial of the Nordic gynaecologic oncology group (chemoradiotherapy versus radiotherapy alone in high risk endometrial cancer) has been prepared. This manuscript will be finalized in early Outcome research project in cancer pain In the context of a wide multidisciplinary project (J. Ambulatory Care Manage 29: ,2006), a nationwide multicenter, prospective outcome research study was launched in Italy in 2006 to investigate the epidemiology of cancer pain, the pattern and quality of analgesic-drug therapy, and the evolution of health outcomes over time. In a large, prospective, cohort of advanced cancer patients reporting pain, investigators collected predictive and prognostic variables, information about type of care, as well as several patientreported-outcomes, such as pain, quality of life, satisfaction with analgesic care using standardized questionnaires and data collections forms. 110 centers recruited 1801 patients from February 2006 to March Subjects were monitored for 28 days and then with a simplified scheme for further 8 weeks. At inclusion, 50% had bone metastasis, 73% a level of pain classified as moderate-severe, 48% reported episodes of breakthrough pain, 49% were still on active anti-cancer treatments and 60% were already on treatment with strong opioids. When the Index of Pain Management was computed to provide a rough estimate of how pain was treated (Cleeland et al, NEJM 330: , 1994), up to 45% had negative values suggesting a possible analgesic under-treatment, with large variations according to a selected list of clinical variables. In the sub-sample of patients with a complete follow-up at 28 days (no.=1461), all pain and palliative outcomes did significantly improve on average, with variations according to case mix, type of treatments and type of recruiting centers. Outcomes based on worst and average pain intensity showed the higher effect size estimates (0.84 and 0.69) when compared to patients' satisfaction and quality of life (0.44, 0.35). Up to 26% of patients were classified as non-responders. This outcome research study carried out at national level produced data to help implement future educative and research activities in Italy. Other research activities During 2008, other activities on patient-oriented clinical translational research in oncology were carried out. The focus was on the improvement of transferring information from the pre-clinical to clinical and research setting, and from clinical research to clinical practice and public health. Most of this work involves data-entry, storage and other computerized applications for the management of large and complex databases of biological and clinical data. In addition to the methodological and bio-informatics issues that are relevant in this area, particular attention was also given to issues related to the ethical and legal issues pertaining the collection, storage and utilization of biological samples from patients and citizens. Finally, we would like to mention two research projects that deal with the problem of testing the effectiveness of complex clinical programs in the context of public health using formal RCT. Evalaution of the effectiveness of long-term follow-up for early stage breast cancer Despite the lack of evidence that post-operative surveillance programs improve the quality of care and eventually the outcomes of breast cancer patients after primary curative therapy, variations in practice patterns exist (between and across nations and clinical settings), with a significant use of quite intensive programs. Most physicians favor intensive surveillance programs assuming that detecting disease recurrence as its earliest stage would offer the chance 52

54 of cure and improve survival and quality of life. As a final result, in addition to frequent visits and useless routine blood tests, complex and costly interventions such as imaging studies, tumor markers, PET and breast RMI are required for unselected women, thus consuming medical resources, increasing costs and creating long waiting lists. The Project is based on the hypothesis that to change the current attitude to use intensive follow-up programs, it is necessary to involve as many as possible stakeholders and to produce new pieces of evidence addressing the issues related to all relevant barriers, including nonscientific or non-medical drivers, such as organization and health-care factors. In particular, the Project has the main objective to develop specific follow-up/surveillance strategies that are targeted to specific sub-groups of patients having different risk of experiencing disease recurrence, and to assess, using a comparative randomized approach, their yield in terms of diagnostic anticipation, recurrence-related clinical events, quality of life and satisfaction. In this context, in addition to carry out a RCT to assess the yield of a very intensive follow-up regime in a high risk women with breast and endometrial cancer, the Project will test in the Italian setting the hypothesis that a routine minimal follow-up program carried out by family physicians (General Practitioner,) is a safe and effective alternative to follow-up carried out by specialist (basically, oncologists), in low risk women. The project, sponsored by The Italin Ministry of Health, involve 5 teams in 5 Italian Regions, started in 2008 and is coordinated by the Laboratory of Translational and Outcome Research. Laboratory of Medical Research and Consumer Involvement This Laboratory promotes activities of research in the field of involvement of citizens and patients and their associations in decision making in health and medical issues. Moreover the activities of this laboratory include: researches on information conveyed to patients on illness and treatment, implementation of web site on the topics of the health and the information ( strategy of involvement of groups of patients for the publication of educational material; research on the evaluation of the quality of the life and satisfaction with care through studies on ad hoc selected groups, and implementation on validated ad hoc questionnaires. PartecipaSalute ( Participate in Health Care ) - fostering a strategic alliance between consumers associations and medical community The project was born in It is coordinated by Mario Negri Institute, with the collaboration of the Italian Cochrane Centre and Zadig, an editorial and publishing company. It is supported by Compagnia di San Paolo, a non profit private-law foundation. During 2008, training courses for consumers, surveys and the website were developed. Training courses: The 3 th edition of the training course Informed decision making in healthcare, targeted to patients associations representatives started in September Thirty people attended the course. A course for lay members of ethic committees was also organized, targeted to the ethic committees of the Regione Lombardia. Twenty people attended the course. The duplicated lecture notes of the course were published and printed. Surveys: In 2007 a 14 items questionnaire has been posted to 147 patients associations entitled Does clinical research answer to patients needs? Considering the results of this survey, a questionnaire specifically targeted to pediatric associations about the relevance of the research was developed in 2008 together with the Laboratorio per la salute materno infantile. 53

55 The website: Three areas with access registration restricted to different working groups were set up. The working groups are: the jury of the Consensus Conference Informing women about hormone replacement therapy ; lay members of ethic committees attending the course; patients associations representatives attending the course. New tools to critically evaluate healthcare information and issues were discussed and developed: the misura-campagne, to evaluate public awareness campaigns; the misura-medico, to evaluate medical practitioners. This tool will be reviewed by a sample of medical practitioners and lay people. We set up a channel on You tube ( and another on Slideshare.net ( in order to share videos and power point presentations. The monthly visits to the websites were on average Consensus conference Informing women about hormone replacement therapy The Partecipasalute project together with the National Guidelines System (SNLG) based at the Istituto Superiore di Sanità, organized the Consensus conference Informing women on hormone replacement therapy in order to assess the current status of the quality of information on hormone replacement therapy (HRT) and re-visit recent research findings on its risks/benefits. We chose a structured approach based on the traditional consensus conference method combined with a structured preparatory work supervised by an organizing committee and a scientific board. The organizing committee and scientific board chose the members of the CC s jury and appointed three multidisciplinary working groups composed by healthcare professionals, journalists, citizens and patients representatives. Before the CC, the three working groups carried out: a literature review on the risk/benefit profile of HRT and two surveys on the quality of information on lay press and booklets targeted to women. A population survey on women s knowledge, attitude and practice was also carried out. The jury received the documents in advance, listened the presentations during the two-day meeting of the CCs, met immediately after in a closed-door meeting and prepared the final document, available at The public session of the conference took place in Turin on 16 and 17 May The project was supported by Compagnia di San Paolo, a non profit private-law foundation. Programma 1, WP5 -Alleanza contro il cancro Servizio nazionale di accoglienza e informazione sul cancro. Gruppo per l'informazione ACCP1 WP5 The project is coordinated by the Istituto superiore di sanità. Other partners are the Centro di Riferimento Oncologico - Aviano, AIMaC Associazione italiana malati di cancro, Istituto Nazionale dei Tumori, Istituto Europeo di Oncologia. The Laboratory of Medical research on consumer involvement planned to evaluate the quality of websites dealing with breast cancer, colon rectal cancer, cervical cancer. The focus on internet is due to the increasing number of people searching for information about cancer, treatments and diagnosis on the web. The protocol was draft on June 2007 and the evaluation form was defined on October The websites included in the sample were collected through a research by key words on the search engine google (November 2008). Each website will be evaluated by two reviewers. SNAP project - Smoke, Nutrition, Alcohol and Physical Activity SNAP is a campaign for the health addressed to employees in a firm in Brianza. This project, for want of FSE - Frontier Science & Technology Research Foundation, Southern Europe, a foundation for the support to the independent search - in collaboration with Istituto Mario Negri, has been launched with the attempt to increase knowledge and to modify the opinions, the attitudes and the behaviors of the people on the four topics examined, with an active process of information addresses to increase to the knowledge and the information. The formationinformation plan is structured through the circulation of paper material and a public event of 54

56 discussion. In order to estimate the effectiveness of this plan, a form on knowledge, opinions and behaviors has been carried out before and three months after the discussion. The data of the first survey have been already analyzed and have been introduced during the event in October 2008, to notice the high rate of participation: 313 employees on 522 have answered the form. The data on the follow-up of the second form will be analyzed and confronted with a before-after analysis methodology. A population based evaluation of an intervention to improve cancer pain management In collaboration with the ASUR of the Marche region, it has been promoted a project that aimed to improve the use of opioids cancer patients. The project - carried out in collaboration with the laboratory of Giovanni Apolone - has been sponsored by the Italian Agency for Drug evaluation (AIFA). The aim of this project is to pull down the existing barriers on the use of these drugs and to help patients to deal, in the better way, with cancer pain. The population, practitioners, oncologists, chemists, psychologists, nurses, patients and cancer voluntary associations will be involved in this project. In the course of the project, formative and informative participations and distribution of popular and scientific material (depending on the target) are previewed and surveys on opinions, attitudes and behaviors of people will be organized. Throughout this year, all the operative phases have been activated: from the draft of informative pamphlets addressed to health workers and those for patients and their families, as well as patients organizations, to the preparation of training courses, to the draft of a pocket book addressed to doctors, concerning interactions between opioids and other drugs, to draft of forms for surveys, involving patients and general practitioners. Follow-up in oncology setting Two studies on follow-up have been designed and carried out in collaboration with the laboratory of Giovanni Apolone. The first in collaboration with the Network Oncologica Piemontese regards the follow-up of patients with endometrial cancer organization for which the evidence available is not sufficient to draw a path of sure effectiveness. TOTEM study that has the characteristics of an open randomized multicenter study comparing two different modulations of visits and examinations and in 2008 was discussed and developed the protocol and all related forms, including from the point of fair and comprehensive information to patients who will be invited to participate in the study. The second study that takes place in the context of the 6th Integrated Project Oncology (Health Ministry) provides for the comparative assessment of two follow-up for women at moderatelow risk with a diagnosis of breast cancer and lead to a randomization minimalist follow-up coordinated by the oncologist or by general practitioner. The study will be operational from Quality of life projects No specific research projects have been carried out on quality of the life evaluation. However are on going the activities of support and coordination of other groups using the instruments of quality of life translated and validated by our research group, SF-36, SF-12, PGWBI. During the year it has been periodically up-to-date the specific website 55

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58 DEPARTMENT OF ENVIRONMENTAL HEALTH SCIENCES STAFF Head Roberto FANELLI, Biol.Sci.D. Laboratory of Analytical Biochemistry Head Chiara CHIABRANDO, Biol.Sci.D. Laboratory of Environmental Chemistry and Toxicology Head Emilio BENFENATI, Chem.D. Industrial and Environmental Health Unit Head Laboratory of Food Toxicology Marco LODI, Chemist Head Ettore ZUCCATO, M.D. Laboratory of Mass Spectrometry Head Enrico DAVOLI, Anim.Sci.D. Laboratory of Molecular Toxicology Head Protein and Gene Biomarkers Unit Head Luisa AIROLDI, Pharm.D. Roberta PASTORELLI, Biol.Sci.D Department s Units Environmental Pollutants Risk Assessment Unit Head Elena FATTORE, Biol.Sci.D Analytical Instrumentation Unit Head Renzo BAGNATI, Chem.D. 57

59 CURRICULA VITAE Roberto Fanelli, Head of the Environmental Health Sciences Department since 1997, Laboratory Head , Researcher , Research fellow at the Mario Negri Institute. Doctoral Degree in Biological Sciences (University of Milan, 1973), Assistant Professor in Biochemistry at Baylor College of Medicine (Houston, Texas). Member of the Commissione Consultiva Prodotti Fitosanitari (Ministero Salute), Member of the Scientific Panel on Contaminants in the Food Chain (European Food Safety Authority, ), Certified Italian Toxicologist. Member of the Comitato Scientifico Ente Risi. Research areas: Sources, diffusion, toxicology, human exposure and risk assessment of persistent environmental pollutants. Environmental risk of plant protection products. Development of analytical methods for identification and measurement of biomarkers in toxicology. Mechanisms of toxic action by proteomic techniques. Selected publications: Castiglioni S, Zuccato E, Chiabrando C, Fanelli R, Bagnati R. Mass spectrometric analysis of illicit drugs in wastewater and surface water. Mass Spectrom Rev 2008 ; 27 : Zuccato E, Chiabrando C, Castiglioni S, Bagnati R, Fanelli R. Estimating community drug abuse by wastewater analysis. Environ Health Perspect 2008 ; 116 : Hodgson S, Thomas Laura, Fattore E, Lind P M, Alfven T, Hellstrom L, Hakansson H, Carubelli G, Fanelli R, Jarup L. Bone mineral density changes in relation to environmental PCB exposure. Environ Health Perspect 2008 ; 116 : Pastorelli R, Carpi D, Campagna R, Airoldi L, Pohjanvirta R, Viluksela M, Hakansson H, Boutros P C, Moffat I D, Okey A B, Fanelli R. Differential expression profiling of the hepatic proteome in a rat model of dioxin resistance: correlation with genomic and transcriptomic analyses. Mol Cell Proteomics 2006; 5: Zuccato E, Chiabrando C, Castiglioni S, Calamari D, Bagnati R, Schiarea S, Fanelli R. Cocaine in surface waters: a new evidence-based tool to monitor community drug abuse. Environ Health 2005; 4: 14 ( 2005) Pastorelli R, Carpi D, Airoldi L, Chiabrando C, Bagnati R, Fanelli R, Moverare S, Ohlsson C. Proteome analysis for the identification of in vivo estrogen-regulated proteins in bone. Proteomics 2005; 5: Luisa Airoldi, Head of the Molecular Toxicology Laboratory since 1994, Unit Head , Researcher , Technician at the Mario Negri Institute. Doctoral Degree in Pharmacy (University of Milan, 1975), Postdoctoral fellow at the Massachusetts Institute of Technology (Cambridge, MA, 1976) and at the Northwestern University Medical School (Chicago, Il, 1977), Researcher at the Yale University Medical School (New Haven, CT, ). Research areas: Proteomics in toxicology with particular interest on the study of proteome changes in tissues and biological fluids from animals and humans after exposure to toxic compounds; clinical proteomics aimed at the identification of protein biomarkers as diagnostic tools; molecular epidemiology focused on the identification and measurement of biomarkers of exposure to environmental carcinogens and disease susceptibility. Selected publications: Peluso M, Airoldi L, Colombi A, Munnia A, Veglia F, Autrup H, Dunning A, Garte S, Gormally E, Malaveille C, Matullo G, Overvad K, Raaschou-Nielsen O, Clavel-Chapelon F, Linseisen J, Boeing H, Trichopoulou A, Palli D, Krogh V, Tumino R, Panico S, Bueno-De-Mesquita H B, Peeters P H, Kumle M, Gonzalez C A, Martinez C, Dorronsoro M, Barricarte A, Tormo M J, Quiros J R, Berglund G, Janzon L, Jarvholm B, Day N E, Key T J, Saracci R, Kaaks R, Riboli E, Bingham S, Vineis P. Bulky DNA adducts, 4-aminobiphenyl hemoglobin adducts, diet and air pollution in a healthy European population. Br J Nutr : Veglia F, Loft S, Matullo G, Peluso M, Munnia A, Perera F, Phillips D H, Tang D, Autrup H, Raaschou-Nielsen O, Tjonneland A, Vineis P, Genair-EPIC Investigators. DNA adducts and cancer risk in prospective studies: a pooled analysis and a meta-analysis. Carcinogenesis 2008 ; 29 : Vineis P, Hoek G, Krzyzanowski M, Vigna-Taglianti F, Veglia F, Airoldi L, Overvad K, Raaschou-Nielsen O, Clavel- Chapelon F, Linseisen J, Boeing H, Trichopoulou A, Palli D, Krogh V, Tumino R, Panico S, Bueno-De-Mesquita HB, Peeters PH, Lund E E, Agudo A, Martinez C, Dorronsoro M, Barricarte A, Cirera L, Quiros JR, Berglund G, Manjer J, Forsberg B, Day NE, Key TJ, Kaaks R, Saracci R, Riboli E. Lung cancers attributable to environmental tobacco smoke and air pollution in non-smokers in different European countries: a prospective study. Environ Health ; 6:7. Pastorelli R, Saletta F, Campagna R, Carpi D, Dell'Osta C, Schiarea S, Vineis P, Airoldi L, Matullo G Proteome characterization of a human urothelial cell line resistant to the bladder carcinogen 4-aminobiphenyl Proteome Sci : 6 58

60 Pastorelli R, Carpi D, Campagna R, Airoldi L, Pohjanvirta R, Viluksela M, Hakansson H, Boutros P C, Moffat I D, Okey A B, Fanelli R. Differential expression profiling of the hepatic proteome in a rat model of dioxin resistance: correlation with genomic and transcriptomic analyses. Mol Cell Proteomics 2006; 5: Airoldi L, Vineis P, Colombi A, Olgiati L, Dell'Osta C, Fanelli R, et al. 4-Aminobiphenyl-hemoglobin adducts and risk of smoking-related disease in never smokers and former smokers in the European Prospective Investigation into Cancer and Nutrition Prospective study. Cancer Epidemiol Biomarkers Prev 2005; 14: Emilio Benfenati, Head of the Laboratory of Environmental Chemistry and Toxicology since 1997, Unit Head , Researcher , Research fellow at the Mario Negri Institute. Researcher at Istituto Biochimico Italiano Doctoral Degree in Chemistry (University of Milan, 1979). Member of Commissione Consultiva Prodotti Fitosanitari (Ministero Salute ), Certified Italian Chemist. Resarch areas: Computer-based models for chemistry and toxicology; Molecular descriptors; QSAR; Toxicity prediction; Metabolism studies; Characterization and assessment of wastes, industrial effluents, emissions from landfill and incinerator; Integration of chemical analysis and eco-toxicological data; Chemical analysis of organic compounds by mass spectrometry. Principali pubblicazioni: Zhao C, Boriani E, Chana A, Roncaglioni A, Benfenati E, A new hybrid QSAR model for the prediction of bioconcentration factors (BCF), Chemosphere : Fjororova N, Novich M, Vrachko M, Kharchevnichova N, Zholdakova Z, Sinitzyna O, Benfenati E, Regulatory assessment of chemicals within OECD Member Countries, EU and in Russia, J Environ Sci Heal C : Roncaglioni A, Benfenati E, In silico-aided prediction of biological properties of chemicals: oestrogen receptor-mediated effects, Chem Soc Rev : Porcelli C, Boriani E, Roncaglioni A, Chana A, Benfenati E, Regulatory perspectives in the use and validation of QSAR. A case study: DEMETRA model for daphnia toxicity, Environ Sci Technol : Pandelova M, Henkelmann B, Roots O, Simm M, Jarv L, Benfenati E, Schramm K-W, Levels of PCDD/F and dioxin-like PCB in Baltin fish of different age and gender, Chemosphere : Benfenati E (Ed.), Quantitative Structure-Activity Relationships (QSAR) for Pesticide Regulatory Purposes, Elsevier Science Ltd, Amsterdam, The Netherlands (2007), Chiara Chiabrando, Head of the Analytical Biochemistry Laboratory since 1997, Unit Head , Researcher , Research fellow at the Mario Negri Institute. Doctoral degree in Biological Sciences (University of Milan, 1974), Postdoctoral fellow at the Baylor College of Medicine (Houston, Texas, ). Postgraduate degree in Pharmacological Research, Mario Negri Institute (1977). Research areas: Development and application of bio-analytical methods based on mass spectrometry in the fields of biochemistry, metabolism, clinical chemistry and pharmacology. Identification and characterization of proteins and peptides of biomedical interest by proteomic approaches and mass spectrometry. Proteomics in oncology. Selected publications Macconi D, Chiabrando C, Schiarea S, Aiello S, Cassis L, Gagliardini E, Noris M, Buelli S, Zoja C, Corna D, Mele C, Fanelli R, Remuzzi G, Benigni A. Proteasomal processing of albumin by renal dendritic cells generates antigenic peptides. J Am Soc Nephrol. 2009; 20: Epub 2008 Dec 17. Zuccato E, Chiabrando C, Castiglioni S, Bagnati R, Fanelli R. Estimating community drug abuse by wastewater analysis. Environ Health Perspect 2008; 116: Castiglioni S, Zuccato E, Chiabrando C, Fanelli R, Bagnati R. Mass spectrometric analysis of illicit drugs in wastewater and surface water. Mass Spectrom Rev 2008; 27: Castiglioni S, Zuccato E, Crisci E, Chiabrando C, Fanelli R, Bagnati R. Identification and measurement of illicit drugs and their metabolites in urban wastewaters by liquid chromatography tandem mass spectrometry (HPLC-MS-MS). Anal Chem 2006;78: Pastorelli R, Carpi D, Airoldi L, Chiabrando C, Bagnati R, Fanelli R, Moverare S, Ohlsson C. Proteome analysis for the identification of in vivo estrogen-regulated proteins in bone. Proteomics. 2005; 5: Chiabrando C, Rivalta C, Bagnati R, Valagussa A, Durand T, Guy A, Villa P, Rossi JC, Fanelli R. Identification of metabolites from type III F2-isoprostane diastereoisomers by mass spectrometry. J Lipid Res. 2002;43:

61 Enrico Davoli, Head of the Mass Spectrometry Laboratory since 1997, Unit Head , Researcher , Research Fellow at the Mario Negri Institute. Fellow at USDA, Beltville, MD Doctoral Degree in Animal Sciences (University of Milan, 1983), Postdoctoral fellow at the University of Nebraska (Lincoln, NE, 1987) and at the University of Colorado Health Sciences Center (Denver, CO, 1988). Postgraduate degree in Pharmacological Research, Mario Negri Institute (1988). Member of the American Association for Mass Spectrometry (ASMS) of the Environment and Safety Commission of IGQ and of the ETS (Emission Trading System) commission. Member of the National Biomass Research Center Scientific Committee. Environmental Applications Interest Group Coordinator (ASMS). Research areas: Development of methodology, instrumentation and software for environmental research. Studies of urban air pollution and characterization of environmental odor annoyance. Selected publications Davoli E, Bianchi G. Odour emission rates from a waste treatment plant: results from a multi year follow-up study. Chemical Engineering Transactions 2008; 15 : Zuccato E, Grassi P, Davoli E, Valdicelli L, Wood D, Reitano G, Fanelli R. PCB concentrations in some food from European countries. Food Chem Toxicol 2008, 46: Bagnati R, Bianchi G, Marangon E, Zuccato E, Fanelli R, Davoli E. Direct analysis of isopropylthioxanthone (ITX) in milk by high-performance liquid chromatography/tandem mass spectrometry. Rapid Commun Mass Spectrom 2007 ; 21 : Riservato M, Rolla A, Davoli E. An isotopic dilution approach for 1,3-butadiene tailpipe emissions and ambient air monitoring. Rapid Commun Mass Spectrom 2004; 18: Davoli E, Gangai L, Morselli P, Tonelli D. Characterisation of Odorant emissions from Landfills by SPME and GC/MS. Chemosphere 2003; 51: Davoli E, Cappellini L, Fanelli R, Bonsignore M, Gavinelli M. On-Site Analysis of World War II Cylinders and Barrels with Unknown Contents. Field Anal. Chem. Technol. 2001; 5: Ettore Zuccato, Head of the Food Toxicology Laboratory since 2005, Unit Head , Researcher , Technician at the Mario Negri Institute. Doctoral degree in Medicine (University of Milan, 1986), Postdoctoral degree in Human Nutrition (1999), Postdoctoral fellow at the King s College School of Medicine (London, UK, ). Member of the ANSISA, EMEA expert, member of the Commissione Consultiva per i Prodotti Fitosanitari, and expert for the evaluation of plant protection products for registration within the EU. Research areas: Food safety, including the study of dietary chemical contaminants, safety assessment of GMO in human nutrition, food allergens and toxicants, emerging issues in food toxicology, risk perception and risk communication to the consumers, and evaluation of plant protection products for registration within the European Union. Environmental pollution by pharmaceuticals, and monitoring of illicit drugs in surface waters to estimate community drug abuse. Selected publications Zuccato E, Chiabrando C, Castiglioni S, Bagnati R, Fanelli R. Estimating community drug abuse by wastewater analysis. Environ Health Perspect 2008, 116: Castiglioni S, Zuccato E, Chiabrando C, Fanelli R, Bagnati R. Mass spectrometry analysis of illicit drugs in wastewater and surface water. Mass Spectrom Rev, 2008, 27: Zuccato E, Grassi P, Davoli E, Valdicelli L, Wood D, Reitano G, Fanelli R. PCB concentrations in some food from European countries. Food Chem Toxicol 2008, 46: Castiglioni S, Zuccato E, Crisci E, Chiabrando C, Fanelli R, Bagnati R. Identification and measurement of illicit drugs and their metabolites in urban wastewaters by liquid chromatography tandem mass spectrometry (HPLC-MS-MS). Anal Chem 2006, 78: Zuccato E, Calamari D, Castiglioni S, Chiabrando C, Bagnati R, Fanelli R. Cocaine in surface water: a new evidencebased tool to monitor community drug abuse. Environmental Health: A Global Access Science Source 2005, 4:14 Zuccato E, Calamari D, Natangelo M, Fanelli R. Presence of therapeutic drugs in the environment. Lancet 2000; 355:

62 Renzo Bagnati, Head of the Analytical Instrumentation Unit since 2005, Researcher , Research fellow at the Mario Negri Institute. Doctoral degree in Chemistry (University of Turin, 1985), Postgraduate degree in Pharmacological Research, Mario Negri Institute (1989). Research areas: Mass spectrometry applied to the analysis of biological and environmental relevant substances (proteins, peptides, hormones, pharmaceuticals, drugs of abuse, pesticides). Selected publications Zuccato E, Castiglioni S, Bagnati R, Chiabrando C, Grassi P, Fanelli R. Illicit drugs, a novel group of environmental contaminants. Water Res 2008 ; 42 : Analysis of phosphoinositides and their aqueous metabolites. Berrie CP, Iurisci C, Piccolo E, Bagnati R, Corda D. Methods Enzymol. 2007;434: Bagnati R, Bianchi G, Marangon E, Zuccato E, Fanelli R, Davoli E. Direct analysis of isopropylthioxanthone (ITX) in milk by high-performance liquid chromatography/tandem mass spectrometry. Rapid Commun Mass Spectrom 2007 ; 21 : Castiglioni S, Zuccato E, Crisci E, Chiabrando C, Fanelli R, Bagnati R. Identification and measurement of illicit drugs and their metabolites in urban wastewater by liquid chromatography-tandem mass spectrometry. Anal Chem 2006; 78: Zuccato E, Chiabrando C, Castiglioni S, Calamari D, Bagnati R, Schiarea S, Fanelli R. Cocaine in surface waters: a new evidence-based tool to monitor community drug abuse Pastorelli R, Carpi D, Airoldi L, Chiabrando C, Bagnati R, Fanelli R, Moverare S, Ohlsson C. Proteome analysis for the identification of in vivo estrogen-regulated proteins in bone. Proteomics 2005; 5: Elena Fattore, Head of the Environmental Pollutants Risk Assessment Unit since 2005, Researcher , Research fellow at the Mario Negri Institute. Doctoral Degree in Biological Sciences (University of Milan, 1991), Postgraduate degree in Pharmacological Research, Mario Negri Institute (1994), Postdoctoral fellow at the National Institute of Environmental Medicine, Karolinska Institutet, Stockholm ( ). Research areas: Environmental chemistry, toxicology, assessment of human exposure and risk from environmental pollutants with emphasis on dioxins and dioxin-like compounds. Selected publications Fattore E, Fanelli R, Dellatte E, Turrini A, Di Domenico A. Assessment of the dietary exposure to non-dioxin-like PCBs of the Italian general population. Chemosphere 2008, 73: S278-S283. Hodgson S, Thomas L, Fattore E, Lind P M, Alfven T, Hellstrom L, Hakansson H, Carubelli G, Fanelli R, Jarup L Bone mineral density changes in relation to environmental PCB exposure. Environmental Health Perspective 2008, 116: Carubelli G, Fanelli R, Mariani G, Nichetti S, Crosa G, Calamari D, Fattore E. PCB contamination in farmed and wild sea bass (Dicentrarchus labrax L.) from a coastal wetland area in central Italy. Chemosphere 2007 ; 68 : Fattore E, Fanelli R, Turrini A, Di Domenico A. Current dietary exposure to polychlorodibenzo-p-dioxins, polychlorodibenzofurans, and dioxin-like polychlorobiphenyls in Italy. Mol Nutr Food Res 2006; 50: Fattore E, Guardo A, Mariani G, Guzzi A, Benfenati E, Fanelli R. Polychlorinated Dibenzo-p-Dioxins and Dibenzofurans in Air of Seveso, Italy, 26 years after the accident. Environ Sci Technol 2003; 37: Fattore E, Fanelli R, La Vecchia C. Persistent organic pollutants in food: Public health and implications. J Epidemiol Community Health 2002; 56:

63 Marco Lodi, Head of the Industrial and Environmental Unit since 2002, Consultant at the Mario Negri Institute. General Certificate of Education in Industrial Chemistry (Milan, 1974). Member of AIDII (Italian Industrial Hygiene Association), certified by ACGIH (American Conference of Governmental Industrial Hygienist). Research areas: Emission sources, environmental diffusion, toxicology, human exposure and risk assessment of persistent environmental pollutants. Environmental risk of chemical pollution products. Development of sampling methods for environmental toxic compounds. Selected publications Benfenati E, Azimonti G, Auteri D, Lodi M Environmental and ecological toxicology: computational risk assessment. Computational toxicology. Risk assessment for pharmaceutical and environmental chemicals John Wiley, Hoboken, NJ, 2007; Grosso M, Cernuschi S, Palini E, Lodi M, Mariani G. PCDD/Fs release during normal and transient operation of a full scale MSWI plant. Organohalogen Compounds 2004; 66: Benfenati E, Mariani G, Lodi M, Reitano G, Fanelli R. Is bioexsiccation releasing dioxins? Organohalogen Compounds 2004; 66: Fattore E, Mariani G, Guzzi A, Di Guardo A, Benfenati E, Lodi M, Fanelli R. Air dioxin concentrations in Seveso area. In: Halogenated Environmental Organic Pollutants, 18th. Symp., Stockholm, Sweden, august 17-21, : Benfenati E, Mariani G, Schiavon G, Lodi M, Fanelli R. Diurnal, weekly and seasonal air concentrations of PCDD and PCDF in an industrial area. Fresenius Journal Analytical Chemistry 1994; 348: Benfenati E, Pastorelli R, Castelli M G, Fanelli R, Carminati A, Farneti A, Lodi M. Studies on the tetrachlorodibenzo-pdioxins (TCDD) and tetrachlorodibenzofurans (TCDF) emitted from an urban incinerator. Chemosphere 1986; 15: Roberta Pastorelli, Head of Protein and Gene Biomarkers Unit since 2004, Researcher , Research fellow at the Mario Negri Institute. Doctoral Degree in Biological Sciences (University of Milan, 1982), Postgraduate degree in Pharmacological Research, Mario Negri Institute (1986), Postdoctoral fellow at the Massachusetts Institute of Technology, Cambridge, MA ( and 1991). Research areas: Toxicoproteomic investigation of global protein expression profiles and their modulation evoked by environmental compounds in different biological compartments. Critical targets and pathways in toxicology. Pharmacogenetics: effects of genetic polymorphisms in the human population on the individual susceptibility to environmental xenobiotic and carcinogen effects. Selected publications: Pastorelli R, Saletta F, Campagna R, Carpi D, Dell Osta C, Schiarea S, Vineis P, Airoldi L, Matullo G. Proteome characterization of a human urothelial cell line resistant to the bladder carcinogen 4-aminobiphenyl. Proteome Science Moretti M, Dell'omo M, Villarini M, Pastorelli R, Muzi G, Airoldi L, Pasquini R. Primary DNA damage and genetic polymorphisms for CYP1A1, EPHX and GSTM1 in workers at a graphite electrode manufacturing plant. BMC Public Health : 270 Pastorelli R, Carpi D, Campagna R, Airoldi L, Pohjanvirta R, Viluksela M, Hakansson H, Boutros P C, Moffat I D, Okey A B, Fanelli R. Differential expression profiling of the hepatic proteome in a rat model of dioxin resistance: correlation with genomic and transcriptomic analyses. Mol Cell Proteomics 2006; 5: Pastorelli R, Carpi D, Airoldi L, Chiabrando C, Bagnati R, Fanelli R, Moverare S, Ohlsson C.Proteome analysis for the identification of in vivo estrogen-regulated proteins in bone. Proteomics 2005; 5: Airoldi L, Magagnotti C, Pastorelli R, Fanelli R. Enzyme polymorphisms influencing the metabolism of heterocyclic aromatic amines. J Chromatogr B Analyt Technol Biomed Life Sci 2004; 802: Vineis P,V eglia F, Anttila S, Benhamou S, Clapper M L, Dolzan V, Ryberg D, Hirvonen A, Kremers P, Le Marchand L, Pastorelli R, Rannug A, Romkes M, Schoket B, Strange R C, Garte S, Taioli E. CYP1A1, GSTM1 and GSTT1 polymorphisms and lung cancer: a pooled analysis of gene-gene interactions. Biomarkers 2004; 9:

64 INTRODUCTION TO THE DEPARTMENT'S ACTIVITIES The Department works to investigate environmental factors and their effects on human health. The main research lines focus on the survey of environmental contaminants, the assessment of human exposure with related health risks, and toxicity mechanisms of pollutants. The assessment of environmental contamination is carried out not only for well-known and widespread compounds, like dioxins and PCBs, but also for new classes of "unconventional" pollutants, e.g., endocrine disruptors, potentially toxic "natural" compounds, and drugs entering the environment after human or veterinary use. The identification for the first time of illicit drugs in urban waste and river waters, led to a new original tool for the evidence-based monitoring of community drug abuse. For all these survey activities sophisticated analytical methods based on advanced mass spectrometric techniques are developed. The Department is active in the assessment of human exposure to toxic compounds in the atmosphere and the diet, which is the main source of priority pollutants (PCBs, dioxins and other endocrine disruptors). Assessment of the risk associated to contamination in real-life scenarios has recently gained much importance. In order to respond to the growing demand for information, the Department is more and more involved in toxicological and ecotoxicological risk analysis, based on studies in field and predictive models of toxicity. Molecular epidemiology studies are used to identify genetic and/or environmental factors posing risks to human health. By this approach, we search for new useful biological markers" to identify susceptible subjects, in view of finding appropriate preventive strategies. The Department has implemented an advanced technological proteomic platform, in order to identify proteins differentially expressed in biological compartments in various experimental and clinical conditions. This approach is particularly relevant in toxicology, since it can contribute to find new biomarkers of toxicity or pathology, and to identify molecular targets and toxic effect mechanisms of pollutants and drugs. To integrate our proteomic studies, we have now introduced among our activities metabolomics, i.e., the study of small molecules, such as amino acids, carbohydrates, lipids, hormones etc., the final products of protein expression and activity which contribute to define the biochemical phenotype of a biological system. Mass spectrometry (MS) is a central analytical technique at the Department, where a complete set of state-of-the-art instrumentation is available, from GC-MS and LC-MS to MALDI-TOF- MS. These instruments are provided with modern solutions for sample introduction (chip-based nanolc), sample ionization (ESI, DESI and MALDI), tandem MS (MSn) by triple quadrupole and TOF-TOF instruments, high mass resolution analysis (hybrid ion trap/orbitrap). FINDINGS/MAIN RESULTS The lack of retinoic acid (knock-out mice for the retinal dehydrogenase 1 gene, RALDH1) modifies the proteome profile of the bone,through changes in the expression of proteins involved in chondrogenesis and osteoclastogenesis. Resistance to the bladder carcinogen 4-aminobiphenyl in human urothelial cells is mediated by deregulation of apoptosis and membrane trafficking proteins. Bone protein profile in a murine model of osteoporosis. Identification of novel protein targets responsive to the effects of estrogens in bone. TCDD's effect on the liver proteome profile of exposed rats. Determination of a subset of rat hepatic proteins indicative of differences in dioxin susceptibility. The presence of 4-aminobiphenyl-hemoglobin adducts may help identify nonsmokers at high risk of cancers related to environmental tobacco smoke exposure. 63

65 Reference values of allele and genotype frequency of several metabolic genes in 15,000 control subjects. CYP1A1 polymorphism affects lung tumor risk. Identification of CYP2C9 genetic polymorphism as a determinant of severe adverse reactions to phenytoin. On-line in silico models to predict ecotoxicity of pesticides for regulatory purposes. A new model for identification of endocrine disruptors using molecular docking. A method aimed at characterizing environmental odors to identify odor sources in complex environments. Albumin can become a source of potentially antigenic peptides in proteinuric nephropaties. An acid protease released by rat renal proximal tubular cells selectively cleaves an albumin N- terminal fragment (ALB1-24). Kidney infiltrating dendritic cells (DC) can capture ALB1-24 and process it, through the proteasome, to shorter peptides capable of activating syngeneic CD8+ T cells. Moderate-to-high fish consumption can result in exceeding the daily intake safety levels of PCBs and dioxin-like compounds established by the European Commission. The same food type may contain significantly different concentrations of PCBs and dioxin-like compounds, depending on the geographic origin (this may help lower the risk for the consumers by understanding and controlling the causes of the differences). The environmental levels of several drugs exceed the safety limits established for them. Environmental pollution from pharmaceuticals is a general phenomenon that can be described by controllable variables (environmental load and mass balance). Illicit drug residues and their metabolites were found in urban waste and river waters. Environmental levels can be used as a new tool to estimate illicit drugs consumption in the population. The distribution of dietary intake values of dioxins, dioxin-like PCBs and non dioxin-like PCBs was characterized for the general Italian population. The higher intake of PCBs due to consumption of farmed fish vs. wild fish is mainly due to the higher fat content in farmed fish. Development of novel mass spectrometric methods for the selective measurement of therapeutic and illicit drugs in environmental samples. NATIONAL COLLABORATIONS ARPA Emilia Romagna ARPA Veneto ASL di Brescia ASL di Como ASL di Cagliari ASL di Napoli CNR - IRSA Comune di Rosignano Marittimo (LI) CSRA-Asti Fondazione 'S. Maugeri' Fondazione ISI, Torino INRAN-Istituto Nazionale di Ricerca sugli Alimenti e la Nutrizione ISPO, Firenze Istituto Clinico Humanitas, Milano 64

66 Istituto Superiore di Sanità I.Z.S.L.T - Istituto Zooprofilattico Sperimentale del Lazio e Toscana Ministero dell'ambiente Politecnico di Milano Politecnico di Torino Provincia di Vercelli Provincia Pordenone Università degli Studi di Cagliari Università degli Studi di Genova Università degli Studi di Milano Università degli Studi di Napoli "Federico II" Università degli Studi di Palermo Università degli Studi di Pavia Università degli Studi di Perugia Università degli Studi di Roma "La Sapienza" Università degli Studi di Torino Università dell Insubria, Varese Università degli Studi di Verona 65

67 INTERNATIONAL COLLABORATIONS BASF Agricultural Centre, Limburgerhorf, Germany Central Science Laboratory, York, UK Centre for Environmental Policy, Imperial College, London, UK Danish Institute of Agricultural Sciences, Research Centre Foulum, Tjele, Denmark Department of Analytical and Pharmaceutical Chemistry, The Royal Danish School of Pharmacy, Denmark Department of Anatomy and Cell Biology, University of Oulu, Oulu, Finland Department of Chemistry, Loyola University, Chicago, IL, USA Department of Computer Science and Engineering, University of Galati, Romania Department of Electrical and Computer Engineering, University of Patras, Greece Department of Environmental Health, National Public Health Institute, Kuopio, Finland Department of Epidemiology & Public Health, Imperial College, London, United Kingdom Department of Inland Fisheries, Institute of Freshwater Ecology and Inland Fisheries, Germany Department of Molecular Biology, University of Bergen, Bergen, Norway Department of Organic Chemistry, Universidad de Cadiz, Spain Division of Endocrinology, Department of Internal Medicine, Sahlgrenska University Hospital, Gothenburg, Sweden Environmental Chemistry, IIQAB-CSIC, Barcelona, Spain Environmental Hygiene and Chemistry Department, Institute of Environmental Medicine and Hospital Epidemiology, University of Freiburg, Germany Environmental Protection Agency, US EPA - National Risk Management Research Laboratory (NRMRL), Cincinnati OH, USA European Monitoring Centre for Drugs and Drug Addiction (EMCDDA), Lisbon, Portugal Faculté de Médicine et de Pharmacie, Université de Mons-Hainaut, Mons, Belgium Faculty of Veterinary Medicine, Utrecht University, Utrecht, The Netherlands Forschungzentrum Jülich Gmbh, Jülich, Germany Gruppo Collaborativo sulla Suscettibilità Genetica ai Cancerogeni Ambientali (GSEC), Italia Institute of Environmental Medicine. Karolinska Institute, Stockholm, Sweden Institute of Pharmaceutical Chemistry, University of Pécs, Pecs, Hungary Institute of Phytomedicine, Biological Control, Horticulture and Nematology, Wien, Austria Institute of Soil Science and Plant Cultivation, Pulawy, Poland Interuniversitaeres Forchunginstitut fuer Agrarbiotechnologie, Tulln, Austria Istituto di Chimica di São Carlos, Università di São Paulo, Brazil KnowledgeMiner Software, Berlin, Germany In Vitro Testing Industrial Platform, Tres Cantos (Madrid), Spain Laboratory of Chemometrics & Bioinformatics, University of Orléans, Orléans, France Laboratory of Neurobiology, Centro de Investigation Principe Felipe, Valencia, Spagna Lithuanian Institute of Agricultrure, Vilnius, Lithuania Liverpool John Moores University, Liverpool, United Kingdom National Institute of Chemistry, Kemijski Institut Ljubljana, Ljubljana, Slovenia Natural Resources Research Institute, University of Minnesota, Duluth, MN, USA National Institute for Public Health and the Environment (RIVM), Bilthoven, The Netherlands Pesticide Safety Directorate, York, UK Plant Protection Institute, Hungarian Academy of Sciences, Budapest, Hungary School of Biomedical Sciences, University of Ulster, Coleraine, Northern Ireland Syngenta Crop Protection AG, Basel, Switzerland UFZ Leipzig, Germany University of Tartu, Tartu, Estonia 66

68 EDITORIAL BOARD MEMBERSHIP Journal of Environmental Science and Health, Part B (Emilio Benfenati), Journal of Environmental Science and Health, Part C (Emilio Benfenati), International Journal of Computational Intelligence (Emilio Benfenati), International Journal of Information Technology (Emilio Benfenati), International Journal of Signal Processing (Emilio Benfenati), Chemistry Central Journal (Emilio Benfenati), Current Computer Aided Drug Design (Emilio Benfenati), Advances in Chemoinformatics and Computational Methods (Emilio Benfenati), The Open Biomarkers Journal (Luisa Airoldi). PEER REVIEW ACTIVITIES Addiction, Analytical and Bioanalytical Chemistry, Chemical Biology & Drug Design, Chemical Research Toxicology, Chemometrics and Intelligent Laboratory Systems, CHEMOLAB, Chemosphere, Clinical Chemistry and Laboratory Medicine, Environment International, Environmental Pollution, Environmental Modeling & Software, Environmental Science & Technology, Journal of Chemical Information and Modeling, International Journal of Molecular Science, Journal Computer-Aided Molecular Design, Journal of Hazardous Materials, Molecular Diversity, Proteomics, Royal Society's Philosophical Transactions, Stroke, Toxicology Letters, Waste Management, Water Research. NATIONAL AND INTERNATIONAL COMMITTEE MEMBERSHIP CCPF - Commissione Consultiva Prodotti Fitosanitari (Ministero della Salute, Ministero dell'ambiente) ECCO - European Commission Coordination EFSA - European Food Safety Authority IGQ - Commissione Ambiente e Sicurezza IGQ - Commissione ETS EVENT ORGANIZATION Workshop Il triplo quadrupolo: trent'anni di successi!, Istituto di Ricerche Farmacologiche Mario Negri, Milan, Italy, December 18, Thermo Proteomics Seminar, Istituto di Ricerche Farmacologiche Mario Negri, Milan, Italy, November 13, Seminario Procedure per l analisi e la caratterizzazione delle proteine: panoramica delle soluzioni, Istituto di Ricerche Farmacologiche Mario Negri, Milan, Italy, October 27, NOSE2008. International Conference on Environmental Odour Monitoring and Control. Rome, Italy, July 6-8,

69 Workshop I modelli predittivi per il REACH: Istruzioni per l uso, Istituto di Ricerche Farmacologiche Mario Negri, Milan, Italy, July 1, Workshop Il Monitoraggio dei Microinquinanti, Istituto di Ricerche Farmacologiche Mario Negri, Milan, Italy, May 29, Workshop From classical Qualitative Proteomics to Imaging and Quantitative Proteomics: an Overview of Technologies and Applications, Istituto di Ricerche Farmacologiche Mario Negri, Milan, Italy, April 8, Workshop of the SCARLET EC project on in silico methods for carcinogenicity and mutagenicity, Istituto di Ricerche Farmacologiche Mario Negri, Milan, Italy, April 2-4, st Assembly of the OSIRIS EC project, Istituto di Ricerche Farmacologiche Mario Negri, Milan, Italy, March 11-13, PARTICIPATION IN EVENTS IN WHICH THE DEPARTMENT WAS INVOLVED The Belgian Society for Toxicology & Ecotoxicology (BELTOX) and The Luxembourg Toxicology Society (BLT) Joint Meeting, Brussels, Belgium, November 28, Venice Second International Symposium on Energy from Biomass and Waste. Venice, Italy, November 17-20, EPAA Annual Conference "Research into alternative approaches (3Rs) in regulatory testing: Are we on the right track?", Brussels, Belgium, November 3, st SETAC Europe Special Science Symposium - "Integrated Testing Strategies for REACH: From science to practical implementation", Brussels, Belgium, October 23-24, th European Conference on Pesticides and Related Organic Micropollutants in the Environment, Marseille, France, October 22-25, Workshop of the ATHON EC project, Midterm review, Amsterdam, The Netherlands, September 28-30, Convegno Interferenti endocrini: valutazione e prevenzione dei possibili rischi per la salute umana. Istituto Superiore di Sanità, Rome, Italy, October 15, Dioxin 2008, 28 th International Symposium on Halogenated Persistent Organic Pollutants (POPs), Birmingham, UK, August 17-22, HUPO 2008, 7 th world congress, Amsterdam, The Netherlands, August 16-20, th American Society for Mass Spectrometry Conference, Denver, CO, USA, June 1-5, Workshop Il Monitoraggio dei Microinquinanti, a c. di Environnement Italia Spa, Istituto di Ricerche Farmacologiche Mario Negri, Milan, Italy, May 29, SETAC Europe 18th Annual Meeting, Warsaw, Poland, May 25-29,

70 Corso Problems & approaches in computational chemistry, Politecnico di Milano, Milan, Italy, April 21-22, Workshop of the SCARLET EC project on in silico methods for carcinogenicity and mutagenicity, Istituto di Ricerche Farmacologiche Mario Negri, Milan, Italy, April 2-4, KNAPPE International Conference, Nimes, France, February19-20, th International Workshop on Liquid Chromatography in Environmental and Food samples, Barcelona, Spain, February 7-8, GRANTS AND CONTRACTS A2A Brescia ACEGAS S.p.A, Trieste AIIPA (Associazione Italiana Industrie Prodotti Alimentari) AMA, Roma ASL Como ASL Mantova ASL Napoli 2 Associazione Italiana Ricerca sul Cancro COOP Italia CSRA Comune di Lomello (PV) Comune di Mazzano e Rezzato (BS) Consorzio Quadrifoglio S.p.A. ECODECO, Pavia European Commission (MEBFOOD, HERBICBIOREM, ATHON, CASCADE, CAESAR, CHEMOMENTUM, CHEMPREDICT, OSIRIS, SCARLET) Federchimica, Milano FIAT Auto S.p.A. Fondazione CARIPLO, Milano Fondazione Italo Monzino, Milano HERA S.p.A. (Holding Energia Risorse Ambiente) I.Z.S.L.T - Istituto Zooprofilattico Sperimentale del Lazio e Toscana Lucart, Cartiera Lucchese S.p.A. Porcari, Lucca Ministero della Salute, Italia Ministero dell'ambiente, Italia Oxon Italia S.p.A., Pero (MI) Provincia di Pordenone Provincia di Vercelli SO.GE.NU.S. S.p.A Telethon Tenacta Group TM.E. S.p.A 69

71 SELECTION OF SCIENTIFIC PUBLICATIONS FROM 2008 Peluso M, Airoldi L, Colombi A, Munnia A, Veglia F, Autrup H, Dunning A, Garte S, Gormally E, Malaveille C, Matullo G, Overvad K, Raaschou-Nielsen O, Clavel-Chapelon F, Linseisen J, Boeing H, Trichopoulou A, Palli D, Krogh V, Tumino R, Panico S, Bueno-De-Mesquita H B, Peeters P H, Kumle M, Gonzalez C A, Martinez C, Dorronsoro M, Barricarte A, Tormo M J, Quiros J R, Berglund G, Janzon L, Jarvholm B, Day N E, Key T J, Saracci R, Kaaks R, Riboli E, Bingham S, Vineis P. Bulky DNA adducts, 4-aminobiphenyl hemoglobin adducts, diet and air pollution in a healthy European population. Br J Nutr : Veglia F, Loft S, Matullo G, Peluso M, Munnia A, Perera F, Phillips D H, Tang D, Autrup H, Raaschou-Nielsen O, Tjonneland A, Vineis P, Genair-EPIC Investigators. DNA adducts and cancer risk in prospective studies: a pooled analysis and a meta-analysis. Carcinogenesis 2008 ; 29 : Roncaglioni A, Piclin N, Pintore M, Benfenati E. Binary classification models as screening tools for endocrine disrupter effects mediated through the estrogen receptor. SAR QSAR Environ Res : Viganò L, Benfenati E, van Cauwenberge A, Eidem J K, Erratico C, Goksøyr A, Kloas W, Maggioni S, Mandich A, Urbatzka R. Estrogenicity profile and estrogenic compounds determined in river sediments by chemical analysis, ELISA and yeast assays. Chemosphere : Casalegno M, Sello G, Benfenati E. Definition and detection of outliers in chemical space. J Chem Inf Model : Zhao C, Boriani E, Chana A, Roncaglioni A, Benfenati E. A new hybrid QSAR model for the prediction of bioconcentration factors (BCF). Chemosphere : Toropov A A, Toropova A P, Benfenati E. QSPR modeling for enthalpies of formation of organometallic compounds by means of SMILES-based optimal descriptors. Chemical Physics Letters : Slavov S, Gini G, Benfenati E. QSAR trout toxicity models on aromatic pesticides. J Environ Sci Heal B : Colombo A, Benfenati E, Karelson M, Maran U. Definition of a univocal architecture for chemical splitting to optimise QSAR models for aquatic toxicity. Chemosphere : Fjodorova N, Novic M, Vracko M, Smirnov V, Kharchevnikova N, Zholdakova Z, Novikov S, Skvortsova N, Filimonov D, Poroikov V, Benfenati E. Directions in QSAR Modeling for Regulatory Uses in OECD Member Countries, EU and in Russia. J Environ Sci Heal C : Toropov A A, Benfenati E. Additive SMILES-based optimal descriptors in QSAR modelling bee toxicity: Using rare SMILES attributes to define the applicability domain. Bioorg Med Chem : Bursztyka J, Perdu E, Tulliez J, Debrauwer L, Delous G, Canlet C, De Sousa G, Rahmani R, Benfenati E, Cravedi J-P. Comparison of genistein metabolism in rats and humans using liver microsomes and hepatocytes. Food Chem Toxicol : Porcelli C, Roncaglioni A, Chana A, Benfenati E. A comparison of DEMETRA individual QSARs with an index with an index of evaluation of uncertainty. Chemosphere : Fjororova N, Novich M, Vrachko M, Kharchevnichova N, Zholdakova Z, Sinitzyna O, Benfenati E. Regulatory assessment of chemicals within OECD Member Countries, EU and in Russia. J Environ Sci Heal C : Roncaglioni A, Benfenati E. In silico-aided prediction of biological properties of chemicals: oestrogen receptormediated effects. Chem Soc Rev : Porcelli C, Boriani E, Roncaglioni A, Chana A, Benfenati E. Regulatory perspectives in the use and validation of QSAR. A case study: DEMETRA model for daphnia toxicity. Environ Sci Technol : Pandelova M, Henkelmann B, Roots O, Simm M, Jarv L, Benfenati E, Schramm K-W. Levels of PCDD/F and dioxin-like PCB in Baltin fish of different age and gender. Chemosphere :

72 Macconi D, Chiabrando C, Schiarea S, Aiello S, Cassis L, Gagliardini E, Noris M, Buelli S, Zoja C, Corna D, Mele C, Fanelli R, Remuzzi G, Benigni A. Proteasomal processing of albumin by renal dendritic cells generates antigenic peptides. J Am Soc Nephrol. 2009; 20: Epub 2008 Dec 17. Davoli E, Bianchi G. Odour emission rates from a waste treatment plant: results from a multi year follow-up study. Chemical Engineering Transactions 2008 ; 15 : Castiglioni S, Pomati F, Miller K, Rossetti C, Zuccato E, Calamari D and Neilan BA. Exchange of the multiple resistance gene mara in antibiotic-contaminated environments involving Bacillus sp. Water Research, 2008, 42: Zuccato E, Chiabrando C, Castiglioni S, Bagnati R, Fanelli R. Estimating community drug abuse by wastewater analysis. Environmental Health Perspectives, 2008, 116: Castiglioni S, Zuccato E, Chiabrando C, Fanelli R, Bagnati R. Mass spectrometry analysis of illicit drugs in wastewater and surface water. Mass Spectrometry Reviews, 2008, 27: Farré M, Petrovic P, Gros M, Kosjek T, Martinez E, Heath E, Osvald P, Loos R, Le Menach K, Budzinski H, De Alencastro F, Müller J, Knepper T, Fink G, Ternes TA, Zuccato E, Kormali P, Gans O, Quintana JB, Pastori F, Gentili A, Barceló D. First interlaboratory exercise on non-steroidal anti-inflammatory drugs analysis in environmental samples. Talanta, 2008, 76: Pomati F, Orlandi C, Clerici M, Luciani F, Zuccato E. Effects and interactions in an environmentally relevant mixture of pharmaceuticals. Toxicol Sci 2008, 102(1): Zuccato E, Castiglioni S, Bagnati R, Chiabrando C, Grassi P, Fanelli R. Illicit drugs, a novel group of environmental contaminants. Water Research 2008, 42: Zuccato E, Grassi P, Davoli E, Valdicelli L, Wood D, Reitano G, Fanelli R. PCB concentrations in some food from European countries. Food Chem Toxicol, 2008, 46: Fattore E, Fanelli R, Dellatte E, Turrini A, Di Domenico A. Assessment of the dietary exposure to non-dioxin-like PCBs of the Italian general population. Chemosphere 2008, 73: S278-S283. Hodgson S, Thomas Laura, Fattore E, Lind P M, Alfven T, Hellstrom L, Hakansson H, Carubelli G, Fanelli R, Jarup L Bone mineral density changes in relation to environmental PCB exposure. Environmental Health Perspective 2008, 116: LAY PRESS SELECTION PUBLISHED IN 2008 Colombo A, Benfenati E, Lodi M. Diossine, fonti di emissione e conseguenze. Laboratorio : Fattore E, Paiano V. Il rischio sanitario in relazione alla qualità dell'aria. Ricerca & Pratica 2008, 144: OTHER PRODUCTS PUBLISHED IN 2008 Zuccato E, Chiabrando C, Castiglioni S, Bagnati R, Fanelli R. Estimating community drug use. In: EMCDDA Insight 9: Assessing illicit drugs in wastewater; potential and limitation of a new monitoring approach. European Monitoring Centre for Drug and Drug Addiction, Lisbon 2008: Davoli E, Fattore E, Paiano V, Fanelli R, Rossi AN, Il Grande M. Integrated Risk Assessment of Waste Management: A Case Study of a Solid Waste Landfill in South Italy. Proceedings Venice 2008, Second International Symposium on Energy from Biomass and Waste, Venice, Italy, November Available only on CD. 71

73 RESEARCH ACTIVITIES Laboratory of Molecular Toxicology Proteome Analysis Proteome analysis includes protein separation by one- and two-dimensional gel electrophoresis, protein excision from the gel, their digestion with proteolytic enzymes and their identification by mass spectrometry (MALDI-TOF-MS, LC-ESI-MS/MS) coupled to the use of existing databases. Alternatively, peptides resulting from the digestion of protein mixtures with specific proteases are separated by two-dimensional liquid chromatography. Toxicoproteomics Studies are ongoing on the characterization of changes in the proteome profile induced by environmental toxic compounds, with the aim of obtaining protein biomarkers with the ability to differentiate two or more biological states. Proteome changes in tissues and target organs of animals, and cells treated with endocrine disruptors, estrogens, or environmental carcinogens, are related to functional changes during toxicological processes. Clinical Proteomics Qualitative and quantitative proteome changes resulting from the exposure to environmental toxic compounds or in pathological conditions are monitored in human biological plasma and urine. Ongoing studies aim at the characterization of protein biomarkers for early diagnosis of diseases and for the identification of therapeutic targets. Metabolomics Metabolites are the biological endpoints of gene expression and enzyme activity and include small molecules such as amino acids, carbohydrates, fatty acids, hormones, etc., that provide the metabolic phenotype of individuals. Metabolomic research focuses on the quantitative analysis of metabolites in biological fluids to link human metabolic profile variations to endogenous or exogenous pathophysiological stimuli and to genetic modifications. Selected metabolites are extracted from plasma or urine by solid phase extraction and analyzed by HPLC coupled to high-resolution mass spectrometry. The integration of proteomic and metabolomic studies will provide information that can help to better understand disease development and to identify preventive interventions. Molecular Epidemiology The laboratory works mainly on the measurement of biological markers used to assess human exposure to environmental toxic compounds. Our studies include DNA- and blood proteinadduct formation by several environmental carcinogens. In addition, we study whether the polymorphism of genes coding for enzymes involved in the activation and detoxification of carcinogens are determinants of adduct formation. Genotypes are detected by restriction fragment length polymorphism analysis, after the amplification by polymerase chain reaction of specific nucleotide sequences of the genes under study. The laboratory participates in an international cooperation study aimed at the collection of reference values on allele and genotype frequency of the most common metabolic enzyme polymorphisms in control populations. 72

74 Laboratory of Analytical Biochemistry Identification and characterization of proteins by mass spectrometry Our laboratory is developing different analytical and instrumental techniques, based on chromatography, electrophoresis and mass spectrometry, for the identification and characterization of proteins and peptides in biological samples. This activity is mainly aimed at 1) global characterization and comparison of secretomes from human cancer cell lines to identify proteins that may be functionally relevant for tumor growth and spread; 2) profiling proteins in biological fluids for discovery and identification of biomarkers of physiopathological and toxicological relevance, 3) identifying and characterizing endogenous degradation products of proteins, 4) identifying proteins produced by cells in vitro in response to given stimuli, 5) selectively isolating biologically relevant proteins by immunoaffinity-based micro-techniques. Ongoing projects include the study of exogenous protein degradation in renal tubular cells in relation to antigen presentation mechanisms, and the characterization of the secretome of cancer cell lines in vitro to identify factors affecting immune cells. Method development in proteomics The laboratory works on the optimization of various analytical methodologies for proteomics, i.e. various complementary techniques for protein isolation and identification by mass spectrometry (MALDI-TOF-MS, LC-ESI-MS/MS). Laboratory of Environmental Chemistry and Toxicology Development and use of analytical methods to evaluate contamination in water bodies, soil, biota, human samples in exposed population Analytical methods are developed to study environmental pollutants in water ecosystems, landfills, contaminated sites. Qualitative and quantitative analyses of organic pollutants are done by mass spectrometry (GC-MS, LC-MS, LC-MS/MS). Typical analyses include PCDD/F, PCB, PAH, polybrominated diphenylethers, pesticides, endocrine disruptor chemicals, and industrial pollutants. Studies on environmental, toxicological and ecotoxicological properties of chemicals Research is carried out on pollutant properties, searching literature data, comparing and evaluating different sources, and mainly developing predictive models to cope with the lack of experimental data. Thus, we develop models starting merely from the chemical structure. The research addresses the different kinds of chemical descriptors and chemical fragments, obtained with different software. Then, we develop models using algorithms such as neural network, fuzzy logic, genetic algorithms, classifiers, multivariate analysis, etc. Different methods are compared and integrated within a structured ensemble. Standardized methods for pesticides were developed and validated according to OECD guidelines. Risk assessment of pollutants Studies are aimed at assessing the risk of pollutants for human population and environment. For this we model transport and diffusion of pollutants, to obtain a predicted concentration on given space and time scales. Such an activity is integrated with those above described on chemical analyses and toxicity prediction, to achieve a continuous transfer of data and research. 73

75 Research on pollutants emitted in the atmosphere (Unit of Industrial and Environmental Hygiene) Studies address different aspects of atmospheric pollution. Research deals with: sampling areas around the pollution source, chemical analyses, transport modeling depending on meteorological conditions and orography, risk assessment for population and environment. Qualitative and quantitative analyses are done by gas chromatography-mass spectrometry using high resolution for PCDDs/PCDFs, and negative ion-chemical ionization for PCBs. Laboratory of Mass Spectrometry Particulate air pollution Epidemiological studies consistently show an association between an increasing number of pathologies, both acute and chronic, and particulate air pollution. This has been shown not only in respiratory, but in cardiovascular diseases as well. Airborne particulate sampling and analysis strategies are developed to characterize both adsorbed compounds and exposition in different activities. Method development in environmental sciences Methods, analytical methodologies, instrumentation and software for data acquisition and reduction, are developed for environmental studies. High-sensitivity instrumentation, mainly based on mass spectrometry, is developed for trace and ultra-trace analysis. Also, transportable instrumentation is developed for field studies or continuous monitoring. Characterization of environmental odor annoyance Characterization of odors poses several analytical problems because they result from a complex mixture of compounds (odorants) stimulating receptors in the nasal cavity. Most odorants are volatile organic compounds (VOC) generated by bacterial degradation of organic matter. They are often present at trace levels, while numerous sources can contribute to the total odor. Using sampling techniques specifically developed for olfactometry, solid phase microextraction and GC/MS analysis, we can detect traces (low ppb to high ppt) of a wide polarity/volatility range of airborne VOC odorant compounds. With a chemometric approach, we can characterize the sources of emissions, assess odor control methods, and identify emissions that contribute to odors in ambient air. Laboratory of Food Toxicology Chemical contaminants in food We are studying human exposure to dietary PCBs and dioxins. PCBs were measured in food items in different European countries, showing differences in PCBs exposure of European consumers. Further studies were aimed at measuring PCBs and dioxins in samples of fish caught in Italy and in food items from an Italian area at high risk of contamination. Ongoing studies are focused to assess levels of PCBs and dioxins in samples of human milk collected from mothers living in highly contaminated areas. Therapeutic and illicit drugs in the environment Pharmaceuticals are a class of emerging environmental pollutants. We have organized a campaign to detect the presence of pharmaceuticals and their metabolites in Italian rivers and sewage treatment plants, with the aim of better characterizing the contamination and assessing related risks. 74

76 Human and environmental risks are evaluated by studying the toxic effects of pharmaceuticals at environmental levels, on cultures of human and zebra fish cells. Further ongoing studies are aimed at investigating a possible relationship between antibiotic occurrence and resistance in environmental bacteria. The possible presence of illicit drugs in water samples from sewage treatment plants and rivers was investigated, starting with cocaine and its metabolites. Their levels, used to estimate drug abuse in the local population, revealed that cocaine consumption greatly exceeds official estimates. This approach has been subsequently extended to include other common drugs of abuse such as cannabis, opiates (heroin, morphine), and amphetamines (amphetamine, methamphetamine, ecstasy). Consumption of these drugs have been subsequently estimated in some European cities. Our evidence-based method allows monitoring of patterns and trends of drug abuse in local communities, and is able to detect qualitative and quantitative consumption changes in real time. This tool can therefore complement survey methods in more realistically describing the drug abuse phenomenon. Regulatory activities On behalf of the Ministry of Health, we carried on the evaluation of the dossiers required for pesticide registration within the European Union. Unit of Environmental Pollutants Risk Assessment Exposure to environmental pollutants Research activities include focus both on quantitative measurement of contaminants in environmental samples, and assessment of human exposure. Specific projects projects are the following: Measurements of persistent organic pollutants such as polybrominated dioxins and furans, perfluorooctanoic acid (PFOA), perfluorooctane sulphonate (PFOS), and identification of new pollutants, in farmed and wild fish coming from Mediterranean sea. Exposure assessment to these persistent pollutants for human population due to fish consumption. on dietary exposure of the general Italian population include : an exposure assessment to PCBs thorough dietary farmed and wild fish coming from Mediterranean sea; a study combining available data from food consumption surveys in Italy with data on contamination by polychlorinated dibenzo-p-dioxins (PCDDs), furans (PCDF), dioxin- and non-dioxin-like PCBs in European foodstuffs; an investigation of the contamination level by PCDD, PCDF and PCB in different food coming from a suspected polluted area. A study on occupational exposure deals with measurements of PCBs and DDE in human blood with the aim of assessing the correlation between exposure to these organochlorine compounds and the bone mineral density of the population investigated. Toxicological risk assessment The unit activities also include risk assessment studies related to specific environmental conditions or human activities which can pose a risk for human health. During 2008 studies of risk assessment related to air quality in a industrialised area, and to the emissions from an incinerator and a landfill have been carried out.new methods for exposure assessment are developed, employing probabilistic approaches and more refined statistical models, starting from real cases of contamination. A current study deals with the exposure of the Seveso population to dioxin from contaminated soil. Evaluation of toxicological data Toxicological data resulting from in vivo sub-chronic studies in rats exposed to individual dioxin-like and non dioxin-like PCBs are evaluated in detail, in order to investigate the dose- 75

77 response relationship and the applicability of the benchmark dose approach. Unit of Analytical Instrumentation Development and application of analytical methods for compounds of biological and environmental interest. Methods are developed mainly using solid phase extraction (SPE) followed by liquid chromatography - mass spectrometry (LC-ESI-MS/MS) or gas chromatography - mass spectrometry (GC-MS). Available intruments include mass spectrometers equipped with different analyzers: magnetic fields, time of flight (TOF), quadrupoles (single and triple), ion traps and high resolution orbitrap. The main ionization techniques are electron ionization (EI), chemical ionization (CI), MALDI, ACPI and Electrospray (conventional and nanospray). Substances of interest include proteins, peptides, hormones, pharmaceuticals, drugs of abuse, pesticides, and other environmental contaminants (PCBs, hydroxy-pcbs, perfluorinated compounds). Work has been started for the development of imaging methods in biological tissues with the MALDI-TOF technique. 76

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79 DEPARTMENT OF NEUROSCIENCE STAFF Head Gianluigi FORLONI, Biol.Sci.D. Laboratory of Biology of Neurodegenerative Disorders Head Cell Death and Neuroprotection Unit Head Gianluigi FORLONI, Biol.Sci.D. Tiziana Borsello, Biol.Sci.D. Laboratory of Drug Metabolism Head Silvio CACCIA, Farm.D. Laboratory of Experimental Neurology Head Annamaria VEZZANI, Biol.Sci.D. Laboratory of Experimental Psychopharmacology Head Luigi CERVO, Ph.D. Laboratory of Geriatric Neuropsychiatry Head Ugo LUCCA, MSc Epidemiology and Social Psychiatry Unit Head Barbara D AVANZO, Philos.D. Geriatric Epidemiology Unit Head Geriatric Pharmacology Unit Head Mauro TETTAMANTI, Biol.Sci.D. Emma RIVA, M.D. Laboratory of Inflammation and Nervous System Diseases Head Pharmacology of septic shock Unit Head Maria Grazia DE SIMONI, Biol.Sci.D. Pia VILLA, Biol. Sci. D Laboratory of Molecular Neurobiology Head Caterina BENDOTTI, Farm.D. 78

80 Laboratory of Neurochemistry abd Behavior Head Roberto William INVERNIZZI, Biol. Sci D Pharmacology of Cognitive Behavior Unit Head Mirjana CARLI, Ph.D. Laboratory of Neurological Disorders Head Ettore BEGHI, M.D. Laboratory of Quality Assessment of Geriatric Services Unit Head Alessandro NOBILI, M.D. 79

81 CURRICULA VITAE Gianluigi Forloni, obtained the Degree of Biological Science at the University of Milan in After two years of post doc at the Department of Neuroscience and Psychiatry at Johns Hopkins University in Baltimore, USA, he came back to the Mario Negri Institute and between 1992 and 1996 he was the head of the Neurobiology of Alzheimer's disease Unit; since 1996 he is the Head of the Biology of Neurodegenerative Diseases Lab and since 2002 the Head of the Neuroscience Department. His scientific interest is focused on the biological and genetic bases of aging-related disorders in particular Alzheimer s disease, Prion-related encephalopathies and Parkinson s disease. He has been member of several European committees for the examination of projects in the neuroscience field. He is now member of the coordination group of the European IMI Consortium Pharmacog. He is President of the Italian Association on Brain Aging Research (AIRIC) and member of the European Academy of Sciences. He is the author of more than 160 peer-reviewed scientific articles and about 30 reviews or book chapters. Selected publications Forloni G. Angeretti N., Chiesa R., Monzani E., Salmona M., Bugiani O.,Tagliavini F. Neurotoxicity of a prion protein fragment. Nature 362: (1993) Forloni, G., Tagliavini, F.,Bugiani, O. and Salmona, M. Amyloid in Alzheimer s disease and prion-related encephalopathies: Studies with synthetic peptides. Progr. Neurobiol. 49: (1996) Forloni, G., Bertani, I. Calella, AM., Thaler, F.Invernizzi. R. Alpha-synuclein and Parkinson's disease selective neurodegeneration effect of alpha synuclein fragment on dopaminergic neurons in vitro. Ann. Neurol. 47: (2000) Forloni G. Iussich, S. Awan T. Colombo L. Angeretti, N. Girola, L. Bertani, I. Poli, G. Caramelli, M. Bruzzone, MG.Farina, L. Limido, L. Rossi, G. Giaccone G. Ironside, JW. Bugiani, O.Salmona M. and Tagliavini, F. Tetracyclines affect prion infectivity Proc. Natl. Acad. Sci. New York 99: (2002) Pesaresi M, Lovati C, Bertora P, Mailland E, Galimberti D, Scarpini E, Quadri P, Forloni G, Mariani C. Plasma levels of beta-amyloid (1-42) in Alzheimer's disease and mild cognitive impairment. Neurobiol Aging., 27:904-5 (2006) Fioriti, L. Angeretti, N.. Colombo, L., De Luigi A., Manzoni, C., Colombo A., Morbin, M., Tagliavini, F., Salmona, M. Chiesa, R. Forloni, G. Neurotoxic and gliotrophic activity of a synthetic peptide homologous to Gerstmann-Sträussler- Scheinker disease amyloid protein. J. Neurosci. 27: (2007) Dossena S, Imeri L, Mangieri M, Garofoli A, Ferrari L, Senatore A, Restelli E, Balducci C, Fiordaliso F, Salio M, Bianchi S, Fioriti L, Morbin M, Pincherle A, Marcon G, Villani F, Carli M, Tagliavini F, Forloni G, Chiesa Mutant prion protein expression causes motor and memory deficits and abnormal sleep patterns in a transgenic mouse model. Neuron ; 60 : (2008) Ettore Beghi (EB) graduated in Medicine in 1972 and received his specialty in neurology in 1976 at the University of Milan. He trained in epidemiology with a fellowship at the Department of statistics and Epidemiology of the Mayo Clinic in Rochester, MN (USA). He is Head of the Laboratory of Neurological Disorders at the Mario Negri Institute, Director of the Neurophysiology/Epilepsy Unit and Professor of Neuroepidemiology at the University of Milano-Bicocca, Monza. He is member of the editorial board of the journals Epilepsia, Neuroepidemiology, Inpharma, Drugs in R & D, Clinical Drug Investigation, Neurological Sciences and is a referee of several national and international medical journals. The main areas of interest and research include studies on the descriptive, analytic, and experimental epidemiology in the field of epilepsy, peripheral neuropathies, headache, and amyotrophic lateral sclerosis. Selected publications Leone, MA. Solari, A.,Beghi, E. for the FIRST Group. Treatment of the first tonic-clonic seizure does not affect longterm remission of epilepsy. Neurology 2006; 67: Millul, A., E. Beghi, G. Logroscino, A. Micheli, E. Vitelli, A. Zardi, for the Registro Lombardo SLA (SLALOM). Survival of patients with amyotrophic lateral sclerosis in a population-based registry. Neuroepidemiology 2005; 25: Tonini, C., E. Beghi, A.T. Berg, G. Bogliun, L. Giordano, R.W. Newton, A. Tetto, E. Vitelli, D. Vitezic, S. Wiebe. Predictors of epilepsy surgery outcome: a meta-analysis. Epilepsy Res 2004; 62: Van den Broek, M., and Beghi E., for the RESt-1 Group. Morbidity in patients with epilepsy: type and complications. A European Cohort Study. Epilepsia 2004; 45: Van den Broek, M. and Beghi E. for the RESt-1 Group. Accidents in patients with epilepsy: type and complications. A European Cohort Study. Epilepsia 2004; 45:

82 Musico, M., E. Beghi, A. Solari, F. Viani for the First Seizure Trial Group. Treatment of the first tonic-clonic seizure does not improve the prognosis of epilepsy. Neurology 1997; 49: Caterina Bendotti, got her degree in Pharmacy at the University of Milano in 1984; In she was post doc at the Genetic developmental Lab, Dept. of Physiology of the Johns Hopkins University, Baltimore, USA. In she was research fellow in the laboratory of Neuropharmacology and in the 1992, she became head of the Molecular Neurobiology Unit in Institute, since 1998 she is head of laboratory. The major research interest is the study of pathogenetic mechanisms of familial Amyotrophic Lateral Sclerosis.. Since 2002 she is a member of the editorial board of Journal of Neurochemistry. In has been Member of Scientific Committees of the International Symposia on ALS held in Milano, Novembre,2003. In has been member of the Italian Ministry of Health Committees for the diagnosis, cure, care and assistance of patients with ALS. Since 2005 is member of the Board of Directors of the Italian Society of Neuroscience. Since 2006 is member of the Research Advisory Panel of the MND Association, UK. Scientific reviewer of 11 international scientific journals. In 2007 she has co-organised the first international meeting on Mutant SOD1 and familial ALS:from the molecule to man held in Milano(13-16 September). She is author and co-author of 110 articles 100 of which with peer-review. Rapporteur of many communications in national and international meetings. Selected publications Sau D, De Biasi S, Vitellaro-Zuccarello L, Riso P, Guarnieri S, Porrini M, Simeoni S, Crippa V, Onesto E, Palazzolo I, Rusmini P, Bolzoni E, Bendotti C, Poletti A. Mutation of SOD1 in ALS: a gain of a loss of function. Hum Mol Genet. 16(13): , Massignan T, Casoni F, Basso M, Stefanazzi P, Biasini E, Tortarolo M, Salmona M, Gianazza E, Bendotti C, Bonetto V. Proteomic analysis of spinal cord of presymptomatic amyotrophic lateral sclerosis G93A SOD1 mouse.biochem Biophys Res Commun. 353(3):719-25, 2007 Peviani M, Cheroni C, Troglio F, Quarto M, Pelicci G, Bendotti C. Lack of changes in the PI3K/AKT survival pathway in the spinal cord motor neurons of a mouse model of familial amyotrophic lateral sclerosis.mol Cell Neurosci. 34: , 2007 Carri MT, Grignaschi G, Bendotti C. Targets in ALS: designing multidrug therapies. Trends Pharmacol Sci. 27(5):267-73, 2006 Cheroni C., Peviani M., Cascio P., Debiasi S., Monti C. and Bendotti C. Accumulation of human SOD1 and ubiquitinated deposits in the spinal cord of SOD1G93A mice during motor neuron disease progression correlates with a decrease of proteasome. Neurobiol. Disease. 18(3): , 2005 Bendotti C and MT Carri. Lessons from models of SOD1-linked familial ALS. Trends Mol Med. 10(8): , 2004 Bendotti C., Tortarolo M., Suchak S.K., Calvaresi N., Carvelli L., Bastone A., Rizzi M., Rattray M. and Mennini T. Transgenic SOD1 G93A mice develop reduced GLT-1 in spinal cord without alterations in cerebrospinal fluid glutamate levels. J. Neurochem.,79, , 2001 Migheli A., Atzori C., Piva R., Tortarolo M., Girelli M., Schiffer D. and Bendotti C. Lack of apoptosis in mice with ALS. Nature Medicine: 5, , Silvio Caccia. Laurea in Pharmacy at the University of Milan. Diploma in Industrial Chemistry at the L. Cobianchi Technical Institute (Verbania, NO) and Diploma in Biochemical Research at the Istituto di Ricerche Farmacologiche Mario Negri (Milan). Research fellow, Laboratory of General Pharmacology at the Mario Negri Institute, ; Permanent Researcher, Laboratory of Neuropharmacology, 1975; Head of Pharmacokinetic Unit, 1983; Head of Drug Metabolism Laboratory, 1988 to date, doing research in the field of pharmacology and toxicology with particular focus on pharmacokinetic aspects, both at the pre-clinical and clinical level. He has been member of the scientific assessment teams (acting as expert) for the evaluation of marketing authorisation applications submitted to the European and Italian Agencies.He is author and co-author of more than 200 articles, including reviews, monographs and book chapters. Selected publications Caccia S. N-dealkylation of arylpiperazine derivatives: disposition and metabolism of the 1-aryl-piperazines formed. Curr Drug Metab 2007 ; 8 : Caccia S. Main active components of St. John's Wort (Hypericum Perforatum) extracts: current analytical procedures for pharmacokinetics and concentration-response studies. Curr Pharm Anal 2006; 2: Caccia S. Antidepressant-like components of Hypericum perforatum extracts: An overview of their pharmacokinetics and metabolism. Curr Drug Metab 2005; 6: Caccia S. Metabolism of the newest antidepressants: Comparisons with related predecessors IDrugs 2004; 7: Caccia S. Biotransformation of post-clozapine antipsychotics. Pharmacological implications. Clin Pharmacokinet 2000; 38: 39. Caccia S. Metabolism of the newer antidepressants. An overview of the pharmacological and pharmacokinetic implications. Clin Pharmacokinet 1998; 34:

83 Luigi Cervo was awarded the degree of Doctor of Philosophy (Ph.D.) from the Open University, Milton Keynes, U. K. in Since 2006 he has been the head of the Experimental Psychopharmacology Laboratory. From 1978 to 2001 he has been a research fellow and then Chief of the Behavioural Pharmacology Unit in the laboratory of Neuropharmacology and in 1981 he was awarded the degree in Biochemical Research from the M. Negri Institute. Between 1981 and 1983 he spent two years as a research fellow in the Department of Psychiatry at the Chicago University, Illinois, U.S.A. His main research interests concern Neuropsychopharmacology and the mechanism of action of psychotropic drugs. In particular the role of receptors subtypes for serotonin, dopamine, noradrenaline and glutamate in drug dependence and drug craving, depression, anxiety. Author and co-author of several peer-review articles, author of communications in international meetings, he is scientific reviewer of several international scientific journals. Member of Society for Neuroscience and Italian Society of Neuropsychopharmacology. Selected publications Cervo L, Carnovali, F, Stark JA, Mennini T. Cocaine-seeking behavior in response to drug-associated stimuli in rats: involvement of D 3 and D 2 dopamine receptors. Neuropsychopharmacology 2003; 28: Cervo L, Cocco A, Carnovali F. Effects on cocaine and food self-administration of (+)-HA-966, a partial agonist at the glycine/nmda modulatory. Psychopharmacology (Berl) 2004; 173: Grignaschi G, Burbassi S, Zennaro E, Bendotti C, Cervo L. A single high dose of cocaine induces behavioural sensitization and modifies mrna encoding GluR1 and GAP-43 in rats. Eur J Neurosci 2004; 20: Cervo L, Canetta A, Calcagno E, Burbassi S, Sacchetti G, Caccia S, Fracasso C, Albani D, Forloni G, Invernizzi R. Deficits of serotonin synthesis cause resistance to antidepressants, J Neuroscience 2005; 25: Cervo L, Cocco A, Putrella C, Heidbreder CA. Selective antagonism at dopamine D3 receptors attenuates cocaineseeking behaviour in the rat. Int J Neuropsychopharmacol. 2007; 10: Burbassi S, Cervo L. Stimulation of serotonin(2c) receptors influences cocaine-seeking behavior in response to drugassociated stimuli in rats. Psychopharmacology (Berl). 2008; 196: Burattini C, Burbassi S, Aicardi G, Cervo L. Effects of naltrexone on cocaine- and sucrose-seeking behaviour in response to associated stimuli in rats. Int J Neuropsychopharmacol. 2008; 11, Marchesi F, Piemonti L, Fedele G, Destro A, Roncalli M, Albarello L, Doglioni C, Anselmo A, Doni A, Bianchi P, Laghi L, Malesci A, Cervo L, Malosio M, Reni M, Zerbi A, Di Carlo V, Mantovani A, Allavena P. The chemokine receptor CX3CR1 is involved in the neural tropism and malignant behavior of pancreatic ductal adenocarcinoma. Cancer Res. 2008; 68, Guzzetti S, Calcagno E, Canetta A, Sacchetti G, Fracasso C, Caccia S, Cervo L, Invernizzi RW. Strain differences in paroxetine-induced reduction of immobility time in the forced swimming test in mice: role of serotonin. Eur J Pharmacol. 2008; 594, Maria Grazia De Simoni got the Doctoral Degree in Biological Sciences in 1977 at the University of Milano, Italy. 1981: Research Specialist in Pharmacology (PhD), Mario Negri Institute, Milan, Italy : European Community fellowship for "Advanced Professional Training", INSERM U 171, Universitè Claude Bernard, Lyon, France; 1984 Department of Histology, Karolinska Institute, Stockholm. Working experience: : Chief of the Neurochemistry Unit, Mario Negri Institute, Milano; 1998-present: Chief of the Laboratory of Inflammation and Nervous System Diseases, Mario Negri Institute. Scientific interests: pathogenesis of cerebral ischemia/reperfusion and traumatic brain injury; inflammatory response and apoptotic mechanisms as targets of therapeutic strategies; animal models and clinical studies. She is member of the board of Master in Tecnologie Avanzate Applicate alle Patologie Neurodegenerative", University of Milan and member of the board of Associazione Italiana per la Ricerca sull Invecchiamento Cerebrale (AIRIC). Selected pubblications De Simoni MG, Storini C, Barba M, Catapano L, Arabia AM, Rossi E, Bergamaschini L. Neuroprotection by complement (C1)-inhibitor in mouse transient brain ischemia. J Cereb Blood Flow Metab, 23: , De Simoni M G, Rossi E, Storini C, Pizzimenti S, Echart C, Bergamaschini L. The powerful neuroprotective action of C1-inhibitor on brain ischemia-reperfusion injury does not require C1q. Am J Pathol., 164: , Bergamaschini L, Rossi E, Storini C, Pizzimenti S, Distaso M, Perego C, De Luigi A, Vergani C and De Simoni MG. Peripheral treatment with enoxaparin, a low-molecular weight heparin, reduces plaques and β-amyloid accumulation in a mouse model of Alzheimer s disease. J. Neurosci. 24: ,

84 Troglio F, Echart C, Gobbi A, Pawson T, Pelicci PG, De Simoni MG & Pelicci G. The neuron-specific Rai (Shc C) adaptor regulates the PI3K-Akt pathway in vivo and protects against cerebral ischemia. Proc Natl Acad Sci U S A 101(43): , Storini C, Bergamaschini L, Gesuete R, Rossi E, Maiocchi D, De Simoni MG. Selective inhibition of plasma kallikrein protects brain from reperfusion injury. JPET 318: , 2006 Capone C, Fabrizi C, Piovesan P, Principato MC, Marzorati C, Ghirardi O, Fumagalli L, Carminati P and De Simoni MG. 2-Aminotetraline derivative protects from ischemia/reperfusion brain injury with a broad therapeutic window, Neuropsychopharmacology, 32: , 2007 Capone C, Frigerio S, Fumagalli S, Gelati M, Principato M C, Storini C, Montinaro M, Kraftsik R, De Curtis M, Parati E, De Simoni MG. Neurosphere - derived cells exert a neuroprotective action by changing the ischemic microenvironment. PLoS ONE 2 e373, Pastori C, Librizzi L, Breschi GL, Regondi C, Frassoni C, Panzica F, Frigerio S, Gelati M, Parati E, De Simoni MG, de Curtis M.Arterially perfused neurosphere-derived cells distribute outside the ischemic core in a model of transient focal ischemia and reperfusion in vitro.plos ONE. 3(7):e Roberto W. Invernizzi started his career in the laboratory of Neuropharmacology of the Istituto di Ricerche Farmacologiche Mario Negri in 1976, where, at present, he heads the Laboratory of Neurochemistry and Behavior. In 1986 he got his degree in Biological Sciences at the Università Statale di Milano and in 1996 he was nominated head of the Intracerebral Microdialysis Unit. Of particular interest to Invernizzi s research team is the study of the neurochemical mechanisms and neuronal circuitries involved in the pathology of the main psychiatric diseases, such as depression and schizophrenia and in the mechanism of action of psychotropic drugs. Since 1987 he applied the intracerebral microdialysis technique to study the in vivo release of monoamines. Using this technique, Invernizzi s team first contributed to clarifying the role of serotonergic and adrenergic autoreceptors in the effect of antidepressant drugs suggesting new hypotheses on their mechanism of action. Currently, Invernizzi s laboratory is involved in two main collaborative projects aimed at clarifying the neurochemical mechanisms involved in the resistance to antidepressant drugs and the role of glutamatergic and serotonergic mechanisms in attentional processes. Reviewer for various international journals in the field of pharmacology and neurochemistry. Author and co-author of more than 60 peer-reviewed articles. Member of the Italian Society of Neuroscience and the Italian Society of Pharmacology. Selected publications Baviera M, Invernizzi RW, Carli M. Haloperidol and clozapine have dissociable effects in a model of attentional performance deficits induced by blockade of NMDA receptors in the mpfc. Psychopharmacology 2008; 196: Guzzetti S, Calcagno E, Canetta A, Sacchetti G, Fracasso C, Caccia S, Cervo L, Invernizzi RW Strain differences in paroxetine-induced reduction of immobility time in the forced swimming test in mice: Role of serotonin. Eur. J. Pharmacol. 2008; 594: Calcagno E, Canetta A, Guzzetti S, Cervo L, Invernizzi RW. Strain differences in basal and post-citalopram extracellular 5-HT in the mouse medial prefrontal cortex and dorsal hippocampus: relation with tryptophan with tryptophan hydroxylase-2 activity. J Neurochem 2007; 103 : Invernizzi RW, Pierucci M, Calcagno E, Di Giovanni G, Di Matteo V, Benigno A, Esposito E. Selective activation of 5HT2C receptors stimulates GABA-ergic function in the rat substantia nigra pars reticulata: a combined in vivo electrophysiological and neurochemical study. Neuroscience : Calcagno E, Carli M, Invernizzi RW The 5-HT1A receptor agonist 8-OH-DPAT prevents prefrontocortical glutamate and serotonin release in response to blockade of cortical NMDA receptors J Neurochem 2006; 96: Cervo L, Canetta A, Calcagno E, Burbassi S, Sacchetti G, Caccia S, Fracasso C, Albani D, Forloni G, Invernizzi RWGenotype-dependent activity of tryptophan hydroxylase-2 determines the response to citalopram in a mouse model of depression J Neurosci 2005; 25: Greco B, Invernizzi RW, Carli M Phencyclidine-induced impairment in attention and response control depends on the background genotype of mice: reversal by the mglu2/3 receptor agonist, LY Psychopharmacology (Berl) 2005; 179: Ugo Lucca got his Master of Science, University of Aberdeen - UK, At the Mario Negri Institute he was investigator from , head of the "Clinical Evaluation of Antidementia Drugs Unit" ( ) and, since 1996, head of the "Laboratory of Geriatric Neuropsychiatry". The main areas of interests include epidemiology and clinic features of dementia; natural history of dementia; neuropsychiatric disorders of the elderly; instruments for the screening diagnosis and clinical course assessment of dementia; clinical evaluation of anti dementia treatments and CNS active drugs (phase I, II, III, IV and observational studies). 83

85 Selected publications Spagnoli A, Lucca U, Menasce G, Bandera L, Cizza G, Forloni G, Tettamanti M, et al. Long-term acetyl-l-carnitine treatment in Alzheimer's disease. Neurology 1991; 41: Lucca U, Comelli M, Tettamanti M, Tiraboschi P, Spagnoli A. Rate of progression and prognostic factors in Alzheimer s disease: a prospective study. J Am Geriats Society 1993; 41: Lucca U, Tettamanti M, Forloni G, Spagnoli A. Nonsteroidal anti-inflammatory drug use in Alzheimer s disease. Biological Psychiatry 1994; 36: Imbimbo BP, Martelli P, Troetel WM, Lucchelli F, Lucca U, Thal LJ, and the Eptastigmine Study Group. Efficacy and safety of eptastigmine for the treatment of patients with Alzheimer s disease. Neurology 1999; 52: Quadri P, Fragiacomo C, Pezzati R, Zanda E, Forloni G, Tettamanti M, Lucca U. Homocysteine, folate, and vitamin B- 12 in mild cognitive impairment, Alzheimer s disease and Vascular Dementia. Am J Clinical Nutr 2004; 80: Lucca U, Tettamanti M, Quadri P. Homocysteine lowering and cognitive performance. New England Journal of Medicine 2006; 355: Lucca U, Tettamanti M, Mosconi P, Apolone G, Gandini F, Nobili A, Tallone MV, Detoma P, Giacomin A, Clerico M, Tempia P, Guala A, Fasolo G, Riva E.Association of mild anemia with cognitive, functional, mood and quality of life outcomes in the elderly: the "Health and Anemia" study. PLoS ONE. 3(4):e1920 (2008) Alessandro Nobili got his degree in Medicine (Milan, 1990). Master in Biotechonological Research, Regione Lombardia, Milan International School of Pharmacology, 31 Course on: Drug Epidemiology and Post-marketing Surveillance, Erice, September Course on: Methods in Epidemiological Research, Milan, October Course: Long Term Clinical Trials, Cogne January Main areas of interest Methodology of Randomized Clinical Trials; Pharmacoepidemiology and postmarketing surveillance research; Drug utilization studies; Quality assessment of geriatric services; Qualitative studies on caregiver role in the care of patients with dementia; Methodological evaluation of the Special Care Unit for Alzheimer Disease patients; Methodology of drug information. Employment and research experience Chief of the Unit of Quality Assessment of Geriatric Services Chief of the Drug Information Services for the Elderly, Laboratory of Geriatric Neuropsychiatry, Istituto di Ricerche Farmacologiche Mario Negri, Milan. Editorial Board of the MICROMEDEX Inc., Englewood, Colorado USA. National Expert accredited by Italian Ministry of Health for The Italian (AIFA) and European Agency for the Evaluation of Medicinal Products (EMEA). Head of the Laboratory of the Quality Assessment of Geriatric Services at the Mario Negri Institute since Selected publications Nobili A, Tettamanti M, Frattura L, et al. Drug use in the elderly in Italy. Ann Pharmacother 1997; 31: Nobili A, Gebru F, Rossetti A, et al. Doctorline a private toll-free telephone medical information service. Ann Pharmacother 1998; 32: Nobili A, Riva E, Tettamanti M, et al. The effect of a structured intervention on caregivers of patients with dementia and problem behavior: a randomized controlled pilot study. Alzheimer Dis Assoc Disord 2004; 18: Lucca U, Nobili A, Riva E, Tettamanti M. Low level of B vitamins and the risk of cognitive and functional decline in the very-old: results from the Monzino 80-Plus Study. Neurobiol Aging 2004; 25: 31. Lucca U, Nobili A, Riva E, Tettamanti M Cholinesterase inhibitor use and age in the general population Arch Neurol 2006; 63: Tettamanti M, Garri' M T, Nobili A, Riva E, Lucca U Low folate and the risk of cognitive and functional deficits in the very old: The Monzino 80-plus study J Am Coll Nutr 2006; 25: Nobili A, Piana I, Balossi L, Pasina L, Matucci M, Tarantola M, Trevisan S, Riva E, Lucca U, Tettamanti M.Alzheimer special care units compared with traditional nursing home for dementia care: are there differences at admission and in clinical outcomes?alzheimer Dis Assoc Disord. 22:352-6 (2008). Annamaria Vezzani got her Degree in Biological Science at the University of Milan in 1978 and she specialized in Neuropharmacology at the Mario Negri Institute in She spent her post-doctoral period in Baltimore at the University of Maryland in working on the mechanisms of epileptogenesis in experimental models of epilepsy. She spent additional post-doctoral periods at the University of Stockholm and at the Karolinska Institute between 1985 and She was on sabbatical at the Albert Einstein College of Medicine in 2002 in the laboratory of Developmental Epilepsy. She is involved in studies on the biochemical and molecular mechanisms involved in the etiopathogenesis of seizures disorders using experimental models of epilepsy. The present research is focused on the functional role of neuroactive peptides and inflammatory mediators in the modulation of neuronal excitability and seizure-related neurodegeneration. Focus of the research is also on the mechanisms of pharmacoresistance. Since 1997 she is the Head of the Laboratory of Experimental Neurology at the Mario Negri Institute. She is member of the Editorial Board of Epilepsy Currents and Neuroscience and 84

86 Associate Editor for Exp Models of Epilepsia. She is appointed of the Chair of the Commission on Neurobiology of International League Against Epilepsy which is promoting initiatives for improving translational research in epilepsy. Selected publications Balosso S, Maroso M, Sanchez-Alavez M, Ravizza T, Frasca A, Bartfai T, Vezzani A. A novel non-transcriptional pathway mediates the proconvulsive effects of interleukin-1 beta (2008) Brain, 131: Balosso S, Ravizza T, Perego C, Peschon J, Campbell I, De Simoni MG, Vezzani A. TNF-alpha inhibits kainic acidinduced seizures in mice via p75 receptors (2005) Ann Neurol, 57, 804 Dube C., Vezzani A., Behrens M., Bartfai T., Baram TZ. (2005) Interleukin-1beta contributes to the generation of experimental febrile seizures. Ann Neurol, 57,152. Richichi C, E-J. D. Lin, D. Stefanin, D. Colella, T. Ravizza,G. Grignaschi, G. Sperk, M. J. During and A. Vezzani Anticonvulsant and antiepileptogenic effects mediated by adeno-associated virus vector neuropeptide Y expression in the rat hippocampus (2004) J Neurosci, 24,3051 Rizzi M, Caccia S, Guiso G, Richichi C, Gorter JA, Aronica E, Aliprandi M, Bagnati R, Fanelli R, D'Incalci M, Samanin R, Vezzani A. Limbic seizures induce P-glycoprotein in rodent brain: functional implications for pharmacoresistance (2002) J Neurosci, 22, 5833 Vezzani A., Moneta D., Conti M., Richichi C., Ravizza T., De Luigi A., De Simoni M.G., Sperk, Andell-Jonsson S., Lundkvist J., Iverfeldt K. and Bartfai T. (2000) "Powerful anticonvulsant action of IL-1 receptor antagonist upon intracerebral injection and astrocytic overexpression in mice" Proc. Natl. Acad. Sci. USA, 97, Vezzani A., Moneta D., Conti M., Richichi C., Ravizza T., De Luigi A., De Simoni M.G., Sperk, Andell-Jonsson S., Lundkvist J., Iverfeldt K. and Bartfai T. (2000) "Powerful anticonvulsant action of IL-1 receptor antagonist upon intracerebral injection and astrocytic overexpression in mice" Proc. Natl. Acad. Sci. USA, 97, Tiziana Borsello got her Degree in Biological Science at the University of Torino in 1990 and she then obtained a PhD in Neuroscience at the University of Turin Medical School. She won 1 year fellow of the European Science Foundation scholarship for work at the Netherlands Research Institute of Amsterdam. From 1997 to 1999 she was a Researcher at the Institute of Neurobiology, CNR, Rome Italy. In the period she was Premier Assistant, Département de Biologie Cellulaire et de Morphologie, Université de Lausanne, Switzerland, and then became Maitre Assistant and group leader in the same institute. In 2004 joined the Biol. Neurodeg. Disorders Lab at the "Mario Negri Institute. In 2005 won the Prize of the Pfizer Foundation, Neuroscience and Diseases Nervous System. Since 2006 she is the Head of the Unit: Neuronal Death and Neuroprotection. Her main scientific interest is understanding the role of signaling pathways in neuronal death after different stress-stimuli and the neuroprotection. In particular, the present research is focused on the study of the mechanisms of excitotocic stress, ischemia, Traumatic Brain Injury and the cell death pathways in neurodegenerative diseases as Alzheimer, with the challenge to define the neuronal death pathways to design more specific methods of neuroprotection. Selected pubblications Repici M, Centeno C, Tomasi S, Forloni G, Bonny C, Vercelli A, Borsello T.Activation After Cerebral Ischemia And Effect Of D-Jnki1 On C-Jun And Caspase-3 Activation. Neuroscience. 2007, 150: 40-9 Borsello T., Centeno C., Riederer IM, Haeflinger JA and Riedere BM. Phosphorylation-dependent dimerization and subcellular localization of islet-brain 1/c-Jun N-terminal kinase-interacting protein 1. J Neurosci Res. 2007, 85: Borsello T Ed Neuroprotection: Methods In Molecular Biology Published By Humana Press, Usa Humana Press, USA, Methods in Molecular Biology, June 2007 Colombo A, Repici M, Pesaresi M, Santambrogio S, Forloni G, Borsello T. The Tat-Jnk Inhibitor Peptide Interferes With Beta Amyloid Protein Stability Cell Death Differ. 2007, 14: Borsello T and Forloni G. JNK signalling: a possible target to prevent neurodegeneration. Current Pharmaceutical Design 2007, 13, Centeno C., Repici M., Chatton J. Y., Riederer B. M., Bonny C., Nicod P., Price M., Clarke P. G., Papa S., Franzoso G. and Borsello T. Role of the JNK pathway in NMDA-mediated excitotoxicity of cortical neurons. Cell Death Differ, 2007, 14: Borsello T. and Bonny C.Use of cell-permeable peptides to prevent neuronal degeneration. Trend in Mol. Med. 2004, 10: Borsello T, Clarke PG, Hirt L, Vercelli A, Repici M, Schorderet DF, Bogousslavsky J, Bonny C. A peptide inhibitor of c-jun N-terminal kinase protects against excitotoxicity and cerebral ischemia. Nature Med. 2003, 9: Mirjana Carli started his scientific career in the laboratory of Neuropharmacology of the Istituto di Ricerche Farmacologiche Mario Negri Milan in 1977, where, at present, she is head of the Pharmacology of Cognitive Behaviour Unit. She spent a few years in the laboratory of Cognitive Neuroscience, Dept. of Experimental Psychology, University of Cambridge (UK) directed by Prof. 85

87 Trevor W. Robbins. Here she took interest in the role of brain monoamines in attention, and for this purpose developed several behavioral tests for rats. In 1986 she returned to the laboratory of Neuropharmacology of the Istituto di Ricerche Farmacologiche Mario Negri. Here she devoted her efforts to the study of the role played by neuronal mechanisms in cognitive processes such as memory, attention and executive functions. Her work has improved the knowledge of the role played by some serotonin receptors in cognitive processes. Selected publications Carli M, Baviera M, Invernizzi R, Balducci C Dissociable contribution of 5-HT1A and 5-HT2A receptors in the medial prefrontal cortex to different aspects of executive control such as impulsivity and compulsive perseveration in rat Neuropsychopharmacology 2006; 31: Greco B, Carli M Reduced attention and increased impulsivity in mice lacking NPY Y2 receptors: Relation to anxiolyticlike phenotype Behav Brain Res 2006; 169: Carli M, Baviera M, Invernizzi R, Balducci C The serotonin 5-HT2A receptors antagonist MI00907 prevents impairment in attentional performance by NMDA receptor blockade in the rat prefrontal cortex Neuropsychopharmacology 2004; 29: Balducci C, Nurra M, Pietropoli A, Samanin R, Carli M Reversal of visual attention dysfunction after AMPA lesions of the nucleus basalis magnocellularis (NBM) by the cholinesterase inhibitor donepezil and by a 5-HT(1A) receptor antagonist WAY Psychopharmacology (Berl) 2003; 167: Carli M, Balducci C, Samanin R. Stimulation of 5-HT1A receptors in the dorsal raphe ameliorates the impairment of spatial learning caused by intrahippocampal 7-chloro-kynurenic acid in naive and pretrained rats Psychopharmacology (Berl) 2000; 158: Carli M, Samanin R The 5-HT1A receptor agonist 8-OH-DPAT reduces rats' accuracy of attentional performance and enhances impulsive responding in a five-choice serial reaction time task: Role of presynaptic 5-HT1A receptors Psychopharmacology (Berl) 2000; 149: Barbara D Avanzo obtained her master in Philosophy in 1989 at the University of Milan. Her main field of interest is epidemiologic research in mental health. She was involved in the analysis of the implementation of the psychiatric reform in Italy and quality evaluation of services and their recent modifications with specific attention to the role of psychiatric residential facilities in the community service networks; evaluation of effectiveness of the most common psychosocial interventions; suicide trend monitoring and study of suicide prevention programs and initiatives. More recently, she is working on issues like recovery-oriented services, consumers empowerment, and methods of participation of consumers to evaluation of services, and acknowledgment of the value of the consumers point of view about psychiatric treatments and services. She worked as researcher in the Laboratory of General Epidemiology between 1991 and 1996, and she is Chief of the Unit of Epidemiology and Social Psychiatry since Member of the Scientific National Board of WAPR Italy and of the World Head Office of the WAPR. Selected publications Barbato A, D'Avanzo B. Efficacy of couple therapy as a treatment for depression: a meta-analysis. Psychiatr Q. 2008, 79: Barbato A, D'Avanzo B. Marital therapy for depression. Cochrane Database Systematic Reviews 2006; Issue 2. Parabiaghi A, Barbato A, D'Avanzo B, Erlicher A, Lora A. Assessing reliable and clinically significant change on Health of the Nation Outcome Scales: method for displaying longitudinal data. Aust N Z J Psychiatry 2005; 39: Barbato A, D'Avanzo B. Involuntary placement in Italy. Br J Psychiatry 2005; 186: Guaiana G, Andretta M, Corbari L, Mirandola M, Sorio A, D'Avanzo B, Barbui C. Antidepressant drug consumption and public health indicators in Italy, J Clinical Psychiatry 2005; 66: D'Avanzo B, Battino R N, Gallus S, Barbato A. Factors predicting discharge of patients from community residential facilities: A longitudinal study from Italy. Aust N Z J Psychiatry 2004; 38: D'Avanzo B, Barbato A, Barbui C, Battino N, Civenti G, Frattura L. Discharges of patients from public psychiatric hospitals in Italy between 1994 and Int J Social Psychiatry 2003; 49: 27-3 Emma Riva, Medical Doctor degree in 1984 University of Milan, PhD in 1990 in Cardiovascular Pathophysiology at the University of London (UK) Training: Research Assistant, Department of Pharmacology, Medical School, University of Ottawa, Canada; Internship in Internal Medicine, Ospedale Luigi Sacco, Milan; Cardiac Fellow, St Thomas' Hospital, London, UK. Field of interest: Prevalence and effects of anemia on cognitive, functional and clinical variables in the elderly; Problem behaviors in dementia; Burden for care-givers of Alzheimer Disease patients; End of life care. Present and past roles in Institute Head of the Geriatric Pharmacology Unit, Istituto "Mario Negri", Milan; Scientific Director of the hospice Via di Natale Franco Gallini, Aviano, Italy; Consultant Istituto Geriatrico Pio Albergo 86

88 Trivulzio, Milan: Project member of PREDICT (Policy Review and Evaluation of Dementia and Institutional Care Trends): a Transnational Comparison. Selected publications Lucca U, Tettamanti M, Mosconi P, Apolone G, Gandini F, Nobili A, Tallone MV, Detoma P, Giacomin A, Clerico M, Tempia P, Guala A, Fasolo G, Riva E.Association of mild anemia with cognitive, functional, mood and quality of life outcomes in the elderly: the "Health and Anemia" study. PLoS ONE. 3(4):e1920 (2008) Tettamanti M, Garrì MT, Nobili A, Riva E, Lucca U. Low folate and risk of cognitive and functional deficits in the very old: The Monzino 80-plus study. J Am Coll Nutr 2006;25: Lucca U, Nobili A, Riva E, Tettamanti M. Cholinesterase inhibitor use and age in the general population. Arch Neurol 2006;63: Nobili A, Riva E, Tettamanti M, Lucca U, Liscio MR, Petrucci B, Salvini Porro G. The effect of a structured intervention on caregivers of patients with dementia. Results of a Randomized Controlled Study. Alzheimer Dis and Associated Disorders 2004;18:75-82 Il malato terminale oncologico. Esperienze dall hospice. Ed. Emma Riva. Il Pensiero Scientifico, 2001 Riva E, Tettamanti M, Gallini C. Il ruolo del medico di medicina generale nella gestione dei malati terminali oncologici. Indagine svolta tra i medici di medicina generale in Friuli Venezia Giulia. Ricerca & Pratica 2001 Riva E, Nobili A, Trecate F. Per un impiego "ragionato" dei neurolettici, per la gestione dei disturbi del comportamento in corso di Malattia di Alzheimer. Rec Prog Med 1998;89: Mauro Tettamanti got his Biology Degree at the Università degli Studi di Milano in 1986, and the specialisation in Epidemiology and Medical Statistics in 1993, at the Università degli Studi di Pavia. Teaching experience Introduction course to statistics, Master in Ergonomy, Politecnico di Milano, years Areas of interest: Planning, conduction and analysis of clinical trials and epidemiologic researches in the geriatric field: Phase I, II, III and observational studies on the efficacy of drugs on neurologic disorders, with special emphasis on dementia; Effects of multi-disciplinary interventions on geriatric/dementia patients; Epidemiology and risk factors of dementia; Care of patients with terminal illness; Association of anemia with prevalence of diseases and cognitive problems Scholarship between 1989 and 1998, Senior Researcher since 1999 and Head of the Unit of Geriatric Epidemiology at the Mario Negri Institute since Selected publications Spagnoli A, Lucca U, Menasce G, Bandera L, Cizza G, Forloni G, Tettamanti M, et al. Long-term acetyl-l-carnitine treatment in Alzheimer's disease. Neurology 1991; 41: Lucca U, Comelli M, Tettamanti M, Tiraboschi P, Spagnoli A. Rate of progression and prognostic factors in Alzheimer's disease: A prospective study. J Am Geriatr Soc 1993; 41:45-49 Quadri P, Fragiacomo C, Pezzati R, Zanda E, Forloni G, Tettamanti M, Lucca U. Homocysteine, folate, and vitamin B- 12 in mild cognitive impairment, Alzheimer disease, and vascular dementia. Am J Clin Nutr 2004; 80: Lucca U, Nobili A, Riva E, Tettamanti M. Cholinesterase inhibitor use and age in the general population. Arch Neurol 2006; 63: Lucca U, Tettamanti M, Quadri P. Homocysteine lowering and cognitive performance. N Engl J Med 2006; 355:1390 Tettamanti M, Garri' M T, Nobili A, Riva E, Lucca U. Low folate and the risk of cognitive and functional deficits in the very old: The Monzino 80-plus study. J Am Coll Nutr 2006; 25: Nobili A, Piana I, Balossi L, Pasina L, Matucci M, Tarantola M, Trevisan S, Riva E, Lucca U, Tettamanti M.Alzheimer special care units compared with traditional nursing home for dementia care: are there differences at admission and in clinical outcomes? Alzheimer Dis Assoc Disord. 22:352-6 (2008). 87

89 INTRODUCTION TO THE DEPARTMENT'S ACTIVITIES The Department of Neuroscience is formed by ten Laboratories; the activities of research are devoted to the study of neurological and psychiatric diseases, evaluated by the biological point of view, clinical and epidemiological aspects and the quality of care. Together with these activities, in the Department other more general expertise are present. Pharmacokinetics studies, drug information service and preparation of protocols for clinical trial and epidemiological studies are activities in charge of the Neuroscience Department. Traditionally part of the Department was devoted to the creation of experimental models for the pharmacological, neurochemical and pathogenetic studies in Alzheimer or prion's diseases, epilepsy, depression and cognitive impairment. More recently, consolidated expertise were created in the pathogenesis of amyotrophic lateral sclerosis (ALS), cerebral stroke and drug abuse. Some of these disorders, like epilepsy, ALS and Alzheimer's disease are investigated from the clinical and epidemiological points of view for the evaluation of drug and care efficacy. The activities of the Department are aimed to an integration of the different expertise to develop multidisciplinary approaches. The purpose is to address at different levels, knowledge, therapy and clinical practice to the numerous questions, largely unresolved, proposed by the disorders of nervoussystem. FINDINGS/MAIN RESULTS 2008 In a cellular model it has been shown that the peptide of D-JNK-1-TAT inhibiting the JNK phosphorylation activity mediated by the enzyme JNK, exerts a control on β amyloid production, these data indicate possible therapeutic perspectives in Alzheimer s disease The intracerebral application of synthetic β amyloid 1-40 e 1-42 in oligomeric form is associated with a cognitive damage that does not occur when the pepetides are applied in monomeric form of fibrils species. The exposure of cultured hippocampal cells to β amyloid 1-42 in oligomeric form induces an alteration of dendritic spines. In the transgenic mice overexpressing a mutated form of human prion protein associated to CJD generated in the Institute exhibit some behavioral features reminiscent of the clinical condition in humans carrying the same mutation. In the same model has been found a significant alteration of endoplasmic reticulum in specific neuronal cells In cellular model the presence of α synuclein mutations associated to Parkinson s disease, does not influence the neuroprotective activity of the protein, this indicates that these mutations affected the aggregation state more than physiological activity of α synuclein. Polymorphisms in gene encoded for serotonin transporter protein influenced the risk to develop Parkinson s disease 88

90 In a prospective population-based study in the very old (Monzino 80-plus Study), behavioral disturbances are uncommon prior to clinical dementia onset. After dementia diagnosis, behavioral disturbances affect a large proportion of subjects, increasing with disease severity while decreasing in the oldest-olds. In the same prospective population-based study (Monzino 80-plus Study), use of antipsychotics in the very old did not seem to be associated with an increased risk of mortality over a follow up period of four years. In a prospective ambulatory population of cognitively normal or mildly cognitively impaired elderly, individuals with baseline elevated homocysteine concentrations have an almost 3-fold increased risk of developing dementia in the following 3-4 years. More than 1 out of 10 elderly persons (65-84 years) were anemic and most of the cases had a mild grade anemia. Hemoglobin concentrations decreased and prevalence of (mild) anemia increased with increasing age. After controlling for many potential confounders, mild anemia was found to be associated with poorer health conditions and with increased risk of clinically relevant outcome such as hospitalization and mortality. According to the survey conducted in the frame of GISAS Study the most important difficulty psychiatrists face in participating to clinical trials is related to application of experimental protocols to daily clinical practice. Patients with severe mental disorders included in the Natural Social Networks improved moderately but significantly on general wellbeing according to the HoNOS, and personal and social functioning according to the GAF, whereas improvements in quality of life were absent or not significant, particularly in relational and social dimensions. In a sample of family members of patients followed by the mental health services there was an overall moderately good evaluation of the services. Most areas related to accessibility and support offered were satisfying according to 60-80% of the respondents, whereas the efficacy in improving health conditions of the patient was satisfying according to less than 50%. Other aspects needing more efforts were: information about the treating plan, possibility of treatments delivered by the service and other agencies; family active involvement in the evaluation of the course of treatment and its effects; difficulties in reaching the service by phone and waiting lists; effective management of poor compliance of the patient; answer to emergency needs during closing hours of the service. Loss of ambulation, percutaneous gastrostomy and non-invasive mechanical ventilation are outcome measures to be used in epidemiological surveys and therapeutic trials. The probability of late remission of epilepsy is not uncommon. Partial seizures and having more seizures prior to treatment predict late remission The long-term prognosis of epilepsy is the same in patients treated at the first seizure and those treated at the recurrence. These findings suggest that treatment should be started at the first seizure only on a case-by-case basis. The use of a third drug in children refractory to two anticonvulsants does not affect the chance of seizure remission, suggesting that drug resistance in epilepsy can be identified at the time of failure of two drugs. 89

91 The incidence of acute symptomatic seizures in patients with a firstly diagnosed stroke followed prospectively is low. Cerebral hemorrhage is the only independent predictor of acute symptomatic seizures. We have demonstrated the crucial involvement of some pro- and anti-inflammatory cytokines in seizures using experimental models of epilepsy in rodents, thus describing a new etiopathological mechanism which may be relevant for human epilepsy The membrane-bound drug transport proteins are functionally activated by seizures and have a significant role in decreasing the brain concentrations of antiepileptic drugs in experimental models. Pharmacological intervention to block the activity of these proteins may contribute to reverse multidrug resistance in epilepsy Gene therapy studies highlight the possibility to significantly reduced spontanerous seizure that are refractory to anticonvulsant drugs opening the perspective of using gene therapy in pharmacoresistant forms of epilepsy. Complement C1-inhibitor (C1-INH) has powerful neuroprotective actions in brain ischemia/reperfusion injury and in traumatic brain injury Microglia can explain protective actions in the ischemic environment Neural stem cell reduce ischemia/reperfusion injury by changing the ischemic environment Blood-brain barrier cells can be preconditioned and induced to express a protective phenotype A dysfunction of proteasome is found in the motor neurons of SOD1 mutant mice, which might contribute to the accumulation of intracellular protein aggregates. This discovery open a route for a possible therapeutic strategy based on the application of substance that may increase the activity of proteasome. Genetic differences in serotonin synthesis contribute to the efficacy of SSRIs in mice 5-HT 1A and 5-HT 2C receptor antagonists restore the antidepressant-like effect in mice nonresponder to the SSRI alone SSRI-induced serotonin release is strongly suppressed in mice carrying the allele 1473G of TPH-2, the limiting step enzyme in brain serotonin synthesis The blockade of NMDA receptors of the rat prefrontal cortex induces an increase of glutamate release and is deleterious for prefrontal cortex-dependent cognitive functions 5-HT 2A receptor antagonists and 5-HT 1A and 5-HT 2C receptor agonists prevent the increase of glutamate release and attentional deficits caused by NMDA receptors blockade suggesting that some serotonin receptor subtypes might constitute a molecular target for the development of drugs for the treatment of cognitive deficits of schizophrenia 90

92 We show that in a glutamate NMDA model of cognitive deficit of schizophrenia antipsychotics may be differentiated by a selective effect of typical antipsychotics on compulsive perseveration, and atypical antipsychotics on impulsivity. A single session of cocaine self-administration is sufficient to shape rat behavior towards goaldirected behaviors and selectively up-regulate Arc expression in mpfc. This is the first evidence that the mpfc's function is already profoundly influenced by the first voluntary cocaine exposure Naltrexone, a non-selective opioid receptor antagonist, selectively modulates, in abstinent rats, the drug seeking behaviour induced by the environmental stimuli predictive of cocaine availability NATIONAL COLLABORATIONS Associazione Familiari Insonnia Familiare Fatale malattie da prioni, Treviso Associazione Italiana GIST A.I.G. Associazione per la Ricerca Neurogenetica, Lamezia Terme (CS) e ASL 6, Regione Calabria Agenzia di Sanità Pubblica del Lazio, Regione Lazio Assessorato alla Salute, Comune di Milano Azienda Ospedaliera Ospedali Riuniti di Bergamo Azienda Sanitaria Locale di Bergamo CEND, Centro Eccellenza per le Malattie Neurodegenerative, Università di Milano Centro Fatebenefratelli San Giovanni di Dio, Cernusco sul Naviglio Centro di Neurofarmacologia, Dipartimento di Scienze Farmacologiche, Università di Milano Centro Studi in Psichiatra, ASL 2, Torino Centro Parkinson-Istituti Clinici di Perfezionamento Clinica IRCSS S. Maria Nascente, Milano Clinica Neurologica III Università di Milano, Azienda Ospedaliera S. Paolo, Milano Clinica Psichiatrica, Università Milano Bicocca Consorzio Ricerche Luigi Amaducci, CRIC, Arcugnano (Vc) Consorzio MIA, Milano DIBIT, San Raffaele Scientific Insitute, Milano. Dipartimento di Chimica Biologica, Università di Padova Dipartimento di Chimica, Università egli Studi di Firenze Dipartimento Endicronologia, Università di Milano Dipartimento Farmaco Chimico Tecnologico, Università di Siena Dipartimento di Farmacologia Medica, Università di Milano Dipartimento di Fisiologia Umana, Facoltà di Medicina, Università di Milano Dipartimento di Medicina e Sanità Pubblica, Sezione di Psichiatria e Psicologia Clinica, Università di Verona Dip. di Morfofisiologia, Scuola di medicina Veterinaria, Università di Torino, Grugliasco (TO). Dip. Neurologia, IRCCS Fondazione Maugeri, Pavia Dipartimento Neurologia, Ospedale Molinette, Torino Dipartimento di Neurologia Università di Milano, Ospedale Luigi Sacco. Dipartimento di Neuroscienze, Università di Parma, Parma Dipartimento di Salute Mentale di Niguarda, Milano 91

93 Dipartimento di Salute Mentale ASL 3 Genovese, Genova Dipartimento di Salute Mentale ASL 4, Torino Dipartimento di Salute Mentale San Carlo, Milano Dip. di Scienze Biomolecolari e Biotecnologie, Università di Milano Dipartimento Scienze Neurologiche, Università di Genova, Genova Dipartimento Scienze Neurologiche, Ospedale Maggiore Policlinico di Milano Direzione Generale Famiglia e Solidarietà Sociale, Regione Lombardia, Milano Direzione Generale Sanità, Regione Lombardia, Milano Direzione Regionale Sanità e Servizi Sociali, Regione Umbria Divisione Neurologica, Università di Bologna Evidentia Medica, Grottaferrata, Roma Federazione Alzheimer Italia, Milano Franco Calori Cell Factory, Centro Trasfusionale e di Immunologia dei Trapianti, IRCCS Ospedale Maggiore, Milano Fondazione Clelio Angelino Fondo Edo Tempia Hospice Via di Natale Franco Gallini, Aviano (PN) IRCSS "Casa Sollievo della Sofferenza", San Giovanni Rotondo IRCCS Istituto Auxologico Italiano, Milano Istituto di Ricovero e Cura a Carattere Scientifico IRCCS (I.N.R.C.A.), Ancona IRCSS "San Raffaele", Milano Istituto Europeo di Oncologia, IRCCS, Milano Istituto di Farmacologia e Farmacognosia, Università di Urbino Istituto di Farmacologia, Università di Milano Istituto G. Ronzoni, Milano Istituto Nazionale Neurologico Carlo Besta, Milano Istituto Scientifico Humanitas Istituto di Scienze e Tecnologie della Cognizione, CNR, Roma Istituto "Stella Maris", IRCCS, Calambrone (PI) Istituto Superiore di Sanità, Roma Istituto Zooprofilattico Piemonte Liguria Val D'Aosta,Torino Laboratorio di Immunopatologia Renale, Ospedale San Carlo, Milano Laboratorio di Neuroscienze, Centro Dino Ferrari, Università di Milano Lega Italiana per la Lotta contro i Tumori Ospedale del Bambin Gesù, Roma Ospedale Regionale Ca Fondello, Treviso Ospedale "Molinette", Torino Polo Oncologico, ASL 12, Biella Provincia Lombardo-Veneta Ordine Ospedaliero San Giovanni di Dio, Fatebenefratelli di Cernusco sul Naviglio Scuola di Specializzazione in Psicoterapia IRIS-Insegnamento e Ricerca Individuo e Sistemi, Milano Scuola di Terapia Cognitiva Studi Cognitivi, Milano Società Italiana Medicina Interna, Roma Unione Nazionale delle Associazioni per la Salute Mentale (UNASAM), Milano Unità di Geriatria, Ospedale Maggiore IRCCS, Università di Milano Unità Operativa di Psichiatria, Azienda Ospedaliera Luigi Sacco di Milano, Milano Unità Operativa di Psichiatria, Azienda Ospedaliera San Gerardo di Monza, Monza Unità Operativa di Psichiatria di Garbagnate, Azienda Ospedaliere Salvini di Garbagnate, Garbagnate Milanese 92

94 Unità Operativa di Psichiatria, Fondazione IRCCS Ospedale Maggiore Policlinico, Mangiagalli e Regina Elena di Milano, Milano Università degli Studi di Foggia Università Cattolica del Sacro Cuore di Roma Università del Piemonte Orientale, Novara Università di Milano, IRCCS Ospedale Maggiore, Milano Università Milano-Bicocca, Monza Università La Sapienza, Roma U.O. Neurologia, Clinica S. Maria, IRCCS, Castellanza (VA). UNASAM, Unione Nazionale delle Associazioni per la Salute Mentale Unità Operativa di Psichiatria, Azienda Ospedaliera Luigi Sacco di Milano, Milano 93

95 INTERNATIONAL COLLABORATIONS Albert Eistein College of Medicine, Bronx, NY, USA Atomic Energy Commission, Service de Neurovirologie, Fontenay-aux-Roses, France Beaumont Hospital, Dublin, Ireland Brain Repair Centre, University of Cambridge, Cambridge, UK Cambridge Centre for Brain Repair, University of Cambridge, UK Centre for Neuroscience Research and Division of Biomolecular Sciences, GKT School, King s College, London, UK Centre National de la Recherche Scientifique, Paris. France Chorley & South Ribble General Hospital, Chorley, Columbia Univ, Haverstraw, NY, USA Department of Anatomy and Physiology, Laval University, Quebec Department of Cell Biology, Washington University, St Louis, USA Department of Chemistry,The Australian National University, Canberra City, Australia Department of Experimental Psychology, University of Cambridge, UK Department of Pathology and Infectious Diseases Royal Veterinary College, Herts, UK Department of Psychiatry, Medical Center University of Mississippi, Jackson, USA Directorate General for the Health and Consumer Protection, European Commission, Luxembourg Division of Medical Genetics, CHUV Lausanne, Switzerland European Union of Family Associations of People with Mental Illness (EUFAMI) Geriatric Division and Department of Metabolic Diseases, Ospedali Regionali of Lugano and Mendrisio, Switzerland HSPH Harvard University, Boston, USA IBCM, University of Lausanne, Lausanne, Switzerland INSERM U 751, Marseille, France Institut de Génétique Humaine du CNRS, Montpellier, France Jefferson Med Coll, Philadelphia, USA Karolinska Institutet, Stockholm, Sweden King s College Hospital, London, UK Lancaster University, Lancaster, UK. Max-Delbrück-Center for Molecular Medicine, Berlin, Germany National Insitute on Aging, NIH, Baltimore, USA Neuroprion, Network of Excellence, WP VI, EC Neurological Department of the University of Tirana, Albania Neurology, GlaxoSmithKline, New Frontiers Science Park North, Harlow UK Ninewells Hospital and Medical School, Dundee, Scotland UK Northern Illinois University, DeKalb, IL, USA Novartis Pharma, Basel, Switzerland NYU, NY, USA Ohio State Univ, Columbus, Ohio, USA Robarts Research Institute, London, Ontario, Canada Royal Manchester Children's Hospital, Manchester, UK Royal Preston Hospital, Preston, UK Sergievsky Center, Columbia University, New York, NY, USA Servizio di Geriatria, Ospedale della Beata Vergine, Mendrisio, Switzerland The Scripps Research Institute, Jupiter, Florida, USA University of Alberta, Canada University of Bristol, Frenchay Hospital, Frenchay, Bristol, UK. 94

96 University of Bristol, School of Medical Sciences, UK University of California at Irvine, Irvine, CA, USA University of Cardiff, United Kingdom University of Chicago, Chicago, IL, USA. University of Colorado, Denver, USA University Hospital, London, ON, Canada University of Innsbruck, Innsbruck, Austria University of Lausanne, Lausanne Switzerland University of Maryland, Baltimore, USA University of Maastricht, the Netherlands University of Rijeka Medical School, Rijeka, Croatia University of Szeged, Ungary Université de la Méditerranée Hôpital de la Timone Marseille, France Université Victor Segalen, Bordeaux, France Unit of Molecular Genetics, CHUV Lausanne, Switzerland Virtanen Institute for Molecular Sciences, University of Kuopio, Finland Vrije Universiteit Medical Center, Amsterdam, The Netherlands Walton Hospital, Liverpool, UK WAPR (World Association for Psychosocial Rehabilitation) Washington University, St Louis, MI,USA Weill Cornell Medical College, New York, USA World Mental Health, Department of Mental Health and Substance Abuse, Geneva, Switzerland EDITORIAL BOARD MEMBERSHIP Annals Pharmacotherapy (Nobili) Biochemical Journal (Chiesa) Brain Aging (Forloni) Clinical Drug Investigation (Beghi) Clinical Neurology and Neurosurgery (Beghi) Cochrane Collaboration, Epilessia (Beghi) Dialogo sui Farmaci (Nobili) Drugs in the R&D (Beghi) Early Intervention in Psychiatry (Barbato) Epidemiologia e Prevenzione (Lucca) Epilepsia (Beghi, Vezzani. Assistant editor) Epilepsy Current (Vezzani) Epilepsy Research (Vezzani) Inpharma (Beghi) International Journal of Mental Health (Barbato) Journal of Neurochemistry (Bendotti) Neurological Sciences (Beghi) Neuroepidemiology (Beghi) Neuroscience (Vezzani) Open Aging Journal (Forloni) Open Drug Metabolism Journal (Caccia) Open Geriatric Medicine Journal (Forloni) Psichiatria di Comunità (Barbato) 95

97 Quality of Life Research (Barbato) Ricerca & Pratica (Nobili) Stroke (De Simoni, Associate editor) PEER REVIEW ACTIVITIES Acta Neurologica Scandinavica Acta Psychiatrica Scandinava Alzheimer Disease and Associated Disorders American Journal of Clinical Nutrition American Journal of Human Genetics American Journal of Pathology American Journal of Physiology Annals of Neurology Annals of Pharmacotherapy Behavioural Brain Research Behavioural Neuroscience Biochimica et Biophysica Acta Biochemical Journal Biochemistry BioMed Central Neurology Biological Psychiatry Brain Research Brain Research Bulletin Brain Research Review Clinical Drug Investigation Clinical Neurology and Neurosurgery Clinical Pharmacokinetics Clin Pharm Therapy CNS Drugs Dialogo sui farmaci Drugs Epidemiologia e Psichiatria Sociale Epilepsia Epilepsy & Behavior European Journal of Immunology European Journal of Neuroscience European Journal of Pharmacology European Journal of Public Health Experimental Neurology European Neuropsychopharmacology Expert Opinion on Pharmacotherapy FASEB Journal FEBS letters Fundamental Clinical Psychopharmacology Future Drugs Giornale di Neuropsichiatria dell Età Evolutiva Glia International Journal of Neuropsychopharmacology 96

98 JAMA Journal of the American Board of Family Practice Journal of Biological Chemistry Journal of Cell. Biology Journal of Cerebral Blood Flow and Metabolism Journal of Chemical Neuroanatomy Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Science Journal of Headache and Pain Journal of Histochemistry and Cytochemistry Journal of Immunology Journal of Internal Medicine Journal of Neurochemistry Journal of Neuroimmunology Journal of Neurology, Neurosurgery and Psychiatry Journal of Neuroscience Journal of Pharmacy and Pharmacology Journal of Psychopharmacology Journal of Psychosomatic Research Journal of Structural Biology Life Sciences Lancet Lancet Neurology Molecular Brain Research Molecular and Cellular Neuroscience Molecular Therapy Nature Neuroscience Neuroepidemiology Neurology Neurological Sciences Nerobiology of Aging Neurobiology of Diseases Neuropharmacology Neuropsychopharmacology Neuroscience Neuroscience Letters N.S. Archives Pharmacology Parkinsonism & Related Disorders Pharmacological Research Pharmacoepidemiology and Drug Safety Pharmacology Biochemistry & Behavior PlosONE Proc Natl Acad Sci, USA Psychopharmacology Synapse Trends Molecular Medicine 97

99 NATIONAL AND INTERNATIONAL COMMITTEE MEMBERSHIP Advisory Board of the Italian League Against Epilepsy Board of "Master in Advanced Technologies for the Study of Neurodegenerative Diseases", University of Milan Commission on Health Care Policy of the International League Against Epilepsy (ILAE) Commission of Italian Minister of Health for the study of the problems associated to diagnosis, therapy and assistance ALS patients Coordination Group of NoE Neuroprion EC Coordination Group of the European project Quelles professionnalités en santé mentale. Perspectives croisées, usagers, élus professionnels. Council of Italian Association on Brain Aging Research (AIRIC) Expert for European Agency for the Evaluation of Medicines (EMEA) Expert reviewer for the Medical Research Council (MRC), UK Expert of the Minister of Health to EMEA Italian Association of Neuroepidemiology (Past President) Italian Association on Brain Aging Research (AIRIC, President) Italian Society of Neuroscience (Council) International Committee on Epilepsy and the Law International Organizing Committee e coordinator della segreteria al Global Forum for Community Mental Health, Department of Mental Health,WHO International Subcommittee of the American Academy of Neurology Medical Research Council Strategic Grant Application Mental Health Working Party of DG-SANCO, Directorate General Public Health and Consumer Protection of the European Union, Brussels, Belgium National expert accredited by AIFA (Italian Medicines Agency) for the European Agency for the Evaluation of Medicinal Products (EMEA) Neurobiology Commission of the International League against Epilepsy Neuroepidemiology Section of the American Academy of Neurology (past Chair) Research Advisory Panel, MND Association, UK Scientific Advisory Board of Sheffield Institute Foundation for MND 98

100 EVENT ORGANIZATION 6 a Giornata di studio sulla malattia di Alzheimer La depressione nella persona anziana 1 March 2008, Ateneo Veneto, Venezia ASL Provincia di Bergamo, Bergamo Interazioni tra farmaci: una valutazione della rilevanza clinica. 1 April 2008 ASL Bergamo (BG) ASL Provincia di Bergamo, Bergamo La prescrizione di FANS e IPP nel rispetto dell appropriatezza e della continuità terapeutica ospedale-territorio 23 April ASL Bergamo. 59th Annual Meeting of American Academy of Neurology Breakfast Seminar How to manage a patient with a first epileptic seizure: An evidence-based approach 28 April 5 May 2007, Boston, MA, USA. Join Meetin AINP-AIRIC, Istituto di Ricerche Farmacologiche Mario Negri, Milano June th European Congress on Epileptogenesis, Berlin, September th Neuroprion Meeting, Madrid, September 2008 ASL Provincia di Bergamo, Bergamo La prescrizione di antibiotici nel rispetto dell appropriatezza e della continuità terapeutica 22 October ASL Bergamo. Primo GiSAS Investigators Meeting 18 November 2008, Istituto di Ricerche Farmacologiche Mario Negri, Milano GRANTS AND CONTRACTS Abbott GmbH & Co. KG Amgen, Milano ASL 2 Piemonte. Assessorato alla Salute, Comune di Milano Association pour la recherché sur la SLA, France Bristol-Myers Squibb Boehringer Ingelheim CURE Epilepsy Dipartimento di Salute Mentale, Azienda Ospedaliera Niguarda Ca Granda, Milano Dana Foundation Evidentia Medica, Grottaferrata (Roma) Fondazione Cariplo, Milano Fondazione Mariani, Milano Fondazione Italo Monzino, Milano 99

101 FP6, European Union Glaxo-SmithKline, Italy Hospice "Via di Natale Franco Gallini", Aviano (PN) Human Frontiers Scientific Programme IMPHA II, DG-SANCO, Public Health and Consumers' Protection (Directorate General) Istituto Comprensivo Statale "G.D. Romagnosi", Carate Brianza, Milano I.R.I.S Istituto Superiore di Sanità Janssen-Cilag H. Lundbeck A/S, Danimark Ministero della Ricerca Scientifica Ministero della Salute MND Association, UK Newron Ospedale Casa Sollievo di San Giovanni Rotondo Pharming Regione Lombardia, Assessorato alla Famiglia e Solidarietà Sociale e Assessorato alla Sanità, Milano Rimoldi e Bergamini Sanofi-Aventis SELECTA MEDICA, Pavia Servier Laboratories, Parigi Sigma-Tau Telethon Unione Nazionale Associazioni per la Salute Mentale UNASAM Vertex 100

102 SELECTION OF SCIENTIFIC PUBLICATIONS FROM 2008 Albani D, Prato F, Fenoglio C, Batelli S, Dusi S, De Mauro S, Polito L, Lovati C, Galimberti D, Mariani C, Scarpini E, Forloni G. Association study to evaluate the serotonin transporter and apolipoprotein E genes in frontotemporal lobar degeneration in Italy. J Hum Genet. 2008; 53: Balosso S, Maroso M, Sanchez-Alavez M, Ravizza T, Frasca A, Bartfai T, Vezzani A. A novel non-transcriptional pathway mediates the proconvulsive effects of interleukin-1 beta 2008 Brain, 131: Batelli S, Albani D, Prato F, Polito L, Franceschi M, Gavazzi A, Forloni G. Early-onset Alzheimer disease in an Italian family with presenilin-1 double mutation E318G and G394V. Alzheimer Dis Assoc Disord. 2008; 22: Barbato A, D'Avanzo B. Efficacy of couple therapy as a treatment for depression: a meta-analysis. Psychiatr Q 2008; 79: Baviera M, Invernizzi RW, Carli M Haloperidol and clozapine have dissociable effects in a model of attentional performance deficits induced by blockade of NMDA receptors in the mpfc Psychopharmacology (Berl), 2008; 196: Beghi, E. The management of a first seizure Still a major debate. Epilepsia 2008; 49 (Suppl.1): 1. Beghi, E. Management of a first seizure. General conclusions and recommendations. Epilepsia 2008; 49 (Suppl.1): Beghi, E. Beghi, M. Epidemiology of epilepsy in women. In: Educational Kit on the Epilepsies 2008; Vol. 4: Beghi, E., Millul, A., Logroscino, G., Vitelli, E., Micheli, A. for the SLALOM Group. Outcome measures and prognostic indicators in patients with amyotrophic lateral sclerosis. Amyotrophic Lateral Sclerosis 2008; 9: Beghi, E:, Atzeni, L. Garattini, L. Economic analysis of newer antiepileptic drugs. CNS Drugs 2008; 22: Bernardino L, Balosso S, Ravizza T, Marchi N, Ku G, Randle JC, Malva JO, Vezzani A. Inflammatory events in hippocampal slice cultures prime neuronal susceptibility to excitotoxic injury: a crucial role of P2X(7) receptormediated IL-1beta release (2008) J Neurochem 106: Biasini E, Seegulam ME, Patti BN, Solforosi L, Medrano AZ, Christensen HM, Senatore A, Chiesa R, Williamson RA, Harris DA. Non-infectious aggregates of the prion protein react with several PrP(Sc)-directed antibodies. J Neurochem. 2008, 106: Biasini E, Medrano AZ, Thellung S, Chiesa R, Harris DA. Multiple biochemical similarities between infectious and non-infectious aggregates of a prion protein carrying an octapeptide insertion. J Neurochem. 2008, 104: Cheroni C, Marino M, Tortarolo M, Veglianese P, De Biasi S, Fontana E, Vitellaro Zuccarello L, Maynard CJ, Dantuma NP, Bendotti C. Functional alterations of the ubiquitin proteasome system in motor neurons of a mouse model of familial Amyotrophic Lateral Sclerosis. Hum Mol Genet Sep 29. [Epub ahead of print] Chiesa R, Piccardo P, Biasini E, Ghetti B, Harris DA. Aggregated, wild-type prion protein causes neurological dysfunction and synaptic abnormalities. J Neurosci. 2008, 28: Colovic M, Caccia S. Liquid chromatography-tandem mass spectrometry of I3,II8-biapigenin, the major biflavone in Hypericum perforatum extracts. J Chromatogr B Biomed Appl 2008 ; 863 : Colovic M, Fracasso C, Caccia S. Brain-to-plasma distribution ratio of the biflavone amentoflavone in the mouse. Drug Metabolism Letters 2008, 2 : Cosentino U, Pitea D, Moro G, Saracino GA, Caria P, Varì RM, Colombo L, Forloni G, Tagliavini F, Salmona M. The anti-fibrillogenic activity of tetracyclines on PrP : a 3D-QSAR study. J Mol Model. 2008, 14: Del Bo R, Ghezzi S, Scarlato M, Albani D, Galimberti D, Lucca U, Tettamanti M, Scarpini E, Forloni G, Bresolin N, Comi GP. Role of VEGF gene variability in longevity: a lesson from the Italian population. Neurobiol Aging. 2008, 29:

103 de Falco, FA., Sterzi, R.,Toso, V., Consoli, D. Guidetti, D., Provinciali, L., Leone, MA., Beghi, E. The neurologist in the emergency department. An Italian nationwide epidemiological survey. Neurol Sci 2008; 29: De Luigi A, Colombo L, Diomede L, Capobianco R, Mangieri M, Miccolo C, Limido L, Forloni G, Tagliavini F, Salmona M. The efficacy of tetracyclines in peripheral and intracerebral prion infection. PLoS ONE. 2008, 3(3):e1888. Dossena S, Imeri L, Mangieri M, Garofoli A, Ferrari L, Senatore A, Restelli E, Balducci C, Fiordaliso F, Salio M, Bianchi S, Fioriti L, Morbin M, Pincherle A, Marcon G, Villani F, Carli M, Tagliavini F, Forloni G, Chiesa Mutant prion protein expression causes motor and memory deficits and abnormal sleep patterns in a transgenic mouse model. Neuron 2008; 60 : Ferrarin M, Rabuffetti M, Tettamanti M, Pignatti R, Mauro A, Albani G. Effect of optical flow versus attentional strategy on gait in Parkinson's Disease: a study with a portable optical stimulating device. J. NeuroEnging. & Rehabil. 2008; 5: 3. Gironi, M., Guerini, FR., Beghi, E., Antonimi, G., Martinelli-Boneschi, F., Ceresa, L., Morino, S., Agliardi, C., Ferrante, P., Nemni, R. HLA-DRB1 polymorphisms distribution in chronic dysimmune polyneuropathy. Neuromuscul Disord 2008; 18: Guzzetti S, Calcagno E, Canetta A, Sacchetti G, Fracasso C, Caccia S, Cervo L, Invernizzi R Strain differences in paroxetine-induced reduction of immobility time in the forced swimming test in mice: role of serotonin. Eur J Pharmacol 2008 ; 594 : Hauser, WA ande Beghi, E.. First seizure definitions and worldwide incidence and mortality. Epilepsia 2008; 49 (Suppl.1): Lehnert M, Relja B, Sun-Young Lee V, Schwestka B, Henrich D, Czerny C, Froh M, Borsello T, Marzi I. A peptide inhibitor of C-jun N-terminal kinase modulates hepatic damage and the inflammatory response after hemorrhagic shock and resuscitation. Shock. 2008, 30: Leopoldo M, Lacivita E, De Giorgio P, Fracasso C, Guzzetti S, Caccia S, Contino M, Colabufo N A, Berardi F, Perrone R Structural modifications of N-(1,2,3,4-tetrahydronaphthalen-1-yl)-4-Aryl-1-piperazinehexa namides: Influence of lipophilicity and 5-HT7 receptor activity. Part III. J Med Chem 2008; 51 : Lucca U, Tettamanti M, Quadri P. The Italian version of Consortium to Establish a Registry of Alzheimer s Disease (CERAD). Alzheimer s & Dementia 2008; 4: 310. Lucca U, Tettamanti M, Mosconi P, Apolone G, Gandini F, Nobili A, Tallone MV, Detoma P, Giacomin A, Clerico M, Tempia P, Guala A, Fasolo G, Riva E. Association of Mild Anemia with Cognitive, Functional, Mood, and Quality of Life Outcomes in the Elderly: The Health and Anemia Study. PLoS ONE 2008; 3(4): e1920. Logroscino, G., Traynor, BJ., Hardiman, O., Chiò, A:, Couratier, P., Mitchell, JD., Swingler, RJ., Beghi, E. and for EURALS. Descriptive epidemiology of amyotrophic lateral sclerosis: new evidence and unsolved issues. J Neurol Neurosurg Psychiatry 2008; 79: Montes, J., Bendotti, C., Tortarolo, M., Cheroni, C., Hallak, H., Speiser, Z., Gore, S:, Blaugrund, E., and Gordon, PH. Translational Research in ALS. in :Animal and Translational Models of Behavioral Disorders. Volume 2 Neurologic Disorders RA McArthur and F Borsini (Eds.), Elsevier, NY, 2008, , Natalello A, Prokorov VV, Tagliavini F, Morbin M, Forloni G, Beeg M, Manzoni C, Colombo L, Gobbi M, Salmona M, Doglia SM. Conformational plasticity of the Gerstmann-Sträussler-Scheinker disease peptide as indicated by its multiple aggregation pathways. J Mol Biol Sep 19;381(5): Nobili A, Piana I, Balossi L, Pasina L, Matucci M, Tarantola M, trevisan S, Riva E, Lucca U, Tettamnti M. Alzheimer Special Care Units compared wuth traditional nursing home for dementia. Are there differences at admission and in clinical outcomes? Alzheimer Dis Assoc Disord 2008; 22:

104 Noè F, Pool AH, Nissinen J, Gobbi M, Bland R, Rizzi M, Balducci C, Ferraguti F, Sperk G, During MJ, Pitkänen A, Vezzani A. Neuropeptide Y gene therapy decreases chronic spontaneous seizures in a rat model of temporal lobe epilepsy 2008 Brain, 131: Pastori C, Librizzi L, Breschi GL, Regondi C, Frassoni C, Frigerio S, Gelati M, Parati E, De Simoni MG, de Curtis M. Arterially perfused neurosphere-derived cells do not cluster in the ischemic area in a model of transient focal ischemia and reperfusion in vitro PLoS ONE 3(7):e2754, Piazzini, A., Beghi, E., Turner, K., Ferraroni, M. the LICE Quality of Life Group. Health-related quality of life in epilepsy: findings with a new Italian instrument. Epilepsy & Behavior 2008; 13: Pugliatti M., Sobocki, P., Beghi, E., Pini, S., Cassano, GB., Altamura, AG, Pozzoli, S., Rosati, G. Cost of disorders of the brain in Italy. Neurol Sci 2008; 29: Repici M, Zanjani HS, Gautheron V, Borsello T, Dusart I, Mariani J. Specific JNK inhibition by D-JNKI1 protects Purkinje cells from cell death in Lurcher mutant mouse. Cerebellum. 2008, 7: Ravizza T, Noé F, Zardoni D, Vaghi V, Sifringer M, Vezzani A. Interleukin Converting Enzyme inhibition impairs kindling epileptogenesis in rats by blocking astrocytic IL- 1β production Neurobiol Dis, 2008, 31: Ravizza T, Gagliardi B, Noè F, Boer K, Aronica E and A. Vezzani. Innate and adaptive immunità during epiletogenesis and spontaneous seizures: evidence from experimental models and human temporal lobe epilepsy (2008) Neurobiol. Dis, 29: LAY PRESS SELECTION PUBLISHED IN 2008 Barbato A. La riabilitazione in psichiatria: cosa è cambiato e cosa dovrebbe cambiare. Psichiatria di Comunità 2008, 7: Barbato A. Mettere tra parentesi la malattia mentale. Nuove (e vecchie) ipotesi per la cura della sofferenza psichica. Animazione Sociale 2008; 8/9: Barbato A, D'Avanzo B, Ferrannini L, Parabiaghi A, Vaggi M. Un'occasione per la ricerca clinica in Italia: lo studio GISAS su aripiprazolo, olanzapina e aloperidolo nel trattamento dei disturbi schizofrenici. Psichiatria di Comunita 2008; 7: Barbato A. Confrontation entre professionels et usagers: vers une modification de la pratique in Italie. Rhizome 2008; 33: D'Avanzo B, Aliprandini E, Beghi M, Cornaggia C M, Erlicher A, Frova M, Mascarini A, Miragoli P, Righi A. Strutture residenziali e semiresidenziali nei servizi di salute mentale. Dove sta la differenza? Epidemiologia e Psichiatria Sociale 2008; 17: Invernizzi R, Caccia S. Gli antidepressivi. In: Le interazioni dei farmaci neuropsichiatrici tra farmacologia e rilevanza clinica. Selecta Medica, Pavia, 2008; Mennini T, Caccia S. Le benzodiazepine. In: Le interazioni dei farmaci neuropsichiatrici tra farmacologia e rilevanza clinica. Selecta Medica, Pavia, 2008; Pasina L, Caccia S. Gli antipsicotici. In: Le interazioni dei farmaci neuropsichiatrici tra farmacologia e rilevanza clinica Selecta. Medica, Pavia, 2008; Pasina L, Caccia S, Nobili A. Gli analgesici oppioidi. In: Le interazioni dei farmaci neuropsichiatrici tra farmacologia e rilevanza clinica. Selecta Medica, Pavia, 2008;

105 Pasina L, Caccia S, Nobili A. Antiparkinsoniani In: Le interazioni dei farmaci neuropsichiatrici tra farmacologia e rilevanza clinica. Selecta Medica, Pavia, 2008; Pasina L, Caccia S, Nobili A. I farmaci antiepilettici. In: Le interazioni dei farmaci neuropsichiatrici tra farmacologia e rilevanza clinica. Selecta Medica, Pavia, 2008; RESEARCH ACTIVITIES Laboratory of Biology of Neurodegenerative Disorders Alzheimer's disease: genetic studies and clinical investigations In collaboration with different neurological centers and the laboratory of Geriatric Neuropsychiatry it has been created a bank of blood samples for DNA of patients with Alzheimer s disease (AD), in familial (FAD) or sporadic form (SAD), and patients with vascular dementia (VD). In all subjects the diagnosis of dementia is performed according to the international guidelines. Since 2005 we started also the collection of blood samples from subjects with fronto-temporal dementia. The genetic studies are aimed to the identification of causal factors in FAD and risk factors in SAD. Mutations on genes encoding proteins involved in the physiopathology of AD were investigated. The pathogenic role of these mutations is under investigation using fibroblasts obtained from skin biopsy. Furthermore, we continued the screening of FAD samples for the genes encoding for presenilin 1 and 2 (PS-1 and PS-2) and APP, missense mutations in these three genes were associated with AD. Alzheimer's disease: preclinical studies The formation of β amyloid (Aβ) deposits in brain parenchyma and on the wall of cerebral blood vessels is an early event in AD and there are now numerous genetic, biochemical and neuropathological studies pointing to a causal role of Aβ in the pathogenesis of AD. Thus, prevention the formation of Aβ aggregates or their elimination once formed is a potential therapeutic approach to the disease. This aim is strongly persecuted with different strategies including the regulation of enzymes responsible of the synthesis and degradation of Aβ and the enzymes influencing the metabolism of amyloid precursor protein (APP). In the lab, we developed the idea to interfere directly with the Aβ deposits formation using anti-amyloidogenic drugs. The experimental studies have shown the potential therapeutic activity of these drugs in AD, and now they will be tested in a clinical setting. The role of oligomers in the Alzheimer pathogenesis Recent data have shown the essential role plays by oligomers, small and soluble aggregates of Aβ, in the Alzheimer pathogenesis and in particular in the cognitive decline associated to the disease. In collaboration with the Department of Biochemistry an Molecular Pharamacology we developed some in vivo models to analyze the neuronal dysfunction induced by Aβ 1-40 e 1-42 but not in monomeric or fibrillar species. The intracerebral application of these different forms confirmed that Aβ oligomers induced behavioral impairment while monomeric or fibrillar forms of Aβ did not affect the cognitive behavior, The intracellular signaling pathways, by which Aβ oligomers induce synaptic failure and consequently neuronal degeneration are poorly understood. Nevertheless increasing evidence indicate the involvement of kinases-dependent signalling pathways, and more specifically the JNK signalling pathway in these early degenerative events. 104

106 The JNK kinase phosphorylates APP (amyloid precursor protein) and its relevance in both neuronal death and brain plasticity is well established. We recently demonstrated, by using the specific cell penetrating JNK inhibitor peptide (D-JNKI1) charcterized by dr. Borsello, that JNK is responsible for APP phosphorylation at Thr668, and that its specific inhibition reduced the βapps and Aβ fragments production in primary cortical neurons. In addition, we could show that JNK inhibition leads to a shift from the amyloidogenic to the non-amyloidogenic pathway, a result with potentially important therapeutical implications. Synaptic Dysfunction Nowadays it is assumed that AD is a synapse-related pathology leading to synaptic dysfunction and loss, a phenomenon that precedes extensive amyloid deposition in the brain. At the same time, soluble diffusible forms of Aβ can perturb in an early stage of the disease the synaptic function causing a reduction of dendritic spines density in the cortex and hippocampus, an acute inhibition of long term potentiation (LTP) and the loss of critical spine proteins (e.g. membrane expression of NMDA receptors). However, the relationship between Aβ and synapses loss remains unclear and more efforts are necessary to better understand the mechanisms underlying Aβ synaptic toxicity. Our aim is to study the effects of Aβ oligomers on MAPKs pathways and elucidate the link between synaptopathies and the activation/inhibition of the JNK, p38 and ERK cascades. This study can potentially be a breakthrough in the comprehension of AD pathogenesis: understanding the cellular and molecular alterations that lead to AD will help in developing effective and preventive therapeutic strategies in order to counteract or nullify the degenerative processes activated by Aβ. MAPKs (ERK, p38 and JNK) are also implicated in the regulation of the immediate early signaling events that modulate synaptic plasticity by controlling LTP, LTD and the recycling of glutamate receptors (NMDAR and AMPAR) as well as their expression. Nevertheless, MAPKs involvement in the regulation of synaptic function and dysfunction and the mechanisms by which they trigger the synaptic loss induced by Aβ oligomers are largely unknown. The cargo strategy as a key tool in neuroprotection The possibility to target protein complexes and enzymes involved in intracellular signaling pathways by means of cell permeable peptide (CPP) conjugates represents a novel, versatile and extremely powerful way of blocking the propagation of intracellular signaling events or intracellular processes, with an unprecedented specificity allowing for reduction of side effects. D-JNKI1 is a cell-permeable, biologically-active peptide consisting of a carboxyl terminal sequence derived from the JNK binding domain of the scaffold protein JIP-1/IB1 (JBD20), and an amino terminal portion containing the HIV-TAT transporter sequence. D-JNKI-1 has been designed to block the interaction, mediated by JBD domain, between JNK and its targets. D-JNKI1 afforded powerful protection against NMDA excitotoxicity in cortical neurons and against cerebral ischemia in vivo, as well as in two other neurodegenerative models (hair-cell loss in animal models of sudden deafness and retinal ganglion cell loss following optic nerve crush in vivo. The peptide progressed in clinical trails for preventing brain ischemia/stroke (see CHUV/Xigenpharma Lausanne web-link). Among the possible targets for neuroprotection there is MKK7, that is activated, unlikely MKK4, during NMDA-stress,. To inactivate MKK7 we use lentiviruses because of the particular capability of integrating genetic material into the genome of non-dividing cells stably. Our lentiviral vector will carry a single si-rna duplex of MKK7. A second approach is to test in vitro the specific inhibitor: Gadd45, a molecule active on MKK7. Gadd45β binds to MKK7 directly and blocks its catalytic activity, the binding between Gadd45β/MKK7 being tighter than JIP1/MKK7. The endogenous Gadd45 interacts to MKK7 through direct, high-affinity contact but not with the other JNK upstream kinase, MKK4. For this study we initially plan to use a viral system that will allow us to observe the role of this 105

107 pathway in excitotoxicity, with a final goal of producing a cell-permeable peptide with a more specific effect in the prevention of neuronal death. SUMOlization in acute and chronic diseases Small ubiquitin-like modifier (SUMO) is a group of proteins responsible for post translational modifications influencing protein function, localization and stability. Recently, protein Sumoylation has attracted neuroscientists since it is implicated in the altered protein dynamics that are associated with various aspects of neurodegenerative disease, including stroke amd Alzheimer's Disease (AD). Our hypothesis is that SUMOylation confers neuroprotection against stressful stimuli through regulation of important stress signaling pathways. The aim of this study is to investigate the role of SUMOylation in models of acute (ischemia) and chronic brain diseases (AD). At present we are characterising the changes in expression and localisation of SUMO1-2/3 in an in vitro model of ischemia (NMDA application) as well as in a model of AD (oligomers application). Genetics of aging In collaboration with Geriatric Neuropsychiatry Lab for the Monzino 80-plus study and with dr. Maurizio Gallucci from the ARGel Association in Treviso for Trelong study we collected a large number of blood samples from subjects over seventy. In these samples we are performing a genetic analysis to identify genetic profiles associate to the longevity and /or to the agingassociated pathologies with specific attention to the dementias. The aim is to cross the genotype/phenotype profile with pathologies and environmental aspects including style of life, diet and economical conditions to identify risks and protective factors. Initially the subjects were genotypized for ApoE, whom allele E4 is a well-known risk factor for Alzheimer s disease and several other disorders and sirt-1 a gene codified for protein member of a enzymatic family of sirtuins associated to the longevity in several experimental models. The results are interesting but before drawing any conclusion we need to considere the numerous other parameters collected in our database. Prion's disease: in vitro studies Prion s diseases (TSE) are neurodegenerative disorders of sporadic inherited origin but also transmissible, they have different clinic and neuropathological features but all are characterized by the cerebral accumulation of an altered form of prion protein (PrPsc). TSE are rare diseases but the transmission from bovine (BSE) to humans induced a public health alarm in UK and successively in all Europe. PrPsc is involved in the pathogenesis of the disease and it is also an essential component of the infective agent. In the lab numerous projects were developed to understand the association between the presence of PrPsc and the neurodegenerative process. The pathogenetic role of PrP was investigated not only through the external application of peptides but also by the evaluation of intracellular mechanisms potentially involved in the formation of PrP aggregates. Prion s diseases: studies in vivo The lab has the facilities to study the experimental scrapie, mice and hamsters are inoculated with infected brain homogenate. The hamsters after intracerebral inoculation develop the disease in days and died within a month. The histopathological analysis of the brain show the presence of PrPsc deposits, neuronal damage, diffuse astrogliosis and the typical spongiosis at cortical and thalamic level. The anti-amyloidogenic activity of tetracyclines has been investigated also in this experimental contest. After the ex-vivo approach, where the homogenate was treated before the inoculation, the curative effect of tetracyclines was tested in collaboration with the lab of Biochemistry and Chemistry of Proteins by treatment the experimental scrapie in hamsters with intramuscular doxycycline, the treatment prolonged the 106

108 survival of the animals. Other drugs are now under investigation to verify the curative effects in TSE. In the lab are available transgenic mouse developed by dr. Chiesa. These mice express the mutated forms of PrP associated to the familial TSE. The homozygotes exhibit at different level of severity the symptoms reminiscent of the pathologies associated with the mutations. From the neuropathological point of view some lines exhibit an accumulation of PrP and clear neurodegeneration of the cerebellar granular cells, in the other brain regions no evident alterations were found. In the brain tissue of the mice was found a PrP with some of the characteristic of PrPsc, however the brain material is not infected. The pathogenesis of TSE is investigated in these models through different approaches including proteomics and electronic microscopy (EM). The EM analysis have shown that in transgenic mice there was an alteration at the neuronal level of the Golgi apparatus and/or of endoplasmic reticulum associated to the different distribution of mutated PrP compared to native protein. Furthermore, there are in progress studies to investigate the molecular mechanisms responsible of the neurodegenerative processes occurring in cerebellar granular cells of the insertional prion transgenic mice (PG14). Parkinson s Disease: genetic studies Parkinson s disease (PD) is the second more diffuse neurodegenerative disorder with an unknown pathogenesis, however for PD several therapies are available and, although at the symptomatic level, their efficacies is well-established. In the etiological studies on PD the genetic component has been traditionally considered with scarce interest whereas the environmental causes were carefully evaluated. This orientation was based on the evidence that the exposure to several toxins can mimic the PD pathology. However the genetic studies in the last few years have completely changed the perspective with the identification of mutations on two genes, encoding for alpha-synuclein and parkin, associated to the juvenile forms of the disease. A mutation on alpha synuclein gene is an event extremely rare, only three mutations identified until now, the parkin mutations are numerous ether in puntiform or in deletion form. The mutations on alpha-synuclein gene are dominant while the parkin mutations are associated with PD in recessive form. We collected, in collaboration with several neurological centers, blood samples from PD subjects and the screening of the samples involved genes like alphasynuclein, parkin, DJ-1 and other factors potentially involved in PD. Recently, an assocation between polymorphisms occurring in gene for serotonin transporter and the appearance of depression in PD subjects has been investigated. The results indicate no association between the serotonin traporter gene polymorphisms and depression in PD, but a direct association between these polymorphisms and PD itself. This indicate a more relevant involvement o serotonergic system in PD pathogenesis compared to whom is generally considered. Parkinson s disease: studies in vitro The identification of the mutations associated to Parkinson s disease (PD) gave a substantial contribute to understand the disease and allowed the develop of cellular models to investigate the pathogenesis of the disease. In past we showed the potential neurotoxic activity of alphasinuclein using the synthetic peptide homologous to the fibrillogenic fragment (NAC) of the protein. Successively with help of dr. Negro at the Department of Biochemistry at the University of Padova we prepared cdna vectors including the sequence of wild type and mutated alpha-synuclein Their transfection to the PC12 cells induced in specific conditions a cellular damage. More recently alpha-synuclein was associated to a TAT sequence capable to transport inside the cells the protein. With this method the intracellular concentration of alphasinuclein was better controlled. In a micromolar range alpha-synuclein was toxic, but in nanomolar range, it exerted neuroprotective effect against oxidative stress induced by hydrogen peroxide. This double effect dose-dependent was confirmed in an inducible model. More 107

109 recently again in collaboration with Dr. Negro (Padua University), we obtained the recombinant form of DJ-1 associated with TAT (TAT-DJ-1). This protein is similar to alpha-synuclein, mutations of its sequence has been associated to PD. TAT-DJ-1 silencing by small interference RNA (sirnai) were used to study the interaction between DJ-1 and alpha synuclein.. Laboratory of Neurological Disorders Epidemiological studies on amyotrophic lateral sclerosis (ALS) Included are studies on the incidence, risk factors and mortality of ALS. The data are obtained from a regional registry of the disease activated in 1998 and including all patients with newly diagnosed ALS identified in eight provinces of Lombardy. Using similar study protocols, the same data are collected in two additional regional registries (from Piemonte and Puglia) included in a network with the Lombard registry. Information obtained from patients enrolled in the Lombard registry and from cases examined by members of the Italian ALS Study Group has been used to assess the validity and reliability of diagnostic criteria for ALS and selected disability scales. Based on the data recorded, the annual incidence of ALS is comparable to that obtained in other Western countries where ALS registries have been activated, and is among the highest ever published (1.9 per 100,000). Mortality of ALS has been found to be comparable to that of studies from similar populations studied with the same protocol. The study on the validation of the current diagnostic criteria for ALS (the El Escorial criteria) showed that to be considered valid and reliable, the criteria should be used after proper training of the investigators. In October 2004, the Laboratory of Neurological Disorders has started a European collaborative group for the ALS registries (EURALS) with the intent to create a common database (completed in the year 2005) with the participation of the existing regional and national disease registries. Three major scientific activities are in course: 1. A comparative study of the clinical characteristics of patients with ALS enrolled in the registries as compared to those seen in secondary and tertiary health care facilities; 2. A meta-analysis of the incidence of ALS, performed by pooling data from the cohorts of patients enrolled in the population-based registries. The scientific reports of these studies have been submitted to international scientific journals for publication. Other studies are ongoing under the coordination of the Laboratory of Neurological Disorders: 1. A case-control study on trauma and risk of ALS (in collaboration with the Italian registries); 2. A case-control study on ALS, physical exercise and sport (in collaboration with the EURALS Consortium). Study # 3 is ongoing only in Italy; A survey of the prevalence of cognitive impairment and extrapyramidal signs in patients with newly diagnosed ALS (Italian registries); 4. A study of the mortality of ALS in the cohort of patients from the European population-based registries (EURALS Consortium). In 2008 a scientific report was published on the progression of ALS (marked by loss of ambulatiom, percutaneous gastrostomy and non-invasive assisted ventilation). Early and late remission of epileptic seizures During the calendar year 2008, the analysis of the data was completed for a retrospective observational study in a cohort of adolescents and adults with newly diagnosed epilepsy seen in two epilepsy centers (Monza, Bari) and followed for several years. The aim of the study was twofold: 1. To assess the long-term prognosis of epilepsy defined by the number of cases with late remission (ie, 2+ year seizure freedom with onset after 24 months of treatment; 2. Identification of the predictors of late remission as compared to early remission (ie, 2+ year remission immediately after treatment start). The cohort included 372 cases, 23% of which achieved early remission and 11% late remission. The chance of late remission increased significantly in patients with partial seizures and an increasing number of seizures prior to starting treatment 108

110 Innovative therapeutic strategies in patients with epilepsy A cohort of patients with a first unprovoked seizure, randomised since 1988 by several Italian centers to immediate treatment or to treatment only at the time of a seizure relapse, was followed to verify the impact of the two therapeutic strategies on the long-term prognosis of epilepsy, measured by the chance of achieving 5-year remission. To provide a pragmatic definition of drug resistance in childhood epilepsy, children refractory to two antiepileptic drugs (in sequence or in combination) were randomised to the use of a third drug or to the optimization of the existing treatment and followed for up to three years. Therapeutic response was measured by the achievement of a six-month period of remission. The study has been conducted in collaboration with the IRCCS Stella Maris of Calambrone (PI). The study has been concluded prior to completing patient recruitment because the eligible patients could not be easily traced. The available data have processed and analyzed and a scientific manuscript is in preparation Epidemiology of neurological disorders in Albania With the collaboration of the Fondazione Mariani and the Neurological Department of the University of Tirana, an epidemiological survey has been started to assess the prevalence and incidence of several neurological conditions (stroke, epilepsy, headache, dementia, peripheral neuropathy, multiple sclerosis) comparing an urban and a rural community (Tirana and Saranda). In 2005, a study on the validation of the diagnostic criteria was conducted and the recruitment of the cases to include in the incidence and prevalence studies is in course. Quality of life in children with neuromuscular disorders With a financial support of the Telethon organization, a study has been completed on the validation of a questionnaire on the quality of life in children and adolescents with different neuromuscular disorders. This is an Italian multicenter study coordinated to the Child Neurology Clinic of Pavia. The data analysis has been completed and two scientific manuscripts have been prepared and are ready for submission to ad-hoc scientific journals. Cerebrovascular disorders and risk of epilepsy Epilepsy is a frequent complication of stroke. Acute symptomatic seizures (i.e. seizures occurring in the seven days after stroke) can occur in up to two-thirds of cases and epilepsy (i.e. repeated unprovoked seizures) in 2-4%. There are no consistent findings on the risk factors for acute symptomatic seizures, unprovoked seizures and epilepsy in patients with stroke. For these reasons, in 2007 a multicenter national prospective survey has been started to assess the risk of seizures and epilepsy (and the main risk factors) in a cohort of patients with a first ischemic or hemorrhagic stroke followed for a maximum period of 24 months. The study was also implemented to assess the feasibility of a pragmatic therapeutic trial on the prophylaxis of seizures and epilepsy in stroke. Based on a preliminary analysis of 583 patients, the incidence of acute symptomatic seizures (5.7%) and the independent predictors of seizures (cerebral hemorrhage). Therapeutic trials in neurological disorders During the year 2008 three therapeutic trials sponsored by the Italian Drug Agency (AIFA) and a therapeutic trial sponsored by the Italian Ministry of Health were started or continued. Included are: 1. A randomized double-blind parallel-group placebo-controlled trial on the efficacy and tolerability of L-acetylcarnitine in ALS; 2. A randomized open-label parallel-group trial comparing Erythropoietine to Metyl-prednisolone in patients with acute spinal cord injury; 3. A randomized double-blind parallel-group placebo-controlled trial on the efficacy and safety of valproate in medication-overuse headache; 4. A randomized trial of the efficacy of a comprehensive rehabilitation program for the prevention of falls in Parkinson s disease. The 109

111 first trial aims at finding a potentially effective drug in a clinical condition for which there is only one product (Riluzole) with at best modest efficacy on survival. L-acetylcarnitine has been found to improve survival in experimental models of motor neuron disease. The second trial intends to verify the efficacy of erythropoietin, a drug shown to mitigate the effects of traumatic spinal shock and accelerate recovery in experimental animals. The drug chosen for comparison (Methylprednisolone at high doses) has been selected for being the present gold standard in clinical practice. The third trial aims at verifying whether valproate (a drug commonly used for the prophylaxis of migraine) abates symptoms occurring in drug-overuse headache, a common and frequently invalidating variety of chronic idiopathic headache. The fourth trial aims at assessing whether a comprehensive rehabilitation program compared to usual care is follone by a reduction in the incidence of falls in patients with Parkinson s disease at risk of falls. The first three trials are multicenter and nationwide. The laboratory of neurological disorders is the coordinator of the first trial and a partner in the other two trials, where the main tasks include protocol and CRF preparation, statistical analysis, and preparation of the final scientific report. The fourth trial is coordinated by the Laboratory and is conducted in Lombardy in conjunction with the Fondazione Don Gnocchi. Last, a randomized trial is in advanced preparation on the efficacy of an educational program for physicians working in nursing homes. The aim of the study is to verify a reduction in the number of inappropriate prescriptions compared to usual care. Laboratory of Drug Metabolism 1-Aryl-piperazine as active metabolites of centrally acting drug Starting from the previously reported 5-HT 7 receptor compounds with N-(1,2,3,4- tetrahydronaphthalen-1-yl)-4-aryl-1-piperazineexanamide structure (Leopoldo et al., J. Med. Chem., 2007, 50, 4214), a new series of 1-(2-methythiophenyl)-, 1-(2-biphenyl)-, 1-(2-isopropylphenyl)- and 1-(2-methoxyphenyl)-piperazine derivatives were designed with the aim to obtain new potent derivatives endowed with suitable physicochemical properties for rapid and extensive penetration into the brain. The newly synthesized compounds underwent radioligand binding assays to assess their affinities for 5-HT 7, 5-HT 1A, and D 2 receptors. The intrinsic activities at 5-HT 7 receptor as well as those for serotonin 5-HT 1A and dopamine D 2 of the most potent compounds were determined (Leopoldo et al., J. Med. Chem., 2008; 51: 5813). The proposed structural modifications on the tetrahydronaphthalenyl ring of the original compounds were, in most cases, detrimental for 5-HT 7 receptor, whereas had limited impact on the affinity for 5- HT 1A and D 2 receptors. Nonetheless, the pursued strategy led to the identification of the 1-(2- biphenyl)piperazine derivatives which retained nanomolar affinity at 5-HT 7 receptor, still showing lipophilicity within the target range. Among these, the N-(4-cyanophenylmethyl)-4-(2-diphenyl)-1- piperazinehexanamide derivative also demonstrated high selectivity over 5-HT 1A and D 2 receptors (324- and 245-fold, respectively) and showed full competitive agonism at 5-HT 7 receptor in an isolated guinea-pig ileum assay. It rapidly reached the systemic circulation and entered the brain, achieving concentrations in the low micromolar range after intraperitoneal doses in mice. Its brain concentration-time profile paralleled that in plasma, indicating that it rapidly and freely distributes across the blood-brain barrier. Similar studies were performed on the most potent and selective 5-HT 7 agonist derivative of the previous series - which had lipophilicity comparable to the new derivative - which, however, was cleared more rapidly from mouse plasma. Moreover, the potential formation of the 1-aryl-piperazine metabolite and its brain-to-plasma concentration ratio were also examinated, because arylpiperazine derivatives generally undergo CYP3A-mediated N-dealkylation of the aliphatic chain attached to the piperazine nitrogen. While the plasma concentrations of the metabolites were always below the detection limit of the analytical procedure, their brain concentrations exceeded that of their parent compound, by about 1.6 and 4 times for the new and original derivative, respectively. This was not surprising as previous studies have shown that 1-arylpiperazines concentrate in brain 110

112 tissue. Future studies will focus on characterization of the neuropharmacological profile of the 1-arylpiperazine metabolite compared with its parent compound. Pharmacological role of the constituents of Hypericum perforatum extracts The chemical composition of extracts of hypericum perforatum L. (St. John s wort) is essentially known but is still not clear which constituent(s) account, wholly or in part, for the antidepressant activity of the extracts, and through what neurochemical mechanism(s). The phloroglucinol hyperforin shares most of the in vitro and in vivo pharmacological properties of the extracts, and is possibly a main antidepressant component, but there is also evidence for other pharmacologically active components. However, identifying the roles of the various derivatives and the mechanism(s) of their activity is complicated by the scarcity of information about their ability to cross the blood-brain barrier and the concentrations reached in brain after administration of the extracts. This is also true for the biflavone biapigenin and particularly its I3,II8 analog amentoflavone which, although present in smaller amounts in extracts, shows a multitude of pharmacological actions in vitro and in vivo in animal models. The lack of pharmacokinetic data in man and animals and questions about the brain uptake of amentoflavone and biapigenin prompted us to examine their brain uptake and concentrations and the relationships with plasma concentrations after pharmacologically effective doses in mice. After doses of Hypericum perforatum extracts the brain concentrations of biapigenin and amentoflavone were below the limit of quantification. The same was true for amentoflavone after a biflavone-enriched extract of Ginkgo biloba. Levels were consistently detected only after intraperitoneal biapigenin or amentoflavone but were low and mostly related to the residual biflavone in the circulation. Poor brain-to-blood permeability is common to other polar components of Hypericum perforatum, resulting in brain concentrations generally too low for any direct interaction with neurotransmitter transporters and receptors which are obviously important for the action of conventional antidepressants. Likewise, the in vitro interactions of biapigenin and amentoflavone with known central mechanisms are apparently not relevant for the in vivo effects of the extracts because they occur at biflavone concentrations far exceeding those found in the brain after pharmacologically effective doses. However, this does not exclude that tissues other than brain may concentrate biapigenin or amentoflavone sufficiently to exert beneficial effects after daily intake of the extracts. Resistence to antidepressant drugs: studies in animal models The selective serotonin reuptake inhibitors are the drugs of choice in the treatment of depression. However, they are not or only partially effective in a fraction of depressed patients. The reasons are substantially unknown, though pharmacogenetic studies have linked the response to serotonin reuptake inhibitors to polymorphisms in various genes coding for serotonin mechanisms, particularly the promoter of the serotonin transporter molecule. These studies are therefore aimed to investigate the neurobiological mechanism(s) of resistance to antidepressant drugs in strains of mice carrying different isoforms of tryptophan hydroxylase-2, the enzyme responsible for the synthesis of brain serotonin. These studies are conducted in collaboration with the laboratory of Experimental Psycopharmacology (L. Cervo) and the laboratory of Neurochemistry and Behaviour (R.W. Invernizzi), who will provide a brief description of recent results with the potent serotonin reuptake inhibitors citalopram and paroxetine. 111

113 Laboratory of Experimental Neurology Role of inflammatory molecules in ictogenesis and epileptogenesis We are studying the role of IL-1beta and TNF-alpha systems in the genesis and propagation of seizures and in the associated neurodegenerative phenomena. We have demonstrated that epileptic activity induces the synthesis of these cytokines and related molecules involved in inflammatory processes. IL-1beta has proconvulsant actions while its naturally occurring antagonist (IL-1Ra) or its syntheis inhibitors have anticonvulsant activities. We are actively studying the role of these molecules in epilepsy models with the intent of promoting their clinical applications in drug-resistant epileptic patients. This possibility is encouraged by their clinical application in chronic inflammatory and autoimmune diseases in humans (e.g. anakinra, the IL-1R antagonist). We are studying pharmacological approaches to block IL-1beta-signaling involved in the proconvulsant effects of this cytokine. Epilepsy and postnatal development. Seizure susceptibility is higher in early infancy although the immature brain appears to be less susceptible to epileptogenesis. Using experimental models of seizures induced during postnatal development in rodents, we study the mechanisms involved in age-dependent seizure susceptibility and the associated neuronal injury. Our studies are primarily focused on inflammatory pathways, angiogenic processes and blood-brain barrier damage. Blood-brain barrier and epileptogenesis We are studying BBB permeability and microvasculature changes induced in the brain by seizures or by neurotrauma or infection and how these modifications may affect the process of epileptogenesis. Experimental models of symptomatic epilepsy are used. New therapeutic approaches of In vivo gene transfer This study concerns the use of adeno-associated viral vectors to introduce genes with therapeutic potential in the brain, thus increasing the synthesis of specific proteins to produce long-lasting anticonvulsant effects. We have demonstrated that adeno-associated viral vector carrying the human neuropeptide Y gene, significantly increases the brain concentration of this peptide after its intrahippocampal injection for a prolonged time (at least up to 5 months after a single intracerebral injection). The rats overexpressing this peptide are less susceptible to limbic seizures and to epileptogenesis. Future development of this study concerns the optimization of the transgene transfer technology to inhibit spontaneously recurring seizures and envisaging a possible clinical application. Laboratory of Geriatric Neuropsychiatry Population study on the prevalence of dementias in the older-old Parallel to the progressive increase of individuals aged 80 years or older within the elderly population (65+), the number of demented patients of 80 years or older makes up an ever increasing fraction of the total population affected by dementia. As very often happens, the exclusion from studies of subjects in the oldest age classes tends to inevitably underestimate the total number of individuals affected by dementia present in the population. To fill this gap, a door-to-door population study on the prevalence, incidence, risk factors and evolution of dementias and age-associated cognitive deficits has been set up in an elderly population aged 80 years or older living in eight small towns of Varese Province. The study is funded by a grant from the Fondazione Italo Monzino, Milano. 112

114 Effects of anemia in the elderly A previous large survey in old resident of Biella (65-84 years old) has been conducted in collaboration with the Local Health Authority of Biella (ASL 12) to determine the prevalence of anemia. We have now extended the investigation to the oldest old residents (about years or older individuals) in order to estimate the prevalence and impact of mild anemia also in this segment of the elderly population. Evaluating risk profiles in hospitalised elderly subjects In collaboration with the Geriatric Division of the Ospedali Regionali of Lugano and Mendrisio, Switzerland, hospitalized and ambulatory patients are evaluated from a neuropsychological, functional and mobility point of view to estimate the impact of these factors on heath-related outcomes and disease progression. Longitudinal follow-up of individuals with mild cognitive impairment (MCI) In collaboration with the Geriatric Unit of Ospedali Regionali of Lugano and Mendrisio, Switzerland, the follow-up study of all Mild Cognitive Impairment or Questionable Dementia (CDR 0.5) patients seen at the Memory Clinic of the Hospitals is continuing to estimate the rate of conversion to dementia and to evaluate the possible risk factors associated with conversion. Quality of care of terminally ill oncological subjects In 1999 we started a collaborative programme with the hospice via di Natale Franco Gallini in Aviano (PN). The aim of the research project was to assess the quality of care given in hospice to terminally ill oncological patients at the end of life. Present aim of the collaboration is the assessment of the hospice activities after its opening and to provide nurses with continuous training on use of databank Global Forum for Community Mental Health The Unit of Epidemiology and Social Psychiatry is in charge of the general bureau of the Global Forum for Mental Health, a working group promoted by the Department of Mental Health of the World Health Organization and aiming at creating a network supporting those who are active in mental health and care for people with mental disorders in low income countries, moving from asylum-centred to community-centred models of care. The Global Forum identifies experiences and practices which deserve to be known and promote and spread them; gives information and technical assistance to those interested in delivering and evaluating sustainable and promising interventions in underserved or disadvantaged populations; identifies barriers to the implementation of good quality interventions and discuss how to remove them, supports the development of research expertise based on real needs of the target populations, and through integration of service deliverers, researchers, policy makers, users and their families, citizens. Randomised controlled trial of the Italian Group for the Study of the Second Generation Antipsychotics (GISAS) The study compares three antipsychotics, aloperidole, olanzapine, aripiprazole, according to efficacy and tolerability through a pragmatic experimental design on 800 patients, identified and randomized in mental health centres of all Italian regions, and representing the first attempt to conduct a study on such large numbers in Italy on this topic. A toital of 35 mental health centres were recruited, among which 26 are active and have randomised 80 patients. Ibn the framework of this study, a survey was conducted on the point of view of the psychiatrists of the participating services on usefulness and feasibility of clinical trials in 113

115 psycopharmacology, degree of uncertainty in the use of antipsychotics and more important sources of information supporting their prescriptive attitudes. Natural Social Networks The project connects patients to identified volunteers willing to accompany the patients in part of their everyday activities. The evaluation of this intervention is based on the longitudinal employment of tools measuring quality of life, general wellbeing and social and personal functioning. Suicides In the framework of a wider project financed by the Ministry of Health, various literature reviews were conducted of topic related to the relationship between suicide attempt/deliberate self-harm and suicide, and treatment and assessment of suicide attempt and self-harm in the Emergency Department. A form for the systematic registration of these events filled by the psychiatrist during the consultation of the patient in the Emergency Department or in hospital was prepared and introduced in the ordinary assessment of suicide attempts of various hospitals and health trusts in North and Central Italy. UNASAM Project Quality improvement and evaluation in mental health with the active participation of the associations During the second year of the project, data collection on family members satisfaction with mental health services was completed in 41 services in four Italian regions. The questionnaire was developed with the participation of groups and associations of family members, who also organized the questionnaires distribution and data collection. Data were analysed and results were presented to the associations. European Network of Bipolar Research Export Centres EMBREC Bipolar disorder is characterised by recurrent depressive and manic episodes, and affects about 1% of the European population. Delay and difficulties in diagnosis have consequences on treatment and care, and it is worth integrating efforts and expertises of various groups. In this framework, the Unit of Epidemiology and Social Psychiatry has the following tasks: review evidences of effective strategies in information delivery, self-help and psychoeducation; development, in collaboration with a sample of users, of a package of practical strategies and tools for users information on relevant topics and self-management. The package will be tested and implemented in one or more mental health services in Italy. Self-accreditation and quality of the housing facilities in the Health Trusts 1 and 2 in Turin In two Health Trusts groups composed by users, family members, professionals working in the housing facilities and professionals working in other services have developed a tool for the systematic evaluation and accreditation of the housing facilities. The tool was tested. Lessons were held throughout the year to teach and train the participants to evaluation principles and organization of the visits and evaluating activities. 114

116 Laboratory of Inflammation and Nervous System Diseases Inhibition of selected aspects of the inflammatory response powerfully reduces ischemia/reperfusion injury Previous studies of ours have indicated that complement and related inflammatory systems such as contact/kinin system may represent novel targets for reducing ischemia/reperfusion injury. We have shown that C1-INH, a serine-protease inhibitor that acts as a major regulator of both complement and kinin systems, markedly improves neurological deficits and reduces infarct volume in mice with focal transient as well as permanent ischemia induced by middle cerebral artery occlusion. We have further extended this finding defining its effectivness on different strains of mice (displaying different levels of complement expression), the time-window and the dose-response curves. Since C1-INH may act on several substrates, we have also evaluated the specific involvement of the different complement pathways and of the other inflammatory systems. To explore the mechanisms of C1-INH neuroprotection, we have also investigated the expression (protein and mrna) of inflammatory cytokines, adhesion molecules, NO synthase isoforms, apoptosis markers. The results obtained show that: i) C1-INH effectively and markedly reduces brain ischemia/reperfusion injury, inducing a decrease up to 90% of the ischemic lesion; ii) C1-INH actions lead to inhibition of cell recruitment, inflammation and apoptosis; iii) C1-INH neuroprotection is independent from the complement classical pathway and inhibition of other complement pathways or other inflammatory systems such as the contact/kinin system are involved in its powerful protective action. Moreover recent data have shown C1-INH protective actions also in a model of traumatic brain injury. Thus C1-INH, which is presently used as replacement therapy in patients with C1-INH deficiency, possesses potent, multi-faceted neuroprotective actions that may be beneficial in acute brain injury. Ongoing studies are aimed at clarifying the mechanism of neuroprotection by C1-INH. Stem cells as a therapeutic approach in stroke and traumatic brain injury The aim of the project is to verify the conditions for the effectivenss of stem cells (SC) in reducing acute brain injury and to investigate the mechanisms triggered by their infusion in the lesioned brain. SC, isolated from newborn mice or obtained from human stem banks, are infused to mice in which transient ischemia is induced by middle cerebral artery occlusion or traumatic brian injury by controlled cortical impact. At different time points several parameters are evaluated: distribution and phenotype of injected cells, neurodegeneration, behavioral deficits, cytokine and trophic factor gene expression, microglia activation. The results obtained up to now have shown that: i) SC effectively counteract brain injury: they can decrease neuronal loss and reverte functional impairments related to exploratory behaviour and sensory/motor activity; ii) they may elicite an early response: in selected conditions 24 h after infusion, chemokines, angiogenic and neurotrophic factor transcripts are activated; iii) while in selected experimental conditions the protective mechanism of SC seems to be mainly due to the induction of beneficial factors, in others a longer time is required for effective neuroprotective effects; iv) activation of microglial cells is required for SC protective action; v) SC presence in the brain tissue is favoured by the ischemic environment. Thus the reciprocal interaction between SC and ischemic environment is crucial for stem cells protective actions Ongoing studies include: 1) definition of the mechanisms of homing of stem cells in the injuried brain; 2) analysis of the protective/toxic role of microglia; 3) investigation of the early events triggered by stem cells in the ischemic brain. (Capone et al, 2007; Pastori et al. 2008). 115

117 Identification of ischemic tolerance mediators in cerebral ischemia Recent studies indicate that cell death resulting from ischemic injury can be reduced when a sublethal ischemic episode occurs hours or days before a severe ischemic insult. This phenomenon is known as Ischemic PreConditioning (IPC) and the induced neuroprotection is called Ischemic Tolerance (IT). Several models of induction and maintenance of tolerance have been described, but the molecular mechanisms of the IPC-induced neuroprotection are not identified yet and a clear view of the mechanisms responsible for ischemic preconditioning is still lacking. Specific aims of the project are: i) to define the characteristics of IT induced by small transient ischemic attack (TIA) in experimental models of cerebral ischemia; ii) to elucidate the pathways and/or the mediators involved in ischemic tolerance; iii) to identify endogenous protective molecules/pathways that may represent potential therapeutic targets for stroke; iv) to assess if IPC can be induced also in traumatic brain injury (TBI). The project plan includes the characterization of TIA model in mouse, the study of the effect of TIA on subsequent ischemia and time window of ischemic tolerance, the identification of mediators involved in IT by protein and mrna assays, the study of direct effect of relevant protective molecules in in vitro and in vivo ischemia models, the set up of protocols for IPC in TBI brains. Thus ischemic preconditioning provides an opportunity to identify the putative candidate that can confer neuroprotection against acute brain injury. A major goal is to identify the underlying endogenous protective cellular receptor/signaling cascades, with the long-term goal to allow therapeutic augmentation of the endogenous protective mechanisms in cerebral ischemia. Blood-brain barrier and ischemic preconditioning The endothelial cells belonging to the cerebral microvasculature are the main component of the Blood-Brain Barrier (BBB). These cells are attached to each other by intercellular Tight Junctions and are closely associated with the endfeet of astrocytes, with pericytes and with microglia, resulting in a complex network of cellular interactions. The integrity of this structure is essential for the maintenance of the ischemic environment. We have recently established a bidimensional BBB model by means of co-cultures of mouse brain endothelial cells and mixed glial cells. These coltures retain the BBB typical features, namely they express thight junctions, they present typical transendothelial electrical resistence (TEER), and paracellular and transcellular permeability values. To mimic the ischemic insult, the BBB coltures are exposed to oxygen-glucose deprivation (ODG) protocols. The ongoing project is aimed at studying the involvement of BBB in ischemic preconditiong (IPC, see previous research topic). Albeit IPC has been reported to reduce edema formation and thus BBB disruption following ischemia, no information about a direct role of BBB cells in IPC is presently available. The major goal of the research has been to assess if BBB cells may be preconditioned. To this purpose a brief OGD stimulus was delivered before a severe OGD exposure. The results obtained have shown that a mild OGD stimuls may actually dampen the effects of a subsequent severe OGD exposure showing that BBB cells can be effectively preconditioned. Ongoing studies are presently aimed at identifying mediators and/or pathways involved in activation and maintenance of IPC in the cerebrovascular unit with the final aim of selecting new pathways and molecular targets useful for a therapeutical stroke strategies. 116

118 Laboratory of Molecular Neurobiology Study on pathogenic mechanisms of Amyotrophic Lateral Sclerosis Role of protein aggregation A pathological feature of ALS is the accumulation of protein aggregates in the perykaria and axons of motor neurons. Our hypothesis is that this may be due to an impairment of the ubiquitin/proteasome system (UPS). To functionally investigate the UPS in ALS motor neurons in vivo, we crossed SOD1G93A mice with transgenic mice that express a fluorescently-tagged reporter substrate of the UPS (Ub G76V -GFP, from now on indicated as GFP mice). In double transgenic GFP/SOD1G93A mice an increase in Ub G76V -GFP reporter, indicative of UPS impairment, was detectable in a few spinal motor neurons and not in reactive astrocytes or microglia, at symptomatic stage but not before symptom onset. These data suggest that UPS impairment occurs in motor neurons of mutant SOD1-linked ALS mice and may play a role in the disease progression. Another major route for intracellular protein degradation is the autophagy lysosomal pathway. The rat microtubule-associated protein 1 light chain 3 (LC3), plays a critical role in the formation of autophagosomes and its conversion from LC3I into LC3II is accepted as a simple method for monitoring authophagy. Recently, we have observed that levels of LC3II which is known to be correlated with the extent of autophagosome formation, was increased in SOD1G93A mice with at an advanced stage of disease compared with non transgenic mice. This indicates that the activation of autophagy may be an alternative mechanism of cell defense to eliminate the proteins misfolded when the proteasome is partially inhibited as demonstrated above. We are now examining the autophagy at earlier times of the pathology. In collaboration with the Molecular Biochemistry and Pharmacology department we carried out a proteomic analysis of the protein composition of the Triton-insoluble fraction (TIF), as a model of protein aggregates, from the SOD1G93A mice at different disease stages. We identified several proteins enriched in TIF of ALS mice already at preclinical stage, including intermediate filaments, chaperones and mitochondrial proteins. Some of them, HSP90, aconitase, HSC70 and cyclophilin A, were also analyzed in TIF of spinal cord of ALS patients and found significantly enriched. Interestingly, the majority of proteins in mice and at least HSP90 in patients were tyrosine nitrated. We therefore investigated the role of nitrative stress in aggregate formation in a cellular model of ALS. We could demonstrate that by inhibiting nitric oxide synthesis it is possible to substantially reduce the amount of insoluble proteins and in particular of aconitase, HSC70, cyclophilin A and SOD1. In conclusion, the analysis of the insoluble fractions from cellular/mouse models and human tissues could reveal novel aggregateprone proteins in ALS and suggest that nitrative stress may contribute to protein aggregate formation. These results are in a manuscript submitted to Brain. Role of glutamate AMPA receptors in the pathogenesis of ALS To further study the role of glutamate AMPA receptors in the susceptibility of motor neurons we have examined the expression and distribution of GluR2 subunit in motor neurons still maintaining a functional connection with muscle fibers. We found a significant decrease of Glur2 suggesting that this is a very early event that may play an important role in the pathogenesis of the disease. The analysis of all these data are under completion. In vitro studies on neuron-glia interaction To investigate further the role of inflammatory mechanisms, we have set up an in vitro coculture of spinal neurons and astrocytes derived from SOD1G93A mice embryos. We are verifying in this model the alterations observed in vivo such as the activation of TNFalphap38MAPK pathway. This model, hopefully, will allow to examine more rapidly the protective 117

119 effect of various strategies interfering with this pathway and will provide a model to test new pathogenic mechanisms. Altered axonuclear communication in motor neurons of a mouse model of familial amyotrophic lateral sclerosis (European collaborative project FP6 program EU-NES AXON SUPPORT) Impairment in the maintenance of axon-nuclear communication, and viceversa, due to motor proteins defects may play a primary role in the ALS. To assess this hypothesis we examined the expression and the distribution of different components of the nucleocytoplasmic-transport system, such as importins and vimentin, in the ventral horn spinal cord and in the peripheral nerves of transgenic mice and rats carrying human SOD1G93A. We found reduction of importin beta and slight increase of vimentin protein levels in homogenates from ventral horn spinal cord concomitantly with the appearance of first symptoms of the pathology. Using confocal miscroscopy, we detected abnormal accumulation of vimentin in the perikaria of motor neurons at the presymptomatic stage; moreover we observed a reduction of importin beta staining in the cytoplasm of motor neurons showing accumulation of phosphorylated neurofilaments, a hallmark of neuronal damage, and defects in retrograde transport. Based on these evidences we suggest that alterations of nucleocytoplasmic transport-related proteins are linked with the degenerative process of motor neurons and may play a role in ALS pathology. Therapeutical interventions in mouse model of ALS Development of target genes-based therapies for the protection of motor neurons It is emerging evidence that in the motoneurons of patients with sporadic ALS and in animal models of the disease, there is a remarkable activation of pro-degenerative pathways (like p38 MAP-Kinase). On the other side, the mechanisms involved in the modulation of cell survival (like PI3K/Akt pathway) are not activated, thus suggesting an impairment in the induction of neuroprotective responses.these pathways may be considered as potential therapeutic targets. Based on these evidences, with this project we propose: 1) to develop gene-targeted strategies aimed at counteracting p38 pro-degenerative pathway and activating Akt pro-survival cascade inside motorneurons of spinal cord; 2) to evaluate efficacy and safety of these potential therapeutic interventions in an animal model of ALS. We have developed lentiviral vectors expressing candidate p38-targeted shrna sequences. These shrnas were tested in primary mouse astrocyte-motoneuronal cell co-cultures or in cultures of rat cortical neurons. They were able to prevent activation of p38 and its downstream targets after TNFalpha stimulation, and to reduce neuronal loss after toxic stimuli. In parallel, we have developed constructs that express constitutively active forms of Akt1 and Akt3 (caakt1 and caakt3). Preliminary experiments in cell lines showed that either caakt1 or caakt3 efficiently phosphorylates and inhibits downstream pro-apoptotic targets, such as GSK3beta. Current studies aim to selectively drive the expression of p38-shrna and Akt1 to motor neurons and to increase the efficiency of their cellular expression. Studies on the effects of the long-chain omega-3 polyunsaturated fatty acids eicosapentaenoic acid and docosahexaenoic acid in the G93A SOD1 mouse (This is a project in collaboration with Dr. A. Michael-Titus del Queen Mary University of London supported by MND Association) Based on the observations by Dr. A. Michael-Titus that a diet enriched of omega 3 is neuroprotective in a model of spinal cord trauma in rat we 118

120 decided to investigate if such treatment could have a beneficial effect on SOD1G93A mice. The study is in progress. Treatment with lithium carbonate does not improve disease progression in two different strains of SOD1 mutant mice Female SOD1G93A mice on different genetic background and different phenotype of disease severity were treated daily with Li 2 CO 3 37mg/kg (1 meq /kg) i.p. starting from age 75 days until death. We observed a significant anticipation of the onset and reduced survival in 129Sv/G93A and no effect in C57G93A mice treated with lithium as compared to vehicle treated mice. Moreover, lithium neither exerted neuroprotective effects not increased the expression of LCII and the activity of mitochondrial complex IV in the spinal cord. The present study does not identify any therapeutic or neuroprotective effect of lithium in SOD1G93A female mice. This study is in press in Amyotrophic Lateral Sclerosis. Studies aimed to identify biomarkers for the diagnosis and progression of the disease in ALS patients In collaboration with the department of Neurology of the Fondazione Salvatore Maugeri, IRCCS, of Pavia, we have started a series of studies aimed to investigate the immune system and the oxidative stress products in the PBMC of sporadic ALS patients in comparison to healthy age matched controls. The results show that sporadic ALS patients exhibit immunological alterations in their blood, in respect to healthy controls. This study strengthens the hypothesis of an involvement of the adaptive immune system associated with a neuroinflammatory process in the pathobiology of ALS. The manuscript describing these data is under revision for the J. of Neuroimmunology. In parallel, we are examining the levels and the characterisation of the nitrated protein in the PBMC of ALS patients compared to healthy controls. We also evaluated the PBMC of SOD1G93A transgenic rats. The protein nitration on tyrosine is an oxidative mechanism that alters the function of proteins inducing their inactivation or a gain of toxic functions. Using a proteomic approach we have observed that a series of proteins are overnitrated in ALS patients and in SOD1G93A rats in respect to controls. Some proteins are the same in rat and patients suggesting that they could be a reliable biomarkers for the diagnosis and prognosis of the disease. These data are in a manuscript under revision for the Antioxidant and Redox Signalling. Another approach to identify potential specific diagnostic and prognostic markers of the disease has been set up in collaboration with the department of neurology of the Istituto Auxologico, IRCCS of Milano. In particular, we have used genomic and proteomic analyses to identify and characterize genes and proteins specifically modified in the muscles of ALS transgenic mouse models, at the onset of disease, in respect to control mice. We have completed the examinations and the data are now under analysis. 119

121 Laboratory of Experimental Psychopharmacology Drug Abuse Neural basis of drug self-administration To separate the direct pharmacological effects of cocaine from those associated with active drug self-administration we employed a yoked control-operant paradigm and investigated the expression of well established markers of the rapid action of cocaine, i.e. the inducible early genes and trophic factors, in rats after a single intravenous (i.v.) cocaine self-administration session. Animals self-administering cocaine did more active lever-presses than yoked-cocaine (YC) and yoked-vehicle (YV) animals. This goal-oriented behavior was accompanied by a selective increase in Arc mrna levels in the medial prefrontal cortex (mpfc). These findings demonstrate that a single session of cocaine i.v. self-administration is sufficient to shape rat behavior towards goal-directed behaviors and selectively up-regulate Arc expression in mpfc (of SA animals), providing the first evidence that the mpfc's function is already profoundly influenced by the first voluntary cocaine exposure. Neural basis of drug craving and relapse in the drug abuse assumption Drug craving, defined as the desire to experience the effect(s) of a previously experienced psychoactive substance is a cardinal feature of drug addiction and is clinically significant because of its potential link to relapse. To provide useful indications to the development of novel therapeutic approaches to prevent the use and abuse and the relapse of drug assumption following the outcome of craving, we elaborated experimental models of self-administration and relapse induced by cocaine, nicotine and alcohol-associated cues, after a period of abstinence. It was found that naltrexone, a non selective antagonist at opioids receptors, selectively modulate rats seeking behaviour induced by cocaine-associated cues after a long period of abstinence and in the absence of any further cocaine. Ongoing studies are evaluating whether this modulation is peculiar for cocaine or could be generalized to other abused drugs. The role of several other neurochemical mechanisms potentially involved in the drug-seeking behaviour are also in progress. Resistance to antidepressant drugs: experimental and clinical studies This project arises from a collaboration between the laboratories of Neurochemistry and Behavior (R.W. Invernizzi), Drug Metabolism (Silvio Caccia), Biology of Neurodegenerative Disorders (GianLuigi Forloni) and focus on behavioral and biochemical characterization of an experimental model of resistance to the antidepressant drugs. Using an animal model predictive of the antidepressant activity, the effects of selective serotonin reuptake blockers (SSRI) was evaluated in several mice strains. It was found that DBA/2J and BALB/c do not respond to the antidepressant-like activity of the SSRI. The lack of effect was attributed to genotypedependent impairment of 5-HT synthesis since DBA/2J and BALB/c carring a single nucleotide polymorphism (C1473G mice) in the gene for the brain-specific isoform of tryptophan hydroxylase-2, the rate-limiting enzyme in the synthesis of serotonin are characterized by a decreased serotonin synthesis. This hypothesis seems to be supported by the observation that DBA/2J and BALB/c mice had less dialysate 5-HT in the medial prefrontal cortex and dorsal hippocampus than C57BL/6J mice. Moreover, in DBA/2J and BALB/c the SSRI raised significantly less extracellular 5-HT when compared to C57BL/6J mice. More recently it was found that 5-HT 1A and 5-HT 2C receptor antagonists restored the SSRIs effect on either the antidepressant-like activity and the extracellular 5-HT. 120

122 Laboratory of Neurochemistry and Behavior Resistance to antidepressant drugs Despite the wide range of antidepressant drugs available for the treatment of mood disorders the delayed onset of the antidepressant effect and the partial or no response in a considerable portion of patients still limits their efficacy. Ongoing studies in collaboration with the Laboratories of Experimental Psychopharmacology and Drug Metabolism are aimed at assessing the neurobiological mechanisms involved in the resistance to antidepressant drugs in mice. The gene for the brain-specific isoform of tryptophan hydroxylase-2 (TPH-2), the rate-limiting enzyme in the synthesis of serotonin, is mutated in DBA/2J and BALB/c mice (C1473G). These mice have a low 5-HT synthesis rate and do not respond to antidepressants inhibiting selectively the reuptake of serotonin (SSRI) in the forced swimming test, a procedure used to screen compounds for antidepressant activity. In addition, the effect of SSRI on the extracellular concentrations of brain serotonin is attenuated in mice carrying the 1473G allele of TPH-2. 5-HT 1A and 5-HT 2C receptor antagonists enhanced the effect of SSRI on extracellular 5-HT and restored the antidepressant-like effect of SSRI. These results suggest that genetic differences in serotonin synthesis contribute to determine the efficacy of SSRI and identify pharmacological strategies that may enhance the response in treatment-resistant depressed patients. Animal model of cognitive deficit of schizophrenia; typical and atypical antipsychotics The cognitive deficit is a core symptom of schizophrenia, which has been linked to functional outcome and is relatively independent of psychotic symptoms. The antipsychotics, either typical or atypical, are able to control positive symptoms such as delirium, hallucinations and paranoia. However, the currently available atypical antipsychotics when compared to conventional antipsychotics show somewhat superior efficacy for the management of cognitive deficits in patients with schizophrenia. The cognitive deficit of schizophrenia was modeled in rats and mice, by using a test of attention such as the 5-choice serial reaction time task (5-CSRTT) and injections of glutamate NMDA receptor antagonists into the medial prefrontal cortex (mpfc). This model makes clear links with psychopathology as dysfunctional glutamate neurotransmission in the mpfc has been implicated in cognitive deficits of schizophrenia and the 5-CSRTT is the rat analogue of the continuous performance test used to assess attention and vigilance in schizophrenic patients. Antipsychotics possess a complex pharmacology across the biogenic amine receptor families as shown by affinity constants derived from radioligand-binding techniques. The ability to antagonise the DA D 2 receptor function is shared by the conventional and by the atypical antipsychotics. However, atypical antipsychotics show a high affinity also for serotonin 5- HT 2A, 5-HT 2C and 5-HT 1A receptors. Our studies compared the effects of conventional and atypical antipsychotics in this model of cognitive deficit of schizophrenia. The results show that antipsychotics may be differentiated by a selective effect of typical antipsychotics on compulsive perseveration, and atypical antipsychotics on impulsivity. Intracerebral microdialysis studies show that attentional deficits induced by NMDA receptor antagonists is associated with excessive glutamate in the medial prefrontal cortex of the rat and this effect was prevented by atypical antipsychotics. 121

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124 DEPARTMENT OF CARDIOVASCULAR RESEARCH STAFF Head Maria Grazia FRANZOSI, Biol.Sci.D. Laboratory of Cardiovascular Clinical Pharmacology Head Roberto LATINI, M.D. Bio-imaging Unit Head Cardiovascular Endocrine Unit Head Tissue Culture Unit Head Fabio FIORDALISO, Biol.Sci.D. Serge MASSON, Ph.D. Giovanna BALCONI, Univ.Dipl. Laboratory of Clinical Drug Evaluation Head Maria Grazia FRANZOSI, Biol.Sci.D. Bioinformatics Unit Head Enrico NICOLIS, Comp.Sci.Stud. Laboratory of General Practice Research Head Maria Carla RONCAGLIONI, Biol.Sci.D. Laboratory of Medical Statistics Head Simona BARLERA, Dr.Sci.Pol., MSc. Laboratory of Clinical Pharmacology Head Nursing Research Unit Head Gianni TOGNONI, M.D. Paola DI GIULIO, R.N., MSc 123

125 CURRICULA VITAE Maria Grazia Franzosi got her Biological Science degree in 1972 at the University of Milan. Education 1972 Doctoral degree in Biological Sciences, University of Milan, Italy 1978 Postdoctoral degree in Pharmacological Research, Istituto di Ricerche Farmacologiche "Mario Negri di Milano, Italy Main fields of activity Coordination of multicentric randomised clinical trials. Relationship between genetic and environmental risk factors in coronary events. Pharmacogenetics. Pharmacoeconomics. Drug Epidemiology and Post-Marketing Surveillance. Position from 2002 Director of the Department of Cardiovascular Research, Istituto di Ricerche Farmacologiche "Mario Negri", Milano, Italy from 2004 Member of Steering Committee, Studio GISSI-AF Study, Milano, Italy from 2001 from 1998 Member of Steering Committee, Studio GISSI-HF Study, Milano, Italy Member of Steering Committee of the PROCARDIS Research Programme - A genome-wide strategy to identify susceptibility loci in precocious coronary artery disease - University of Oxford, UK from 1997 Member of Antithrombotic Trialists Collaboration, Oxford, UK from 1996 Membro dello Steering Committee e National Coordinator per l Italia della Organization to Assess Strategies for Ischemic Syndromes (OASIS-2, OASIS-4 CURE, Michelangelo OASIS-5 e OASIS 6,, CURRENT OASIS-7, FUTURA OASIS-8), dello studio INTER- HEART e degli studi ACTIVE, RELY, AVERROES, Population Health Research Institue, McMaster University, Hamilton, Canada Director of European Coordinating Centre and Member of Steering Committee, Collaborative from 1993 from 2002 Organization for RheothRx Evaluation (CORE), McMaster University, Hamilton, Canada Member of Steering Committee, Studio GISSI-Prevenzione, Milano, Italy Member of Fibrinolytic Therapy Trialists s Collaboration, Oxford, UK e del Collaborative Group on Angiotensin Converting Enzyme Inhibitors Trials, National Institutes of Health, Bethesda, Washington, USA Head of the Laboratory of Clinical Drug Evaluation, Istituto di Ricerche Farmacologiche "Mario Negri" Head of the Clinic Drug Evaluation Unit of the Laboratory of Clinical Pharmacology, Istituto di Ricerche Farmacologiche "Mario Negri" from 1984 Member of the Scientific and Organising Secretariat, Gruppo Italiano per lo Studio della Sopravvivenza nell'infarto Miocardico (GISSI-1, GISSI-2, GISSI-3 studies) Milano, Italy Researcher at the Laboratory of Clinical Pharmacology, Istituto di Ricerche Farmacologiche "Mario Negri" and at the Regional Center for Drug Information of the Lombardy Region Selected publications Franzosi MG. Should we continue to use BMI as a cardiovascular risk factor? Lancet 2006; 368: Farrall M, Green FR, Peden JF, Olsson PG, Clarke R, Hellenius ML, Rust S, Lagercrantz J, Franzosi MG, Schulte H, Carey A, Olsson G, Assman G, Tognoni G, Collins R, Hamsten A, Watkins H, on behalf of the PROCARDIS Consortium. Genome-wide mapping of susceptibility to coronary artery disease identifies a novel replicated locus on chromosome 17. PLoS Genet Chiodini B, Franzosi MG, Barlera S, Signorini S, Lewis CM, D'Orazio A, Mocarelli P, Nicolis E, Marchioli R, Tognoni G, GISSI, SIBioC-GISSI Prevenzione Group. Apolipoprotein E polymorphisms influence effect of pravastatin on survival after myocardial infarction in a Mediterranean population: the GISSI-Prevenzione study. Eur Heart J 2007 ; 28: Anand SS, Islam S, Rosengren A, Franzosi MG, Steyn K, Yusufali AH, Keltai M, Diaz R, Rangarajan S, Yusuf S, INTERHEART Investigators. Risk factors for myocardial infarction in women and men: insights from the INTERHEART study. Eur Heart J : Broadbent HM, Peden JF, Lorkowski S, Goel A, Ongen H, Green F, Clarke R, Collins R, Franzosi MG, Tognoni G, Seedorf U, Rust S, Hamsten A, Farrall M, Watkins H, PROCARDIS Consortium. Susceptibility to coronary artery disease and diabetes is encoded by distinct, tightly linked, SNPs in the ANRIL locus on chromosome 9p. Hum Mol Genet 2008; 17: GISSI-HF Investigators (Writing Committee: Tavazzi L, Maggioni AP, Marchioli R, Barlera S, Franzosi MG, Latini R, Lucci D, Nicolosi GL, Porcu M, Tognoni G). Effect of n-3 polyunsaturated fatty acids in patients with chronic heart failure (the GISSI-HF trial): a randomised, double-blind, placebo-controlled trial. Lancet 2008; 372:

126 GISSI-HF Investigators (Writing Committee: Tavazzi L, Maggioni AP, Marchioli R, Barlera S, Franzosi MG, Latini R, Lucci D, Nicolosi GL, Porcu M, Tognoni G). Effect of rosuvastatin in patients with chronic heart failure (the GISSI-HF trial): a randomised, double-blind, placebo-controlled trial. Lancet 2008; 372 : Simona Barlera got her degree in Political Science, area Statistics at the Università degli Studi di Milano in Milano in 1992, followed by a master in Medical Statistics at the London School of Hygiene and Tropical Medicine, University of London in Education Degree in Political Science, area Statistics at the Università degli Studi di Milano in Milano Master post-lauream in Medical Statistics at the London School of Hygiene and Tropical Medicine, University of London, London Visiting Scientist in the Department of Statistical Genetics, Wellcome Trust Centre for Human Genetics, University of Oxford (UK). Main fields of activity Methodology of Clinical Trials in the cardiovascular field. Preparation and viewing of research protocols, planning and conduct of statistical analyses and the reporting of findings on scientific journals. Genetic epidemiology: genome-wide strategies (linkage analysis) to identify susceptibility genes in coronary artery disease; case-control studies in order to identify candidate genes involved in the cardiovascular pathology. Position from Oct 2006 Head of the Laboratory of Medical Statistics, Department of Cardiovascular Research, Istituto di Ricerche Farmacologiche "Mario Negri", Milano, Italy May 99-Sept 06 Head of the Medical Statistics Unit, Department of Cardiovascular Research, Istituto di Ricerche Farmacologiche "Mario Negri", Milano, Italy Visiting Scientist in the Department of Statistical Genetics, Wellcome Trust Centre for Human Genetics, University of Oxford (UK) Researcher in the Unit of Applied Statistics and Information Technology, Istituto di Ricerche Farmacologiche "Mario Negri", Milano, Italy Selected publications Chiodini B, Barlera S, Franzosi MG, Labarta V, Introna M, Tognoni G.APO B gene polymorphisms with coronary artery disease: A meta-analysis. Atherosclerosis 2003; 167: Latini R, Masson S, Anand I, Salio M, Hester A, Judd D, Barlera S, Maggioni AP, Tognoni G, Cohn J N, Val-HeFT Investigatore. The comparative prognostic value of plasma neurohormones at baseline in patients with heart failure enrolled in Val-HeFT. Eur Heart J 2004; 25: Maggioni AP, Latini R, Carson PE, Singh SN, Barlera S, Glazer R, Masson S, Cere` E, Rognoni G, Cohn JN. Valsartan reduces the incidence of atrial fibrillation in patients with heart failure: Results from the Valsartan Heart Failure Trial (Val-HeFT). Am Heart J 2005; 149: 1-10 Masson S, Latini R, Anand IS, Vago T, Angelici L, Barlera S, Missov ED, Clerico A, Tognoni G, Cohn JN, Val-HeFT Investigators. Direct comparison of B-type natriuretic peptide (BNP) and amino-terminal probnp in a large population of patients with chronic and symptomatic heart failure. The Valsartan Heart Failure (Val-HeFT) data. Clin Chem 2006; 52: Barlera S, Specchia C, Farrall M, Chiodini BD, Franzosi MG, Rust S, Green F, Nicolis E, Peden J, Assmann G, Collins R, Hamsten A, Tognoni G, PROCARDIS Consortium. Multiple QTL influence the serum Lp(a) concentration: a genome-wide linkage screen in the PROCARDIS study. Eur J Hum Genet 2007; 15: GISSI-HF Investigators (Writing Committee: Tavazzi L, Maggioni AP, Marchioli R, Barlera S, Franzosi MG, Latini R, Lucci D, Nicolosi GL, Porcu M, Tognoni G). Effect of n-3 polyunsaturated fatty acids in patients with chronic heart failure (the GISSI-HF trial): a randomised, double-blind, placebo-controlled trial. Lancet 2008; 372:

127 Roberto Latini got his Medical Doctor degree in 1978 at the University of Milan. Education University of Milan School of Medicine, degree in Medicine Merck Sharp & Dohme International Fellow in Clinical Pharmacology Main fields of activity Mechanisms of cardiac damage following ischemia, with focus on eurohumoral activation. Use of stem cells for cardiac repair. Biohumoral investigations within large scale clinical trials in heart failure and atrial fibrillation. Positions from 1990 Head of the Cardiovascular Clinical Pharmacology Laboratory (Cardiovascular Research Department) Istituto di Ricerche Farmacologiche Mario Negri, Milan, Italy from 2001 Member of the GISSI-HF Steering Committee from 2004 Member of the GISSI-AF Steering Committee from 2005 Member of the CandHeart Steering Committee from 1999 Visiting Professor Dept of Medicine, New York Medical College, Valhalla, NY, USA Cardiology Fellow (Dr. R. E. Kates, Laboratory) Stanford University Medical Center, CA, USA Member of the Sub-Group RMs for Drugs (Community Bureau of Reference, Commission of the European Communities) Fellow at the Laboratory of Clinical Pharmacology of the Istituto di Ricerche Farmacologiche "Mario Negri", Milano, Italy Selected publications Latini R, Masson S, Anand I S, Missov E, Carlson M, Vago T, Angelici L, Barlera S, Parrinello G, Maggioni A P, Tognoni G, Cohn J N, Val-HeFT Investigators. Prognostic value of very low plasma concentrations of troponin T in patients with stable chronic heart failure. Circulation 2007; 116: Sarto P, Balducci E, Balconi G, Fiordaliso F, Merlo L, Tuzzato G, Pappagallo G L, Frigato N, Zanocco A, Forestieri C, Azzarello G, Mazzucco A, Valenti M T, Alborino F, Noventa D, Vinante O, Pascotto P, Sartore S, Dejana E, Latini R. Effects of exercise training on endothelial progenitor cells in patients with chronic heart failure. J Card Fail 2007; 13: Galvez BG, Sampaolesi M, Barbuti A, Crespi A, Covarello D, Brunelli S, Dellavalle A, Crippa S, Balconi G, Cuccovillo I, Molla F, Staszewsky L, Latini R, DiFrancesco D, Cossu G. Cardiac mesoangioblasts are committed, self-renewable progenitors, associated with small vessels of juvenile mouse ventricle. Cell Death Differ 2008; 15: Masson S, Latini R, Anand IS, Barlera S, Angelici L, Vago T, Tognoni G, Cohn J N, for the Val-HeFT Investigators. Prognostic value of changes in N-termianl pro-brain natriuretic peptide in the Val-HeFT (Valsartan Heart Failure Trial). J Am Coll Cardiol 2008; 52: Salio M, Chimenti S, De Angelis N, Molla F, Maina V, Nebuloni M, Pasqualini F, Latini R, Garlanda C, Mantovani A. Cardioprotective function of the long pentraxin PTX3 in acute myocardial infarction. Circulation 2008; 117: Maria Carla Roncaglioni got her Biological Science degree in 1987 at the University of Milan. Education 1987 Doctoral degree in Biological Sciences, University of Milan, Italy Research Fellow at the Dept. of Biochemistry, Faculty of Medicine, Rijksuniversiteit of Limburg, Maastricht, The Netherland (Prof. C.Hemker); Visiting Scientist at the Cardiovascular Research Unit, Hammersmith Hospital, London, UK (Prof. A. Maseri) Main fields of activity Coordination of multicenter clinical trials and observational studies in different cardiovascular areas (neurological, angiological, cardiological). Coordination of a network of more than 1000 GPs actively involved in epidemiological and experimental studies in the prevention of cardiovascular diseases. Position from 2001 Head of the Laboratory for General Practice Research, Istituto di Ricerche Farmacologiche "Mario Negri", Milano, Italy from 1989 Senior Researcher in the Clinical Pharmacology Laboratory, Istituto di Ricerche Farmacologiche "Mario Negri", Milano, Italy from 1974 Researcher in the Laboratory for the Study of Haemostasis and Thrombosis, Istituto di Ricerche Farmacologiche "Mario Negri", Milano, Italy 126

128 Selected publications Tognoni G, Avanzini F, Pangrazzi J, Roncaglioni M C, Bertele V, de Gaetano G, Caimi V, Tombesi M, Colombo Fabio, Barlera S, PPP - Primary Prevention Project. Low-dose aspirin and vitamin E in people at cardiovascular risk: A randomized trial in general practice. Lancet 2001; 357: Sacco M, Pellegrini F, Roncaglioni MC, Avanzini F, Tognoni G, Nicolucci A, PPP - Primary Prevention Project. Primary prevention of cardiovascular events with low-dose aspirin and vitamin E in type 2 diabetic patients. Results of the Primary Prevention Project (PPP) trial. Diabetes Care 2003; 26: Roncaglioni MC, Avanzini F, Roccatagliata D, Monesi L, Tamayo-Benitez D, Tombesi M, Caimi V, Longoni P, Lauri D, Barlera S, Tognoni G, Collaborative Group Risk Prevention Study. How general practitioners perceive and grade the cardiovascular risk of their patients Eur J Cardiovasc Prev Rehabil 2004; 11: Monesi L, Avanzini F, Barlera S, Caimi V, Lauri D, Longoni P, Roccatagliata D, Tombesi M, Tognoni G, Roncaglioni MC. Appropriate use of antiplatelets: is prescription in daily practice influenced by the global cardiovascular risk? Eur J Clin Pharmacol 2005; 61: Berger JS, Roncaglioni MC, Avanzini F, Pangrazzi J, Tognoni G, Brown DL. Aspirin for the primary prevention of cardiovascular events in women and men: A sex-specific meta-analysis of randomized controlled trials. JAMA 2006; 295: Montalvo G, Avanzini F, Anselmi M, Prandi R, Ibarra S, Marquez M, Armani D, Moreira J M, Caicedo C, Roncaglioni MC, Colombo Fabio, Camisasca P, Milani V, Quimi' S, Gonzabay F, Tognoni G. Diagnostic evaluation of people with hypertension in low income country: cohort study of "essential" method of risk stratification. BMJ 2008; 337: a1387 Gianni Tognoni got his Medical Doctor in 1970, University of Milan. Main areas of methodology Randomized clinical trials; outcomes studies; pharmacoepidemiology; pharmacoeconomics; epidemiological monitoring and assessment of health care systems, drug policy; genetic epidemiology; community epidemiology; transfer of technology; health and human rights. Main clinical areas Acute and chronic CV diseases; psychiatry; aging; intensive care; neurodegenerative disordes; hematooncology. Position from 2004 Member, Commission of Human Experimentation of the Italian Drug Agency (AIFA) Member, Commissione Unica del Farmaco (CUF), Ministry of Health from 2002 Director, Consorzio Mario Negri Sud, S. Maria Imbaro, Chieti Coordinator, Department of Cardiovascular Research, Istituto di Ricerche Farmacologiche "Mario Negri", Milano from 1990 Co-Director, Scuola Superiore di Ricerca in Medicina Generale (CSeRMEG) from 1976 Founding member of the International Society of Drug Bulletins (ISDB) Coordinator, Commission of Human Experimentation, Regione Lombardia from 1983 Founder and in the Editorial Board of the nursing research Journal Rivista dell'infermiere/assistenza Infermieristica e Ricerca from 1977 Consultant to WHO and other UN agencies for drug selection and policy; training in methods of clinical and epidemiological research in developing countries mainly in Latin America and Africa Head, Laboratory of Clinical Pharmacology of the Istituto di Ricerche Farmacologiche from 1975 "Mario Negri", Milano Head, Regional Centre for Drug Information (CRIF), Regione Lombardia, Istituto di Ricerche Farmacologiche "Mario Negri", Milano Research Assistant, Laboratory of Clinical Pharmacology, Istituto di Ricerche Farmacologiche "Mario Negri", Milano Selected publications Landolfi R, Marchioli R, Kutti J, Gisslinger H, Tognoni G, Patrono C, Barbui T, for the European Collaboration on Low- Dose Aspirin in Polycythemia Vera Investigators. Efficacy and safety of low-dose aspirin in polycythemia vera. N. Engl. J. Med. 2004; 350: Eden T, Pui CH, Schrappe M, Tognoni G, Masera G, et al. All children have a right to full access to treatment for cancer. Lancet 2004; 364: Marchioli R, Finazzi G, Landolfi R, Kutti J, Gisslinger H, Patrono C, Marilus R, Villegas A, Tognoni G, Barbui T. Vascular and neoplastic risk in a large cohort of patients with polycythemia vera. J. Clin. Oncol. 2005; 23: Berger JS, Roncaglioni MC, Avanzini F, Pangrazzi I, Tognoni G, Brown DL. Aspirin for the primary prevention of cardiovascular events in women and men: a sex-specific meta-analysis of randomized controlled trials. JAMA Jan 18;295(3): Erratum in: JAMA May 3;295(17):2002 Strippoli GF, Tognoni G, Navaneethan SD, Nicolucci A, Craig JC. Haemoglobin targets: we were wrong, time to move on. Lancet 2007;369:

129 GISSI-HF Investigators (Writing Committee: Tavazzi L, Maggioni AP, Marchioli R, Barlera S, Franzosi MG, Latini R, Lucci D, Nicolosi GL, Porcu M, Tognoni G). Effect of n-3 polyunsaturated fatty acids in patients with chronic heart failure (the GISSI-HF trial): a randomised, double-blind, placebo-controlled trial. Lancet 2008; 372: Giovanna Balconi got her degree at the School for Technicians of Biomedical Institutes of the University of Milan, with a specialisation in Histology in the Pathological Anatomy Laboratory of the same University (1968). Main fields of interest Isolation, culture and characterization of peripheral blood circulating progenitor cells of patients with heart failure. In vitro culture and characterization of stem cells for repair of myocardial infarction in experimental animal models. Positions from July 2005 Head of Tissue Culture Unit, Cardiovascular Clinical Pharmacology Laboratory, Istituto di Ricerche Farmacologiche "Mario Negri", Milano, Italy Oct June 2005 Head of Tissue Culture Unit, Vascular Biology Laboratory, Istituto di Ricerche Farmacologiche "Mario Negri", Milano, Italy Dec Oct 1995 Head of Tissue Culture Unit, Anticancer Chemotherapy Laboratory, Istituto di Ricerche Farmacologiche "Mario Negri", Milano, Italy Oct Nov 1983 Researcher, Anticancer Chemotherapy Laboratory, Istituto di Ricerche Farmacologiche "Mario Negri", Milano, Italy Selected publications Condorelli G, Borello U, De Angelis L, Latronico M, Sirabella D, Coletta M, Galli R, Balconi G, Follenzi A, Frati G, Cusella De Angelis MG, Gioglio L, Amuchastegui CS, Adorini L, Naldini L, Vescovi A, Dejana E, Cossu G. Cardiomyocytes induce endothelial cells to trans-differentiate into cardiac muscle: Implications for myocardium regeneration. Proc Natl Acad Sci USA 2001; 98: Cattelino A, Liebner S, Gallini R, Zanetti A, Balconi G, Corsi A, Bianco P, Wolburg H, Moore R, Oreda B, Kemler R, Dejana E. The conditional inactivation of the beta-catenin gene in endothelial cells causes a defective vascular pattern and increased vascular fragility. J Cell Biol 2003; 162: Cusella De Angelis MG, Balconi G, Bernasconi S, Zanetta L, Boratto R, Galli D, Dejana E, Cossu G. Skeletal myogenic progenitors in the endothelium of lung and yolk sac. Exp Cell Res 2003; 290: Galli D, Innocenzi A, Staszewsky L, Zanetta L, Sampaolesi M, Bai A, Martinoli E, Carlo E, Balconi G, Fiordaliso F, Chimenti S, Cusella G, Dejana E, Cossu G, Latini R. Mesoangioblasts, vessel-associated multipotent stem cells, repair the infarcted heart by multiple cellular mechanisms. A comparison with bone marrow progenitors, fibroblasts, and endothelial cells. Arterioscler Thromb Vasc Biol 2005; 25: Sarto P, Balducci E, Balconi G, Fiordaliso F, Merlo L, Tuzzato G, Pappagallo GL, Frigato N, Zanocco A, Forestieri C, Azzarello G, Mazzucco A, Valenti M T, Alborino F, Noventa D, Vinante O, Pascotto P, Sartore S, Dejana E, Latini R. Effects of exercise training on endothelial progenitor cells in patients with chronic heart failure. J Card Fail 2007; 13: Galvez BG, Sampaolesi M, Barbuti A, Crespi A, Covarello D, Brunelli S, Dellavalle A, Crippa S, Balconi G, Cuccovillo I, Molla F, Staszewsky L, Latini R, DiFrancesco D, Cossu G. Cardiac mesoangioblasts are committed, self-renewable progenitors, associated with small vessels of juvenile mouse ventricle. Cell Death Differ 2008; 15: Paola Di Giulio got her Nursing Diploma in the Nursing School of Istituto Nazionale dei Tumori in Milano and her Master in Oncology Nursing at Guildford University (UK) in Main fields of activity Coordination of multicentre and observational studies studies in cardiology and palliative care. Coordination of nursing networks. Position from March 2001 Associated professor at the Turin University. from 1997 Responsible of the Nursing Research Unit from 1995 Senior researcher of the Cardiovascular Research Department from 1989 Consultant of the Clinical Phrmacology Laboratory since 1998 Coordinator of the Editorial Board of Assistenza Infermieristica e Ricerca Selected publications Laquintana D, Di Giulio P: Per un ruolo infermieristico nella farmacovigilanza. Assistenza Infermieristica e ricerca 2002; 21: Di Giulio P, Saiani L, Laquintana D, Palese A, Gruppo PARI-ETLD. Studio clinico randomizzato controllato in doppio cieco sull efficacia dei trattamenti delle lesioni da decubito. Assistenza Infermieristica e Ricerca 2004; 23:

130 Toscani F, Di Giulio P, Brunelli C, Miccinesi G. Laquintana D. How people die in hospital general wards: a descriptive study. J Pain Symptom Manage 2005; 30: Saiani L, Di Giulio P, Gruppo PARI-FV (Percorsi Assistenziali e Ricerca Infermieristica-Farmaco Vigilanza) Epidemiologia dei problemi assistenziali legati a farmaci e presidi in RSA e distretto. Assistenza Infermieristica e Ricerca 2007; 26: Lepore V, Cecchetto G, Di Giulio P, Saiani L, Samarelli V, Saugo M, Romero M, Scurti V, Tognoni G, Valerio M. Età anziana-molto-anziana, e aspettativa di vita? Assistenza Infermieristica e Ricerca 2007; 26: Di Giulio P, Toscani F, Villani D, Brunelli C, Gentile S, Spadin P. Dying with advanced dementia in long-term care geriatric institutions: a retrospective study. J Palliat Med 2008; 11: Fabio Fiordaliso got his Biological Science degree in 1995 at the University of Milan. Education 1998 Postdoctoral degree in Pharmacological Research, Istituto di Ricerche Farmacologiche Mario Negri, Milan, Italy 1995 Doctoral degree in Biological Sciences, University of Milan, Italy Main fields of activity Therapeutical potential of stem cell and antioxidant treatments in experimental model of diabetic cardiomyopathy and in primary myocyte cultures exposed to hyperglycemia. Morphological and structrural analysis of cells and tissue by optical, confocal and electron microscopy. Positions from 2007 from 2006 from 2005 from 2005 from 2001 Head of Bio-imaging Unit, Department of Cardiovascular Research, Istituto di Ricerche Farmacologiche Mario Negri, Milan Member of the Heart Failure Association (HFA) of the European Society of Cardiology Member of the Working group on myocardial function (WG 4) of the European Society of Cardiology Member of the steering committee of the Consorzio of Microscopy and Image Analysis (MIA) Senior Research Scientist, Laboratory of Cardiovascular Clinical Pharmacology (Department of Cardiovascular Research), Istituto di Ricerche Farmacologiche Mario Negri, Milan Post-Doctoral Research Fellow at Cardiovascular Research Institute (Department of Medicine), New York Medical College, Valhalla, New York Research Fellow, Laboratory of Cardiovascular Clinical Pharmacology (Department of Cardiovascular Research), Istituto di Ricerche Farmacologiche Mario Negri, Milan Research training, Institute of General Pathology, University of Milan (Italy) Selected publications Fiordaliso F, Bianchi R, Staszewsky L, Cuccovillo I, Doni M, Laragione T, Salio M, Savino C, Melucci S, Santangelo F, Scanziani E, Masson S, Ghezzi P, Latini R. Antioxidant N-Acetyl-L-Cysteine attenuates hyperglycemia-induced cardiomyocyte death in rats. J Mol Cell Cardiol 2004; 37: Fiordaliso F, Chimenti S, Staszewsky L, Bai A, Carlo E, Cuccovillo I, Doni M, Mengozzi M, Tonelli R, Ghezzi P, Coleman T, Brines M, Cerami A, Latini R. A non-erythropoietic derivative of EPO effectively ameliorates experimental cardiac ischemia with reperfusion. Proc Natl Acad Sci USA 2005; 102: Fiordaliso F, Cuccovillo I, Bianchi R, Bai A, Doni M, Salio M, De Angelis N, Ghezzi P, Latini R, Masson S. Cardiovascular oxidative stress is reduced by an ACE inhibitor in a rat model of streptozotocin-induced diabetes. Life Sci 2006; 79: Fiordaliso F, De Angelis N, Cuccovillo I, Bai A, Salio M, Serra DM, Bianchi R, Razzetti R, Latini R, Masson S. Effect of β-adrenergic and renin-angiotensin system blockade on myocyte apoptosis and oxidative stress in diabetic hypertensive rats. Life Sci 2007; 81: Latini R, Brines M, Fiordaliso F. Do non-hemopoietic effects of erythropoietin play a beneficial role in heart failure? Heart Fail Rev 2008; 13: Dossena S, Imeri I, Mangieri M, Garofoli A, Ferrari L, Senatore A, Restelli E, Balducci C, Fiordaliso F, Salio M, Bianchi S, Fioriti L, Morbin M, Pincherle A, Marcon G, Villani F, Carli M, Tagliavini F, Forloni G, Chiesa R. Mutant prion protein expression causes motor and memory deficits and abnormal sleep patterns in a transgenic mouse model. Neuron 2008; 60:

131 Serge Masson obtained his doctorate (PhD) in Biochemistry and Cellular Biology in 1990 at the University of Marseilles (France), followed by a postdoctoral stay at the Panum Institute in Copenhagen (Denmark) Education Doctorate fellow, Faculty of Medicine, University of Aix-Marseilles, France Post-doctoral Researcher, Panum Institute and Assistant Lecturer, University of Copenhagen, Denmark 1993 Research Scientist, NMR Laboratory, Hospital San Raffaele, Milan, Italy from 1994 Research Scientist, Department of Cardiovascular Research, Istituto di Ricerche Farmacologiche "Mario Negri", Milano, Italy Main fields of activity Physiopathology, diagnostic and prognostic role of the activation of neuroendocrine systems in cardiovascular disease Position from 2002 from 2002 from 2002 Head of the Cardiovascular Endocrine Unit, responsible for Quality Assurance for the Department of Cardiovascular Research, Istituto di Ricerche Farmacologiche "Mario Negri", Milano, Italy Tutor of fellows of the School of Specialists in Pharmacological Research, Istituto di Ricerche Farmacologiche "Mario Negri", Milano, Italy Fellows of the American Heart Association (Basic Council) and the Working Group on Myocardial Function of the European Society of Cardiology Selected publications Anand IS, Latini R, Florea VG, Kuskowski MA, Masson S, Signorini S, Mocarelli P, Hester A, Glazer R, Cohn JN, for the Val-HeFT Investigators. C-reactive protein in heart failure: prognostic value and the effect of valsartan. Circulation 2005; 112: Masson S, Latini R, Anand I S, Barlera S, Judd D, Salio M, Perticone F, Perini G, Tognoni G, Cohn JN, on behalf of the Val-HeFT Investigators. The prognostic value of Big endothelin-1 in more than 2,300 patients with heart failure enrolled the Valsartan in Heart Failure Trial (Val-HeFT). J Card Fail 2006; 12: Latini R, Masson S, Anand I S, Missov E, Carlson M, Vago T, Angelici L, Barlera S, Parrinello G, Maggioni AP, Tognoni G, Cohn J N, Val-HeFT Investigators. Prognostic value of very low plasma concentrations of troponin T in patients with stable chronic heart failure. Circulation 2007; 116: Masson S, Latini R, Anand IS, Vago T, Angelici L, Barlera S, Missov ED, Clerico A, Tognoni G, Cohn JN, on behalf of the Val-HeFT Investigators. Direct comparison of B-type natriuretic peptide (BNP) and amino-terminal probnp in a large population of patients with chronic and symptomatic heart failure. The Valsartan Heart Failure (Val-HeFT) data. Clin Chem 2006; 52: Staszewsky L, Wong M, Masson S, Barlera S, Carretta E, Maggioni AP, Anand IS, Cohn JN, Tognoni G, Latini R, Valsartan Heart Failure Trial Investigators. Clinical, neurohormonal, and inflammatory markers and overall prognostic role of chronic obstructive pulmonary disease in patients with heart failure: data from the Val-HeFT Heart Failure Trial. J Card Fail 2007; 13: Masson S, Latini R, Anand IS, Barlera S, Angelici L, Vago T, Tognoni G, Cohn J N, for the Val-HeFT Investigators. Prognostic value of changes in N-termianl pro-brain natriuretic peptide in the Val-HeFT (Valsartan Heart Failure Trial). J Am Coll Cardiol 2008; 52: Enrico Bjørn Nicolis has attended the courses in Computer Science at the University of Milan. Education Research fellow, Istituto di Ricerche Farmacologiche "Mario Negri", Milano, Italy Main fields of activity Data management and analysis of randomized clinical trials. Developing of database and tools for studies of population genetics, particularly for linkage analysis. Position from 2001 Head of the Bioinformatics Unit, Istituto di Ricerche Farmacologiche "Mario Negri", Milano, Italy from 1999 Research fellow of the Laboratory of Clinical Drugs Evaluation from 1997 System administrator at the EDP center, Istituto di Ricerche Farmacologiche "Mario Negri", Milano, Italy from 1991 Research fellow at the Medical Informatics and Applied Statistics Unit, Istituto di Ricerche Farmacologiche "Mario Negri", Milano, Italy 130

132 Selected publications Nobili A, Gebru F, Rossetti A, Schettino F, Zahn R W, Nicolis E, Macario G, Celani L, Acik V O, Farina ML, Naldi L. Doctorline: A private toll-free telephone medical information service. Five years of activity: Old problems and new perspectives. Ann Pharmacother 1998; 32: Santoro E, Nicolis E, Franzosi MG.Telecommunication technology for the management of large scale clinical trials: The GISSI experience. Comput Methods Programs Biomed 1999; 60: Santoro E, Nicolis E, Franzosi MG, Tognoni G. Internet for clinical trials: Past, present, and future. Control Clin Trials 1999; 20: Tognoni G, Franzosi MG, Nicolis E, Barlera S, Specchia C, Chiodini B, Crociati L, Ferrario L, PROCARDIS Consortium. A trio family study showing association of the lymphotoxin-alfa N26 (804A) allele with coronary artery disease. Eur J Hum Genet 2004; 12: Barlera S, Specchia C, Farrall M, Chiodini BD, Franzosi MG, Rust S, Green F, Nicolis E, Peden J, Assmann G, Collins R, Hamsten A, Tognoni G, PROCARDIS Consortium. Multiple QTL influence the serum Lp(a) concentration: a genome-wide linkage screen in the PROCARDIS study. Eur J Hum Genet 2007; 15: Specchia C, Barlera S, Chiodini BD, Nicolis EB, Farrall M, Peden J, Collins R, Watkins H, Tognoni G, Franzosi MG, PROCARDIS Consortium. Quantitative trait genetic linkage analysis of body-mass index in familial coronary artery disease. Hum Hered 2008; 66: INTRODUCTION TO THE DEPARTMENT'S ACTIVITIES The areas of interest of the Department of Cardiovascular Research include the experimental, clinical, genetic, epidemiological aspects of acute myocardial infarction, cardiac failure, cardiac arrhythmias, as well as the clinical and epidemiological investigation of cardiovascular prevention, hypertension and stroke. Following the successful experience of the GISSI-trials (Gruppo Italiano per lo Studio della Sopravvivenza nell'infarto), the activation of large collaborative networks in the setting of the National Health Service hospitals and in general practice has become a key characteristics of the Department, which can now rely on the permanent collaboration of over 300 clinical groups and of several hundred general practitioners. Over the years, firm links have also been established with international leading research groups. The experimental research activity concerns the physiopathology, the pharmacological modulation and the prognostic role of the activation of the renin-angiotensin-aldosterone system, as well as other neurohormonal systems, in myocardial infarction and heart failure, the physiopathology, the pharmacological modulation and prognostic role of the activation of the inflammatory processes in myocardial infarction and heart failure; a more recent research topic is the cell therapy of myocardial infarction. The activity in clinical research includes the clinical assessment of therapeutic strategies with large scale clinical trials in the field of acute coronary syndromes, congestive heart failure and atrial fibrillation. A recently developing area is the genetic epidemiology of myocardial infarction and heart failure. Several studies have been conducted in the area of clinical epidemiology and risk factors assessment of myocardial infarction. The collaboration with a large network of General Practitioners in the area of cardiovascular prevention allowed to test new hypotheses through large scale clinical trials and to evaluate the actual transferability of evidence based interventions in the every day practice through epidemiological or outcome research studies. Pharmacoepidemiological studies through the analysis of a large sample of Local Health Units drug prescriptions were also performed. A research network of nurses has been developed with the main focus on the assessment of healthrelated quality of life of patients and on the epidemiology of nursing interventions and their implications for patients' well being and outcomes. 131

133 FINDINGS/MAIN RESULTS The GISSI-HF Study, a randomized multicenter trial conducted in nearby 7000 patients from 357 cardiology sites in Italy, showed that a simple, safe, one-a-day capsule of n-3 polyunsaturated fatty acids (PUFA) can reduce mortality and admission to hospital for cardiovascular reasons in patients with heart failure. Patients received either n-3 PUFA in a capsule once daily (3494 patients) or placebo (3481). 955 patients in the PUFA group (27%) died, compared with 1014 (29%) in the placebo group, meaning a relative risk reduction of 9% in the PUFA group. A higher proportion of patients in the placebo group (2053/59%) died or were admitted to hospital for cardiovascular reasons than in the PUFA group (1981/57%) a relative reduction of 8% in the PUFA group. In absolute terms, 56 patients needed to be treated with PUFA for just under four years to avoid one death, or 44 patients to avoid one event of either death or admission to hospital for cardiovascular causes. Statin treatment with rosuvastatin does not affect clinical outcomes in patients with chronic heart failure. In 2500 patients with chronic heart failure, enrolled to the GISSI-HF multicenter trial we showed that microalbuminuria, a marker of vascular and renal injury, is one of the strongest predictors of death and hospitalization for cardiovascular reasons. The relation with outcomes is still strong after adjustment for major markers of risk, including creatinine, BNP, CRP. It's the largest study showing the prognostic value of microalbuminuria in heart failure. GISSI-AF, a randomized, prospective, parallel group, placebo-controlled, multicenter study on the use of valsartan, an angiotensin II AT1-receptor blocker (ARBs) in the prevention of Atrial Fibrillation (AF) recurrence, has recruited and treated for 12 months 1400 patients with a history of recent AF associated with cardiovascular diseases/comorbidities. In GISSI-AF, the largest trial ever conducted with ARBs in pts with AF, Valsartan did not reduce AF recurrence. A trend favoring valsartan was observed only in the small group of pts with heart failure and/or left ventricular dysfunction. The PROCARDIS consortium was conceived to study the complex genetics of coronary artery disease (CAD) susceptibility in Europeans and has assembled clinical resources to undertake genetic association studies. Recently PROCARDIS has undertaken a case-control study in 4,251 CAD cases and 4,443 controls using previously reported and tagging SNPs (Single Nucleotide Polymorphisms) of a region on chromosome 9p that is associated with CAD and with type 2 diabetes (T2D). PROCARDIS replicated the SNPs and showed that the strong consistent association detected by these SNPs is a consequence of a yin-yang of markers spanning 53 kb. There was no evidence of additional CAD susceptibility alleles over the major risk haplotype. CAD patients without myocardial infarction (MI) showed a trend towards stronger association than MI patients. The CAD susceptibility conferred by this locus did not differ by sex, age, smoking, obesity, hypertension or diabetes. A simultaneous test of CAD and diabetes susceptibility with CAD and T2D-associated SNPs indicated that these associations were independent of each other. A genetic study conducted in the patients of the GISSI-Prevenzione (GISSI-P) trial showed that Apolipoprotein E polymorphisms influence effect of pravastatin on survival after myocardial infarction. We analyzed 3304 Italian patients with MI randomized to pravastatin or no treatment +with a median follow-up time of 24 months and we found that epsilon-4 allele is a determinant of the response to pravastatin in terms of survival. Though in the entire population investigated we found a beneficial effect of pravastatin in terms of survival, only the epsilon-4 carriers seemed to have gained a significant benefit from this treatment. We suggest that the effect of 132

134 statins is of particular interest in this fraction of the population and that genetic markers can help in identifying patients that benefit more from statin treatment. NATIONAL COLLABORATIONS Angiogenesis and Tumor Targeting Research Unit & Telethon Institute for Gene Therapy, Ospedale San Raffaele, Milano ANMCO (Associazione Nazionale Medici Cardiologi Ospedalieri) Centro Cardiologico Monzino IRCCS, Milano CINECA (Consorzio Interuniversitario per il Calcolo Automatico dell'italia Nord-Orientale) CSeRMEG (Centro Studi e Ricerche in Medicina Generale) Dipartimento di Cardiologia e UTIC, Istituto Clinico Humanitas IRCCS, Milano Dipartimento Cardio-Vascolare ed Endocrino-Metabolico, Ospedale Casa Sollievo della Sofferenza IRCCS, San Giovanni Rotondo Ematologia, Ospedale Sant Anna, Torino Fondazione Don Gnocchi IRCCS, Milano Gruppi organizzati di MMG (FIMMG, CoS, Ass.Cu.M.I., AMISI) IEO (Istituto Europeo di Oncologia), Milano IFOM-FIRC, Milano Istituto di Anestesiologia e Rianimazione IRCCS, Ospedale Maggiore Policlinico, Mangiagalli, Regina Elena, Milano Istituto di Ricerca in Cure palliative Lino Maestroni, Cremona Laboratorio di Endocrinologia, Ospedale Luigi Sacco, Milano Regione Lombardia SIBioC (Società Italiana di Biochimica Clinica e Biologia Molecolare) SIFO (Società Italiana di Farmacia Ospedaliera) Stem Cell Research Institute, Ospedale San Raffaele, Milano Unità Operativa Semplice di Neuroanestesia e Neurorianimazione, Dipartimento di Medicina Perioperatoria e Terapie Intensive, Ospedale San Gerardo, Monza Università degli Studi di Milano, Dipartimento di Medicina Interna Università degli Studi di Milano, Dipartimento di Scienze Farmacologiche Università degli Studi di Torino, Dipartimento di Anatomia, Farmacologia e Medicina Forense Università degli Studi di Torino, Dipartimento di Sanità Pubblica e Microbiologia Università degli Studi di Verona, Dipartimento di Sanità Pubblica Università degli Studi di Verona, Istituto di Anatomia Umana INTERNATIONAL COLLABORATIONS Cecomet (Centro de Epidemiologia comunitaria y Medicina tropical, Esmeraldas) Ecuador Cochrane Collaboration, Oxford, UK Clinical Trial Research Unit, Auckland University, New Zealand CTSU (Clinical Trial Service Unit) /ISIS (International Studies on Infarct Survival), Oxford, UK Division of Genetics and Development, Guy's, King's and St Thomas' School of Medicine, King's College, London, UK DSAN SUPSI (Scuola Universitaria Professioni Sanitarie), Lugano CH ECLA (Estudios Cardiologicos de Latino-America) 133

135 EVGN (European Vascular Genomics Network) VI Framework Program, European Union JW Goethe University, Department of Cardiology, Frankfurt, Germany Karolinska Institutet, Stockholm, Sweden PHRI (Population Health Research Institute), McMaster University, Hamilton, Ontario, Canada SIOP (International Society of Paediatric Oncology) University of Cambridge, UK University of Minnesota, Minneapolis, USA University of Oslo, Norway Wellcome Trust Centre for Human Genetics, University of Oxford, UK WONCA (World Organization of Family Doctors) EDITORIAL BOARD MEMBERSHIP Circulation, International Journal of Health Services, European Heart Journal (Gianni Tognoni) Journal of Cardiac Failure, Journal of Cardiovascular Medicine (Roberto Latini) Assistenza Infermieristica e Ricerca, European Journal of Cancer Care, European Journal of Oncology Nursing, International Nursing Perspectives (Paola Di Giulio) PEER REVIEW ACTIVITIES American Heart Journal, Atherosclerosis Thrombosis and Vascular Biology, Cardiovascular Research, Circulation, Circulation Research, Clinical Pharmacology and Therapeutics, European Heart Journal, European Journal of Cancer Care, European Journal of Cardiovascular Nursing, European Journal of Oncology Nursing, International Journal of Cardiology, International Journal of Obesity, Italian Journal of Cardiology, Journal of Cardiac Failure, Journal of Internal Medicine, Journal of Cardiovascular Medicine, Life Sciences, The Lancet, PharmacoEconomics. NATIONAL AND INTERNATIONAL COMMITTEE MEMBERSHIP Ethical Committee of the ASL of Milan Ethical Committee of Lombardy Region Ethical Committee of the Provincia of Verona Agenzia Italiana del Farmaco, Direzione Generale Farmaci e Dispositivi Medici EVENT ORGANIZATION Investigator's Meeting - Stato di avanzamento dello studio Prono/Supino II 16/01/08, Istituto di Ricerche Farmacologiche Mario Negri, Milano Investigator's Meeting Presentazione dei risultati dello studio GISSI-AF 134

136 01/06/08, Fortezza da Basso, Firenze Investigator's Meeting GISSI-HF: un modello di ricerca collaborativa 06/09/08, Aula Magna Santa Lucia, Bologna Investigator's Meeting Stato di avanzamento dello studio "NeuroMorfeo" 08/10/08, Istituto di Ricerche Farmacologiche Mario Negri, Milano Investigator's Meeting - Stato di avanzamento dello studio Rischio & Prevenzione 18/10/08, Istituto di Ricerche Farmacologiche Mario Negri, Milano PROCARDIS Research Program PROCARDIS Ethics Workshop - Ethical and regulatory aspects of the Italian participation in the PROCARDIS program - Differences in attitudes and common concerns on genetic research within Europe , Istituto di Ricerche Farmacologiche Mario Negri, Milano Master di I Livello in Ricerca Clinica dell Università degli Studi di Milano, Facoltà di Medicina e Chirurgia, Dipartimento di Medicina Interna (Anno Accademico ) 15/02/08 La ricerca clinica oggi: profit e no-profit. Il protocollo dello studio 18/02/08 Il disegno degli studi clinici 28/02/08 Legislazione sulla sperimentazione clinica e ruolo dei Comitati Etici 10/03/08 Monitoraggio degli studi clinici 20/03/08 Calcolo della dimensione campionaria 28/03/08 Dalla preclinica alla clinica: sviluppo di nuovi farmaci cardiovascolari 02/04/08 Studi di fase 2: Obiettivi, disegno e stima del campione in oncologia 09/04/08 Le revisioni sistematiche e metanalisi 08/05/08 Ricerca clinica nel campo dell'epilessia 14/05/08 Farmacovigilanza 15/05/08 Ricerca clinica nell'ictus 23/05/08 Interazione tra farmaci 13/06/08 Internet e le nuove tecnologie per l aggiornamento medico 18/06/08 Regolamentazione dei farmaci in Europa 09/09/08 Studi di fase 3: obiettivi, disegno e indicatori 16/09/08 Scienza ed etica 17/09/08 Studi osservazionali: obiettivi, disegno ed aspetti regolatori 23/09/08 Studi osservazionali: obiettivi, disegno ed aspetti regolatori - II^ parte Istituto di Ricerche Farmacologiche Mario Negri, Milano Master di I Livello in Ricerca Clinica dell Università degli Studi di Milano, Facoltà di Medicina e Chirurgia, Dipartimento di Medicina Interna (Anno Accademico ) 10/11/08 Esercitazioni di statistica descrittiva Il disegno dello studio in epidemiologia Il disegno degli studi clinici 11/11/08 Esercitazioni di inferenza statistica Studi di fase 2: obiettivi, disegno e stima del campione in oncologia 12/11/08 Legislazione sulla sperimentazione clinica e ruolo dei Comitati Etici Misure di rischio in epidemiologia Internet e le nuove tecnologie per l'aggiornamento medico-scientifico 17/11/08 Analisi della sopravvivenza Scienza ed etica 18/11/08 Farmacovigilanza Monitoraggio degli studi clinici profit. Report delle Reazioni Avverse 135

137 19/11/08 Monitoraggio degli studi clinici no-profit Report delle Reazioni Avverse negli studi clinici no profit Dalla preclinica alla clinica: sviluppo di nuovi farmaci cardiovascolari 24/11/08 Studi di fase 3: obiettivi, disegno e indicatori in oncologia Le revisioni sistematiche e metaanalisi 25/11/08 Ricerca clinica nel campo dell'epilessia. Ricerca clinica nell'ictus Interazioni tra farmaci La ricerca bibliografica oggi 26/11/08 Ricerca in medicina generale Medicina di domani, etica di ieri? Istituto di Ricerche Farmacologiche Mario Negri, Milano PARTICIPATION IN EVENTS IN WHICH THE DEPARTMENT WAS INVOLVED European Society of Cardiology. Winter Meeting on Translational Cardiology - Heart Failure and Regeneration, 23-26/01/08, Garmisch-Partenkirchen, Germany - Cardioprotective function of the long pentraxin PTX3 in acute myocardial infarction Università di Modena e Reggio Emilia Master Gestione delle Sperimentazioni Statistica Medica II, 25-28/2/08, Dipartimento di Oncologia ed Ematologia, Università di Modena e Ferrara, Italy Il concetto di prognosi nei diversi tipi di studi in epidemiologia: - Outcome negli studi osservazionali - Outcome negli studi sperimentali - Definizione di Endpoint surrogato Metodi statistici (1) per l analisi della prognosi: - Modelli di regressione lineare - Modello logistico Guida alla lettura critica di un articolo scientifico Metodi statistici (2) per l analisi della prognosi: - Modello a rischi proporzionali di Cox - Analisi della sopravvivenza Guida alla lettura critica di un articolo scientifico Istituto Clinico Humanitas. IV Corso di specializzazione in emodinamica e cardiologia interventistica, 04/03/08, Auditorium Istituto Clinico Humanitas, Rozzano (MI), Italy - La cardiomiopatia postinfartuale: il futuro delle cellule staminali American College of Cardiology. 57th Annual Scientific Session, 29/03-01/04/08, Chicago, USA - Multiple RAAS inhibition influences the changes of biohumoral markers in patients with heart failure. Data from Aliskiren Observation of Heart Failure Treatment (ALOFT) Study Istituto di Ricerche Farmacologiche Mario Negri European Clinical Research Infrastructures Network-ECRIN. Donne & Ricerca Clinica, Giornata Internazione della ricerca clinica, 21/05/08, Istituto di Ricerche Farmacologiche Mario Negri, Milano, Italy 136

138 - Fatti, problemi e difficoltà nella ricerca clinica al femminile in cardiologia Dipartimento di Malattie Cardiovascolari e A.S.O. Santa Croce e Carle di Cuneo. Luci ed ombre nell utilizzo dei sartani nelle malattie cardiovascolari: i dati della letteratura, 25/05//08, Salone Costantia Castello Rosso, Costigliole-Saluzzo (CN) - L uso dei sartani nei pazienti con scompenso cardiaco Associazione Nazionale Medici Cardiologi Ospedalieri. XXXIX Congresso Nazionale di Cardiologia, 30/05-02/06/08, Firenze, Italy - Studi clinici - Razionale e disegno dello Studio GISSI-AF - Razionale per l uso di un protocollo dinamico infusionale di insulina - Valutazione dell utilità di un intervento infermieristico di supporto telefonico per migliorare gli stili di vita dopo un ricovero per sindrome coronarica acuta - Valore prognostico delle variazioni di NT-proBNP in pazienti con insufficienza cardiaca coronarica. Dati dallo studio Val-HeFT - Determinanti di disfunzione endoteliale (microalbuminuria) e infiammazione vascolare (Pentraxina-3) in pazienti con insufficienza cardiaca cronica. Dati dallo studio GISSI-HF - La pentraxina lunga PTX3 è un marcatore di comorbidità nei pazienti con insufficienza cardiaca. Dati dallo studio GISSI-HF - Interazione fra un marcatore del turnover del collagene (PIIINP) e un antagonista dell aldosterone (Canrenone) sul rimodellamento ventricolare in pazienti con insufficienza cardiaca lieve. Dati dallo studio AREA IN.CHF - Trattamento dell iperglicemia in corso di sindrome coronarica acuta: efficacia e sicurezza di un protocollo di infusione endovenosa di insulina a gestione infermieristica Roche Diagnostics. 6th International Symposium onnf-probnp, 18-19/06/08, Baveno, Italy - hs Troponin T: The Val-HeFT and GISSI Studies - Serial measurements of NT-proBNP in chornic heart failure European Society of Cardiology. ESC Congress 2008, 30/08-03/09/08, Munich, Germany - Effectiveness and safety of a new nurse-implemented insulin infusion protocol (DDD) for intensive glucose control in diabetic patients with acute coronary syndromes - Clinical determinants of pentraxin-3 (PTX3), a novel marker of vascular inflammation, in patients with chronic heart failure. Data from the GISSI-HF study - Categorial changes of NT-proBNP concentrations predict outcome in ambulatory patients with chronic stable heart failure. Data from the valsartan heart failure trial WONCA EUROPE th Regional Conference. Overcoming the distance - Family doctor, bringing the art of medicine to the patients, 04-07/09/08, Istanbul, Turkey - The optimisation of cardiovascular prevention in everyday practice: preliminary results of the Risk & Prevention Study Clinical Forum S.r.l. Easy future. Nuove prospettive terapeutiche, 12-13/09/08, Istituto di Ricerche Farmacologiche Mario Negri, Milano, Italy - Revisione dell ipotesi metabolico-infiammatoria nello scompenso. Implicazioni terapeutiche Dipartimento Cardiovascolare dell Azienda Ospedaliero Universitaria degli Ospedali Riuniti di Trieste e Centro Cardiovascolare dell Azienda per i Servizi Sanitari N. 1 Triestina, 10-11/10/08, Trieste, Italy - Citochine, infiammazione e BNP nello scompenso cardiaco: aiuteranno realmente a comprendere e curare meglio la sindrome? 137

139 Struttura Complessa di Cardiologia dell Azienda Ospedaliera G Brotzu di Cagliari e Unità Operativa di Cardiologia dell Ospedale SS Annunziata-ASL N. 1 di Sassari. La Sardegna nel cuore. 2 Congresso di cardiologia a carattere nazionale, 23-25/10/08, Cagliari, Italy - Le terapie geniche e cellulari American Society of Nephrology. ASN Congress, 4-9/11/08, Philadelphia, USA - Safety and tolerability profile of aliskiren added to optimized therapy in patients with heart failure and renal impairment American Heart Association. AHA Scientific Session 2008, 8-12/11/08, New Orleans, USA - Elevated albumin excretion is an independent risk factor in patients with chronic heart failure. Data from the GISSI-Heart Failure Trial - Circulating markers of myocyte injury predict first recurrence of atrial fibrillation. Data from the GISSI-Atrial Fibrillation Trial - Role of inflammatory markers in the prediction of first recurrence of atrial fibrillation. Data from the GISSI-Atrial Fibrillation Trial - Stable precursor fragments of vasoactive peptides predict outcome in patients with chronic heart failure. Data from the GISSI-Heart Failure Trial - The role of valsartan in the prevention of atrial fibrillation recurrence: the GISSI-AF results Ospedale Luigi Sacco. La comorbidità scompenso cardiaco e fibrillazione atriale: quali approcci terapeutici? La gestione multidisciplinare, 29/11/08, Milano, Italy - Luci ed ombre nel trattamento farmacologico: cosa ci suggeriscono le nuove linee guida GRANTS AND CONTRACTS AIFA (Agenzia Italiana del Farmaco), AstraZeneca, Azienda Ospedaliera San Gerardo Monza, Chiesi Farmaceutici, Boehringer Ingelheim Italia SpA, BRAHAMS AG, Centro Cardiologico Monzino IRCCS Milano, Cambridge University, CONGENIA, Collegio Interprovinciale Milano-Lodi, Fondazione Don Gnocchi Milano, Fondazione San Raffaele Milano, Heart Care Foundation, International Biomedical System SpA, Istituto Auxologico di Milano, Kinetic Concepts Inc., Ministero della Salute, Novartis Pharma, Ospedale Casa Sollievo della Sofferenza IRCCS San Giovanni Rotondo, Oxford University, Population Health Research Institute-Mc Master University, Pfizer Italia, Regione Lombardia, Roche Diagnostics, Sanofi- Aventis, Sigma Tau, SPA Società Prodotti Antibiotici S.p.A., Takeda Italia S.p.A., Università degli Studi Torino 138

140 SELECTION OF SCIENTIFIC PUBLICATIONS FROM 2008 Amigoni M, Bellani G, Scanziani M, Masson S, Bertoli E, Radaelli E, Patroniti N, Di Lelio A, Pesenti A, Latini R Lung injury and recovery in a murine model of unilateral acid aspiration: functional, biochemical, and morphologic characterization Anesthesiology 2008; 108: Anand SS, Islam S, Rosengren A, Franzosi MG, Steyn K, Yusufali AH, Keltai M, Diaz R, Rangarajan S, Yusuf S, on behalf of the the INTERHEART Investigators Risk factors for myocardial infarction in women and men: insights from the INTERHEART study Eur Heart J 2008; 29: Bertele' V, Angelici L, Barlera S, Garattini S Thrombolysis or nothing for acute myocardial infarction? It's all the same! Br J Clin Pharmacol 2008; 65: Broadbent HM, Peden JF, Lorkowski S, Goel A, Ongen H, Green F, Clarke R, Collins R, Franzosi MG, Tognoni G, Seedorf U, Rust S, Hamsten A, Farrall M, Watkins H, for the PROCARDIS Consortium Susceptibility to coronary artery disease and diabetes is encoded by distinct, tightly linked, SNPs in the ANRIL locus on chromosome 9p Hum Mol Genet 2008; 17: Cholesterol Treatment Trialists' CTT Collaboration Efficacy of cholesterol-lowering therapy in 18,686 people with diabetes in 14 randomised trials of statins: a metaanalysis Lancet : Connolly SJ, Pogue J, Eikelboom J, Flaker G, Commerford P, Franzosi MG, Healey JS, Yusuf S, on behalf of the ACTIVE W Investigators Benefit of oral anticoagulant over antiplatelet therapy in atrial fibrillation depends on the quality of international normalized ratio control achieved by centers and countries as measured by time in therapeutic range Circulation 2008; 118: CPR CHD Genetics Collaboration Collaborative pooled analysis of data on C-reactive protein gene variants and coronary disease: judging causality by Mendelian randomisation Eur J Epidemiol 2008; 23: Dossena S, Imeri L, Mangieri M, Garofoli A, Ferrari L, Senatore A, Restelli E, Balducci C, Fiordaliso F, Salio M, Bianchi S, Fioriti L, Morbin M, Pincherle A, Marcon G, Villani F, Carli M, Tagliavini F, Forloni G, Chiesa R Mutant prion protein expression causes motor and memory deficits and abnormal sleep patterns in a transgenic mouse model Neuron 2008; 60: Galvez BG, Sampaolesi M, Barbuti A, Crespi A, Covarello D, Brunelli S, Dellavalle A, Crippa S, Balconi G, Cuccovillo I, Molla F, Staszewsky L, Latini R, DiFrancesco D, Cossu G Cardiac mesoangioblasts are committed, self-renewable progenitors, associated with small vessels of juvenile mouse ventricle Cell Death Differ 2008; 15: Latini R, Brines M, Fiordaliso F Do non-hemopoietic effects of erythropoietin play a beneficial role in heart failure? Heart Fail Rev 2008; 13: Maggioni AP, Franzosi MG, Latini R Beta-adrenoceptor antagonists and antianginal drugs In: Side effects of drugs, Annual 30 Elsevier, Amsterdam, 2008; Masson S, Latini R Amino-terminal pro-b-type natriuretic peptides and prognosis in chronic heart failure Am J Cardiol 2008; 101 Suppl: 56A-60A 139

141 Masson S, Latini R, Anand IS, Barlera S, Angelici L, Vago T, Tognoni G, Cohn J N, for the Val-HeFT Investigators Prognostic value of changes in N-termianl pro-brain natriuretic peptide in the Val-HeFT (Valsartan Heart Failure Trial) J Am Coll Cardiol 2008; 52: McMurray JJV, Pitt B, Latini R, Maggioni AP, Solomon SD, Keefe DL, Ford J, Verma A, Lewsey J, for the Aliskiren Observation of Heart Failure Treatment (ALOFT) Investigators Effects of the oral direct renin inhibitor aliskiren in patients with symptomatic heart failure Circulation Heart Failure 2008; 1: Montalvo G, Avanzini F, Anselmi M, Prandi R, Ibarra S, Marquez M, Armani D, Moreira J M, Caicedo C, Roncaglioni MC, Colombo Fabio, Camisasca P, Milani V, Quimi' S, Gonzabay F, Tognoni G Diagnostic evaluation of people with hypertension in low income country: cohort study of "essential" method of risk stratification BMJ 2008; 337: a1387 Pedrazzini GB, Masson S, Latini R, Klersy C, Rossi M G, Pasotti E, Faletra FF, Siclari F, Minervini F, Moccetti T, Auricchio A Comparison of brain natriuretic peptide plasma levels versus logistic EuroSCORE in predicting in-hospital and late postoperative mortality in patients undergoing aortic valve replacement for symptomatic aortic stenosis Am J Cardiol 2008; 102: Pedrazzini G, Santoro E, Latini R, Fromm L, Franzosi MG, Moccetti T, Staszewsky L, Barlera S, Tognoni G, Maggioni A P, for the GISSI-3 Investigators Causes of death in patients with acute myocardial infarction treated with angiotensin-converting enzyme inhibitors: Findings from the Gruppo Italiano per lo Studio della Sopravvivenza nell'infarto (GISSI)-3 trial Am Heart J 2008; 155: Salio M, Chimenti S, De Angelis N, Molla F, Maina V, Nebuloni M, Pasqualini F, Latini R, Garlanda C, Mantovani A Cardioprotective function of the long pentraxin PTX3 in acute myocardial infarction Circulation 2008; 117: Specchia C, Barlera S, Chiodini BD, Nicolis EB, Farrall M, Peden J, Collins R, Watkins H, Tognoni G, Franzosi MG, on behalf of the PROCARDIS Consortium Quantitative trait genetic linkage analysis of body-mass index in familial coronary artery disease Hum Hered 2008; 66: GISSI-HF Investigators (Writing Committee: Tavazzi L, Maggioni AP, Marchioli R, Barlera S, Franzosi MG, Latini R, Lucci D, Nicolosi GL, Porcu M, Tognoni G) Effect of n-3 polyunsaturated fatty acids in patients with chronic heart failure (the GISSI-HF trial): a randomised, double-blind, placebo-controlled trial. Lancet 2008; 372: GISSI-HF Investigators (Writing Committee: Tavazzi L, Maggioni AP, Marchioli R, Barlera S, Franzosi MG, Latini R, Lucci D, Nicolosi GL, Porcu M, Tognoni G) Effect of rosuvastatin in patients with chronic heart failure (the GISSI-HF trial): a randomised, double-blind, placebocontrolled trial Lancet 2008; 372: LAY PRESS SELECTION PUBLISHED IN 2008 Latini R Cytokines and heart failure Nova Acta Leopold 2008; n. 351: Latini R, Masson S Prognostic value of circulatory cardiac troponins in heart failure Rev Esp Cardiol 2008; 61:

142 Latini R, Masson S Il valore diagnostico della troponina T nell'insufficienza cardiaca Recenti Prog Med 2008; 99: Masson S, Latini R, Salio M NT-proBNP and prognosis in chronic heart failure In: NT-proBNP as a biomarker in cardiovascular diseases. Prous Science, Barcelona, 2008; Tognoni G Per una cultura-pratica cardiovascolare non-arrogante Informazione sui Farmaci 2008; 32, n. 5: Turazza FM, Masciocco G, Galvanin S, Macera F, Lenardon A, Iannì S, Foti G, Frigerio M Introduzione alla cardiomiopatia ipertrofica: eziologia, presentazione e storia naturale In: Cardiologia Atti del 42 convegno internazionale del dipartimento cardiologico A. De Gasperis settembre 2008, Milano. Fond. Centro Cardiologia e Cardiochirurgia A. De Gasperis, Milano, 2008; RESEARCH ACTIVITIES Laboratory of Cardiovascular Clinical Pharmacology The effects of mesoangioblasts and of different progenitor cells on injury after experimental myocardial infarction in the mouse Many studies have demonstrated that autologous and homologous cells of various origins can repair myocardium damaged due to an acute ischemic insult. Mesangioblasts are potentially interesting when compared with bone marrow precursors because (a) they are easily expanded and (b) obtainable by a biopsy of skeletal muscle in man. Mesoangioblasts isolated from human heart biopsies migrate and home in the heart of immunodeficient mice after coronary ligation, and they survive for at least 4 weeks. Studies are ongoing to establish their protective and regenerative potential, as a basis for possible clinical studies. The same model of coronary ligationin immunodeficient mice is being used for testing in vivo the angiogenic potential of other progenitor cells such as mononuclear cells Tie-2+ (collaboration with Michele De Palma, HSR, Milano), CD34+ (collaboration with Franca Fagioli, Osp S. Anna, Torino), CD133+ (collaboration with Giulio Pompilio, Centro Cardiologico Monzino, Milano). This research is partly done within the sixth EU program, European Vascular Genomics Network, that ended in December 2008 (EVGN, Pulmonary injury by hydrochloric acid in the mouse: a model of Aspiration pneumonitis to test protective interventions Aspiration pneumonitis (AP) occurs when the acid content of the stomach makes his way through the larynx in the lower respiratory tract. Patients with consciousness disturbance are at risk for this event. Specifically, it has been shown that Pulmonary Aspiration can complicate between % general anesthesia procedures. The main injurious mechanism of AP is the chemical insult due to the low ph of gastric secretions, which causes a chemical burn of the airway tree and of the alveolar structures. The course of AP can be extremely variable, ranging from the silent aspiration characterized by a modest desaturation to the dramatic sequelae of Acute Lung Injury (ALI) and Acute Respiratory Distress Syndrome (ARDS), requiring prolonged mechanical ventilation and potentially leading to death. In a murine model of monolateral acid instillation established in our laboratory, we have shown the protective effect of exogenous pulmonary surfactant instillation. We are currently working on a model of ventilation-induced lung injury (VILI) to assess the effect of 141

143 exogenous surfactant and its modulation in transgenic mice for the acute-phase protein pentraxin-3. Conditional transgenic model of cardiac HGF expression to promote neovascularization and to recruit stem cells in the myocardial infarction Growth factors such as hepatocyte growth factor (HGF) through angiogenic and anti-apoptotic effects may promote cardiac repair after myocardial infarction and in heart failure. The cardiacspecific α-myosin Heavy Chain (MHC-α) transactivator was used to direct expression of HGF to cardiomyocytes: by this way the effects of HGF can be tested under cardiac ischemia and reperfusion, without the need for administration of exogenous HGF. The Heart-HGF transgenic model is being used to verify the ability of HGF in vivo to promote neovascularisation, to protect cardiomyocytes from apoptosis, to recruit and activate endogenous or transplanted stem cells and to sustain their cellular replication and differentiation into cardiomyocytes after ischemic damage and reperfusion. Roles of macrophages in cardiac ischemia/reperfusion injury and in cardiac repair Macrophages either resident or from blood-borne monocytes play several key roles in the response of the heart to ischemic injury. They may be useful in particular during cardiac repair, when collagen deposition occurs and neonagiogenesis, stimulated by growth factors produced by macrophages. The aim of the project is to assess the relevance of macrophages in myocardial repair and scar formation after myocardial infarction, in the attempt to dissect the role of inflammatory cells in myocardial injury vs repair. The effects of coronary ligation (with/without reperfusion) is being tested in mice in which machrophages can be ablated by administration of Dyphteria toxin in transgenic mice expressing human diphteria toxin receptor (CD11b-DTR). We established a regimen of administration of dyptheria toxin that allows the halving of monocytes level, and hence of peritoneal macrophages. A preliminary experiment of cardiac ischemia/reperfusion has shown that the number of macrophages was also halved in the infarct area in transgenic mice treated with dyphteria toxin. We are now running experiments to evaluate the structural and functional consequences of this reduction in macrophages number. Effects of dipeptidyl peptidase-4 (DPP-4) inhibition on progenitor cells and cardiac repair in type 2 diabetes In patients and in experimental animal models with type 2 diabetes mellitus, inhibition of plasma dipeptidyl peptidase IV (DPP-4) activity, increased plasma levels of the glucagonlike peptide-1 (GLP-1) and glucose-dependent insulinotropic polypeptide (GIP) improving glucose tolerance and islet cell function. Stromal cell-derived factor 1 (SDF-1α) is also cleaved by DPP- 4 and is crucially important in the mobilization and selective homing of endothelial progenitor cells (EPCs) to ischemic tissues. EPCs have the potential to differentiate into functional mature endothelial cells, which can substitute diseased endothelium and contribute to repair ischemic tissues, including the heart. This regenerative action of EPCs is compromised in both type 1 and type 2 diabetic patients. The present study aimed at evaluating whether the administration of DPP-4 inhibitor to obese/diabetic ob/ob mice improves mobilization/homing of EPCs by increasing levels of SDF- 1, and by this way contributing to the regeneration of myocardium and blood vessels in a model of isoproterenol-induced cardiac infarction. To further increase cardiac injury a group of animals were forced to swim for 5 minutes. Hypoxic areas, detected by Hypoxyprobe -1, were found in the epicardium of mice treated with isoproterenol and forced to swim. The phenomenon was exacerbated in the diabetic ob/ob mice treated in the same way. The hypoxic myocardial damage determined a significative 142

144 mobilization of EPCs in control mice whereas this protective response was compromised in the diabetic ob/ob mice. Whether SDF-1 levels paralleled the EPC mobilization, the hypoxic damage determined a decrease of capillary density and whether forced swim play a role in the cardiovascular damage will be evaluated before beginning with the treatment of these animals with an DPP-4 inhibitor GISSI-HF: biohumoral substudy and microalbuminuria The clinical trial GISSI-HF (see related paragraph) was designed to assess whether two treatments (a statin and n-3 polyunsaturated fatty acids or PUFA) can improve the prognosis of patients with heart failure of any etiology, with preserved or compromised left ventricular ejection fraction. Main results have recently been published (see references). The biomarker substudy aims at exploring the pathophysiology of heart failure and mechanisms of action of the treatments in study. Overall, 1237 patients have been enrolled in the substudy. Blood samples were collected at enrolment and after three months to measure plasma PUFA, and markers of ventricular myocyte stress (brain natriuretic peptides, BNP and N-terminal probnp, MRproANP), myocardial damage (cardiac troponin T, measured with the standard assay or with a newly developed high sensitive method) and inflammation (C-reactive protein, CRP, pentraxin- 3, PTX3). Baseline fraction of n-3 PUFA averages 3.4 and increases by about 50% over 3 months. Baseline BNP concentration in 1223 patients is 141 pg/ml (median) and correlates with the severity of heart failure. The number of circulating endothelial progenitor has been measured in a smaller group of 68 patients, in collaboration with the laboratory of E. Dejana (IFOM). Number of endothelial progenitor cells in patients with heart failure is 30-50% lower than in healthy volunteers was inversely correlated to morbidity/mortality in a population of patients from 5 Italian centers and a Cardiology center in Frankfurt (collaboration with Andreas Zeiher). Microalbuminuria (defined as the ratio between urinary concentrations of albumin and creatinine) is being measured in more than 2000 patients enrolled in the GISSI-HF trial as an indicator of renal endothelial dysfunction. Microalbuminuria (albumin/creatinine = mg/g) is present in 19% of the patients and is associated to inflammation, endothelial dysfunction and neurohormonal activation. Novel and more stable circulating biomarkers, have been successively assayed. They belong to the family of natriuretic peptides (mid-regional proatrial natriuretici peptide, MR-proANP), endothelin (C-terminal pro-endothelin-1, CT-proET-1), vasopressin (C-terminal pro-vasopressin, CT-proAVP) and adrenomedullin (mid-regional proadrenomedullin, MR-proADM). Other markers related to infection and innate immunity are currently measured (cromogranin A, mannose-binding lectin, osteoprotegerin, adiponectin and alpha-defensin). First data on the biomarkers substudy have been presented to national and international scientific meetings and original publications are under preparation. PTX-3, a novel long pentraxin is a marker of severity of disease and of outcome in cardiovascular diseases, independent of C-reactive protein PTX-3 is a novel long pentraxin whose expression is induced by cytokines in endothelial and mononuclear cells, mostly in striated muscle and heart, while C-reactive protein (CRP) is mainly synthesized in the liver. PTX3 was shown to peak in plasma around 7 h after onset of symptoms of MI and to be an independent predictor of 3-month mortality. PTX3 has been assayed with a more accurate method in 1200 patients with symptomatic heart failure (GISSI- HF) and in 380 patients with atrial fibrillation (GISSI-AF) to explore its role in other cardiovascular diseases. First results indicate that PTX3 is associated with different clinical characteristics in patients with heart failure, including advanced age, ventricular dysfunction, functional class (NYHA class), and comorbidities such as atrial fibrillation and diabetes. PTX3 independently predicts mortality and morbidity in the patients with chronic heart failure. 143

145 Echocardiographic and biohumoral substudy GISSI-AF trial The GISSI Atrial Fibrillation trial (GISSI-AF) tests the efficacy of Valsartan, an angiotensin II AT1-receptor blocker, in the prevention of atrial fibrillation recurrence in 1400 patients. A substudy of the GISSI-AF has recently been concluded; it evaluated the potential role of biohumoral factors and cardiac structural remodeling in the reoccurrence and severity of atrial fibrillation. In approx. 400 patients three serial echocardiographic exams (at randomization, 6 months and 1 year) and contemporaneous blood collection were performed. Left ventricular and atrial dimensions were determined by echocardiography, whereas plasma levels of natriuretic peptides (BNP, NT-proBNP and MR-proANP), troponin T (high sensitive method), stable vasopactive peptides (endothelin-1, Adrenomedullin and vasopressin), inflammatory markers (C-reactive protein, interleukin-6 and pentraxin-3) and procalcitonin have been measured. Besides giving clues on the pathophysiology of atrial fibrillation, the most common arrhythmia in elderly, this substudy aims at providing mechanical insights of the potential benefits of the study drug. The study was completed on January 31, 2008 and results being analyzed. First data on circulating biomarkers have been presented to the scientific meeting of the American Heart Association (November 2008). CandHeart: effects of candesartan on BNP and left ventricular function in patients with symptomatic heart failure Candesartan, an antagonist of angiotensin II type 1 receptors, significantly reduces mortality and morbidity in heart failure, as shown by the CHARM trials. The principal objective of the trial is to assess the effects Candesartan on circulating levels of brain natriuretic peptide (BNP) in patients suffering from CHF with depressed or preserved left ventricular (LV) systolic function. The study enrolled 487 patients in 70 clinical centers with a follow-up of 1-year. Serial circulating blood samples and echocardiographic examinations have been performed at baseline and after 3 and 12 months (end of study). Besides BNP, other prognostic biomarkers such as aldosterone and microalbuminuria have been assayed. A statistical plan of analyses has been written and main results are expected for the beginning of Trial Prone/Supine 2: effects of prolonged prone position on survival in severe acute respiratory distress syndrome (ARDS) This trials extends the findings of the previous Prone/Supine trial that showed in 304 patients with ARDS that 6-hour pronation in the first 10 days improved blood gas parameters, but did not reduce mortality (around 50% in the first 6 months). This was one of the rare examples of a multicenter trial in Intensive Care performed entirely in Italy. The same group of Centers (headed by the Istituto di Anestesia e Rianimazione from the Policlinico di Milano, L Gattinoni with data management by Mario Negri) has recently concluded a new study that aims at evaluating the effect of a prolonged pronation (20 hours a day) in patients with severe ARDS, a subgroup that showed a trend towards improved survival on pronation in the previous study. Three hundred and forty two patients have been enrolled in 20 centers and the main results of this study have been submitted for publication. DyDa: left ventricular dysfunction in diabetes. Prevalence and incidence of left ventricular dysfunction in diabetics patients without clinical cardiac disease This is a prospective, multicentric, national and epidemiological trial aimed at evaluating the prevalence of left ventricular dysfunction (systolic or diastolic) in 1000 patients with type 2 diabetes mellitus but no clinical cardiovascular disease at enrolment. The incidence of left ventricular dysfunction is monitored during a 2-year follow-up using ECG and echocardiography. The Biomarker Core Laboratory evaluated the biohumoral profile of these 144

146 patients at study entry, measuring the circulating levels of brain natriuretic peptide, C-reactive protein, microalbuminuria and glycated hemoglobin. Enrolment started in July 2006 and ended in March 2008 with 970 patients recruited. The assays of the biomarkers are concluded and first data on the association between theses markers and baseline clinical characteristics are under analysis. In a subgroup of patients from the study, levels of circulating progenitor cells and their angiogenic potential in vitro will be measured to assess whether they are associated to risk of developing cardiovascular disorders in diabetics. Albumin Italian Outcome Sepsis Study. The ALBIOS Study (AIFA) ALBIOS is a multicenter, controlled, randomized clinical trial that compares the efficacy of human albumin and a crystalloid solution for volume replacement in patients with severe sepsis or septic shock. The primary endpoint is survival at 28 and 90 days after enrolment. Secondary endpoints include the number of organ dysfunctions, severity of organ dysfunction (SOFA scale), and lengths of stay in (intensive care unit) ICU and in hospital. More than 150 ICU in Italy are expected to participate to this large study, coordinated by the Ospedale Maggiore Policlinico in Milan and the Consorzio Mario Negri Sud. A group of 48 ICUs participates to a biomarkers substudy, coordinated by the laboratory of Clinical Cardiovascular Pharmacology, with the aims of enrolling 800 patients. Serial blood samples are collected to measure the possible effects of albumin on markers of inflammation, infection, cardiac function and coagulation. 21 patients have been randomized by the end of Clinical, biochemical and instrumental predictors of outcome in rehabilitation after cardiac surgery (MIUR) Due to the short length of hospital recovery in patients with cardiac disease, the period of rehabilitation becomes crucial and is indicated for almost all the cardiomyopathies, including myocardial revascularization and surgery of cardiac valves. The objective of this study is to evaluate the prognostic role of circulating biomarkers in 250 patients enrolled in 5 Centers for rehabilitation after cardiac surgery. A plasma bank will be collected under the responsibility of the Laboratory of Cardiovascular Clinical Pharmacology from the Mario Negri Institute to assay potential markers of interest. The project is coordinated by the Fondazione Don Gnocchi in Milan. The first patient was enrolled in Evaluation of different anesthesiological strategies for supratentorial neurosurgery. The NeuroMorfeo Study (AIFA) The aim of the study is to evaluate whether an anesthesia with volatile anesthetics is equivalent to endovenous anesthetics for elective supratentorial surgery. This is a multicenter, randomized, controlled and opened study, based on a design of equivalence for comparison of different anesthesiological strategies. Patients to be included (n= 393) will be selected in 14 neurosurgical centers in Italy for elective intracranial surgery with supratentorial lesions without signs of endocranial hypertension (range of age years). The biohumoral response to surgical stress will be measured as an indicator of homeostasis and neurovegetative status. The urinary excretion of catecholamines and cortisol and the plasma concentration of cortisol will be measured in a central laboratory. The study is coordinated by the San Gerardo Hospital in Monza and by the Department of Cardiovascular Research from the Mario Negri Institute. The first patient has been enrolled in December 2007, and since then 314 patients have been randomized. Preliminary data show that the urinary excretion of catecholamines (normalized by creatinine concentration) is within the expected range. Prevalence of asymptomatic cardiac dysfunction and heart failure in a population of elderly subjects from Lazio. The PREDICTOR study This observational study aims at evaluating the prevalence of asymptomatic cardiac dysfunction 145

147 and heart failure in a random sample of elderly subjects from the Lazio area. The secondary objective is to identify clinical, biohumoral (natriuretic peptides) and non-invasive instrumental (echocardiography and ECG) markers of asymptomatic cardiac dysfunction and heart failure. The population under observation is a randomly selected sample of elderly subjects (age ranging from 65 and 84 years) resident in the area of 10 hospital cardiology centers. In a first phase, 1000 subjects have been recruited; a second phase has recently started with the objective of enrolling another 2000 subjects. Blood samples are collected for each subject and stored in the biobank in the Laboratory of Clinical Cardiovascular Pharmacology. Preliminary data obtained in the first 600 subjects show a good pathophysiological agreement between the circulating levels of NT-proBNP and indexes of left or right asymptomatic cardiac dysfunction or the severity of heart failure. Laboratory of Clinical Drug Evaluation PROCARDIS: A genome-wide strategy to identify susceptibility loci in precocious coronary artery disease The PROCARDIS research programme, a genome-wide strategy to identify susceptibility loci in precocious coronary artery disease (CAD) supported by the 5th Framework Programme of the EC, was initiated as a collaboration between the universities of Oxford and Mƒnster, the Karolinska Institute, the Mario Negri Institute with the support of the GISSI group, Oxagen, a biotechnology company, and AstraZeneca. The objectives of the first stage of this programme were to collect a minimum of 2000 affected sibling pairs (ASPs) and families with precocious CAD and to apply genome-wide linkage mapping techniques, by typing anonymous highly informative markers spaced throughout the genome, to identify chromosomal regions which are linked to the susceptibility to early-onset CAD. The study design is based on family ascertainment, to allow non-parametric linkage analyses (in ASPs). The PROCARDIS collected 2,036 CAD families from four European countries, in order to maximise the power of detecting genes that confer modest risks. A genome-wide linkage scan identified three promising regions for intensive study; one of the linked regions (Chromosome 17) was confined to families with multiple cases of myocardial infarction and was replicated in a second independent series of families. In addition the linkage scan confirmed a previously identified locus on Chromosome 2. These results demonstrate that novel CAD susceptibility genes are tractable to positional cloning which promises to lead to the identification of new molecular insights into this condition, and hopefully, new treatments. Additionally, the program has collected nuclear families (e.g. parent-offspring trios) for transmission-based tests of association for fine mapping by linkage disequilibrium analysis and testing of positional candidate genes. Extensive clinical and biochemical intermediate phenotype data have also been collected and assessed. The second stage of PROCARDIS, supported by the EC 6th Framework Programme, is conducting a large GWAS (genome wide association study), where the patients with myocardial infarction enrolled in the first stage will be compared with control subjects to identify novel candidate genes. The results will be replicated in different populations. The collaboration has been extended to include complementary expertise, as proteomics, bioinformatics, functional analysis. GISSI-HF: A large scale clinical trial testing the effects of n-3 PUFA and Rosuvastatin on mortality/morbidity of patients with symptomatic Congestive Heart Failure 146

148 The GISSI (Gruppo Italiano per lo Studio della Sopravvivenza nell'insufficienza cardiaca) is a collaborative group endorsed by ANMCO (Associazione Nazionale Medici Cardiologi Ospedalieri) and by the Istituto Mario Negri, active from 20 years in the cardiovascular research field. The GISSI-HF was the fifth large scale clinical trial conducted by the Group and was a prospective, multicenter, randomized, double blind, placebo controlled study, with randomized allocation of patients with a clinical diagnosis of heart failure to: Randomization 1 (R1): n-3 PUFA vs corresponding placebo; Randomization 2 (R2): rosuvastatin vs corresponding placebo. The primary objective of the GISI-HF was to demonstrate that, in patients with heart failure treated at the best of recommended treatment, long-term administration of n-3 PUFA and/or rosuvastatin is more effective than the corresponding placebo in the reduction of all-cause mortality and all-cause mortality or cardiovascular hospitalizations. The study started in September 2002 with the participation of 357 departments of cardiology and nearby 7000 patients; 3494 patients received n-3 PUFA in a capsule once daily and 3481 received placebo; in a follow-up time of four years, 955 patients in the PUFA group (27%) died, compared with 1014 (29%) in the placebo group, meaning a relative risk reduction of 9% in the PUFA group. A higher proportion of patients in the placebo group (2053/59%) died or were admitted to hospital for cardiovascular reasons than in the PUFA group (1981/57%) a relative reduction of 8% in the PUFA group. In absolute terms, 56 patients needed to be treated with PUFA for just under four years to avoid one death, or 44 patients to avoid one event of either death or admission to hospital for cardiovascular causes. Statin treatment with rosuvastatin did not affect clinical outcomes in patients with chronic heart failure. Several substudies focus on possible mechanistic effects of the study treatments: ventricular remodeling (echo); biohumoral; genetic; arrhythmic and autonomic pattern (Holter monitoring); exercise capacity; cognitive function; burden of care. The analysis of these results are in progress. The project is conducted in collaboration with the Laboratory of Cardiovascular Clinical Pharmacology. GISSI-HF Genetic Substudy The role of genetic factors in causes, evolution, prognosis and treatment of heart failure is largely unexplored, with the exception of heart failure originated by specific cardiomyopathies (such as dilated, hypertrophic, arrhythmogenic right ventricular cardiomyopathies), for which the role of heritable gene mutations is increasingly well understood. Heart failure (HF) is a syndrome with different etiologies, and more than one half is caused by coronary heart disease (CHD). A genetic substudy to GISSI-HF (see "ad hoc" section) offers the opportunity to improve knowledge on the role of genetic factors involved in heart failure, through a collection of blood samples of a large population of patients, involving cases of heart failure of different etiologies, i.e. non-ischaemic and ischaemic heart disease. The objective of the genetic substudy is 1) to assess the relationships between the polymorphysms of various candidate genes and the clinical outcome in patients enrolled in GISSI-HF study; 2) to assess whether these relationships are modified by the experimental treatments. The genetic markers to be studied will initially focus on polymorphisms of genes involved in lipid metabolism and inflammation. However, as new genetic information is being accumulated continually, at present it is not known if other polymorphisms/genes will be important at the time of study completion. The possibility to assay other allelic variants is then foreseen. The GISSI-HF genetic substudy is conducted by nearly 100 Centres that have included 2200 patients. GISSI-Prevenzione-Genetic Study Myocardial infarction is a multifactorial disease. While the role of known risk factors on coronary heart disease susceptibility is well defined, the impact of the genetic components and 147

149 its interaction with environmental factors need investigation. The GISSI-Prevenzione trial investigated the effects of pharmacological treatments with n-3 PUFA and pravastatin on morbidity and mortality after myocardial infarction. During the study more than 8000 samples of a large population of patients affected by this disease has been collected and stored with the collaboration of SIBioC (Societê Italiana di Biochimica Clinica e Biologia Molecolare). The GISSI-Prevenzione-Genetic Study investigates the role of genetic factors in ischaemic heart disease. The objectives of the project are 1) to assess the relationships between the polymorphysms of various candidate genes and the clinical outcome in patients enrolled in the large clinical trial GISSI-Prevenzione study; 2) to assess whether these relationships are modified by the pharmacological treatments. According to these objectives, we investigated the relationship between APOE, mortality and the response to treatment in 3304 myocardial infarction survivors randomized to pravastatin or no treatment. We found that epsilon 4 allele is a determinant of pravastatin response in terms of survival. Though in the entire population investigated we found a beneficial effect of pravastatin in terms of survival, only the epsilon 4 carriers seemed to have gained a significant benefit from this treatment. We suggest that the effect of statins is of particular interest in this fraction of the population and that genetic markers can help in identifying patients that benefit more from statin treatment. Association studies in the same population on the adiponectin gene, the CRP (C-reactive protein) gene variants, some genetic variants on Chromosome 9p21 are in progress. GISSI-AF. Clinical trial testing the efficacy of an angiotensin II AT1- receptor blocker in Atrial Fibrillation The GISSI-AF is a randomized, prospective, parallel group, placebo-controlled, multicenter study on the use of valsartan, an angiotensin II AT1-receptor blocker in the prevention of Atrial Fibrillation (AF) recurrence. Atrial fibrillation is the most frequent form of arrhythmia in clinical practice, affecting 6% of people over 65 years old. The traditional therapies are often able to restore the sinus rhythm but are subject to a very high percentage of relapses, above all when the AF has been present for a long time and when there are structural changes at both atrial and ventricular level. The reninangiotensin-aldosterone system (RAAS) plays a key role in the remodeling phenomenon, which makes increasingly irreversible the electrical, mechanical and structural properties of the atrial tissue. Existing experimental and clinical data do not allow any definite conclusion regarding the efficacy of an angiotensin II AT1-receptor blocker in the prevention of AF. The GISSI group decided to conduct a specific trial of adequate size versus placebo aimed to assess if the addition of valsartan on top of established therapies can reduce the recurrence of atrial fibrillation in patients with a history of recent AF associated with cardiovascular diseases/comorbidities. The study has recruited and treated for 12 months 1400patients with a history of recent AF associated with cardiovascular diseases/comorbidities. In GISSI-AF, the largest trial ever conducted with ARBs in pts with AF, Valsartan did not reduce AF recurrence. A trend favoring valsartan was observed only in the small group of pts with heart failure and/or left ventricular dysfunction. The project was conducted in collaboration with the Laboratory of Cardiovascular Clinical Pharmacology. NeuroMorfeo Study: see Laboratory of Cardiovascular Clinical Pharmacology The Population Health Research Institute (PHRI), McMaster University, Hamilton, Ontario, is the coordinating center of a multinational network of cardiology clinics that collaborate to the conduction of multicenter large scale clinical trials (nearly 40 Countries and more than 600 cardiology clinics). The Laboratory of Clinical Drug Evaluation is responsible for the scientific coordination in Italy of some of these trials (ACTIVE, RE-LY, CURRENT, FUTURA OASIS- 8). 148

150 ACTIVE study (Atrial Fibrillation Clopidogrel Trial with Irbesartan for prevention of Vascular Events) The aims of the study were to evaluate whether clopidogrel plus acetylsalicylic acid (ASA) is superior to ASA alone (A study) and non-inferior to standard oral anticoagulant therapy (W study) in preventing vascular events in patients with atrial fibrillation and to evaluate whether blood pressure lowering with irbesartan is superior to placebo in preventing vascular events in patients with atrial fibrillation (I study). The primary efficacy outcome is the first occurrence of stroke, myocardial infarction, vascular death over the duration of follow-up, (a minimum of 2 and maximum of 4 years approximately). A sample size of patients was planned, 6500 in the W study (testing the efficacy of warfarin vs ASA + clopidogrel) and 7500 in the A study (testing the efficacy of ASA + clopidogrel vs ASA alone). The W study was stopped early by the Data and Safety Monitoring Board in September 2005 after an interim analysis showing a significant difference in favor of warfarin over the combination of ASA + clopidogrel. The details of the ACTIVE W have been published (Lancet 2006; 367: ). The A and the I studies are continuing and final results are expected for RE-LY study (Randomized Evaluation of Long term anticoagulant therapy) Non-valvular atrial fibrillation is implicated in nearly 15% of strokes. Dose-adjusted warfarin decreases the risk of stroke by 62%. However, in practice, the risk of bleeding, the variability of anticoagulation intensity and the need of frequent monitoring and dose adjustments limits treatment with warfarin, leaving patients outside the therapeutic range almost half the time. Underuse of warfarin in patients with atrial fibrillation at high risk of bleeding calls for safer, more reliable alternatives. The direct thrombin inhibitor dabigatran could offer fixed oral dosing without need for coagulation monitoring, rapid onset and offset of action, stable pharmacokinetics with little potential for drug interactions, and no known food interactions. For these reasons an international multicentre, randomized, active controlled, parallel group, noninferiority, clinical trial (RE-LY study) was designed to evaluate the efficacy and safety of dabigatran etexilate compared with open label adjusted warfarin for the prevention of stroke and systemic embolism in patients with non-valvular atrial fibrillation. The recruitment of patients was completed and the long-term follow-up is ongoing. Final results are expected for CURRENT OASIS-7 study OASIS 7 is a randomized, multinational, 2X2 factorial design, parallel-group, double-blind study, comparing a high loading dose regimen of clopidogrel versus standard dose and high dose regimen of aspirin versus standard dose, in patients with acute coronary syndrome managed with an early invasive strategy. The study will involve approximately 25,000 patients in 800 clinical centers worldwide. The primary objective of the study is to determine whether a high dose regimen of clopidogrel is superior to a standard dose of clopidogrel in preventing CV death, myocardial infarction or stroke and to determine if high dose of aspirin is as safe as low dose in terms of TIMI major bleeding rate. Secondary objective is to evaluate the safety of the clopidogrel high dose regimen compared to the standard dose regimen in terms of TIMI major bleeding. Final results are expected for INTER-HEART Study INTER-HEART is an epidemiological study sponsored by the World Health Organization and the World Heart Federation aimed to determine the association between risk factors and acute myocardial infarction within populations defined by ethnicity and/or geographic region, and to assess the relative importance of risk factors across these populations. The project is coordinated 149

151 by the PHRI, McMaster University, Hamilton, Canada. The Clinical Drug Evaluation Laboratory serves as National Coordination Office for Italy. INTER-HEART was conducted in 52 countries in Asia, Europe, Middle East, Africa, Australia, and North and South America, utilizing a standardized protocol. The study evaluated the importance of conventional and emerging risk factors within each geographic region, and whether their impact varies by region in a population of nearly individuals (cases with acute myocardial infarction and matched controls). The study collected data on demographic factors (country of origin, first language), socioeconomic status (education, occupation, income), lifestyle (tobacco use, physical activity, dietary patterns), and personal and family history of cardiovascular disease and risk factors. The INTER-HEART has documented that nine simple modifiable risk factors can account for over 90% of the population attributable risk for heart disease globally. More importantly, these risk factors appear to have similar predictability in all regions of the world, as well as in all ethnic groups. A further analysis published in the 2005 redefined obesity: traditional definitions have been based on Body Mass Index (BMI), but this paper very clearly showed that, irrespective of BMI, the waist-to-hip ratio (WTHR) is a far better predictor of myocardial infarction risk across diverse populations. The observations carry important implications for people and populations hitherto considered to be low risk on the basis of their BMIs. In a further analysis, the INTERHEART study established that all forms of tobacco exposure, such as smoking, chewing tobacco or inhaling second hand smoke, increase the risk of heart attack. Compared to people who had never smoked, smokers had a three-fold increased risk of a heart attack. An ad hoc assessment of the gender influence, comparing risk factors for acute myocardial infarction (MI) between women and men globally, the INTER-HEART showed that women experience their first acute MI on average 9 years later than men. Nine modifiable risk factors are significantly associated with acute MI in both men and women and explain greater than 90% of the PAR. The difference in age of first MI is largely explained by the higher risk factor levels at younger ages in men compared to women. The approach to prevention of MI in men and women can be based on similar principles in all regions of the world. Laboratory of General Practice Research Risk and Prevention Study (R&P) R&P is a study on the optimization of cardiovascular prevention of subjects at high risk performed at national level by General Practitioners. Study objective and design - Controlled clinical trial, double-blind and randomised, of the efficacy of a n-3 PUFA treatment in reducing the incidence of cardiovascular events, both fatal and non-fatal, in a population defined as at high risk by participating GPs. - Practicability and overall yield of the preventive interventions adopted (outcome study) The epidemiological and care history of this population shall form the object of a specific evaluation according to a plan of formal predefined analyses. Study population Inclusion criteria Among the subjects deemed by GPs to be at high cardiovascular risk, patients are selected if presenting: - multiple risk factors (e.g. hypertension, hypercholesterolemia, diabetes, smoking, family history of myocardial infarction, obesity, sex and old age) - previous cardio-cerebrovascular events or clinical manifestations of atherosclerotic disease (stroke, TIA, peripheral arteriopathy, previous arterial revascularisation procedures, angina pectoris). 150

152 Exclusion criteria - serious co morbidity with an unfavorable prognosis over the short term (e.g. cancer); - expected non-compliance over a long period of time; contraindications (known allergies to n- 3PUFA) - indications (previous MI) for treatment with n-3 PUFA. Efficacy measures The primary objective is to evaluate if a long-term administration of n-3 PUFA is more effective than the corresponding placebo in reducing cardiovascular mortality and hospitalization for cardiovascular causes. Randomisation is central, stratified by GP. The experimental treatment consists of one capsule containing 1g of n-3 PUFA, or the corresponding placebo, to be taken daily. The duration of follow-up is 5 years. In order to document with sufficient statistical reliability that the experimental treatment with n- 3 PUFA reduces of 15% the incidence of the events considered in the primary end-point, a total of 12,000 patients is required. Up-date of the study: From February 2004 to March ,521 patients have been enrolled by a network of 860 GPs. The Local Health Authorities involved are 57 and in each one investigator s meeting has been organized. The characteristics of the population so far enrolled are the following: mean age 65 years, males 62%, hypertension 79%, hypercholesterolaemia 62%, diabetes 56%, smokers 16%, obesity 35%, family history of premature myocardial infarction 20%. Twenty five% of patients have a clinical manifestation of atherosclerotic diseases, 50% have diabetes in association with another risk factor and 23% have multiple risk factors. Currently the mean follow-up is 2.5 years. The observed incidence of cardiovascular events (cardiovascular mortality and hospitalizations for cardiovascular causes) in this period was 6%. Five % of patients prematurely withdrawn from the trial and 2.5% of patients discontinued treatment because of side effects (mainly gastrointestinal) that disappeared after discontinuation. After 2 years of follow-up in patients with high blood pressure a statistically significant reduction of blood pressure was observed, as well as significantly reduction of LDL levels in patient with high cholesterol. During this time 20% of smokers stopped smoking. On the contrary no changes were observed in glycated hemoglobin level in diabetic patients, and in BMI in obese patients. This data suggest that GPs interventions have a clinically relevant impact on risk factors control and life styles habits of their patients. More information available on the website The attributability of cardiovascular events in the GP s perception A cardiovascular event is the sudden manifestation - more or less expected - of clinical history, life-style habits, social environment, patient and medical attitudes and health strategies adopted. To carefully examine the reasons why a cardiovascular event occurred is part of a good care in the context of general practice. To explicitate this process, for all the representative cases that occurred during a defined period of care could transform a usual attitude of good clinical practice into an epidemiology of the attributability. The aim of this study is to evaluate the epidemiology of the attributability of cardiovascular events through GPs perception. Clinical, socio-economic, psychological, risk factors and lifestyle variables were investigated. The study started in October 2007and it has been concluded in October GPs were actively involved in study and have reported at least one event. Overall 248 cases have been included during one year. The events were expected by GPs in the 66% of the cases, while in 1/3 were not expected. The majority of patients were known to GP and acute myocardial infarct was the event that occurred 151

153 more frequently. The relevant area most often reported by GPs were: clinical history, lifestyle and the psychological factors. The patients for whom the cardiovascular event was expected, were older, known by GP and had more clinical disease in comparison with those for whom the event was not expected. GPs have reported their consideration on the event in the 58% of the cases; particularly for the younger patients with extra-clinical problems and when the cardiovascular event was not expected. It can be concluded that GPs, in their perception on the attributability of the events, taken into account not only patient s history (clinical and extra-clinical) but also their relationship with the patient and often they feel that they haven t done in half to avoid the event. Epidemiological and clinical profile of diabetic patients in Lombardy Region using administrative databases. The study is part of an ongoing pharmacoepidemiological project in collaboration with the Health Department of the Lombardy Region. Its main objective is the definition of a model to assess and control the use of health resources of diabetic patients by means of integrated administrative database. Specific aims of the study are: To identify the diabetic population by specific drug consumption (ATC group = A10) To assess the co morbidities by co-administered cardiovascular and non-cardiovascular drugs To describe the diagnostic procedures by the assessment of laboratory, instrumental and specialist examinations To estimate the rate and causes of hospitalization by the analyses of hospital admission data Study population Diabetic patients have been identified on the basis of the specific diabetic drugs. A first pilot study has been carried out on the data of the ASL of Bergamo, testing also the feasibility of a case-control study. Those resident that during year 2005 have received at least a prescription of an antidiabetic drug (oral insulin or hypoglycaemic drugs, A10 group of ATC classification) have been considered as diabetic. They then have been followed in the course of the year, linking the registry office database with those of the prescription s and the Hospital Admission s (HD) ones. The sample consisted of inhabitants (10,9% of all the Lombardy Region population). The prevalence of drug-treated diabetes is 4.1% (42,618 subjects); the number of patients treated with antidiabetic drugs increases with increasing of the age, till to reach 15% in those > 75 years. The oral antidiabetics associations (OAD) are more frequently prescribed followed by sulphanylureas, insulins alone and the biguanides. The association between insulins and AOD was prescribed to 10,3% of diabetics. 80% of the diabetic patients received at least a cardiovascular drug. Of the 42,618 diabetic ones, 12,057 (28,3%) have been hospitalized at least once during the same year. Case-control study Case-control methodology has been applied in order to estimate the use of health resources of the cases compared to the control group. The cases are diabetic patients, having more than 50 years. The controls are persons without diabetes, matched to the cases for age, sex and for being in charge to the same general practitioner. As expected, the cases have a greater pharmacologic burden and a greater hospitalization rate. To represent the clinical complexity of the diabetic patients opposed to the controls, co-morbidity diabetes-correlated hospitalizations were used as indicators, in particular: acute myocardial infarction, coronary disease, heart failure, cerebrovascular disease and chronic renal failure. The results show that all these events are much more frequent in the diabetic patients than in the control group. 152

154 The stratification of global cardiovascular risk in hypertensive patients of the district of Borbon Ecuador The Laboratory is involved in a collaborative project with the Cecomet (Centro de Epidemiologia comunitaria y Medicina tropical) in Esmeralda, Ecuador, on the prevalence and treatment of hypertension in the district of Borbon, a rural zone of Ecuador in the northern part of the country. In this area, 36% of the adult population is affected by hypertension and more than half of hypertensive patients present blood pressure levels > 160/110 mmhg. From 2001, in the District is ongoing an intensive follow-up of the hypertensive population with the following aims: to evaluate the global cardiovascular risk of the population, to better control blood pressure levels increasing the number of subjects treated with hypertensive therapy (in particular those at high cardiovascular risk) and monitoring of the clinical complications. Preliminary data show that: Patients treated with hypertensive therapy are increased from 39% to 59% Antihypertensive drugs are mainly prescribed to subjects with high blood pressure levels (80% of those with systolic blood pressure >180mmHg are actually under treatment) or at high cardiovascular risk (82%) Blood pressure control is improved (patients with systolic blood pressure levels > 180mmHg decreased from 33% to 24% and those with levels < increased from 26% to 34%) The fraction of patients at high or very high cardiovascular risk is decreased from 40% to 33% However, the compliance to antihypertensive treatment is still unsatisfactory since only half of the subjects are compliant with the prescribed therapy. Laboratory of Clinical Pharmacology Quality of Life, Depression and Cognitive problems in heart failure patients (QDF-GISSI-HF) The project is a sub-project of the GISSI-HF study. The aims of the study are 1) to describe the evolution of depression, cognitive problems and the quality of life in a sample of 1500 heart failure patients; 2) to assess the use of common instruments that measure QDF variables; 3) to compare the assessment of the instrument (Geriatric Depression scale, Mini Mental State Examination, Kansas City Cardiomiopathy Questionnaire) with the clinical perception of the nurses; 4) to describe if assessed or perceived patients' problems (low quality of life, high depression or compromised cognitive function) lead to any caring intervention. The baseline clinical characteristics of the 1564 patients included in the QDF study are closely comparable with those of the GISSI-HF population. The study instruments could be validly administered to the greatest majority of patients (KCQQ 97.2%, GDS 94.9%, MMSE 80.6% of patients >70 years). The nurses network nested in a major clinical trial, has produced one of the largest prospective cohort of HF patients who are comprehensively assessed and prospectively monitored, to allow an integrated evaluation of the relevance and implications of QDF measurements also on the clinical outcomes of this population. Epidemiology of nursing problems and drug-surveillance in nursing homes and home care The aim of the study is to describe problems that require nursing decisions, and that are encountered by nurses in the care of patients admitted to nursing home and home care, with 153

155 specific attention to drug-related problems. In fact an adverse drug reaction is likely to be the cause of most unexpected problems that arise in elderly patients. The project has been completed and more than 350 nurses have been involved in 95 Nursing Homes and Districts. In the 6 observation days 1500 patients were observed. Overall 2224 problems were identified and 25% attributed to drugs and devices. 154

156 DEPARTMENT OF MOLECULAR BIOCHEMISTRY AND PHARMACOLOGY STAFF Head Mario SALMONA, Sci.Prep.Alim.D., Ph.D. Laboratory of Biochemistry and Protein Chemistry Head Mario SALMONA, Sci.Prep.Alim.D.,Ph.D. Medical Biochemistry Unit Head Valentina BONETTO, Chem.Pharm.D. Synaptic Transmission Unit Head Marco GOBBI, Pharm.D. Laboratory of Molecular Biology Head Enrico GARATTINI, M.D. Pharmacogenomics Unit Head Maddalena FRATELLI, Biol.Sci.D. Gene Structure and Regulation Unit Head Mineko TERAO, Bioch.D., Ph.D. Laboratory of Receptor Pharmacology Head Tiziana MENNINI, Pharm.D. Laboratory of Neuroimmunology Head Pietro GHEZZI, Ph.D. Pharmacology of Septic Shock Unit Head Pia VILLA, Pharm.D. Metabolic Neuropathies Unit Head Roberto BIANCHI, Biol.Sci.D. Laboratory of Molecular Pathology Head Lavinia CANTONI, Biol.Sci.D. Laboratory of Systems Biology Head Gianfranco BAZZONI, M.D. 155

157 CURRICULA VITAE Mario Salmona obtained his doctorate degree in Biochemistry and Food Technology at the University of Milan in His background is in biochemistry, biophysics and pharmacology. His scientific interests relate to problems of human and animal diseases originating from the aberrant folding of proteins. In this context, a major portion of his studies was devoted to the etiopathogenesis and therapy of prion diseases. He has published over 200 articles on peer reviewed scientific journals Ph.D in Pharmacology, Mario Negri Institute 1975 Visiting Fellow in the Department of Biology of the Weizmann Institute of Science, Rehovot, Israel Head, Laboratory of Enzyme Research, Mario Negri Institute 1995 to date Dean of the School of Advanced Pharmacology, Mario Negri Institute 1997 to date Head, Department of Biochemistry and Molecular Pharmacology, Mario Negri Institute 2003 to date Member of the American Society of Biochemistry and Molecular Biology Referee for international scientific journals Selected publications Saracino GA, Villa A, Moro G, Cosentino U, Salmona M. Spontaneous beta-helical fold in prion protein: The case of PrP(82-146). Proteins Oct 29. [Epub ahead of print] Massignan T, Casoni F, Basso M, Stefanazzi P, Biasini E, Tortarolo M, Salmona M, Gianazza E, Bendotti C, Bonetto V. Proteomic analysis of spinal cord of presymptomatic amyotrophic lateral sclerosis G93A SOD1 mouse. Biochem Biophys Res Commun. 2007; 353: De Luigi A, Colombo L, Diomede L, Capobianco R, Mangieri M, Miccolo C, Limido L, Forloni G, Tagliavini F, Salmona M. The efficacy of tetracyclines in peripheral and intracerebral prion infection. PLoS ONE. 2008;3(3):e1888 Cosentino U, Pitea D, Moro G, Saracino GA, Caria P, Varì RM, Colombo L, Forloni G, Tagliavini F, Salmona M. The anti-fibrillogenic activity of tetracyclines on PrP : a 3D-QSAR study. J Mol Model : Fioriti L, Angeretti N, Colombo L, De Luigi A, Colombo A, Manzoni C, Morbin M, Tagliavini F, Salmona M, Chiesa R, Forloni G. Neurotoxic and gliotrophic activity of a synthetic peptide homologous to Gerstmann-Sträussler-Scheinker disease amyloid protein. J Neurosci. 2007; 27: Gobbi M, Colombo L, Morbin M, Mazzoleni G, Accardo E, Vanoni M, Del Favero E, Cantù L, Kirschner DA, Manzoni C, Beeg M, Ceci P, Ubezio P, Forloni G, Tagliavini F, Salmona M. Gerstmann-Sträussler-Scheinker disease amyloid protein polymerizes according to the "dock-and-lock" model. J Biol Chem. 2006; 281:843-9 Gianfranco Bazzoni got his Medicine and Surgery degree in 1988 (at the University of Milan) and the specialisation in Pharmacological Research in 1992 (at the Mario Negri Institute, Milan). His area of expertise is cell biology, with focus on the processes of cell adhesion and migration Research Fellow, Mario Negri Institute Post-doctoral Fellow, Dana Farber Cancer Institute and Harvard Medical School, Boston, MA Research Scientist, Mario Negri Institute 2003 Head, Unit of Cell Adhesion, Mario Negri Institute 2004 to date Head, Laboratory of Systems Biology, Mario Negri Institute 2004 Regular Member of The American Physiological Society, Bethesda, MD Referee for international scientific journals Selected publications Paris L, Bazzoni G The protein interaction network of the epithelial junctional complex: a system-level analysis Mol Biol Cell : Paris L, Tonutti L, Vannini C, Bazzoni G. Structural organization of the tight junction. Biochim Biophys Acta : Huang H, Cruz F, Bazzoni G. Junctional adhesion molecule-a regulates cell migration and resistance to shear stress. J. Cell Physiol 2006; 209; Martinez-Estrada OM, Manzi L, Tonetti P, Dejana E, Bazzoni G. Opposite effects of Tumor Necrosis Factor and soluble fibronectin on Junctional Adhesion Molecule-A in endothelial cells. Am J Physiol (Lung Cell Mol Physiol) 2005; 288: L1081-L1088. Bazzoni G, Tonetti P, Manzi L, Cera MR, Balconi G, Dejana E. Expression of Junction Adhesion Molecule-A prevents spontaneous and random motility. J Cell Sci 2005; 118:

158 Bazzoni G, Dejana E. Endothelial cell-to-cell junctions: molecular organization and role in vascular homeostasis. Physiol Rev 2004; 84(3): Lavinia Cantoni obtained her degree in Biological Sciences in 1973 at the University of Milan. Then she specialized in pharmacological research at the Mario Negri Institute ( ). Research areas 1) biochemical-molecular mechanisms activated by oxidative stress 2) drug metabolism 3) porphyrias Post-doctoral Fellow, Medical Research Council, Toxicology Unit, Carshalton, UK (Winner of a Welcome Trust Research Fellowship) Research Scientist, Mario Negri Institute Visiting Scientist, Toxicology Unit, Carshalton, UK, and Cornell Medical Center, New York, NY (short periods) Head, Unit of Heme and Hemoprotein Metabolism, Mario Negri Institute to date, Head, Laboratory of Molecular Pathology, Mario Negri Institute. Member of the National Roll of Biologists and of the Italian Toxicology Society. Referee for international scientific journals, tutor for university degree thesis and teacher in the Pharmacological Research Specialisation Course for graduates held at the Mario Negri Institute. Selected publications Rizzardini M., Chiesa R., Angeretti N., Lucca E., Salmona M., Forloni G., Cantoni L. Prion protein fragment differentially induces heme oxygenase-1 mrna in cultured neurons and astroglial cells. J. Neurochem. 68: , 1997 Rizzardini M., Zappone M., Villa P, Gnocchi P., Sironi M., Diomede L., Meazza C., Monshouwer M., Cantoni L. Kupffer cell depletion partially prevents hepatic heme oxygenase 1 messenger RNA accumulation in systemic inflammation in mice: role of interleukin 1 beta. Hepatology 27: , 1998 Cantoni L.,Valaperta R., Ponsoda X., Castell, J.V., Barelli D., Rizzardini M., Mangolini A., Hauri L., Villa P. Induction of hepatic heme oxygenase-1 by diclofenac in rodents: role of oxidative stress and cytochrome P-450 activity. J. Hepatology 38: , 2003 Babetto E., Mangolini A., Rizzardini M., Lupi M., Conforti L., Poletti A., Rusmini P., Cantoni L. Tetracycline-regulated gene expression in the NSC-34-tTA cell line for investigation of motor neuron diseases. Mol. Brain Res. 140: 63-72, 2005 Rizzardini M., Lupi M., Mangolini A., Babetto E., Ubezio P., Cantoni L. Neurodegeneration induced by complex I inhibition in a cellular model of familial amyotrophic lateral sclerosis. Brain Res. Bull. 69: , 2006 Raimondi A., Mangolini A., Rizzardini M., Tartari S., Massari S., Bendotti C., Francolini M., Borghese N., Cantoni L., Pietrini G. Cell culture models to investigate the selective vulnerability of motoneuronal mitochondria to familial ALSlinked G93ASOD1. Eur. J. Neurosci. 24: , 2006 Enrico Garattini obtained his degree in Medicine and Surgery with full marks (110/110) in 1982 at the University of Milan. His scientific interests relate to problems of Cellular Biology and Molecular Biology Research Fellow of the National Research Council, Mario Negri Institute Postdoctoral Researcher at the Roche Institute of Molecular Biology, Department of Neurosciences Nutley, New Jersey, US Senior Researcher Regione Lombardia and Head of the Molecular Biology Unit, Mario Negri Institute 1997 to date Head, Laboratory of Molecular Biology, Mario Negri Institute From 2005 Dean, Advanced School of Pharmacology (Philosophy Doctor), Mario Negri Institute Member of the Editorial Board of the European Journal of Cancer and of Current Cancer Therapy Reviews Member of the American Society of Biochemistry and Molecular Biology (ASBMB) Selected publications Gianni M, Boldetti A, Guarnaccia V, Rambaldi A, Parrella E, Raska I Jr, Rochette-Egly C, Del Sal G, Rustighi A, Terao M, Garattini E Inhibition of the peptidyl-propyl-isomerase Pin1 enhances the responses of acute myeloid leukemia cells to retinoic acid via stabilization of RARα and PML-RARα. Cancer Res. 2008, in press Terao M, Kurosaki M, Barzago M M, Fratelli M, Bagnati R, Bastone A, Giudice C, Scanziani E, Mancuso A, Tiveron C, Garattini E Role of the molybdo-flavoenzyme, aldehyde oxidase homolog 2, in the biosynthesis of retinoic acid: generation and characterization of a knock-out mouse, Mol Cell Biol 2008 [Epub ahead of print] Gianni M, Parrella E, Raska I Jr, Gaillard E, Nigro EA, Gaudon C, Garattini E, Rochette-Egly C. P38MAPK-dependent phosphorylation and degradation of SRC-3/AIB1 and RARalpha-mediated transcription. EMBO J Feb 22;25(4):

159 Garattini E, Parrella E, Diomede L, Gianni M, Kalac Y, Merlini L, Simoni D, Zanier R, Ferrara F F, Chiarucci I, Carminati P, Terao M, Pisano C. ST1926, a novel and orally active retinoid-related molecule inducing apoptosis in myeloid leukemia cells: Modulation of intracellular calcium homeostasis. Blood 2004; 103: Kurosaki M, Terao M, Barzago M M, Bastone A, Bernardinello D, Salmona M, Garattini E. The aldehyde oxidase gene cluster in mice and rats: Aldehyde oxidase homologue 3, a novel member of the molybdo-flavoenzyme family with selective expression in the olfactory mucosa. J Biol Chem 2004; 279: Pisano C, Kollar P, Gianni M, Kalac Y, Giordano V, Ferrara F F, Tancredi R, Devoto A, Rinaldi A, Rambaldi A, Penco S, Marzi M, Moretti G, Vesci L, Tinti O, Carminati P, Terao M, Garattini E. Bis-indols a novel class of molecules enhancing the cytodifferentianting properties of retinoids in myeloid leukemia cells. Blood 2002; 100: Pietro Ghezzi Research Areas: Cytokines and inflammation; redox regulation : Researcher, Mario Negri Institute 1991 to date: Head, Laboratory of Neuroimmunology, Mario Negri Institute : Research Associate at Stanford University School of Medicine, Department of Genetics 2000 to date: Member, Kenneth Warren Laboratory, Ossining, NY (USA) Referee for international scientific journals Selected publications Leist M, Ghezzi P, et al.. Derivatives of erythropoietin that are tissue protective but not erythropoietic. Science Jul 9;305(5681): Fratelli M, Goodwin LO, Orom UA, Lombardi S, Tonelli R, Mengozzi M, Ghezzi Gene expression profiling reveals a signaling role of glutathione in redox regulation.proc Natl Acad Sci U S A Sep 27;102(39): Villa P, Bigini P, Mennini T, Agnello D, Laragione T, Cagnotto A, Viviani B, Marinovich M, Cerami A, Coleman TR, Brines M, Ghezzi P. Erythropoietin selectively attenuates cytokine production and inflammation in cerebral ischemia by targeting neuronal apoptosis. J Exp Med Sep 15;198(6): Siren AL, Fratelli M, Brines M, Goemans C, Casagrande S, Lewczuk P, Keenan S, Gleiter C, Pasquali C, Capobianco A, Mennini T, Heumann R, Cerami A, Ehrenreich H, Ghezzi P. Erythropoietin prevents neuronal apoptosis after cerebral ischemia and metabolic stress. Proc Natl Acad Sci U S A Mar 27;98(7): Laragione T, Bonetto V, Casoni F, Massignan T, Bianchi G, Gianazza E, Ghezzi P. Redox regulation of surface protein thiols: identification of integrin alpha-4 as a molecular target by using redox proteomics. Proc Natl Acad Sci U S A Dec 9;100(25): Tiziana Mennini got her degree in Pharmacy at the University of Milano (1975). In the same year she obtained a fellowship from the European Molecular Biology Organization, to learn sub-cellular fractionation techniques and synaptosomes utilization in neurochemistry, at the laboratory of Prof. VP Whittaker ( Stockholm, Sweden). In 1882 she spent a further period in Prof. Whittaker s laboratories (Max-Plank-Institut fur Biophysikalische Chemie, Abteilung Neurochemie Am Fassberg, Gottingen, Germany). She spent all her scientific career at the Mario Negri Institute: Research Assistant in the Laboratory of Drug Metabolism Chief of the Unit of Neurochemical Transmission, 1988 to date Chief of the Laboratory of Receptor Pharmacology Speaker, chairman and organizer at many congresses and courses, author of more than 200 articles published in international journals in the area of receptor pharmacology and neuropharmacology. Selected publications Beghi E, Bendotti C, Mennini T Merits of a new drug trial for ALS? Science 308: Gobbi M, Mennini T Is St John's wort a 'Prozac-like' herbal antidepressant? Trends Pharmacol Sci 22: The Italian ALSSG Ceftriaxone in amyotrophic lateral sclerosis. Eur J Neurol 3: Mennini T, Mocaer E, Garattini S Tianeptine, a selective enhancer of serotonin uptake in rat brain. Naunyn- Schmiedebergs Arch Pharmacol 336: Mennini T, Garattini S Benzodiazepines receptor binding in vivo: pharmacokinetic and pharmacological significance. Advances Biochemical Psychopharmacol 38: Mennini T, Bernasconi S, Manara L, Samanin R, Serra G The effect of intracerebral 6-hydroxy dopamine on 3Hreserpine binding to different brain regions of the rat. Pharmacol Res Commun 9: Roberto Bianchi got his degree in Biological Sciences at University of Milan, Italy in Since 1975 he served as student teacher and supervisor at Mario Negri Institute and from 1989 to 1997 at Houston University. His main interests are diabetic complications (peripheral and autonomic neuropathies) and neurodegenerative disorders (Multiple Sclerosis, drugs induced neuropathies) Technician in the Laboratory of Biochemical Pharmacology, Mario Negri Institute 158

160 Visiting Scientist in the Center for Neurosciences Behavioral Research, The Weizmann Institute of Science, Rehovot, Israel Research Assistant in the Laboratory of Biochemical Pharmacology, Mario Negri Institute Research Fellow in the Dept. Biochemical Biophysical Sciences, University of Houston, US (3 mo. a year) Research Fellow in the Dept. Medicine, Case Western Reserve University, Cleveland, US (1 mo. a year) Since 1996 Head, Unit of Metabolic Neuropathies, Mario Negri Institute Selected publications Bianchi R., Berti-Mattera L.N., Fiori M.G., Eichberg J.:Correction of altered metabolic activities in sciatic nerves of streptozotocin-induced diabetic rats. Effects of ganglioside treatment. Diabetes 39: (1990). Scarpini E., Bianchi R., Moggio M., Sciacco M., Fiori M.G., Scarlato G.: Decrease of nerve Na+,K+-ATPase activity in the pathogenesis of diabetic neuropathy. J. Neurol. Sci. 120: (1993). Conti G., Scarpini E., Baron P.L., Livraghi S., Tiriticco M., Bianchi R, Vedeler C., Scarlato G.: Macrophage infiltration and death in the nerve during the early phases of experimental diabetic neuropathy: a process concomitant with endoneurial induction of IL-1 and p75ntr. J. Nuerol. Sci. 195: (2002) Bianchi R., Buyukakilli B., Brines M., Savino C., Cavaletti G., Oggioni N., Lauria G., Borgna M., Lombardi R., Cimen B., Comelekoglu U., Kanik A., Tataroglu C., Cerami A., Ghezzi P. Erythropoietin both protects from and reverses experimental diabetic neuropathy. Proc Natl Acad Sci USA 101: (2004) Leist M., Ghezzi P., Grasso G, Bianchi R., Villa P., Fratelli M., Savino C., Bianchi M., Nielsen J., Gerwien J., Kallunki P., Larsen A.K., Helboe L., Christensen S., Pedersen L.O., Nielsen M., Troup L., Sager T., Sfacteria A., Erbayktar S, Erbayktar Z., Gokmen N., Yilmaz O., Cerami-Hand C., Xie, Q-W., Coleman T., Cerami A., Brines M. Erythropoietinderived tissue-protective cytokines that do not bind to the classical erythropoietin receptor. Science, 305(5681) , Savino C., Pedotti R., Baggi F., Furlan R., Ubiali F., Gallo B, Nava S., Bigini P., Barbera S., Fumagalli E., Mennini T., Vezzani A., Rizzi M., Coleman T., Cerami A.,Brines M., Ghezzi P., Bianchi R.Delayed administration of erythropoietin and its non-erythropoietic derivatives ameliorates chronic murine autoimmune encephalomyelitis. Journal of Neuroimmunology, 2005, E-pub December 6, 2005 Valentina Bonetto has got the degree in Pharmaceutical Chemistry and Technology at the University of Padua, Italy in She has got the Ph.D. in Medical Biochemistry and Biophysics at Karolinska Institutet, Stockholm, Sweden. Her principal lines of research are: 1) Study of the pathogenetic mechanisms at the basis of amyotrophic lateral sclerosis (ALS); 2) Identification of biomarkers of ALS; 3) Role of the oxidative modification in neurological disorders. These issues are investigated by different experimental approaches, including proteomics and mass spectrometry. Since 2000 she is at the Mario Negri Institute in the Laboratory of Biochemistry and Protein Chemistry, from 2002 is also Assistant Telethon Scientist at Dulbecco Telethon Institute. Since October 2007 she is the Head of the Unit of Medical Biochemistry inside the Laboratory of Biochemistry and Protein Chemistry. She is author of 26 publications from 1994 to 2007, in peer-reviewed journals. Among them she is first author in 11 and last author in 4. She is also author of 2 reviews. She is reviewer for scientific journals in the field of Proteomics and Neuroscience. Selected publications Massignan, T., Casoni, F., Basso, M., Stefanazzi, P., Biasini, E., Tortarolo, M., Salmona, M., Gianazza, E., Bendotti, C., Bonetto V. (2007) Proteomic analysis of spinal cord of presymptomatic amyotrophic lateral sclerosis G93A SOD1 mouse. Biochem. Biophys. Res. Commun., 353: Basso, M., Massignan, T., Samengo, G., Cheroni, C., De Biasi, S., Salmona, M., Bendotti, C., Bonetto, V. (2006) Insoluble mutant SOD1 is partly oligoubiquitinated in amyotrophic lateral sclerosis mice. J. Biol. Chem., 281: Ghezzi, P., Casagrande, S., Massignan, T., Basso, M., Bellacchio, E., Mollica, L., Biasini, E., Tonelli, R., Eberini, I., Gianazza, E., Dai, W.W., Fratelli, M., Salmona, M., Sherry, B., Bonetto. V. (2006) Redox regulation of cyclophilin A by glutathionylation. Proteomics, 6: Casoni, F., Basso, M., Massignan, T., Gianazza, E., Cheroni, C., Salmona, M., Bendotti, C., Bonetto, V. (2005) Protein nitration in a mouse model of familial amyotrophic lateral sclerosis: Possible multifunctional role in the pathogenesis. J. Biol. Chem., 280: Laragione, T., Bonetto, V., Casoni, F., Massignan, T., Bianchi, G., Gianazza, E., Ghezzi, P. (2003) Redox regulation of surface protein thiols: identification of integrin alpha-4 as a molecular target by using redox proteomics. Proc. Natl. Acad. Sci. U S A 100: Casagrande, S.*, Bonetto, V.*, Fratelli, M., Gianazza, E., Eberini, I., Massignan, T., Salmona, M., Chang, G., Holmgren, A., Ghezzi, P. (2002) Glutathionylation of human thioredoxin: a possible crosstalk between the glutathione and thioredoxin systems. Proc. Natl. Acad. Sci. U S A 99, *These authors contributed equally to the study. 159

161 Maddalena Fratelli got her degree in Biological Sciences at the University of Pisa and at the Scuola Normale Superiore di Pisa in Then the specialization in Pharmacological Research at the Mario Negri Institute in Her main fields of interest are: 1. High throughput genomic systems for the study of drug action and pharmacoresistance. 2. Redox regulation of protein function and gene expression: glutathionylation and gene expression profiling of glutathione dependent responses to oxidant challenge Postdoctoral Research Fellow in the Medical Research Council, Neurobiology Unit, Cambridge, UK. Since 1995, Head, Unit of Mediators of inflammation, Laboratory of Neuroimmunology, Mario Negri Institute Since 2005, Head, Unit of Pharmacogenomics, Laboratory of Molecular Biology, Mario Negri Institute Selected publications Fratelli M, Goodwin LO, Orom UA, Lombardi S, Tonelli R, Mengozzi M, Ghezzi P. Gene expression profiling reveals a signaling role of glutathione in redox regulation. Proc Natl Acad Sci U S A. 2005;102: Brines M, Grasso G, Fiordaliso F, Sfacteria A, Ghezzi P, Fratelli M, Latini R, Xie QW, Smart J, Su-Rick CJ, Pobre E, Diaz D, Gomez D, Hand C, Coleman T, Cerami A. Erythropoietin mediates tissue protection through an erythropoietin and common beta-subunit heteroreceptor. Proc Natl Acad Sci U S A. 2004; 101: Leist M, Ghezzi P, Grasso G, Bianchi R, Villa P, Fratelli M, Savino C, Bianchi M, Nielsen J, Gerwien J, Kallunki P, Larsen AK, Helboe L, Christensen S, Pedersen LO, Nielsen M, Torup L, Sager T, Sfacteria A, Erbayraktar S, Erbayraktar Z, Gokmen N, Yilmaz O, Cerami-Hand C, Xie QW, Coleman T, Cerami A, Brines M. Derivatives of erythropoietin that are tissue protective but not erythropoietic. Science. 2004; 305: Fratelli M, Minto M, Crespi A, Erba E, Vandenabeele P, Del Soldato P, Ghezzi P. Inhibition of nuclear factor-kappab by a nitro-derivative of flurbiprofen: a possible mechanism for antiinflammatory and antiproliferative effect. Antioxid Redox Signal. 2003; 5: Fratelli M, Demol H, Puype M, Casagrande S, Eberini I, Salmona M, Bonetto V, Mengozzi M, Duffieux F, Miclet E, Bachi A, Vandekerckhove J, Gianazza E, Ghezzi P. Identification by redox proteomics of glutathionylated proteins in oxidatively stressed human T lymphocytes. Proc Natl Acad Sci U S A. 2002; 99: Siren AL, Fratelli M, Brines M, Goemans C, Casagrande S, Lewczuk P, Keenan S, Gleiter C, Pasquali C, Capobianco A, Mennini T, Heumann R, Cerami A, Ehrenreich H, Ghezzi P. Erythropoietin prevents neuronal apoptosis after cerebral ischemia and metabolic stress. Proc Natl Acad Sci U S A. 2001; 98: Marco Gobbi got his degree in Pharmacy at the University of Milan, Italy, in Currently, his main fields of interest are: 1) the study of presynaptic mechanisms, such as neurotransmitter release/reuptake with a particular focus on plasma membrane transporters; and 2) the study of protein misfolding and aggregation and in particular the characterization of the prion protein amyloid formation, investigated by different approaches including surface plasmone resonance. Since 1981, Researcher in the Laboratory of Neuropharmacology and, from 1988, in the Laboratory of Receptor Pharmacology, Mario Negri Institute Starting from 1989 Chief, Unit of Synaptic Transmission, Mario Negri Institute In Jan 2006, his group joined the Laboratory of Biochemistry and Protein Chemistry, Mario Negri Institute Co-author in more than 70 scientific publications on peer-reviewed international journals. First or last author in more than 40 of them. Reviewer for international scientific journals operating in the Neuroscience/Neuropharmacology fields. Selected publications Gerstmann-Straussler-Scheinker disease amyloid protein polymerizes according to the "dock-and-lock" model. Gobbi M, Colombo L, Morbin M, Mazzoleni G, Accardo E, Vanoni M, Del Favero E, Cantù L, Kirschner D A, Ceci P, Ubezio P, Manzoni C, Forloni G, Tagliavini F, Salmona M J Biol Chem. 2006; 281: Funicello M, Conti P, De Amici M, De Micheli C, Mennini T, Gobbi M Dissociation of [3H]L-glutamate uptake from L-glutamate-induced [3H]D-aspartate release by 3-hydroxy-4,5,6,6a-tetrahydro-3aH-pyrrolo[3,4-d]isoxazole-4- carboxylic acid and 3-hydroxy-4,5,6,6a-tetrahydro-3aH-pyrrolo[3,4-d]isoxazole-6-carboxylic acid, two conformationally constrained aspartate and glutamate analogs. Mol Pharmacol 66: Gobbi M, Moia M, Pirona L, Ceglia I, Reyes-Parada M, Scorza C, Mennini T p-methylthioamphetamine and 1- (m-chlorophenyl)piperazine, two non-neurotoxic 5-HT releasers in vivo, differ from neurotoxic amphetamine derivatives in their mode of action at 5-HT nerve endings in vitro. J Neurochem 82: Gobbi M, DallaValle F, Ciapparelli C, Diomede L, Morazzoni P, Verotta L, Caccia S, Cervo L, Mennini T Hypericum perforatum L. extract does not inhibit 5-HT transporter in rat brain cortex. Naunyn Schmiedebergs Arch Pharmacol 360: Gobbi M, Gariboldi M, Piwko C, Hoyer D, Sperk G, Vezzani A Distinct changes in peptide YY binding to, and mrna levels of, Y1 and Y2 receptors in the rat hippocampus associated with kindling epileptogenesis. J Neurochem 70:

162 Crespi D, Mennini T, Gobbi M Carrier-dependent and Ca(2+)-dependent 5-HT and dopamine release induced by (+)-amphetamine, 3,4-methylendioxymethamphetamine, p-chloroamphetamine and (+)-fenfluramine. Br J Pharmacol 121: Mineko Terao obtained her doctorate degree in Pharmaceutical Science from the Kobe Women s College of Pharmacy, Japan in Her scientific interests relate to problems of Cellular Biology and Molecular Biology Ph.D in Molecular Biology, Kyoto University, Japan Research Fellow, Department of Medical Chemistry, Kyoto University Faculty of Medicine, Japan Postdoctoral Associate of the Institute for Cancer Research, Philadelphia, US From 1987 Visiting Scientist of Mario Negri Institute From 1998 Head of the Unit of Gene Structure and Regulation, Mario Negri Institute Selected publications Terao M, Kurosaki M, Barzago MM, Varasano E, Boldetti A, Bastone A, Fratelli M, Garattini E. Avian and canine aldehyde oxidases. Novel insights into the biology and evolution of molybdo-flavoenzymes. J Biol Chem Jul 14;281(28): Garattini E, Parrella E, Diomede L, Gianni M, Kalac Y, Merlini L, Simoni D, Zanier R, Ferrara F F, Chiarucci I, Carminati P, Terao M, Pisano C. ST1926, a novel and orally active retinoid-related molecule inducing apoptosis in myeloid leukemia cells: Modulation of intracellular calcium homeostasis. Blood 2004; 103: Vila R, Kurosaki M, Barzago M M, Kolek M, Bastone A, Colombo L, Salmona M, Terao M, Garattini E. Regulation and biochemistry of mouse molybdo-flavoenzymes. The DBA/2 mouse is selectively deficient in the expression of aldehyde oxidase homologues 1 and 2 and represents a unique source for the purification and characterization of aldehyde oxidase. J Biol Chem 2004; 279: Kurosaki M, Terao M, Barzago M M, Bastone A, Bernardinello D, Salmona M, Garattini E. The aldehyde oxidase gene cluster in mice and rats: Aldehyde oxidase homologue 3, a novel member of the molybdo-flavoenzyme family with selective expression in the olfactory mucosa. J Biol Chem 2004; 279: Parrella E, Gianni M, Cecconi V, Nigro E, Barzago MM, Rambaldi A, Rochette-Egly C, Terao M and Garattini E. Phosphodiesterase 4 inhibition by piclamilast potentiates the cyto-differentiating action of retinoids in myeloid leukemia cells. J Biol Chem 2004; 279: Garattini E, Gianni M and Terao M. Retinoid related molecules an emerging class of apoptotic agents with promising clinical potential in oncology: pharmacological activity and mechanisms of action. Curr Pharm Design 2004, 10: Pia Emilia Villa got her degree in Pharmaceutical Chemistry and Technology at the University of Pavia in 1975 and the specialisation in Pharmacological Research at the Mario Negri Institute, Milan in Her scientific interests are the physiopathologic factors of sepsis and their pharmacological modulation, the cellular and molecular mechanisms of neurodegeneration and neuroprotection in experimental cerebral ischemia : Research fellow, laboratory of Perfusion of Isolated Organs and Toxicology, Mario Negri Institute : Visiting fellow, laboratory of Cultured Hepatocytes, Toxicology Unit, MRC, Carshalton, England 1983: Visiting fellow, Unité de Recherche Hépatologique, Rennes, France : Research Scientist, laboratory of Neuroimmunology, Mario Negri Institute 1995 to date: Head, Unit of Pharmacology of Septic Shock, Mario Negri Institute 1982 Regular member of the Italian Society of Toxicology 1992 Regular member of Celltox 1996 Regular member of the International Cytokine Society. Selected publications Villa P, Shaklee CL, Meazza C, Agnello D, Ghezzi P, Senaldi G. Granulocyte colony-stimulating factor and antibiotics in the prophylaxis of a murine model of polymicrobial peritonitis and sepsis. J Infect Dis. 1998; 178: Villa P, Saccani A, Sica A, Ghezzi P. Glutathione protects mice from lethal sepsis by limiting inflammation and potentiating host defense. J Infect Dis. 2002; 185: Erbayraktar S, Grasso G, Sfacteria A, Xie QW, Coleman T, Kreilgaard M, Torup L, Sager T, Erbayraktar Z, Gokmen N, Yilmaz O, Ghezzi P, Villa P, Fratelli M, Casagrande S, Leist M, Helboe L, Gerwein J, Christensen S, Geist MA, Pedersen LO, Cerami-Hand C, Wuerth JP, Cerami A, Brines M. Asialoerythropoietin is a nonerythropoietic cytokine with broad neuroprotective activity in vivo. Proc Natl Acad Sci U S A. 2003;100 (11): Villa P, Bigini P, Mennini T, Agnello D, Laragione T, Cagnotto A, Viviani B, Marinovich M, Cerami A, Coleman TR, Brines M, Ghezzi P. Erythropoietin selectively attenuates cytokine production and inflammation in cerebral ischemia by targeting neuronal apoptosis. J Exp Med. 2003;198 (6):

163 Leist M, Ghezzi P, Grasso G, Bianchi R, Villa P, Fratelli M, Savino C, Bianchi M, Nielsen J, Gerwien J, Kallunki P, Larsen AK, Helboe L, Christensen S, Pedersen LO, Nielsen M, Torup L, Sager T, Sfacteria A, Erbayraktar S, Erbayraktar Z, Gokmen N, Yilmaz O, Cerami-Hand C, Xie QW, Coleman T, Cerami A, Brines M. Derivatives of erythropoietin that are tissue protective but not erythropoietic. Science. 2004;305: Garau A, Bertini R, Colotta F, Casilli F, Bigini P, Cagnotto A, Mennini T, Ghezzi P, Villa P. Neuroprotection with the CXCL8 inhibitor repertaxin in transient brain ischemia. Cytokine. 2005;30: INTRODUCTION TO THE DEPARTMENT'S ACTIVITIES The Department comprises six laboratories. Research is heterogeneous in terms of scientific interests and aims, but it is unified by the structural and functional study of specific, pharmacologically important gene products, using a common body of techniques. Classical biochemistry and molecular biology methods are used to define proteins that might be targets for the pharmacological activity of drugs. Potential direct interactions between drugs and proteins are studied at the molecular level by a variety of approaches ranging from animal studies to computer simulation. FINDINGS/MAIN RESULTS Identification of tetracylines as potential anti-prion drugs. Synthesis and physicochemical and biological characterization of peptides deduced from the primary sequence of prion protein. Identification of a relationship between cholesterol synthesis and production of prion protein. Protein identifications by mass spectrometry and data base searching using a combination of techniques. Characterization of the Pentraxin 3 role in the organization of the cumulus oophorus extracellular matrix and in female fertility. System-level analysis of protein interactions in the epithelial junctional complex. Characterization of the role of Junctional Adhesion Molecule-A (JAM-A) in the control of cell motility. Characterization of the effect of inflammatory cytokines on JAM-A function. Development of a constitutive and of an inducible motor neuron cellular model to unravel the toxicity of mutant G93A superoxide dismutase 1 responsible for some forms of familial amyotrophic lateral sclerosis. Conditions of oxidative stress or the presence of compounds impairing the electron transport chain are a risk factor to motor neurons of individuals carrying mutant forms of superoxide dismutase 1. Mitochondrial damage due to mutant G93A superoxide dismutase 1 occurs selectively in motor neurons. Mitochondrial damage by mutant G93A superoxide dismutase 1 in motor neurons is modulated by the level of expression of the mutant protein. Identification and characterization of a novel class of retinoids endowed with strong and selective apoptogenic acivity on the neoplastic cell. Pre-clinical development of these agents for the treatment of acute leukemia. Identification and characterization of novel retinoid-based pharmacological combinations for the treatment of acute myelogenous leukemia. Molecular cloning and characterization of the cdnas and genes of four novel members of the mammalian molybdo-flavoprotein family. Definition of a novel gene cluster on human chromosome 2 and mouse chromosome 1. Development of knok-out animals for molybdo-flavoproteins: AOX1, AOH1, AOH2, AOH3. 162

164 Creation of integrated instruments for the rationalization of Microarray analysis processes. Identification of erythropoietin as a neuroprotective agent and of new molecules with neuroprotective activity. Identification of the pharmacological action of erythropoietin against peripheral neuropathy of diabetes. Identification of erythropoietin derivatives that retain its neuroprotective actions but have lost its hemopoietic ones. Discovery of proteins that are regulated by the redox state through the formation of reversible disulfide bonds with glutathione (protein glutathionylation). Identification of exofacial proteins undergoing thiol redox regulation. The use of antioxidant molecules in models of sepsis and inflammation diminishes the inflammatory response while potentiates the innate immune response. Identification of the gene expression profile regulated by thiols. The treatment with a non haematopoietic derivate of Erythropoietin (CEPO) reduces motor neuron loss and clinical progression in a mouse model of ALS related to alterations in vesicle trafficking, the wobbler mouse. Treatment with a soluble TNF receptor in the wobbler mouse, reduces motor neuron degeneration and the phosphorylation of the two main stress kinases (p38 e JNK) activated by TNF receptors. Riluzole treatment reduces motor neuron loss and clinical progression of wobbler mouse by increasing the endogenous BDNF expression. Oxidative stress, glial activation and inflammation occur in the retinopathy as well as in cerebral and spinal cord dysfunction in the mnd mouse, a model of progressive epilepsy with mental retardation related to mutation in the CLN8 gene. These findings provide further evidence for the implication of TNF death receptor signalling in the pathology of Neuronal Ceroid Lipofuscinosis The affinity of pergolide for human cloned 5-HT 2A and 5-HT 2B receptors is similar and higher that that for the human cloned D 2L receptor. These findings, together with the fact that it acts as agonist at both h5-ht 2A and h5-ht 2B receptors, could explain its potential toxicity mediated by activation of cardio-pulmonary 5-HT 2B receptor. New conformationally constrained aspartate and glutamate analogues dissociate glutamate uptake inhibition and reverse transport-mediated release. Dimethyl sulfoxide, a solvent commonly utilized to dissolve hydrophobic compound for in vitro experiments, interferes with the 5-HT6 agonists activity when the scintillation proximity assay is used for evaluating 35S-GTP-γ-S binding; but does not interfere with the europium labelled GTP binding determined by time-resolved fluorescence. NATIONAL COLLABORATIONS Advanced Biology Center, Genoa Dip. Anatomia, Farmacologia, Medicina Legale, University of Turin Dip. Biotecnologie, Università degli Studi, Milan Dip. Chimica Biochimica e Biotecnologie per la Medicina, Università degli Studi, Milan Dip. Chimica Farmaceutica e Tossicologica, Università degli Studi, Milan Dip. Farmaco-Chimico, Università degli Studi, Messina Dip. Farmaco-Chimico-Tecnologico, University of Siena Dip. Farmacologia Medica, Università degli Studi, Milan Dip. Scienze Biochimiche, University of Florence 163

165 Dip. Scienze Farmaceutiche, University of Catania Dip. Scienze Farmaceutiche, University of Genoa Dip. Scienze Farmacologiche, Università degli Studi, Milan Dip. Scienze Fisiologiche e Farmacologiche, University of Pavia Dip. Scienze Molecolari, University of Milan Dip. Studi pre-clinici, University of Milan Facoltà di Biologia, Università degli Studi, Milan Facoltà di Chimica, Università degli Studi, Milan Facoltà di Chimica, University of Ferrara Fondo Edo Tempia, Biella GlaxoSmithkline, Verona IFOM Fondazione Istituto FIRC di Oncologia Molecolare, Milan IRCCS Fondazione "Istituto C. Mondino", Laboratorio di Neurobiologia Sperimentale, Pavia Istituto di Biologia Molecolare Buzzati Traverso, Naples Istituto di Biomedicina e Immunologia Molecolare CNR, Palermo Istituto di Endocrinologia, Centro di Eccellenza per le Malattie Neurodegenerative, Università degli Studi, Milan Istituto di Clinica Neurologica, Ospedale Maggiore Policlinico, Milan Istituto Clinico Humanitas, Milan Istituto di Neuroscienze C.N.R., Pisa Istituto Nazionale dei Tumori, Milan Istituto Nazionale dei Tumori, Naples Istituto Nazionale Neurologico "C. Besta", Milan Istituto Oncologico Europeo, Milan Istituto Regina Elena, Rome Istituto Toscano Tumori, Florence Newron Pharmaceuticals, Milan Ospedale Maggiore Policlinico, Milan Ospedale Pediatrico Bambino Gesu', Rome Ospedale Pediatrico "Gaslini", Genoa Ospedale S. Gerardo, Monza, Milan Sigma-Tau, Pomezia, Rome Zambon, Milan INTERNATIONAL COLLABORATIONS The Babraham Institute, Cambridge, UK Boston College, Boston, MA, USA Case Western Research University, Cleveland, OH, USA Dept. of Neurology, Keio University, Tokyo, Japan Dept. de Quimica-Fisica de Macromoleculas Biologicas, CSIC, Madrid, Spain Faculdad de Ciencias Medicas, Universidad de Santiago de Chile, Chile Dept. of Biological Chemistry, The Alexander Silberman Institute of Life Sciences, The Hebrew University of Jerusalem, Jerusalem, Israel Flanders Interuniversity Institute for Biotechnology (VIB) University of Gent, Belgium FMP, Berlin, Germany Giessen Polyclinic University, Giessen, Germany Houston University, TX, USA IBSN CNRS, Marseille, France Indiana University, Indianapolis, IN, USA 164

166 Institut de Genetique et Biologie Moleculaire et Cellulaire, Strasbourg, France Institut Pasteur, Paris, France John Innes Centre, Norwich, UK Kenneth S. Warren Institute, Ossining, NY, USA Max-Planck-Institut für experimentelle Medizin, Göttingen, Germany National Institute of Health, Bethesda, MD, USA Nippon University, Tokyo, Japan North Shore University Hospital, Manhasset, NY, USA Pepscan System BV, Lelystad, Holland Polichem S.A., Lugano, Switzerland Stanford University School of Medicine, Stanford, CA, USA Technical University Braunschweig, Germany Trinity College, Dublin, Ireland Universidad de La Laguna, Tenerife, Spain Universidad Nova, Lisbon, Portugal Universitat des Saarlandes, Hamburg, Germany Universitat Freiburg, Germany Université Paris, France Université Victor Segalen Bordeaux 2, Bordeaux, France University of Aberdeen, UK University of Birmingham, UK University of Cardiff, UK University of Colorado, School of Medicine, Denver, CO, USA University of Glasgow, UK University of Gottingen, Germany University of Muenster, Germany University of Southampton, UK University of Sussex, UK University of Vienna, Austria Waring-Webb Institute, University of Colorado, Denver CO, USA Weizmann Institut, Rehovot, Israel Westfaelische Wilhelms-Universitaet Muenster, Germany EDITORIAL BOARD MEMBERSHIP Neurobiology of Lipids (L. Diomede) Neuroimmunomodulation (P. Ghezzi) Newsletters of the International Cytokine Society (P. Ghezzi) European Journal of Cancer (E. Garattini) PEER REVIEW ACTIVITIES American Journal Physiology, Biochemical Journal, Biochemical Pharmacology, Biochimica Biophysica Acta, Brain Research, Cancer Research, Cell Death and Differentiation, Cell Research, Circulation, Drug Investigation, European Journal of Cancer, European Journal of Immunology, European Journal of Neuroscience, International Journal of Cancer, Journal of Cell Biology, Journal of Hepatology, Journal of Immunology, Journal of Investigative Dermatology, Journal of Lipid Mediators, Journal of Neurochemistry, Journal of Neuroimmunology, Journal of Translational Medicine, Neuroscience Letters, Pharmacological 165

167 Research, Physiological Genomics, Proceedings of the National Academy of Sciences, Life Sciences. PARTICIPATION IN EVENTS IN WHICH THE DEPARTMENT WAS INVOLVED Meeting: First European Synapse Meeting, SNAP-25 negatively modulates voltage-gated calcium channels and network activity, 26 March, Bordeaux, France Meeting: Associazione Italiana di Neuropatologia (AINP) e Associazione Italiana per la Ricerca sull Invecchiamento Cerebrale (AIRIC), Analysis of C1-INH to MBL, using Surface Plasmon Resonance, Synaptic dysfunction in a mouse model of a familial prion disease: impairment of glutamate exocytosis, Synaptic dysfunction in a mouse model of a familial prion disease: impairment of calcium dynamics, 18 June, Milan, Italy Conference: ICAD Alzheimer s Associations International Conference on Alzheimer s Disease and Related Disorders, Beta-amyloid interferences with cognitive functions in an aggregation status-dependent manner, 30 July, Chicago, USA Meeting: 8th Siena Meeting, From Genome to Proteome: Integration of proteome completion, Comparison of proteomic approaches for protein biomarker discovery in amyotrophic lateral sclerosis, 31 August, Siena, Italy Meeting: XIX National Meeting on Medicinal Chemistry, New thienopyrimidine derivatives as potential 5-HT7 receptor ligands, Heteroaryloxy analogues of WA4101: synthesis and biological evaluation at alfa1-adrenoreceptor subtypes, 14 September, Verona, Italy Simposium: SFB Symposium, Partial agonism for the induction of SERT-mediated 5-HT release and currents in vitro, Glutamate uptake is dissociated from substrate-induced glutamate release as evidence by conformationally constrained aspartate and glutamate analogues, 26 September Vienna, Austria Congress: Congresso annuale Società Italiana di Neurologia, Analgesic effect of buprenorphine in an experimental model of painful diabetic peripheral neuropathy, 18 October, Napoli, Italy GRANTS AND CONTRACTS Agenzia Italiana del Farmaco, Rome, Italy Biotecnologies BT - Perugia, Italy Dompè, L' Aquila, Italy European Union, Bruxelles, Belgium Istituto Auxologico Italiano, Milan, Italy Istituto Nazionale Neurologico "C. Besta", Milan, Italy Italian Association for Cancer Research (AIRC), Milan, Italy Italian Ministry of University and Research (MIUR), Rome, Italy Kenneth S. Warren Institute, NY, USA 166

168 Lundbeck A/S, Copenhagen, Denmark Cariplo Foundation, Milan, Italy Don Gnocchi Foundation, Milan, Italy Mariani Foundation, Milan, Italy Monzino Foundation, Milan, Italy Ministry of Health, Rome, Italy National Research Council (CNR), Palermo, Italy North Shore University Hospital, NY, USA Perfetti-Van Melle, Lainate (Mi), Italy Sigma Tau, Pomezia (Rome), Italy Telethon, Milan, Italy University of Florence, Italy University of Milan-Bicocca, Italy University of Siena, Italy Weizmann-Pasteur-Negri Foundation, Paris, France Zambon Group, Bresso (Mi), Italy SELECTION OF SCIENTIFIC PUBLICATIONS FROM 2008 Bate C, Marshall V, Colombo L, Diomede L, Salmona M, Williams A Docosahexaenoic and eicosapentaenoic acids increase neuronal death in response to HuPrP and Abeta(1-42 Neuropharmacology : Bate C, Tayebi M, Diomede L, Salmona M, Williams A Docosahexaenoic and eicosapentaenoic acids increase prion formation in neuronal cells BMC Biol : 39 Bazzoni G Signalling pathways and adhesion molecules as targets for antiangiogenesis therapy in tumors Adv Exp Med Biol : Bigini P, Repici M, Cantarella G, Fumagalli E, Barbera S, Cagnotto A, De Luigi A, Tonelli R, Bernardini R, Borsello T, Mennini T Recombinant human TNF-binding protein-1 (rhtbp-1) treatment delays both symptoms progression and motor neuron loss in the wobbler mouse Neurobiol Dis : Brines M, Patel N S A, Villa P, Brines C, Mennini T, De Paola M, Erbayraktar Z, Erbayraktar S, Sepodes B, Thiemermann C, Ghezzi P, Yamin M, Hand C C, Xie Q W, Coleman T, Cerami A Nonerythropoietic, tissue-protective peptides derived from the tertiary structure of erythropoietin Proc Natl Acad Sci USA : Colleoni S, Jensen A A, Landucci E, Fumagalli E, Conti P, Pinto A, De Amici M, Pellegrini-Giampietro D E, De Micheli C, Mennini T, Gobbi M Neuroprotective effects of the novel glutamate transporter inhibitor (-)-3-hydroxy-4,5,6,6a-tetrahydro-3aHpyrrolo[3,4-d]-isoxazole-4-carboxylic acid, which preferentially inhibits reverse transport (glutamate release) compared with glutamate reuptake J Pharmacol Exp Ther : Colombo Alessio, Bastone A, Ploia C, Sclip A, Salmona M, Forloni G, Borsello T JNK regulates APP cleavage and degradation in a model of Alzheimer's disease Neurobiol Dis 2008 [Epub ahead of print] Cosentino U, Pitea D, Moro G, Saracino A A G, Caria P, Vari' M R, Colombo L, Forloni G, Tagliavini F, Salmona M The anti-fibrillogenic activity of tetracyclines on PrP : a 3D-QSAR study J Mol Model :

169 Deban L, Jarva H, Lehtinen M J, Bottazzi B, Bastone A, Doni A, Jokiranta T S, Mantovani A, Meri S Binding of the long pentraxin PTX3 to factor H: interacting domains and Function in the regulation of complement activation J Immunol : De Luigi A, Colombo L, Diomede L, Capobianco R, Mangieri M, Miccolo C, Mangeri M, Limido L, Forloni G, Tagliavini F, Salmona M The efficacy of tetracyclines in peripheral and intracerebral prion infection PLoS One : e1888 De Paola M, Diana V, Bigini P, Mennini T Morphological features and responses to AMPA receptor-mediated excitotoxicity of mouse motor neurons: comparison in purified, mixed anterior horn or motor neuron/glia cocultures J Neurosci Methods : Fumagalli E, Funicello M, Rauen T, Gobbi M, Mennini T Riluzole enhances the activity of glutamate transporters GLAST, GLT1 and EAAC1 Eur J Pharmacol : Garattini E, Fratelli M, Terao M Mammalian aldehyde oxidases: genetics, evolution and biochemistry Cell Mol Life Sci : Ghezzi P, Di Simplicio P Glutathionylation pathways in drug response Curr Opin Pharmacol : Gianni M, Boldetti A, Guarnaccia V, Rambaldi A, Parrella E, Raska I Jr, Rochette-Egly C, Del Sal G, Rustighi A, Terao M, Garattini E Inhibition of the peptidyl-propyl-isomerase Pin1 enhances the responses of acute myeloid leukemia cells to retinoic acid via stabilization of RARα and PML-RARα Cancer Res. 2008, in press Gobbi M, Funicello M, Gerstbrein K, Holy M, Moya P R, Sotomayor R, Forray M I, Gysling K, Paluzzi S, Bonanno G, Reyes-Parada M, Sitte H H, Mennini T N,N-dimethyl-thioamphetamine and methyl-thioamphetamine, two non-neurotoxic substrates of 5-HT transporters, have scant in vitro efficacy for the induction of transporter-mediated 5-HT release and currents J Neurochem : Inforzato A, Rivieccio V, Morreale A P, Bastone A, Salustri A, Scarchilli L, Verdoliva A, Vincenti S, Gallo G, Chiapparino C, Pacello L, Nucera E Structural characterization of PTX3 disulfide bond network and its multimeric status in cumulus matrix organization J Biol Chem : Manzoni C, Colombo L, Messa M, Cagnotto A, Cantù L, Del Favero E, Salmona M Overcoming synthetic Aβ peptide aging: a new approach to an age-old problem Amyloid, 2008 in press Mengozzi M, Cervellini I, Bigini P, Martone S, Biondi A, Pedotti R, Gallo B, Barbera S, Mennini T, Boraso M, Marinovich M, Petit E, Bernaudin M, Bianchi R, Viviani B, Ghezzi P Endogenous erythropoietin as part of the cytokine network in the pathogenesis of experimental autoimmune encephalomyelitis Mol Med : Micale N, Colleoni S, Postorino G, Pellicano' A, Zappala' M, Lazzaro J, Diana V, Cagnotto A, Mennini T, Grasso S Structure-activity study of 2,3-benzodiazepin-4-ones noncompetitive AMPAR antagonists: identification of the 1-(4- amino-3-methylphenyl)-3,5-dihydro-7,8-ethylenedioxy-4h-2,3-benzodiazepin-4-one as neuroprotective agent Bioorg Med Chem : Modica M N, Romeo G, Salerno L, Pittala' V, Siracusa M A, Mereghetti I, Cagnotto A, Mennini T, Gaspar R, Gal A, Falkay G, Palko M, Maksay G, Fulop F Synthesis and receptor binding of new thieno[2,3-d]-pyrimidines as selective ligands of 5-HT(3) receptors 168

170 Arch Pharm (Weinheim) : Natalello A, Prokorov V V, Tagliavini F, Morbin M, Forloni G, Beeg M, Manzoni Claudia, Colombo L, Gobbi M, Salmona M, Doglia S M Conformational plasticity of the Gerstmann-Sträussler-Scheinker disease peptide as indicated by its multiple aggregation pathways J Mol Biol : Paris L, Bazzoni G The protein interaction network of the epithelial junctional complex: a system-level analysis Mol Biol Cell : Paris L, Bononi E, Bazzoni G Network analysis of cell adhesion. Adhesomes as context-defined subnetworks Communicative & Integrative Biology 2008, in press Paris L, Tonutti L, Vannini C, Bazzoni G Structural organization of the tight junctions Biochim Biophys Acta : Pera M, Martinez-Otero A, Colombo L, Ruiz-Molina D, Badia A, Clos M V Acetylcholinesterase as an amyloid enhancing factor in PrP aggregation process Mol Cell Neurosci 2008 [Epub ahead of print] Pinto A, Conti P, De Amici M, Tamborini L, Grazioso G, Colleoni S, Mennini T, Gobbi M, De Micheli C Synthesis of enantiomerically pure HIP-A and HIP-B and investigation of their activity as inhibitors of excitatory amino acid transporters Tetrahedron Asymmetry : Pittala' V, Modica M, Salerno L, Siracusa M, Guerrera F, Mereghetti I, Cagnotto A, Mennini T, Romeo G Synthesis and endothelin receptor binding affinity of a novel class of 2-substituted-4-aryl-3-quinolinecarboxylic acid derivatives Med Chem : Repici M, Mare L, Colombo A, Ploia C, Sclip A, Bonny C, Nicod P, Salmona M, Borsello T. c-jun N-terminal kinase binding domain-dependent phosphorylation of mitogen-activated protein kinase kinase 4 and mitogen-activated protein kinase kinase 7 and balancing cross-talk between c-jun N-terminal kinase and extracellular signal-regulated kinase pathways in cortical neurons. Neuroscience 2008 [Epub ahead of print] Roglio I, Bianchi R, Camozzi F, Carozzi V, Cervellini I, Crippa D, Lauria G, Cavaletti G, Melcangi R C Docetaxel-induced peripheral neuropathy: protective effects of dihydroprogesterone and progesterone in an experimental model J Peripher Nerv Syst 2008, in press Roglio I, Bianchi R, Gotti S, Scurati S, Giatti S, Pesaresi M, Caruso D, Panzica G, Melcangi R C Neuroprotective effects of dihydroprogesterone and progesterone in an experimental model of nerve crush injury Neuroscience : Roglio I, Giatti S, Pesaresi M, Bianchi R, Cavaletti G, Lauria G, Garcia-Segura L M, Melcangi R C Neuroactive steroids and peripheral neuropathy Brain Res Rev : Saracino A A G, Villa A, Moro G, Cosentino U, Salmona M Spontaneous β-helical fold in prion protein. The case of PrP(82-146) Proteins 2008 [Epub ahead of print] Siracusa M A, Salerno L, Modica M N, Pittala' V, Romeo G, Amato M E, Nowak M, Bojarski A J, Mereghetti I, Cagnotto A, Mennini T Synthesis of new arylpiperazinylalkylthiobenzimidazole, benzothiazole, or benzoxazole derivatives as potent and selective 5-HT1A serotonin receptor ligands J Med Chem :

171 Taoufik E, Petit E, Divoux D, Tseveleki V, Mengozzi M, Roberts M L, Valable S, Ghezzi P, Quackenbush J, Brines M, Cerami A, Probert L TNF receptor I sensitizes neurons to erythropoietin- and VEGF-mediated neuroprotection after ischemic and excitotoxic injury Proc Natl Acad Sci USA : Terao M, Kurosaki M, Barzago M M, Fratelli M, Bagnati R, Bastone A, Giudice C, Scanziani E, Mancuso A, Tiveron C, Garattini E Role of the molybdo-flavoenzyme, aldehyde oxidase homolog 2, in the biosynthesis of retinoic acid: generation and characterization of a knock-out mouse Mol Cell Biol 2008 [Epub ahead of print] Valli C, Paroni G, Di Francesco A M, Riccardi R, Tavecchio M, Erba E, Boldetti A, Gianni M, Fratelli M, Pisano C, Merlini L, Antoccia A, Cenciarelli C, Terao M, Garattini E Atypical retinoids ST1926 and CD437 are S-phase-specific agents causing DNA double-strand breaks: significance for the cytotoxic and antiproliferative activity Mol Cancer Ther : RESEARCH ACTIVITIES Laboratory of Receptor Pharmacology Studies on the role of dysfunction of the endoplasmic reticulum (ER) or the Golgi apparatus (GA) in neurodegenerative disease The secretory pathway starts at the endoplasmic reticulum (ER) where proteins are synthesized and folded and chaperone mediated quality control prevents misfolded proteins to reach their destination and interfere with normal metabolism. Protein transport by mean of vesicles continues through the Golgi Apparatus (GA), that is most abundant in neurons, and finishes in the plasma membrane, secretory vesicles or lysosomes. The endocytic pathway enables internalized macromolecules to be delivered via endosomes to lysosomes where they are enzymatically digested. In mammalian cells, the Golgi-associated retrograde protein (GARP) complex is involved in retrograde transport of endosomes to the trans GA network. Defective intracellular membrane trafficking is common to several neurodegenerative diseases. Among the neuronal population, motor neurons, due the their high energy requirement and long axons are, together with retinal cells, the most sensitive ones. The Laboratory of Receptor Pharmacology utilizes two mouse models of neurodegeneration related to cellular transport disruption, carrying mutation in proteins resident in the ER (the mnd mouse) or in the GARP complex (the wobbler mouse). The neuronal ceroid lipofuscinosis (NCLs) are a group of autosomal recessive neurodegenerative diseases and a significant cause of childhood progressive intellectual and neurological deterioration, for which no curative or preventive treatment are available. Among them, the progressive epilepsy with mental retardation is the newest form with mutation in the CLN8 gene encoding a novel ER transmembrane protein with undefined function. An orthologue of CNL8 is mutated in the motor neuron degeneration mouse (mnd) which shows early retinopathy and delayed motor neuron dysfunction without degeneration. How CLN8 mutation leads to NCL defect is unknown. Our laboratory is studying mnd mice since many years: we have already reported decreased spinal cord GLT-1 glial glutamate transporter and increased plasma glutamate concentration already at presymptomatic stage in mnd mice, with increased GluR2 and lowered GluR3 AMPA receptor subunits in the lumbar spinal cord. The AMPA receptor antagonists ZK (non-competitive), like NBQX (competitive), ameliorate motor behavior in mnd mice. We also found that TNF and TNFR1 is increased in the spinal cord of mnd mice already at presymptomatic stage, when intensive astrocytes and 170

172 microglial proliferation occurs. The rate of oxygen consumption (QO2), and mitochondrial functions were decreased in mnd spinal cord. The level of lipid peroxide derivatives reacting with thiobarbituric acid (TBARS) were increased in mnd spinal cord and retina. L-carnitine treatment delayed the onset of motor behaviour impairment, increased the mitochondrial enzyme activities and was effective in enhancing QO 2 and decreasing TBARS levels. At present we are testing the susceptibility to convulsions in mnd mice at pre-symptomatic stage, in order to find a further link to the human pathology. In these studies new non-invasive approaches, like Optical Imaging, MRI, MicroCT and Confocal Angiography are applied, in order to allow a better translation of the results to the human disease. Amyotrophic Lateral Sclerosis (ALS) is a progressive neurodegenerative disorder involving primarily motor neurons in the spinal cord, brainstem and cerebral motor cortex and leading to denervation, muscular atrophy and paralysis. The disease is sporadic in approximately 90% of cases, and the mechanism(s) responsible for the selective motor neuron degeneration are far from being elucidated. A missense mutation in Vps54 has been described in the wobbler mouse, which share many pathological features with ALS patients. In mammalian cells Vps54 forms heterotrimeric complexes with Vps52 and Vps53 to form the GARP complex, involved in retrograde transport of endosomes to the trans GA network. Thus we use the wobbler mouse as a reliable tool to understand the interplay between endosomal dynamics and the selective loss of motor neurons. A series of experiments are in progress in cultured neural cells obtained from wobbler and healthy homozygous mice to investigate the possible effect of Vps54 mutation on intracellular trafficking, mitochondrial activity, and lysosome accumulation. We have already reported that wobbler mice are sensitive to riluzole treatment, without marked changes in AMPA/NMDA receptor subunits expression in motor neurons of early symptomatic mice. In addition we found increased levels of TNF and TNFR1 in the cervical spinal cord and a significant effect of chronic treatment with a soluble h-tnf binding protein, resulting in slower clinical symptoms progression, reduced motor neuron loss and selective inhibition of the two main stress-kinases (p38 and JNK) associated to TNF receptors activation. Finally we investigated two different approaches of cell therapy. Undifferentiated adult neural stem cells (in collaboration with Dr. Parati, Istituto Besta) produced a weak and transient protective effect in clinical progression but significantly reduced motor neuron loss occurring in the wobbler mouse. Transplantation of mononucleate cells from human cord blood (in collaboration with Dott. Lazzari, Policlinico), although did not replace degenerating motor neurons, produced a marked neuroprotective effect by slowing the clinical progression and reducing motor neuron loss, biceps atrophy and neuroinflammation (reactive gliosis). Neuropharmacology of the glutammatergic and serotonergic systems Glutamate is the major excitatory amino acid in the central nervous system; the extracellular glutamate concentration has to be maintained at physiological level by active uptake mediated by specific transporters (Excitatory Amino Acid Transporters, EAATs) located on the plasma membranes of neurons and glia. Alterations in this process might lead to a relevant increase in extracellular glutamate concentrations, that is highly toxic for neurons in the central nervous system. A research project is in progress in order to better characterize the involvement and the role of the neuronal compartment in the general process of glutamate homeostasis. The functional properties are evaluated using biochemical assays and the quantitative characteristics are evaluated by western blot for specific neuronal and glial proteins and flow cytometry analysis (in collaboration with Dr. Bernasconi, Department of Oncology, IRFMN). These evaluations are done on purified preparations from mouse spinal cord. The same characteristics are evaluated on two different animal models of motor neuron degeneration, the wobbler mouse and the transgenic SOD1G93A mouse (in collaboration with Dott. Bendotti, Department of 171

173 Neuroscience, IRFMN), in order to understand the possible involvement in neurodegenerative diseases. Results obtained to date indicate that there is an active neuronal component in the glutamate uptake. The comparison between the two animal models of neurodegeneration suggest that this component has a different importance in the motor neuron death processes. In the wobbler mouse the excitotoxicity related to alterations of glutamate uptake is likely to be not involved, while in the transgenic SOD1G93A mouse we found a significant reduction of glutamate uptake in the neuronal fraction. This alteration probably contribute to the exacerbation of processes leading to motor neuronal death in this animal model. A research project is in progress in collaboration with professor Bonanno (University of Genova) to characterize mechanisms of glutamate release from synaptosomes obtained from spinal cord of wobbler mouse, in order to evaluate if the motor neuronal death might depend on altered processes of neurotransmitter release in the nerve endings. We recently concluded a project focused on the study of the interaction between glutamate transporters and riluzole. Riluzole is a drug with a complex mechanism of action, and is the only drug approved for the treatment of amyotrophic lateral sclerosis (ALS). We used as experimental model cell cultures that stably express the main three glutamate transporters (GLT1, GLAST and EAAC1) and we demonstrated for the first time that riluzole acts on the glutamate uptake mediated by the three EAAT subtypes, significantly increasing the efficiency of the process. This mechanism might be relevant in pathological conditions characterized by glutamate concentrations over the physiological threshold, as occurs in ALS. AMPA receptor-mediated excitotoxicity is one of the main events involved in motor neuron degeneration in ALS pathogenesis. We established a new model of primary cocultures of purified motor neurons over a glial layer. This in vitro model allows to obtain healthier motor neurons maintained in more physiological conditions and was used to demonstrate that AMPA receptor agonists can induce both apoptotic or non-apoptotic death pathways depending on their concentrations. We studied the interaction between excitotoxicity and other potentially neurotoxic factors, such as the inflammatory mediators, and we demonstrated that the proinflammatory chemokine IL-8 induces motor neuron death through the CXCR2 receptor. Preliminary studies on the effect of TNF-α, whose levels were found increased in the spinal cords of animal models of motor neuron degeneration, indicated the pivotal role of glial cells in mediating neurotoxicity of this cytokine. At present, we are studying the main intracellular biochemical pathways involved in neurodegenerative mechanisms (such as calcium influx) and testing new potential treatments to selectively interfere with each of them. We have also started a study on the neurotoxic effect of serum from professional soccer players aimed at identifying potential risk factors present in the blood which could support the high incidence of ALS in this group of athletes. Studies on primary spinal cord or hippocampal neuronal cultures from wild type or mnd mice are in progress in order to investigate their sensitivity to AMPA receptor agonists and antagonists, and to evaluate the role of astrocytes obtained from mnd mice in affecting motor neuron viability. Studies on astrocytes derived from neural stem cells of wobbler or control mice are in progress in order to evaluate whether typical biochemical alterations of wobbler mice are already evident in precursor-derived cells : we study the role of glutamate transporters and possible effects of this kind of cells on healthy motor neurons. Finally, studies on the role of Vsp54 mutation on intracellular trafficking are in progress in primary cultures of astrocytes obtained from the spinal cord of adult wobbler or control mice. The Laboratory of Receptor Pharmacology is maintaining, since many years, collaborations with medicinal chemistry laboratories to characterize the affinity and selectivity of newly synthesized compounds on neurotransmitter receptors using in vitro binding methods. The results are utilized for molecular modelling (QSAR) studies and/or for further pharmacological evaluations. Particularly, a collaboration is ongoing with Prof. S. Grasso 172

174 (University of Messina) and Prof. C. De Micheli (University of Milan), for the development of new non-competitive AMPA receptor antagonists. An useful application of this technique is also the evaluation of the agonist/antagonist activity of compounds acting on G-protein coupled receptors (GPCR), measuring their effects on 35S- GTP-γ-S binding. New non-radioactive methods based on time-resolved fluorescence are under characterization as functional assays to monitor GPCR activity on cell membranes. Recently we have verified the effect of dimethyl sulfoxide, a solvent commonly utilized to dissolve hydrophobic compound for in vitro experiments, on different in vitro assays utilizing HEK 293 cells expressing the 5-HT6 serotonin receptors. Our results indicate that dimethyl sulfoxide interferes with the agonist activity on 5-HT6 receptor when the scintillation proximity assay 35S-GTP-γ-S binding is used. On the contrary, it does not interferes with europium labelled GTP binding determined by time-resolved fluorescence. In addition, the laboratory perform autoradiography binding studies in order to evaluate ex vivo drug-receptor occupancy. Finally, we have ongoing collaborative studies with Dr. Gobbi (Lab. biochemistry and protein chemistry, IRFMN), to characterize the role of the allosteric site at the serotonin transporter on the mechanism of action of SSRI (selective serotonin reuptake inhibitors) antidepressant, like escitalopram. Laboratory of Neuroimmunology Redox regulation The study of the so-called oxidative stress has led to the identification of biochemical events that are modified by antioxidant molecules even in the absence of oxidative stress intended as overproduction of toxic oxygen intermediates (free radicals). We use the term redox regulation to define the pattern of cell functions (gene expression, activity of enzymes or transcription factors) that are in some way modified by the redox state of the cell, defined as the ratio between oxidizing and reducing species (usually: the oxidized glutathione / reduced glutathione ratio). Our work focuses on the molecular mechanisms by which small changes in the redox state can affect proteins, with particular attention to the reversible modification of cysteines to form disulfide bonds (with proteins or with small molecular weight thiols such as glutathione). We recently focused our attention on the identification of the redox state of proteins present on the outside of the plama membrane since these often have key functions (e.g.: transporters, receptors) and are the closest target of extracellular oxidants. We also apply proteomics techniques and gene expression profiling using DNA microarrays to identify the pathways susceptible of redox regulation. Neuroprotection and erythropoietin The pathologies of the central or peripheral nervous systems studied in the lab are: cerebral ischemia, experimental autoimmune encephalomyelitis, and diabetic neuropathy). Using animal models and cell culture, we try to clarify the relationships between neuronal death and inflammation, and to intervene with protective agents. Among the latter, we are studying endogenous molecules that have shown an unexpected anti-apoptotic action on neuronal cells, particularly erythropoietin and anti-inflammatory drugs. Laboratory of Molecular Biology Novel retinoids for the treatment of acute myeloid leukemia The synthetic and natural derivatives of retinoic acid (retinoids) have shown promising activity in the treatment of leukemia and solid cancer. Retinoids exert their therapeutic activity through 173

175 three distinct types of effects: cyto-differentiation, growth inhibition and apoptosis. The three effects can be dissociated, albeit partially, as retinoids endowed primarily with cytodifferentiating or anti-proliferative activities are known. Recently, we identified and characterized a novel class of retinoids with strong and selective apoptotic activity towards the neoplastic cell. These compounds (RRMs, retinoid related molecules), which were originally developed as selective agonists of the gamma-types of the nuclear receptors of retinoic acid (RARy), induce apoptosis in different types of leukemia and solid cancer cells through a largely unknown mechanism. The process of apoptosis set in motion by RRMs is different from that of other known chemotherapeutic agents and does not require the activation of the nuclear retinoic acid receptors. RRMs are active not only in vitro but also in vivo on a number of pre-clinical models of acute myeloid leukemia. Currently, some of these innovative molecules are in an advanced phase of pre-clinical development. Novel retinoic-acid-based pharmacological combinations for the treatment of acute leukemia The clinical use of retinoic acid for the treatment of acute promyelocytic leukemia (APL) is based on the ability of this compound to induce the maturation of the leukemic blast along the normal myelocytic/granulocytic pathway. At present, the clinical use of retinoic acid for the treatment of patients suffering from APL is the sole example of differentiation therapy. Differentiation therapy is worth pursuing as it is theoretically associated with a lower level of toxicity relative to what observed following treatment with the classical cyto-toxic agents. However, the clinical use of retinoic acid is still burdened by a number of problems including natural and induced resistance as well as systemic and local toxicity. One of the possible ways to increase the therapeutic index of retinoic acid is based on the identification of compounds or drugs that potentiate the pharmacological activity of the retinoid. We have recently demonstrated that a series of agents, such as G-CSF, interferons, camp analogs, phosphodiesterase IV inhibitors and a number of other novel compounds sensitize the leukemic blast to the pharmacological activity of retinoic acid. More recently, Pin1, a peptidyl-prolylisomerase, was identified as an important molecular target for the sensitization of leukemia and cancer cells to the affects of retinoic acid and derivatives. In the long run, it is our objective to develop novel combinations and bring some of the above mentioned retinoic-acid-based combinations to the clinic. In addition we intend to use some of the combinations as pharmacological tools to dissect the intricacies of the cyto-differentiation process set in motion by retinoic acid in the leukemic blast. We are widening the scope of our scientific activity to the study of solid tumors with particular reference to breast carcinoma. In this last type of tumor we are investigating the cross-talk between estrogens and retinoids with the development of appropriate cellular models genetically engineered for the expression or silencing of estrogen receptors. The family of molybdo-flavoproteins Molybdo-flavoenzymes are proteins of medical and industrial interest. They are the sole enzymes that require molybdenum, in the form of the molybdenum cofactor, for their catalytic activity. The laboratory has a long-standing and specific interest in the biochemistry and biology of mammalian molybdo-flavoproteins. In the past, the laboratory contributed to the elucidation and characterization of the primary structure of the two mammalian molybdoflavoproteins, aldehyde oxidase (AOX1) and xanthine oxidoreductase (XOR). In the last few years, the group identified and cloned the cdnas and the genes coding for three novel mouse molybdo-enzymes (AOH1, AOH2 and AOH3) belonging to the sub-family of molybdoflavoproteins. The long-term goal of our studies is to define the structure, the substrate specificity, the mechanisms of catalysis as well as the physo-pathological function of the three new proteins. We will also continue the biochemical and functional studies on mammalian 174

176 AOX1 and XOR. To achieve our aims, we have recently developed cell lines over-expressing XOR in a tetracycline inducible fashion. In addition, we have generated knock-out mice for the genes encoding AOH2 and AOH3. The first knock-out model suggests that AOH2 is involved in the synthesis of retinoic acid from retinaldehyde in the skin and the Harderian gland, which is the main intra-orbital exocrine gland present in rodents. Creation of integrated instruments for the rationalization of Microarray analysis processes An interdisciplinary group has been created in the Institute, with several external collaborations, that deals, at different levels and with different professional backgrounds, with microarray data analysis. The procedures adopted by the different groups have been discussed, compared and formally described with the aim of establishing a single workflow (that is a software that provides end users with an easier way to orchestrate and describe complex processing of data in a visual form). The advantages of this approach are manifold. In fact, the highest possible automation of the processes makes them more reliable and facilitates documentation and reproducibility of the entire analysis process. Moreover it allows an easy comparison among the different methodological choices, with the aim of reaching a shared procedure among the collaborative groups. The activity has required the development of a software for the analysis the preparation of a hardware platform (Beowulf cluster), to support the required computational power. Laboratory of Biochemistry and Protein Chemistry Chimico-physical and molecular-biochemical studies on the prion protein and on the peptides deduced from its aminoacid sequence Prion protein fibril formation is associated with neuronal cytotoxicity and astrogliosis observed in prion diseases. The formation of fibrils is the consequence of a conformational switch between the structure of the native and the pathological proteins and it is considered of pivotal importance for the appearance and progression of prion protein diseases. Identifying the molecular determinants responsible for the conformational transition is likely to give insight into the pathogenetic process leading to prion diseases. A reductionist appraoch to the problem calls for the generation of simple experimental models in which the dynamics of the conformational transition can be studied in detail. In our laboratory we developed synthetic peptides that mimic the fibrillogenic properties of pathological prion protein. With the use of various types of biochemical and biophysical techniques we studied the conformation of these peptides through the evaluation of their secondary structure, the resistance to protease digestion as well as the aggregating and amyloidogenic properties. Our approach gave detailed informations on the conformational plasticity of various types of prion protein fragments. To understand the correlation between the chemico-physical properties of prion protein-derived peptides and their biological effects we used appropriate cellular models. These experiments, performed in collaboration with the Laboratory of the Biology of Neurodegenerative Disorders, gave informations on the sub-cellular distribution of the peptides to identify the intracellular biological targets. Development of a therapeutical strategy against prion-related diseases Currently, no therapeutic options for the cure of prion-related diseases are available and the identification of new molecules for the prevention and treatment of the infettivity is of great interest. Althought some compounds gave positive results in prion cellular models, in human 175

177 they exhert low efficacy for toxicity and for their inability to pass the blood brain barrier. A first approach for the development of anti-prion candidates involved molecules capable of interfering with the fibrils formation, directly interacting with the β-sheet conformation of PrPSc causing its disaggregation. In collaboration with the Laboratory of the Biology of the Neurodegenerative Disorders we aimed to identify compounds endowed with anti-fibrillogenic activity and test their efficacy in different in vitro and in vivo pre-clinical models of prion diseases. Our studied indicated that tetracyclines are new good candidates as anti-prion drugs since they act as anti-fibrillogenic compunds increasing the prion sensitivity to protease K digestion. Moreover, they inhibited neuronal cell death and astrogliosis caused by prion peptides and prolonged the survival of prion-infected animals. Another therapeutic approach is based on the development of molecules that inhibited the PrP formation by interacting with the lipid metabolism and destabilizing specific cellular membrane domains. In collaboration with the Department of Infective Pathology and Diseases of the Royal Veterinary School (Hawkshead, UK) we reported that statins, inhibiting cholesterol synthesis, reduced the in vitro prion production. On the other hand, compounds such as polyunsaturated fatty acids, known for their reducing effects on cholesterol levels, increased the PrP production. Our future studies are aimed at understanding the relationship between cholesterol cellular membrane distribution, lipid domain stability and the conversion of cellular prion protein in its pathologic form. Oxidative stress and protein aggregation in amyotrophic lateral sclerosis: a proteomic approach The molecular mechanisms at the basis of neurodegenerative diseases including the geneticallylinked ones, such as amyotrophic lateral sclerosis (ALS), are still unknown. However, there is evidence that oxidative stress and protein aggregation play central roles in the pathogenesis of such diseases. The Unit of Medical Biochemistry, conducted proteome analysis of an animal model of familial ALS. In collaboration with the laboratory of Molecular Neurobiology, we focused the attention on the analysis of protein expression changes and protein modifications, such as tyrosine nitration and ubiquitination, in a transgenic mouse, which over-express human mutated (G93A) superoxide dismutase (SOD1). We analyzed, by proteomic tools, spinal cord of presymptomatic G93A SOD1 mice, we identified nitrated proteins and quantified the level of nitration for each protein in comparison with healthy controls. We have revealed that there is a substantial increase of the nitration level in at least five proteins: actin, alpha and gamma enolase, ATP synthase and a chaperone protein, HSC71. The alteration of the function of these proteins may have important consequences on the cellular metabolism and catabolism, and therefore may be at the basis of the molecular mechanisms leading to neurodegeneration. In addition, by mass spectrometry, we have identified the specific nitrated tyrosines for a number of proteins. We have observed that al least enolase and glyceraldehyde 3-phosphate dehydrogenase are nitrated at the same tyrosine site known to be phosphorylated. This is an important finding, which is worthwhile further studying. In fact, it may indicate a possible involvement of nitration in signaling pathways and phosphorylation cascades. Regarding the aggregation studies, we have isolated detergent insoluble-protein fractions from spinal cord of G93A SOD1 mice and we have completed the comprehensive characterization of all the proteins contained. With this study we have identified the protein constituents of aggregates, still unknown, and therefore have contributed to the comprehension of the role of protein inclusions in ALS pathogenesis. Laboratory of Molecular Pathology In vitro models for investigating motor neuron pathologies 176

178 Presence of mutant forms of specific proteins plays a key role in many neurodegenerative diseases. Experimental models in vivo and in vitro are sorely needed to study the effects of these toxic proteins. We have applied the ptet-off system to control gene expression through the level of tetracyclines to a murine motor neuron-like cell line (NSC-34) establishing new cell lines (called NSC-34 tta) that stably express the transactivating protein tta. These cell lines are suitable to study the pathogenic mechanisms of motor neuron diseases after transient/stable transfection with genes of interest for these pathologies. We used this approach to establish NSC-34 tta cell lines that express in a doxycycline inducible fashion the human G93A mutant superoxide dismutase 1. Mutant forms of superoxide dismutase 1 are responsible for some of the familial forms of amyotrophic lateral sclerosis. These conditional cell lines as well as others previously obtained that constitutively express mutant G93A superoxide dismutase 1 are used to study the pathogenic mechanisms of amyotrophic lateral sclerosis. Novel intracellular targets in the selective degeneration of motor neurons in amyotrophic lateral sclerosis Amyotrophic lateral sclerosis is a rapidly fatal neurodegenerative disease characterized by loss of motor neurons. The management of this disease remains essentially supportive and symptomatic. Understanding the mechanisms underlying the disease is a way to favor more efficient therapeutic strategies. Alterations of mitochondria morphology were observed in the early stages of the disease in the motor nerve terminals of ALS patients and in murine experimental models. For these reasons we addressed our studies to determine biochemicalmolecular alterations involved in the mitochondrial damage utilizing our cellular models. We have studied the toxicity of mutant forms of superoxide dismutase 1, responsible for some familial forms of amyotrophic lateral sclerosis. We showed that mutant superoxide dismutase 1 (G93ASOD1) altered the mitochondrial morphology and that motor neurons are selectively susceptible to this damage. Mitochondrial damage was modulated by the extent of expression of G93ASOD1 protein. Furthermore the expression of G93ASOD1 protein increased the susceptibility of motor neurons to inhibitors of the electron transport chain (ETC) and to oxidants. Exposure to drugs or exogenous compounds impairing the ETC could thus be a risk to motor neurons of individuals carrying mutant superoxide dismutase 1. Cytochrome P-450 superfamily Cytochrome(s) P-450 have evolved into a large superfamily that varies enormously in substrate affinity and product formation. This system plays a major role in the metabolism of drugs and other chemicals. The majority of existing drugs depends on the P-450 system for terminating their biological effects or for side effects or adverse reaction. The laboratory has a long-standing interest in the induction/degradation mechanisms of specific cytochrome P-450 families due to drug administration or to disease states. Our recent research focused on cytochrome P-450 induction by herbal remedies such as Hypericum perforatum extracts (St. John s Wort), which have an alleged activity in mild to moderate depression but interfere with the effect of several drugs. Activation of enzymes of the heme metabolic pathway (heme oxygenase system, biliverdin reductase) as a protective response to stress The enzymatic system of heme oxygenase (HO) is devoted to cellular degradation of heme containing molecules, like cytochromes and hemoglobin, and to recycling of iron. Products formed by the catalytic activity of HO - carbon monoxide and bile pigments - are important regulating factors in the cell. An increase of HO activity (which is usually sustained by activation of the inducible form HO-1) is now considered a protective mechanism against 177

179 untoward stimuli particularly when oxidative stress is involved. In the past, the laboratory of Molecular Pathology identified cytokines as inducers of HO activity and as transcriptional activators of the HO-1 gene. We are currently investigating the functional significance of HO-1 activation in neurodegeneration. Laboratory for the Study of Biological Systems System-level analysis of protein interactions in the epithelial junctional complex Inter-cellular junctions form the apical junctional complex and mediate adhesion between adjacent cells, thus representing the cellular basis for tissue cohesion (for instance, the epithelial lining of the intestine). In order to acquire system-level understanding of the apical junctional complex, we have studied (using a methodological approach of network analysis ) all the protein interactions that have been described at the junctions in epithelial cells of human origin. We also found that proper hubs (i.e., very rare proteins with an exceedingly high number of interactions with other proteins) were absent from the junctional network. Nevertheless, we observed that the most connected (albeit non-hub) proteins were also essential proteins. In addition, we have detected modules within the junctional networks (i.e., densely inter-connected groups of proteins). Analysis of the modules has highlighted general organizing principles of the junctional complex, thus confirming the usefulness of network analysis for studying the components and the interactions of the cell. Novel regulators of cell motility Cell motility plays a central role in several biological processes, under both normal (e.g. embryonic development) and pathological conditions (e.g. tumor cell dissemination). Thus, it is important to identify the molecular mechanisms that regulate cell motility. In recent years, we have characterized Junctional Adhesion Molecule-A (JAM-A), a membrane molecule that localizes to the intercellular tight junctions and binds PDZ-type intracellular proteins. In the course of these studies, we have discovered that JAM-A expression reduces cell motility. In addition, we have found that JAM-A enhances microtubule stability and focal adhesion formation, which are the adhesive points of contact between cells and extracellular matrix. All these functional changes require amino acid residues that mediate binding to PDZ-type intracellular proteins. These findings have highlighted a novel mechanism of motility inhibition that requires the interaction between a membrane protein and PDZ-type intracellular proteins. Effect of inflammatory cytokines on Junctional Adhesion Molecule-A (JAM-A) In the course of inflammatory responses, JAM-A contributes to the leakage of plasma proteins and the transmigration of circulating leukocytes. Although it has been reported that the inflammatory cytokine Tumor Necrosis Factor (TNF) causes the disassembly of JAM-A from the intercellular junctions, the mechanism has not been elucidated fully. Recently, we found that TNF enhances the solubility of JAM-A in non-ionic detergents and increases the amount of detergent-soluble JAM-A at the cell surface. In addition, we found that, upon cell treatment with TNF, higher levels of JAM-A become detectable at the cell surface (by FACS analysis). As these higher levels of JAM-A derive from the intercellular junctions (and not from intracellular stores), we propose that TNF causes not only the disassembly of JAM-A from the junctions and its subsequent redistribution to the cell surface, but also its dispersal in such a way that JAM-A becomes more easily accessible to the antibodies used for FACS analysis. These findings are important to highlight potential mechanisms of permeability regulation during inflammation that might be modulated by inflammatory interventions. 178

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182 DEPARTMENT OF EPIDEMIOLOGY STAFF Head Carlo LA VECCHIA, M.D. LABORATORY OF GENERAL EPIDEMIOLOGY Head Carlo LA VECCHIA, M.D. Cancer Epidemiology Unit Head Cristina BOSETTI, Mat.Sci.D. Lifestyle Habits and Prevention Unit Head Liliane CHATENOUD, Biol.Sci.D. Epidemiology for Clinical Research Unit Head Silvano GALLUS, Comp.Sci.D. LABORATORY OF EPIDEMIOLOGICAL METHODS Head Eva NEGRI, Mat.Sci.D. LABORATORY OF EPIDEMIOLOGY OF CHRONIC DISEASES Head Alessandra TAVANI, Biol.Sci.D. LABORATORY OF MEDICAL INFORMATICS Head Eugenio SANTORO, Comp.Sci.D. 181

183 CURRICULA VITAE Carlo La Vecchia holds a Doctor of Medicine from the University of Milan, a Master of Science in Clinical Medicine (epidemiology) from Oxford University and a Diploma from the Post-Graduate School of Pharmacological Research at the Mario Negri Institute for Pharmacological Research in Milan. Work experiences: He is Head of the Department of Epidemiology at the Mario Negri Institute for Pharmacological Research in Milan, Italy. He is also Associate Professor of Epidemiology at the University of Milan, Adjunct Professor of Epidemiology, University of Lausanne, Switzerland (since 2002) and Adjunct Professor of Medicine, School of Medicine, Vanderbilt University, Nashville, TN ( ). Dr. La Vecchia is a temporary advisor at the International Agency for Research on Cancer IARC/WHO in Lyon and at the WHO in Geneva, and a registered journalist in Milan. He was an Honorary Senior Lecturer in Oral Medicine at the Eastman Dental Institute at the University College of London ( ) and Adjunct Associate Professor of Epidemiology at the Harvard School of Public Health ( ). He is Associate Editor to: European J. of Cancer Prevention and presently serves on the editorial boards of the Jornals: Alimentazione e Prevenzione; Archives of Medical Science; Asian Pacific Journal of Cancer Prevention; Current Cancer Therapy Reviews; Dermatology Research and Practice; Digestive and Liver Disease; Economia Politica del Farmaco; European Journal of Cancer Prevention; European Journal of Nutrition; In Scope Oncology & Haematology; Journal of Nephrology; Maturitas; Nutrition and Cancer; Oncology; Open Cancer Journal; Oral Oncology; Revisiones en Ginecologia y Obstetricia; Revista Española de Nutrición Comunitaria; Revue d'epidémiologie et de Santé Publique; The Lancet, italian edition; Tumori. In 1993, he received the Glaxo Prize for medical publication. He has authored or co-authored over 1,800 publications in peer reviewed journals (1420 included in Pubmed), with over 52,000 quotations. Eva Negri got a degree in Mathematics in 1985 at the University of Milan, School of Mathematics. Awards: EEC scholarship for postgraduate training in Epidemiology (1988). Areas of interest: Design, conduction and analysis of epidemiologic studies on chronic diseases (e.g. cancer and myocardial infarction) and injuries, analysis of mortality of cohorts of workers, analysis of temporal trends and geographic distribution of mortality from cancer, cardiovascular disease, injuries and other selected conditions, analysis of national health surveys, application of linear modeling techniques to the analysis of epidemiological data, collaborative re-analyses and meta-analyses of epidemiological studies. Work experiences: Since 2007: Laboratory Chief, Unit of Epidemiologic Methods, Department of Epidemiology; : Unit Chief, Unit of Epidemiologic Methods, Laboratory Epidemiology; since : Researcher at the Laboratory of Epidemiology; : Collaborator of the Laboratory of Epidemiology. Selected publications Negri E, La Vecchia C, Pelucchi C, Tavani A The risk of acute myocardial infarction after stopping drinking Prev Med 2005; 40: Negri E, Pelucchi C, Talamini R, Montella M, Gallus S, Bosetti C, Franceschi S, La Vecchia C Family history of cancer and the risk of prostate cancer and benign prostatic hyperplasia Int J Cancer 2005; 114: Negri E, Little D, Boiocchi M, La Vecchia C, Franceschi S. B-cell non-hodgkin s lymphoma and hepatitis C virus infection: A systematic review Int J Cancer 2004; 111: 1-8 Negri E, Ron E, Franceschi S, La Vecchia C, Preston-Martin S, Kolonel L, et al. Risk factors for medullary thyroid carcinoma: A pooled analysis Cancer Causes Control 2002; 13: Levi F, La Vecchia C, Boyle P, Lucchini F, Negri E Western and eastern European trends in testicular cancer mortality Lancet 2001; 357:

184 Alessandra Tavani degree in Biological Sciences, University of Milan, Italy (July 1977); Pharmacological Research Specialist, Mario Negri Institute for Pharmacological Research, Milan, Italy (July 1979). Work experiences: : Researcher at the laboratory of Drug Metabolism, Mario Negri Institute for Pharmacological Research. 1981: Researcher at the Unit for Research on Addictive Drugs (director prof. H.W. Kosterlitz), University of Aberdeen, Scotland, U.K : Head of the Unit of Opioid Neuropharmacology, Mario Negri Institute for Pharmacological Research. 1990: Researcher at the Unit of Clinical Perinatal Pharmacology, Mario Negri Institute for Pharmacological Research. From : Head of the Unit of Epidemiology of Chronic Diseases of the Laboratory of Epidemiology, Mario Negri Institute for Pharmacological Research : Head of the Laboratory of Epidemiology of Chronic Diseases of the Department of Epidemiology, Mario Negri Institute for Pharmacological Research. Awards: "Rafaelsen Scholar Award" from the Collegium Internationale Neuro-Psychopharmacologicum (CINP), 16th Meeting, Munich (F.R.G.), Areas of interest: Epidemiology of cancer and coronary heart disease. Organization of case-control studies and color studies on cancer and coronary heart disease, including biological sample collection. Analyses of risk factors related to genetic factors and lifestyles, particularly coffee, diet, physical activity. Selected publications Tavani A, Zucchetto A, Dal Maso L, Montella M, Ramazzotti V, Talamini R, Franceschi S, La Vecchia C. Lifetime physical activity and the risk of renal cell cancer. Int J Cancer 2007; 120: Bosetti C, Gallus S, Peto R, Negri E, Talamini R, Tavani A, Franceschi S, La Vecchia C. Tobacco smoking, smoking cessation, and cumulative risk of upper aerodigestive tract cancers Am J Epidemiol 2008; 167: Tavani A, Pelucchi C, Parpinel M T, Negri E, Franceschi S, Levi F, La Vecchia C. n-3 Polyunsaturated fatty acid intake and cancer risk in Italy and Switzerland. Int J Cancer 2003; 105: Herrero R, Castellsague X, Pawlita M, Lissowska J, Kee F, Balaram P, Rajkumar T, Sridhar H, Rose B, Pintos J, Fernandez L, Idris A, Sanchez M J, Nieto A, Talamini R, Tavani A, et al. Human papillomavirus and oral cancer: The International Agency for Research on Cancer Multicenter Study. J Natl Cancer Inst 2003; 95: Tavani A, Pelucchi C, Negri E, Bertuzzi M, La Vecchia C. n-3 polyunsaturated fatty acids, fish, and nonfatal acute myocardial infarction. Circulation 2001; 104: Eugenio Santoro got his degree in Computer Science in 1990 at the Milan University. He started to work at the Mario Negri Institute in 1985 as a research fellow. He was Head of the Applied Statistics and Informatics Unit and of the Applied Statistics and Informatics laboratory, which was part of the Department of Cardiovascular Research. Since 2001 he is Head of the Laboratory of Medical Informatics that is currently part of the Department of Epidemiology. His main areas of interest have been biostatistics and clinical informatics with the development of software for data management and data analyses of large scale clinical trials in cardiology, such as the GISSI studies (Gruppo Italiano per lo Studio della Sopravvivenza nell Infarto miocardico). His main current area of interest is the Internet, and more recently the web 2.0, and their application in the medical field, in clinical research, and in medical education through the development of health related websites. He is author or co-author of more than 160 scientific papers published in peer reviewed journals, and of more than 70 scientific abstracts submitted to the main international meetings in the cardiology and in the computer science fields. He is also author of three books (available in Italian) about the use of the Internet in medicine ( Web 2.0 and medicine, Guida alla medicina in rete and Internet in medicina. Guida all uso e applicazioni pratiche, published by the Pensiero Scientifico Editore, Rome) and of one section about Internet and medicine, included in one of the most important italian medical encyclopedia ( Enciclopedia Medica Italiana, UTET 2007). He also collaborates to the publication of the Italian National Bioethics Committee s guidelines about ethics, health, and the new information technologies. Selected publications Santoro E. Podcast, wiki e blog: il web 2.0 al servizio della formazione e dell aggiornamento del medico. Recenti Prog Med 2007;98: Santoro E, Rossi Valentina, Pandolfini C, Bonati M. DEC-NET: The development of the European register of clinical trials on medicines for children. Clin Trials 2006; 3: Clivio L, Tinazzi A, Mangano S, Santoro E. The contribution of information technology: Towards a better clinical data management. Drug Dev Res 2006; 67: Santoro E. Internet and information on breast cancer: an overview. Breast 2003; 12: Santoro E, Nicolis E, Franzosi M G, Tognoni G. Internet for clinical trials: Past, present, and future. Control Clin Trials 1999; 20:

185 Franzosi M G, Santoro E, Zuanetti G, Latini R, Maggioni A P, Tognoni G, GISSI. Indications for ACE inhibitors in the early treatment of acute myocardial infarction. Systematic overview of individual data from patients in randomized trial. Circulation 1998; 97: Franzosi M G, Santoro E, De Vita C, Geraci E, Lotto A, Maggioni A P, Mauri F, Rovelli F, Santoro L, Tavazzi L, Tognoni G, GISSI. Ten-year follow-up of the first megatrial testing thrombolytic therapy in patients with acute myocardial infarction. Results of Gruppo Italiano per lo Studio della Sopravvivenza nell Infarto-1 study. Circulation 1998; 98: Franzosi M G, Latini R, Maggioni A P, Barlera S, Negri E, Nicolis E, Santoro E, Santoro L, Tognoni G, Garattini S, GISSI-3. GISSI-3: Effects of lisinopril and transdermal glyceryl trinitrate singly and together on six-week mortality and ventricular function after acute myocardial infarction. Lancet 1994; 343: Maggioni A P, Maseri A, Fresco C, Franzosi M G, Mauri F, Santoro E, Tognoni G, GISSI-2. Age-related increase in mortality among patients with first myocardial infarctions treated with thrombolysis. N Engl J Med 1993; 11: Cristina Bosetti got her degree in Mathematics in 1994 at the University of Milan, School of Mathematics, and the Post-Graduate Diploma in Pharmacological Research in 1999 at the Mario Negri Institute for Pharmacological Research in Milan. Areas of interest: Epidemiology of cancer, cardiovascular diseases and other chronic conditions. In particular case-control studies on cancers of the upper respiratory and digestive sites, thyroid, breast, hormone-related cancers, and on ischemic heart disease. Analysis of risk related to diet, alcohol, tobacco, reproductive and hormonal factors, occupational and environmental exposure to toxic substances, through the application of generalized linear models. She is author/coauthor of more than 130 publications on these issues on peer-reviewed scientific journals. Work experiences: Unit Head, Unit of Cancer Epidemiology, Department of Epidemiology, Istituto di Ricerche Farmacologiche Mario Negri, Milan (since Sept. 2005); Collaboration with the International Epidemiology Institute, Rockville, MD, USA (since ); Visiting scientist at the Unit of Field and intervention studies, International Agency for Research on Cancer (IARC), Lyon, France (sett-2000-giu. 2001); Visiting scientist at the Department of Epidemiology, Harvard School of Public Health, Boston, MA (sett-nov. 1998); Researcher at the Laboratory of Epidemiology, Istituto di Ricerche Farmacologiche Mario Negri, Milan ( ); Researcher at the Laboratory of Mother and Child Health, Istituto di Ricerche Farmacologiche Mario Negri, Milan ( ). Selected publications: Bosetti C, Bertuccio P, Levi F, Lucchini F, Negri E, La Vecchia C. Cancer mortality in the European Union, , with a joinpoint analysis. Ann Oncol. 2008;19: Bosetti C, Gallus S, Peto R, Negri E, Talamini R, Tavani A, Franceschi S, La Vecchia C.Tobacco Smoking, Smoking Cessation, and Cumulative Risk of Upper Aerodigestive Tract Cancers.Am J Epidemiol. 2007; 167: Bosetti C, Malvezzi M, Chatenoud L, Negri E, Levi F, La Vecchia C. Trends in cancer mortality in the Americas, Ann Oncol 2005; 16: Bosetti C, Spertini L, Parpinel M T, Gnagnarella P, Lagiou P, Negri E, et al. Flavonoids and breast cancer risk in Italy. Cancer Epidemiol Biomarkers Prev 2005; 14: Bosetti C, Micelotta S, Dal Maso L, Talamini R, Montella M, Negri E, et al. Food groups and risk of prostate cancer in Italy. Int J Cancer 2004; 110: Smith J S, Herrero R, Bosetti C, Munoz N, Bosch F X, Eluf-Neto J, et al. IARC Multicentric Cervical Cancer Study Group Herpes simplex virus-2 as a human papillomavirus cofactor in the etiology of invasive cervical cancer. J Natl Cancer Inst 2002; 94: Liliane Chatenoud, Doctor in Science Biology, University of Milan (1987); Postgraduate Doctor in Health Statistics University of Milan (1995). Areas of interest: Epidemiological studies on obstetric diseases. Dermato-epidemiology. Cancer epidemiology (case-control studies on cancers of the breast, female genital tract). Analysis of temporal trends and geographical distribution of perinatal, infant mortality, cancer and other selected conditions (over 100 publications on these topics, ). Work experiences: Since Sept. 2005: Unit Head, Lifestyle and Prevention, Department of Epidemiology : Researcher at the Laboratory of Epidemiology; : Staff Statistician Bracco S.p.A., Milan. Selected publications Naldi L, Chatenoud L, Belloni A, Peserico A, Balato N, Virgili A R, Bruni P L, Ingordo V, Lo Scocco G, Solaroli C, Schena D, Di Landro A, Pezzarossa E, Arcangeli F, Gianni C, Betti R, Carli P, Farris A, Barabino G F, La Vecchia C, Parazzini F Medical history, drug exposure and the risk of psoriasis. Evidence from an Italian case-control study Dermatology :

186 Valentini L G, Casali C, Chatenoud L, Chiaffarino F, Uberti Foppa C, Broggi G Surgical site infections after elective neurosurgery: a survey of 1747 patients. Neurosurgery : Chatenoud L, Mosconi P, Malvezzi M, Colombo P, La Vecchia C, Apolone G. Impact of a major thermoelectric plant on self-perceived health status. Prev Med. 2005;41: Naldi L, Chatenoud L, Linder D, Belloni Fortina A, Peserico A, Virgili AR, et al. Cigarette smoking, body mass index, and stressful life events as risk factors for psoriasis: results from an Italian case-control study. J Invest Dermatol. 2005;125:61-7. Bosetti C, Malvezzi M, Chatenoud L, Negri E, Levi F, La Vecchia C. Trends in cancer mortality in the Americas, Ann Oncol 2005; 16: Chatenoud L, Tavani A, La Vecchia C, Jacobs D R, Negri E, Levi F, Franceschi S. Whole grain food intake and cancer risk. Int J Cancer 1998; 77: Silvano Gallus got his degree in Computer Science in 1999 at the Milan University. Areas of interest: Design, data managing, and statistical analyses of case-control studies on the associations between several risk factors (including in particular tobacco smoking, alcohol drinking and Mediterranean diet) and risk of cancer, coronary heart disease and several other conditions. Analyses of occupational cohort studies. Monitoring of prevalence and trends of smoking habit and obesity in Italy. Author/coauthor of over 130 publication in peer-reviewed journals since Work experiences: Chief of the Unit of Epidemiology for Clinical Research of the Department of Epidemiology (since 2006); computer analyst, graphic designer, and statistical and epidemiological consultant, Milan and Bergamo (since 2002); researcher at the Laboratory of Epidemiology (since 1997); creator, designer and webmaster of the website of one of the major Italian public hospital, Milano ( ). Since 2008, Dr Gallus is Associate Editor of the journal BMC Public Health, and is member of the editorial board of The Open Obesity Journal Selected publications Gallus S, Foschi R, Negri E, Talamini R, Franceschi S, Montella M, Ramazzotti V, Tavani A, Dal Maso L, La Vecchia C. Dietary zinc and prostate cancer risk: a case-control study from Italy. Eur Urol. 2007;52: Gallus S, Naldi L, Carli P, La Vecchia C; Italian Group for Epidemiologic Research in Dermatology (GISED). Nevus count on specific anatomic sites as a predictor of total body count: a survey of 3,406 children from Italy. Am J Epidemiol. 2007;166: Gallus S, Scotti L, Negri E, Talamini R, Franceschi S, Montella M, Giacosa A, Dal Maso L, La Vecchia C. Artificial sweeteners and cancer risk in a network of case-control studies. Ann Oncol. 2007;18:40-4. Gallus S, Schiaffino A, La Vecchia C, Townsend J, Fernandez E. Price and cigarette consumption in Europe. Tob Control. 2006;15: Gallus S, Zuccaro P, Colombo P, Apolone G, Pacifici R, Garattini S, La Vecchia C. Effects of new smoking regulations in Italy. Ann Oncol. 2006;17: Clifford GM, Gallus S, Herrero R, Muñoz N, Snijders PJ, Vaccarella S, Anh PT, Ferreccio C, Hieu NT, Matos E, Molano M, Rajkumar R, Ronco G, de Sanjosé S, Shin HR, Sukvirach S, Thomas JO, Tunsakul S, Meijer CJ, Franceschi S; IARC HPV Prevalence Surveys Study Group. Worldwide distribution of human papillomavirus types in cytologically normal women in the International Agency for Research on Cancer HPV prevalence surveys: a pooled analysis.lancet. 2005;366:

187 INTRODUCTION TO THE DEPARTMENT S ACTIVITIES The Department of Epidemiology is involved in the epidemiology of several common cancers (including cancers of the breast, female genital tract, respiratory and digestive sites, prostate and urinary organs, lymphoid malignancies, melanoma, etc.) and of cardiovascular diseases, both through a descriptive and an analytical approach. Among the activities of descriptive epidemiology are the analysis of temporal trends and geographical distribution of mortality from cancer, cardiovascular diseases, and other selected conditions, in Italy and Europe; the analysis of trends in tobacco consumption in the Italian population, and the corresponding effects on incidence and mortality from lung and other tobacco-related neoplasms. The analytic epidemiology activities include the conduction and analysis of case-control studies, aimed at identifying and better quantifying the association between genetic factors (family history), selected lifestyle habits (diet, tobacco, alcohol, etc.), use of exogenous hormones and exposure to various substances and the development of various forms of cancers and cardiovascular diseases. In particular, the Department works on the analysis of dietary correlates of cancer and cardiovascular disease risk; quantification of health effects of tobacco smoking, alcohol consumption and implications for prevention; epidemiological studies on the risk related to oral contraceptive and hormone replacement therapy use; evaluation of the impact of screening in the early diagnosis and prevention of cancer. Other activities include: the conduction of quantitative reviews and meta-analysis of published data; the re-analysis of original data from epidemiological studies of cancers of the oral cavity and pharynx, pancreas, thyroid, breast, ovary, and cervix; epidemiological and clinical studies in dermatology in collaboration with the Gruppo Italiano per gli Studi Epidemiologici in Dermatologia (GISED); the analysis of historical cohort studies of occupational exposures to aromatic amines, asbestos, herbicides and other known or potential carcinogens; monitoring and prevention of injuries. Other Department s activities are related to the development of medical websites, the study of the quality of medical information available on the Internet, and the training and research on issues related to medical informatics and those concerning the use in the medical field of the Internet and web 2.0 applications. FINDINGS/MAIN RESULTS The risk of cancers of the pharynx, larynx and oesophagus is significantly increased for smokers of 2 cigarettes/day as compared to nonsmokers. This highlights the absence of any harmless level for cigarette smoking in relation to upper aero-digestive tract cancers. There is a significant inverse association between diet diversity and risk of cancers of the oral cavity, pharynx and esophagus. Diversity within consumption of vegetables and fruits has also a protective effect on upper aero-digestive tract cancers, including laryngeal cancer. According to the type of epidemiological study, the risk of oral and pharyngeal cancer is decreased by 35% to 48% for high vegetable consumption and by 22% to 45% for high fruit consumption. Subjects with a family history of gastric cancer have a 2.5-fold increased risk of gastric cancer. The risk might be higher when the affected relative is a sibling rather than a parent. 186

188 A diet rich in cereals and potatoes and poor in vegetables and fruit has an unfavorable role on gastric cancer. Among micronutrients, vitamin E, alpha-carotene and beta-carotene intake are inversely associated with gastric cancer, while sodium is directly associated with risk. Vitamin D intake might decrease the risk of colon cancer, particularly of the proximal colon, while it is unrelated to rectal cancer in the Italian population investigated. Intake of vitamin D over 3.57 μg/day or 143 IU appears to have a protective effect against breast cancer. Our analyses of data from a study conducted in Harbin, Northeast China, give further support to a positive association between indoor air pollution, history of selected non-malignant lung diseases, and lung cancer risk in both sexes. Extended information on family history of cancer, including data on second-degree relatives, could be useful to improve the discriminatory power of the Gail model risk factors for breast cancer. Consistently with previous epidemiological data, in our Italian study alcohol is more strongly related to estrogen receptors positive than to estrogen receptors negative breast tumors. In a case-control study on endometrial cancer, we found a significant direct association with consumption of red meat and a moderate increase in risk for consumption of sweets and poultry. On the other hand, a high consumption of cereals and vegetables decreased endometrial cancer risk by 44% and 41%, respectively. Frequent use of allium vegetables, including onions and garlic, might reduce the risk of endometrial as well as of various other cancers. A selection of carotenoids, particularly beta-carotene, beta-cryptoxanthin and lutein/zeaxanthin, is inversely related with endometrial cancer. Thus, all our findings indicate that diet might have an important role on endometrial cancer risk. In our meta-analysis of published studies on coffee and endometrial cancer, the risks are decreased by 13% for low-to-moderate coffee drinkers and by 36% for heavy drinkers, as compared to nondrinkers. Our results rule out a strong relation between physical activity (both occupational and recreational) and endometrial cancer. Women with at least one first-degree relative diagnosed with endometrial cancer have a more than doubled risk of developing the same neoplasm. Also, a family history of uterine and colorectal, but not breast, cancers increases the risk of endometrial cancer in the Italian population. Isoflavones and flavonols may have a distinct role in explaining the effect of fruit and vegetable against ovarian cancer. Using data from a collaborative reanalysis of 45 epidemiological studies from 21 countries, we estimate that oral contraceptives already prevented some 200,000 ovarian cancers and 100,000 deaths from the disease, and that over the next few decades the number of cancers prevented will rise to at least 30,000 per year. 187

189 Our study of renal cell cancer, based on a large dataset, provides further evidence that coffee, decaffeinated coffee and tea consumption are not related to renal cell cancer risk. When 6 major macronutrients are included in the same model of renal cell cancer, the adverse effect of high intake of starch and the protective effect of polyunsaturated fatty acids remain statistically significant. In a meta-analysis of thirty-nine published studies on glycemic index and load, we found a stronger direct association with colorectal and endometrial cancer than with breast cancer. No association was found for pancreatic cancer. Mortality from cancer in the European Union (EU) shows favorable patterns over recent years in both sexes. Mortality from cancer in Italy showed a favorable trend over recent years. The epidemic of non Hodgkin lymphoma observed during the second half of the 20th century has now started to level off in Europe as in other developed areas of the world. Mortality from hepatocellular carcinoma remains largely variable across Europe. Favorable trends were observed in several European countries (including Italy) mainly over the last decade, particularly in women and in young adults. For men in the EU, mortality rates from esophageal cancer were stable around 6/100,000 between the early 1980 s and the early 1990 s, and slightly declined in the last decade (5.4/100,000 in the early 2000s). Oesophageal cancer mortality rates remained comparatively low in European women. Over recent years, most countries showed decreasing trends in bladder cancer mortality. In men in the EU, mortality rates from kidney cancer peaked at 4.8 per 100,000 in , and declined to 4.1 in In women in the EU, the corresponding values were 2.1 in and 1.8 in The Mediterranean diet is rich in fat and starch, and hence may be related to overweight. However, in our study adherence to the major characteristics of the Mediterranean diet is unrelated to body mass index (BMI) and waist-to-hip ratio, confirming previous data from Greece and Spain. Body mass index influences the response to treatment with biological drugs in patients with moderate to severe psoriasis. In a survey conducted in 2008 on a sample of 3,035 individuals, representative of the Italian population aged 15 years or over, 22.0% of Italians described themselves as current cigarette smokers. Subjects with less privileged socio-economic characteristics should be considered target populations for tobacco control. In a sample of 20,800 immigrants, those who are illegal (16,033 subjects), have a higher frequency (10%) of acute infections of the upper respiratory tract, esophagus, stomach and duodenum diseases when compared to those immigrants with a regular residence card. No other major differences emerged between these two subgroups of subjects. 188

190 In a survey on 3406 Italian schoolchildren aged years, 17.4% of children had one or more congenital melanocytic naevi or congenital naevus-like naevi. A higher number of common melanocytic naevi, family history of malignant melanoma and personal history of diabetes were directly associated with congenital nevi. A systematic review of risk factors for falls in community-dwelling elderly people, including relevant studies published up to 2006, showed that the strongest associations were found for history of falls, vertigo, gait problems, use of walking aids, and use of antiepileptics. NATIONAL COLLABORATIONS Agenzia giornalismo scientifico Zadig, Milano Associazione Nazionale dei Medici Cardiologi Ospedalieri (ANMCO) Centro di Riferimento Oncologico, Servizio di Epidemiologia, Aviano (PN) Comune di Milano, Direzione centrale salute, Settore politiche per la salute Fondazione LuVI Fondazione SmithKline, Milano Gruppo Italiano per lo Studio della Sopravivenza nell Infarto miocardico (GISSI) Gruppo Italiano Studi Epidemiologici in Dermatologia GISED, Bergamo International Centre for Pesticides and Health Risk Prevention, Milano Istituto Auxologico Italiano, Divisione Malattie Metaboliche III, IRCCS, Piancavallo (VB) Istituto Auxologico Italiano, Laboratorio Sperimentale di Ricerche Endocrinologiche (LSRE), IRCCS, Milano Istituto Europeo di Oncologia, Divisione di Epidemiologia e Biostatistica, Milano Istituto Europeo di Oncologia, Divisione di Chirurgia Cervico Facciale, Milano Istituto Nazionale di Ricerca per gli Alimenti e la Nutrizione (INRAN), Roma Istituto Nazionale Neurologico Carlo Besta, Milano Istituto Nazionale per lo Studio e la Cura dei Tumori, Oncologia Sperimentale, Unità di Eredità Poligenica, Milano Istituto Tumori Fondazione Pascale, Servizio di Epidemiologia, Napoli Novartis Vaccines SpA, Siena Ordine dei Medici della Provincia di Bari Ospedale Casa Sollievo della Sofferenza San Giovanni Rotondo Ospedale Niguarda Ca Granda, Dipartimento Trapianti di Fegato, Milano Ospedale Niguarda Ca Granda, Istituto di Fisiologia Clinica CNR, Sezione di Milano, Milano Ospedali Riuniti di Bergamo Policlinico di Monza, Unità Operativa di Endoscopia I, Monza (MI) Prima Clinica Ostetrico Ginecologica, Mangiagalli, Milano Società Italiana Attività Regolatorie Università Bocconi di Milano, Dipartimento di Analisi Istituzionale e Management Pubblico Università degli Studi di Milano-Bicocca, Dipartimento di Statistica, Milano Università degli Studi di Milano-Bicocca, I Clinica Otorinolaringoiatria, DNTB, Monza Università di Milano, Clinica Pediatrica De Marchi, Milano Università di Milano, Istituto di Statistica Medica e Biometria G.A. Maccacaro, Milano Università di Milano, Prima Clinica Ostetrico Ginecologica, Milano Università di Pavia, Azienda di Servizi alla Persona, Pavia Università di Torino, Istituto di Medicina del Lavoro, CTO, Torino Università di Verona, Clinica Ostetrico Ginecologica, Verona 189

191 INTERNATIONAL COLLABORATIONS Catalan Institute of Oncology, Institut d Investigaciò Biomédica de Bellvitge (IDIBELL), Cancer Prevention and Control Unit, L Hospitalet de Llobregat, Spagna Center of Oncology, Dept. of Epidemiology and Cancer Prevention, Varsavia, Polonia Centre for Research in Environmental Epidemiology (CREAL) and Municipal Institute of Medical Research (IMIM), Barcellona, Spagna Harvard School of Public Health, Department of Epidemiology, Boston, USA Hôpital Necker - Enfants Malades, Centre of the Association Claude Bernard on Auto-immunes diseases, Parigi, Francia International Agency for Research on Cancer, Lione, Francia International Epidemiology Institute (IEI), Rockville, USA International Life Science Institute (ILSI), Bruxelles, Belgio National Cancer Institute, Environmental Studies Section, Bethesda, USA National Cancer Institute, Radiation Epidemiology Branch, Bethesda, USA National School of Public Health, WHO, Atene, Grecia Registre Vaudois des Tumeurs, Institut Universitaire de Médecine Sociale et Préventive, Losanna, Svizzera Senologic International Society Society for Internet in Medicine UNDP/UNFPA/WHO/WORLD Bank special program of research development and research training in human reproduction, Ginevra, Svizzera. Universitat Pompeu Fabra, Department of Experimental and Health Sciences, Barcellona, Spagna University of Athens Medical School, Department of Hygiene and Epidemiology, Atene, Grecia University of Las Palmas de Gran Canaria, Department of Clinical Sciences, Las Palmas de Gran Canaria, Spagna Vanderbilt University, Department of Medicine, School of Medicine, Nashville, USA EDITORIAL BOARD MEMBERSHIP Advances in Therapy (Eva Negri) Alimentazione e Prevenzione (Carlo La Vecchia) Archives of Medical Science (Carlo La Vecchia) BMC Public Health (Silvano Gallus, Associate Editor) Cancer Causes and Control (Carlo La Vecchia) Cancer Letter (Carlo La Vecchia, Associate Editor) Current Cancer Therapy Reviews (Carlo La Vecchia) Dermatology Research and Practice (Carlo La Vecchia) Digestive and Liver Disease (Carlo La Vecchia) Economia Politica del Farmaco (Carlo La Vecchia) European Journal of Cancer Prevention (Carlo La Vecchia, Associate Editor) European Journal of Clinical Nutrition (Carlo La Vecchia) European Journal of Nutrition (Carlo La Vecchia) Evidence Based Dermatology (Carlo La Vecchia, Liliane Chatenoud) In Scope Oncology & Haematology (Carlo La Vecchia) 190

192 International Journal of Cancer (Carlo La Vecchia) Maturitas (Carlo La Vecchia) Nutrition and Cancer (Carlo La Vecchia) Open Cancer Journal (Carlo La Vecchia) Oral Oncology (Carlo La Vecchia) Portale Partecipasalute.it (Eugenio Santoro) Revisiones en Ginecologìa y Obstetricia (Carlo La Vecchia) Revista Española de Nutriciò Comunitaria (Carlo La Vecchia) Revue d Epidémiologie et de Santé Publique (Carlo La Vecchia) Società Italiana Attività Regolatorie News, SIARNews (Eugenio Santoro) The Lancet, edizione italiana (Carlo La Vecchia) The Open Obesity Journal (Silvano Gallus) Tumori (Carlo La Vecchia) PEER REVIEW ACTIVITIES Acta Psychiatrica Scandinavica, American Journal of Clinical Nutrition, American Journal of Epidemiology, Annals of Epidemiology, Annals of Oncology, Archives of Internal Medicine, BMC Public Health, British Journal of Cancer, British Journal of Nutrition, British Medical Journal, Canadian Journal of Physiology and Pharmacology, Cancer, Cancer Causes and Control, Cancer Detection and Prevention, Cancer Epidemiology Biomarkers and Prevention, Computer Methods and Programs in Biomedicine, Diabetes/Metabolism Research and Reviews, Digestive Liver Disease, Epidemiologia & Prevenzione, Epidemiology, Epidemiology & Biostatistic, European Heart Journal, European Journal of Cancer, European Journal of Cancer Prevention, European Journal of Clinical Nutrition, European Journal of Epidemiology, European Journal of Public Health, Evidence-Based Healthcare and Public Health, Gynecological Endocrinology, Gut, Hepatology, Human Reproduction, International Journal of Cancer, International Journal of Epidemiology, International Journal of Obesity, JAMA, Journal of American College of Nutrition, Journal of Clinical Endocrinology and Metabolism, Journal of Clinical Epidemiology, Journal of Epidemiology and Community Health, Journal of Investigative Dermatology, Journal of Medical Internet Research, Journal of the National Cancer Institute, Lancet Oncology, Maturitas, Melanoma Research, Nicotine & Tobacco Research, Nutrition and Cancer, Obstetric and Gynecology, Oncology, Preventive Medicine, Public Health Nutrition, Radiation Research, Revue d Epidèmiologie et de Santé Publique, The Breast, The Cancer Journal, The Lancet, Tobacco Control, Tumori. NATIONAL AND INTERNATIONAL COMMITTEE MEMBERSHIP Advisory Committee of the Oxford Collaborative group on Aetiological Factors in Cancers of the Female Genital Tract Comitato Scientifico del Gruppo Italiano Studi Epidemiologici in Dermatologia Comitato Scientifico della Società Italiana di Colposcopia e Patologia Cervico Vaginale Data and Safety Monitoring Board of the Phase II therapeutic trial with a humanized nonmitogenic CD3 (ChAgly CD3) monoclonal antibody in recently diagnosed type I diabetic patients IARC/OMS di Lione e OMS di Ginevra Ministero della Salute, Sottocomitato fumo. 191

193 Ministero della Salute, Commissione Oncologica Nazionale. Scientific Review Committee del UND/WHO/World Bank Human Reproduction Programme Società Italiana della Riproduzione EVENT ORGANIZATION I edizione del corso ECM "La ricerca bibliografica su database biomedici", Istituto di Ricerche Farmacologiche Mario Negri, Milano, 13 maggio 2008 I edizione del corso ECM "Internet e l aggiornamento professionale in ambito medico", Istituto di Ricerche Farmacologiche Mario Negri, Milano, 28 maggio 2008 I edizione del corso ECM "Il web 2.0 al servizio della formazione e dell aggiornamento del medico", Istituto di Ricerche Farmacologiche Mario Negri, Milano, 11 giugno 2008 HiWATE Mid-term review meeting (Project EC FP6), Institute of Pharmacological Research Mario Negri, Milan, 3-5 June 2008 II edizione del corso ECM "La ricerca bibliografica su database biomedici", Istituto di Ricerche Farmacologiche Mario Negri, Milano, 24 novembre 2008 II edizione del corso ECM "Internet e l aggiornamento professionale in ambito medico", Istituto di Ricerche Farmacologiche Mario Negri, Milano, 25 novembre 2008 II edizione del corso ECM "Il web 2.0 al servizio della formazione e dell aggiornamento del medico", Istituto di Ricerche Farmacologiche Mario Negri, Milano, 26 novembre PARTICIPATION IN EVENTS IN WHICH THE DEPARTMENT WAS INVOLVED Meeting. FP6 Network of Excellence CCPRB. Assessor. Malmo, Svezia Gennaio Meeting. Pancreatic Cancer Case-control Consortium. New Members and Studies. Rockville, MD. 23 Gennaio International Workshop. Screening for lung cancer and management of early stage disease. Istituto Clinico Humanitas. An update in lung cancer epidemiology. Prevention perspectives. Rozzano (MI). 25 Gennaio Riunione Commissione Droghe Istituto Superiore Sanità. Roma. 30 Gennaio 2008 Seminario a Nerviano Medical Sciences. Cancer mortality and control in the European Union. Nerviano (MI). 1 Febbraio

194 5th International Conference. Cancer Prevention Aspirin and NSAID s in Cancer Prevention: Attempts of an International Consensus. Consensus finding roundtable. St. Gallen, Switzerland. 6-8 Marzo Meeting. IARC Working group on burden of cancer caused by asbestos. Estimates of asbestosrelated cancer burden: Europe, Australia and New Zealand, Asia and North America. Lyon, France. 6-7 Marzo XIV Congresso Nazionale delle Malattie Digestive (FIMAD) Caffe e rischio di tumori con particolare riferimento al tumore colorettale ed epatico Palacongressi della Riviera di Rimini, Rimini marzo 2008 VII Barcelona International Congress on the Mediterranean Diet Grande Covian Awardees Conference. Barcelona, Spagna Marzo Convegno Lega Italiana per la Lotta contro i Tumori. I sapori della salute. Dieta e prevenzione del cancro: dati italiani. Roma. 17 Marzo Epidemiology Specialist Panel Meeting. WHO headquarters. A cluster randomized controller trial to evaluate the effectiveness of the clinically integrated RHL-Evidence-based Medicine Course (New proposal). Ginevra, Svizzera. 31 Marzo Commissione Lega Italiana per la Lotta Contro i Tumori. Alimentazione-Oncologia geriatrica. Roma. 2 Aprile Congresso Nazionale ANDID. Food nutrition, physical activity and the prevention of cancer: a global perspective. Firenze 9-12 Aprile Congresso nazionale del collegio dei docenti di odontoiatria. Epidemiologia e fattori di rischio del carcinoma orale. Roma Aprile rd International meeting methodological issues in oral health research Palazzo Feltrinelli. Survival analysis of orthodonic brackets with clustered observations. Gargnano del Garda Aprile 2008 X Convegno nazionale tabagismo e servizio sanitario nazionale. ISS. L andamento dei tumori tabacco correlati nell ultimo decennio. Roma. 30 Maggio 2008 Meeting Bladder cancer from pathogenesis to prevention. Coffee and alcohol consumption and bladder cancer. Stockholm, Sweden Aprile th Congress of the European Society of Contraception. Epidemiology. Praga, Repubblica Ceca. 30 Aprile 3 Maggio 2008 Corso di epidemiologia descrittiva. Porto, Portogallo. 7-8 Maggio 2008 First International Congress on nutrition and cancer. Diet and cancer with a focus on Mediterranean diet. Antalya, Turchia Maggio 2008 Toxicology course Vienna. Nutrition and disease in epidemiological studies, Epidemiological studies on cancer risk factors. Vienna, Austria 29 Maggio

195 X Convegno nazionale tabagismo e servizio sanitario nazionale. L andamento dei tumori tobacco correlate nell ultimo decennio. Roma 30 Maggio 2008 Convegno. Prevenzione del carcinoma della cervice uterina: incontro-confronto su conoscenze e prospettive. Epidemiologia. Milano 17 Giugno a Conferenza italiana di oncologia toracica. The changing in the Epidemiology of Lung Cancer: the epidemiological evidence. Napoli Giugno 2008 Quinquennial Review of the Cancer Research UK Centre for Epidemiology. Mathematics & Statistics, directed by Professor Jack Cuzick at the Wolfson Institute of Preventative Medicine, Barts & the London School of Medicine & Dentistry. Londra, UK, 25 Giugno 2008 Review on Man-made fibers and cancer risk. Visit to the International Epidemiology Institute. Rockville, MD. 27 Luglio-2 Agosto Meeting. Contraception over 40. Hormonal contraception in aged women: cardiovascular and cancer risk. Capri 31 Agosto 1 Settembre 2008 Corso Partecipasalute: Divieto di fumare nei locali pubblici: prime valutazioni dell effetto della legge. Istituto di Ricerche Farmacologiche Mario Negri, Milano, 15 Settembre 2008 Membro del working group. Atlas of cancer mortality in Europe. Lione, Francia 5 Ottobre nd Congress of the International Society on Nutrigenetics and Nutrigenomics. "Fish consumption, omega-3 FA and the risk of cancer - cohort and case-control studies" Geneva, Svizzera 6-8 ottobre 2008 UNDP/UNFPA/WHO/World bank special programme of research, development and research training in human reproduction. Membro del working group. The safety of quinacrine in humans when administered as intrauterine doses for non-surgical sterilization. Geneva, Svizzera 8-10 Ottobre nd European Conference on injury prevention and safety promotion. Membro del working group. Parigi, Francia Ottobre th APOLLO WP Leaders meeting. WP4: Falls among elderly. Parigi, Francia. 11 Ottobre 2008 X Congresso Nazionale di Oncologia Medica. Controllo del cancro in Europa: attuali andamenti. Verona Ottobre 2008 Meeting Europa Donna. The European breast cancer coalition. Prevention: Lifestyle factors and their impact on breast cancer. Milano. 15 Ottobre 2008 XXXII Congresso annuale. Epidemiologia per la Prevenzione. Consumo di aglio e cipolla e rischio di tumore, Profili dietetici e rischio di tumore dell alto apparato digerente e respiratorio in Italia, Mortalità per tumori in Italia, dal 1970 al 2002, Nutrizione, dieta e attività fisica nell epidemiologia e Prevenzione del cancro. Milano Ottobre

196 7 th International Symposium on Multiple risk factors in cardiovascular diseases. Prevention and Intervention Health Policy. Smoking and other risk factors for cardiovascular disease: a quantification. Venezia (Lido), ottobre 2008 Convegno Extra Virgin Olive Oil International week. Olio d oliva e tumori: i dati italiani. Bari, 4 novembre 2008 Corso di formazione psicologi disassuefazione progetto tabagismo e scuola. Fumo e tumori: I rischi del fumo e i benefici della cessazione. Como, 8 Novembre 2008 II International Conference on olive and health. Olive oil and cancer risk. Jaen & Cordoba, Novembre 2008 Giornata Nazionale sulla Prevenzione primaria e secondaria del cancro del colon-retto. Dieta. Roma, 4 Dicembre 2008 Convegno Nazionale. Cancerogenesi Professionale: della valutazione del rischio e sorveglianza sanitaria, alla diagnosi per il riconoscimento della malattia professionale. Risultati falsopositivi negli studi epidemiologici sulla cancerogenesi occupazionale ed ambientale. Torino, 16 dicembre European Conference of Medical and Health Libraries, Helsinki giugno Titolo relazione: Supporting the evidence: Clinical Trials, Where to Find and How to Search Corso, Corso di Formazione per volontari dei punti di Accoglienza e Informazione in oncologia promosso dall Istituto Superiore di Sanità e da Alleanza Contro il Cancro, Roma Dicembre Titolo relazione: La valutazione di qualità tecnico-formale del materiale cartaceo e dei siti web Corso, PubMed e ClinicalTrials.gov per l aggiornamento professionale del medico promosso dall Ordine dei Medici della Provincia di Bari, Bari 21 novembre III Edizione del corso di formazione per rappresentanti di associazioni di cittadini & pazienti Orientarsi in salute e sanità per fare scelte consapevoli, promosso da Partecipasalute, Milano 18 novembre Titolo della relazione: Come navigare in Internet alla ricerca di informazioni di qualità. Siti utili per le associazioni di pazienti Master Universitario di I livello in Ricerca Clinica, Università di Milano, anno accademico Ruolo di docenze nel modulo Internet e le nuove tecnologie per l aggiornamento medico-scientifico, Milano 12 novembre 2008 Corso "Epidemiologia clinica e metodologia della ricerca" promosso dal Centro per la ricerca e lo studio del dolore dell Istituto Mario Negri, Rimini ottobre Titolo relazione: Internet in medicina Corso Internet e l uso dei nuovi strumenti del web 2.0 in ambito medico: potenzialità e applicazioni, Istituto Nazionale dei Tumori, Milano 8 ottobre 2008 Corso, Medicina basata sulle prove di efficacia promosso dall Azienda Unità Sanitaria Locale n.1 di Sassari, Sassari 23 maggio Titolo della lezione Siti e applicazioni Internet in ambito medico: tipologia di informazione e modalità di reperimento. 195

197 Corso, Medicina basata sulle prove di efficacia. Corso avanzato promosso dall Azienda Unità Sanitaria Locale n.1 di Sassari, Sassari 12 settembre Titolo della lezione Siti e applicazioni web 2.0 in ambito medico: tipologia di informazioni e metodologia d uso. AIFA Associazione Italiana per la Ricerca sul Cancro Comune di Milano Lega Italiana Lotta contro i Tumori European Commission (FP7) GISED Grunenthal-Formenti Ministero della Salute Regione Lombardia Weber Shandwich Consorzio Melinda ISA GRANTS AND CONTRACTS SELECTION OF SCIENTIFIC PUBLICATIONS FROM 2008 Bosetti C, Gallus S, Peto R, Negri E, Talamini R, Tavani A, Franceschi S, La Vecchia C. Tobacco smoking, smoking cessation, and cumulative risk of upper aerodigestive tract cancers. Am. J. Epidemiol., 167: (2008). Bosetti C, Levi F. Boffetta P, Lucchini L, Negri E, La Vecchia C. Trends in mortality from hepatocellular carcinoma in Europe, Hepatology, 48: (2008). Garavello W, Foschi R, Talamini R, La Vecchia C, Rossi M, Dal Maso L, Tavani A, Levi F, Barzan L, Ramazzotti V, Franceschi S, Negri E. Family history and the risk of oral and pharyngeal cancer. Int. J. Cancer, 122: (2008) La Vecchia C. Coffee and liver cancer prevention: is it a fact or just a potential? Hepatology, 48: 7-9 (2008). Pelucchi C, Dal Maso L, Montella M, Parpinel M, Negri E, Talamini R, Giudice A, Franceschi S, La Vecchia C. Dietary intake of carotenoids and retinol and endometrial cancer risk in an Italian case-control study. Cancer Causes Control., 19: (2008). Smedby KE, Vajdic CM, Falster M, Engels EA, Martinez-Maza O, Turner J, Hjalgrim H, Vineis P, Seniori Costantini A, Bracci PM, Holly EA, Willett E, Spinelli JJ, La Vecchia C, Zheng T, Becker N, De Sanjosé S, Chiu BC-H, Dal Maso L, Cocco P, Maynadié M, Foretova L, Staines A, Brennan P, Davis S, Severson R, Cerhan JR, Breen EC, Birmann B, Grulich AE, Cozen W. Autoimmune disorders and risk of non-hodgkin lymphoma subtypes: a pooled analysis within the InterLymph Consortium. Blood, 111: (2008). 196

198 Fustinoni S, Campo L, Liesivuori J, Pennanen S, Vergieva T, van Amelsvoort LGPM, Bosetti C, Vam Loveren H, Colosio C. Biological monitoring and questionnaire for assessing exposure to ethylenebisdithiocarbamates In a multicenter European field study. Human Exp. Toxicol., 27: (2008). Rossi M, Negri E, Bosetti C, Dal Maso L, Talamini R, Giacosa A, Montella M, Franceschi S, La Vecchia C. Mediterranean diet in relation to body mass index and waist-to-hip ratio. Publ. Health Nutr., 11: (2008). Galeone C, Pelucchi C, La Vecchia C, Negri E, Bosetti C, Hu J. Indoor air pollution from solid fuel use, chronic lung diseases and lung cancer in Harbin and Northeast China. Eur. J. Cancer Prev., 17: (2008). Bosetti C, McLaughlin JK, Tarone RE, Pira E, La Vecchia C. Formaldehyde and cancer risk: a quantitative review through Ann. Oncol., 19: (2008). Pelucchi C, Naldi L, Di Landro A, La Vecchia A, on behalf of the Oncology Study Group of the Italian Group for Epidemiologic Research in Dermatology (GISED). Anthropometric measures, medical history and risk of basal cell carcinoma in an Italian case-control study. Dermatology, 216: (2008) Naldi L., Chatenoud L., Belloni A., Peserico A., Balato N., Virgili A.R., Bruni P.L.,Ingordo V., Lo Scocco G., Solaroli C., Schena D., Di Landro A., Pezzarossa E., Arcangeli F., Gianni C., Betti R., Carli P., Farris A., Barabino G.F., La Vecchia C., Parazzini F. Medical history, drug exposure and the risk of psoriasis. Evidence from an Italian case-control study. Dermatology 216: , Lucenteforte E, Talamini R, Montella M, Dal Maso L, Tavani A, Deandrea S, Pelucchi C, Greggi S, Zucchetto A, Barbone F, Parpinel M, Franceschi S, La Vecchia C, Negri E. Macronutrients, fatty acids and cholesterol intake and endometrial cancer. Ann. Oncol., 19: (2008). Gallus S, Negri E, Boffetta P, McLaughlin JK, Bosetti C, La Vecchia C, European studies on long-term exposure to ambient particulate matter and lung cancer Eur. J. Cancer Prev., 17: (2008). La Vecchia C, Zhang ZF, Altieri A. Alcohol and laryngeal cancer: an update. Eur. J. Cancer Prev., 17: (2008). Levi F, Boffetta P, La Vecchia C. High constant incidence rates of second primary neoplasms. Eur. J. Cancer Prev., 17: (2008). Bagnardi V., Zatonski W., Scotti L., La Vecchia C., Corrao G: Does drinking pattern modify the effect of alcohol on the risk of coronary heart disease? Evidence from a meta-analysis. J. Epidemiol. Community Health, 62: (2008). Pelucchi C., Galeone C., Montella M., Polesel J., Crispo A., Talamini R., Negri E., Ramazzotti V., Grimaldi M., Franceschi S., La Vecchia C. Alcohol consumption and renal cell cancer risk in two Italian case-control studies. Ann. Oncol., 19: (2008). Randi G, Ferraroni M, Talamini R, Garavello W, Deandrea S, Decarli A, Franceschi S, La Vecchia C. 197

199 Glycemic index, glycemic load and thyroid cancer risk. Ann. Oncol., 19: (2008). Pandolfini C, Bonati M, RossiV, Santoro E, Choonara I, Naylor C, Sammons H, Jacqz-Aigrain E, Zarrabian S, Arnau JM, Castel JM, Danés I, Fuentes I. The DEC-net European register of paediatric drug therapy trials: its content and their context. Eur.J. Clin. Pharmacol., 64: (2008). Pelucchi C, Gallus S, Garavello W, Bosetti C, La Vecchia C. Alcohol and tobacco use, and cancer risk for upper aerodigestive tract and liver. Eur. J. Cancer Prev., 17: (2008). Bosetti C, Levi F, Ferlay J, Garavello W, Lucchini F, Bertuccio P, Negri E, La Vecchia C. Trends in oesophageal cancer incidence and mortality in Europe. Int. J. Cancer, 122: (2008) Num. Reg.: Pedrazzini G, Santoro E, Latini R, Fromm L, Franzosi M G, Mocetti T, Staszewsky L, Barlera S, Tognoni G, Maggioni A P, GISSI-3 Causes of death in patients with acute myocardial infarction treated with angiotensinconverting enzyme inhibitors: Findings from the Gruppo Italiano per lo Studio della Sopravvivenza nell'infarto (GISSI)-3 trial Am. Heart J., 155 : (2008). Garavello W., Giordano L., Bosetti C., Talamini R., Negri E., Franceschi S., La Vecchia C. Diet diversity and risk of oral and pharyngeal cancer risk. Eur. J. Nutrition, 47: (2008). Hu J, La Vecchia C, DesMeules M, Negri E, Mery L, and the Canadian Cancer Registries Epidemiology Research Group. Meat and fish consumption and cancer in Canada. Nutr. Cancer, 60: (2008). Bidoli E, Talamini R, Zucchetto A, Polesel J, Bosetti C, Negri E, Maruzzi D, Montella M, Franceschi S, La Vecchia C. Macronutrients, fatty acids, cholesterol and renal cell cancer risk. Int. J. Cancer, 122: (2008). Willett EV, Morton LM, Hartge P, Becker N, Bernstein L, Boffetta P,Bracci P, Cerhan J, Chiu BCH, Cocco PL, Dal Maso L, Davis S, De Sanjose S, Smedby KE, Ennas MG, Foretova L, Holly EA, La Vecchia C, Matsuo K, Maynadie M, Melbye M, Negri E, Nieters A, Severson R, Slager SL, Spinelli JJ, Staines A, Talamini R, Vornanen M, Weisenburger DD, Roman E for the Interlymph Consortium. Non-Hodgkin lymphoma and obesity: a pooled analysis from the InterLymph consortium. Int. J. Cancer, 122: (2008). Iacobelli N, Gallus S, Petridou E, Zuccaro P, Colombo P, Pacifici R, La Vecchia C, Negri E. Smoking behaviors and perceived risk of injuries in Italy, Prev. Med., 47: (2008). Petridou ET, Pourtsidis A, Dessypris N, Katsiardanis K, Baka M, Moschovi M, Polychronopoulou S, Koliouskas D, Sidi V, Athanasiadou-Piperopoulou F, Kalmanti M, Belechri M, La Vecchia C, Curado MP, Skalkidis I. Childhood leukaemias and lymphomas in Greece ( ): a nationwide registration study. Arch. Dis. Childhood., 93: (2008). Polesel J, Talamini R, La Vecchia C, Levi F, Barzan L, Serraino D, Franceschi S, Dal Maso L. Tobacco smoking and ther risk of upper aero-digestive tract cancers: A re-analysis of case-control studies using spline models. Int. J. Cancer, 122: (2008). Bosetti C, Bertuccio P, Levi F, Lucchini F, Negri E, La Vecchia C. Cancer mortality in the European Union, , with a jointpoint analysis. Ann. Oncol., 19: (2008). 198

200 Lagiou P, Rossi M, Lagiou A, Tzonou A, La Vecchia C., Trichopoulos D. Flavonoid intake and liver cancer: a case-control study in Greece. Cancer Causes Control, 19: (2008). Lucenteforte E, Scita V, Bosetti C, Bertuccio P, Negri E, La Vecchia C. Food groups and alcoholic beverages and the risk of stomach cancer: a case-control study in Italy. Nutr. Cancer, 60: (2008). Ferketich AK, Gallus S, Colombo P, Fossati R, Apolone G, Zuccaro P, La Vecchia C. Physician-delivered advice to quit smoking among Italian smokers Am. J. Prev. Med., 35: (2008). Gnagnarella P, Gandini S, La Vecchia C, Maisonneuve P. Glycemic index, glycemic load and cancer risk: a meta-analysis. Am. J. Clin. Nutr., 87: (2008) Edefonti V, Decarli A, La Vecchia C, Bosetti C, Randi G. Franceschi S, Dal Maso L, Ferraroni M. Nutrient dietary patterns and the risk of breast and ovarian cancers. Int. J. Cancer, 122: (2008) Levi F, Ferlay J, Galeone C, Lucchini F, Negri E, Boyle P, La Vecchia C. The changing pattern of kidney cancer incidence and mortality in Europe. BJU Int., 101: (2008). Gallus S, Naldi L and the Oncology Study Group of the Italian Group for Epidemiologic Research in Dermatology (GISED). Distribution of congenital melanocytic naevi and congenital naevus-like naevi in a survey of 3406 Italian schoolchildren Br. J. Dermatol., 159: (2008). Corrao G, Zambon A, Conti V, Nicotra F, La Vecchia C, Fornari C, Cesana G, Contiero P, Tagliabue G, Nappi RE, Merlino L. Menopause hormone replacement therapy and cancer risk: an Italian record linkage investigation. Ann. Oncol., 19: (2008). Deandrea S, Talamini R, Foschi R, Montella M, Dal Maso L, Falcini F, La Vecchia C, Franceschi S, Negri E. Alcohol and breast cancer risk defined by estrogen and progesterone receptor status: A case-control study. Cancer Epidemiol. Biomarkers Prev., 17: (2008). Foschi R, Lucenteforte E, Bosetti C, Bertuccio P, Tavani A, La Vecchia C, Negri E. Family history of cancer and stomach cancer risk. Int. J. Cancer, 123: (2008). Pelucchi C, Tavani A, La Vecchia C. Coffee and alcohol consumption and bladder cancer. Scand. J. Urol. Nephrol., 42 (Suppl. 218): Negri E, La Vecchia C, Collaborative Group on Epidemiological Studies of Ovarian Cancer. Ovarian cancer and oral contraceptives: collaborative reanalysis of data from 45 epidemiological studies including women with ovarian cancer and controls. Lancet 371: (2008). Rossi M, Negri E, Lagiou P, Talamini R, Dal Maso L, Montella M, Franceschi S, La Vecchia C. Flavonoids and ovarian cancer risk: A case-control study in Italy. Int. J. Cancer, 123: (2008). Levi F., Randimbison L., Te VC, Maspoli Conconi M., La Vecchia C. Risk of prostate, breast and colorectal cancer after skin cancer diagnosis. Int. J. Cancer, 123: (2008). 199

201 Boffetta P, McLaughlin JK, La Vecchia C, Tarone RE, Lipworth L, Blot WJ False-positive results in cancer epidemiology: a plea for epistemological modesty. JNCI, 100: (2008). Bosetti C, Levi F, Ferlay J, Lucchini F, Negri E, La Vecchia C. Incidence and mortality from non-hodgkin lymphoma in Europe: the end of an epidemic? Int. J. Cancer, 123: (2008). Num. Reg.: Malvezzi M, Bosetti C, Negri E, La Vecchia C, Decarli A. Cancer mortality in Italy, Tumori, 94: (2008). Poli A, Marangoni F, Paoletti R, MannarinoE, Lupattelli G, Notarbartolo A, Aureli P, Bernini F, Cicero A, Gaddi A, Catapano A, Cricelli C, Gattone M, Marrocco W, Porrini M, Stella R, Vanotti A, Volpe M, Volpe R, Cannella C, Pinto a, del Toma E, La Vecchia C, Tavani A, Manzato E, Riccardi G, Sirtori C, Zambon A. Non-pharmacological control of plasma cholesterol levels. Nutr. Metab. & Cardiovasc. Dis. 18: S1-S16 (2008). Bosis S, Esposito S, Niesters HGM, Zuccotti GV, Marseglia G, Lanari M, Zuin G, Pelucchi C, Osterhaus ADME, Principi N. Role of respiratory pathogens in infants hospitalized for a first episode of wheezing and their impact on subsequent recurrences. Clin. Microbiol. Infect., 14: (2008). Zucchetto A, Talamini R, Dal Maso L, Negri E, Polesel J, Ramazzotti V, Montella M, Canzonieri V, Serraino D, La Vecchia C, Franceschi S. Reproductive, menstrual, and other hormone-related factors and risk of renal cell cancer. Int. J. Cancer, 123: (2008). Ju J, La Vecchia C, DesMeules M, Negri E, L. Mery and the Canadian Cancer Registries Epidemiology Research Group., Nutrient and fiber intake and risk of renal cell carcinoma. Int. J. Cancer, 60: (2008). Lucenteforte E, Garavello W, Bosetti C, Talamini R, Zambon P, Franceschi S, Negri E, La Vecchia C. Diet diversity and the risk of squamous cell esophageal cancer. Int. J. Cancer, 123: (2008). Crispo A, D Aiuto G, De Marco MR, Rinaldo M, Grimaldi M, Capasso I, Amore A, Bosetti C, La Vecchia C, Montella M. Gail model risk factors: impact of adding an estended family history for breast cancer. Breast J., 14: (2008). Valentini LG, Casali C, Chatenoud L, Chiaffarino F, Uberti-Foppa C, Broggi G. Surgical site infections after elective neurosurgery: a survey of 1747 patients. Neurosurgery, 61: (2008). Boffetta P, McLaughlin JK, La Vecchia C. Reply: Environment in cancer causation and aetiological fraction: limitations and ambiguities. (by Boffetta, P et al. (2007) Carcinogenesis, 28, ). Carcinogenesis, 29: 1850 (2008). Dal Maso L, Zucchetto A, Talamini R, Serraino D, Stocco CF, Vercelli M, Falcini F, Franceschi S, for Prospective Analysis of Case-control studies on Environmental factors and health (PACE) study group, La Vecchia C, Negri E, Gallus S, Galeone C, Pelucchi C, Effect of obesity and other lifestyle factors on mortality in women with breast cancer. Int. J. Cancer 123: (2008) 200

202 Pelucchi C, Negri E. Bladder cancer. In: Heggenhougen IH, Quah S, eds., International Encyclopedia of Public Health, First Edition, San Diego, Academic Press, 1: (2008). Parazzini F, Cipriani S, Bianchi S, Gotsch F, Zanconato G, Fedele L. Risk factors for deep endometriosis: a comparison with pelvic and ovarian endometriosis. Fertil. Steril., 90: (2008). Naldi L, Addis S, Chimenti S, Giannetti A, Picardo M, Tomino C, Maccarone M, Chatenoud L, Bertuccio P, Caggese E, Cuscito R and the Psocare Study Centres. Impact of body mass index and obesity on clinical response to sistemic treatment for psoriasis. Dermatology,, 217: (2008). Boers D, van Amelsvoort L, Colosio C, Corsini E, Fustinoni S, Campo L, Bosetti C, La Vecchia C, Vergieva T, Tarkowski M, Liesivuori J, Steerenberg P, van Loveren H. Asthmatic symptoms after exposure to ethylenebisdithiocarbamates and other pesticides in the Europit field studies. Human Exp. Toxicol., 27: (2008). Van Almelsvoort LGPM, Mohren DCL, Slangen JJ, Swaen G, Corsini E, Fustinoni S, Vergieva T, Bosetti C, Liesivuori J, Tarkoeski M, Colosio C, van Loveren H. Immune effects and exposure to ethylenebisdithiocarbamate pesticides in re-entry workers in the Netherlands. Human Exp. Toxicol., 27: (2008). Swaen GMH, van Amelsvoort LGPM, Boers D, Corsini E, Fustinoni S, Vergieva T, Bosetti C, Pennanen S, Liesivuori J, Colosio C, van Loveren H. Occupational exposure to ethylenebisdithiocarbamates in agriculture and allergy: results From the EUROPIT field studies. Human Exp. Toxicol., 27: (2008). Negri E. Message 3: Prevent falls. Arch. Ellenic med., 25 (S 1): (2008). 201

203 LAY PRESS SELECTION PUBLISHED IN 2008 Santoro E. I podcast al servizio della formazione e dell'aggiornamento del cardiologo. Recenti Prog Med : Santoro E. Health-e-child e il grid computing in ambito sanitario: un nuovo supporto alla dignostica e alla clinica. SIAR News : Santoro E. Tecnologia RSS e aggregatori di notizie. Ricerca & Pratica 2008 n.139: Santoro E. I podcast nell'aggiornamento professionale del medico (parte I) Ricerca & Pratica 2008 n.140: Santoro E. I podcast nell'aggiornamento professionale del medico (parte II) Ricerca & Pratica 2008 n.141: Santoro E. Le cartelle cliniche personali in rete. Un nuovo modo di controllare le proprie informazioni sanitarie? Ricerca & Pratica 2008 n.142: Santoro E. Social network e ricercatori biomedici. Ricerca & Pratica 2008 n.143: Santoro E. I blog e l aggiornamento medico. Ricerca & Pratica 2008 n.144: Santoro E. Web 3.0 e medicina. CARE : 6-7 Santoro E. Web 3.0 e medicina: un nuovo web all'orizzonte? Partecipasalute 2008; Santoro E. Cartelle cliniche informatizzate: un aiuto alla pratica clinica e alla ricerca medica. Partecipasalute 2008; Santoro E. Le cartelle cliniche personali. Partecipasalute 2008; Santoro E. Google health e le cartelle sanitarie on line: una nuova sfida per Google. Partecipasalute 2008; Santoro E. Cartelle cliniche informatizzate, standard e interoperabilità: un aiuto alla pratica clinica e alla ricerca medica. SIAR News : Santoro E. Google Flu Trends e la previsione di epidemie. Partecipasalute 2008; OTHER PRODUCTS PUBLISHED IN 2008 Pistotti V, Santoro E. Navigare sulla rete alla ricerca di informazioni di salute. In: La dispensa di Partecipasalute: orientarsi in salute e sanità per fare scelte consapevoli. IRFMN, Milano, 2008;

204 RESEARCH ACTIVITIES LABORATORY OF GENERAL EPIDEMIOLOGY Case-control studies on cancer The collection and analysis of data from case-control studies on cancer have continued in This is an integrated research program, designed to identify and quantify the role of environmental, genetic factors, and their interaction on several common neoplasms, with specific reference to the Italian population. The program started in the early 1980's and has generated, during the last five years, over 200 publications. The project has evolved by continuously updating the datasets, by refining and validating the data collection instruments, by developing and integrating a detailed food composition database and introducing a larger number of relevant covariates. Further developments are on-going, including follow-up of subjects through a record-linkage approach and collection and analysis of biological samples. The dataset includes now about: 6,500 cases of colorectal and breast cancers; over 1,000 of oral and pharyngeal, stomach, endometrial, ovarian, prostate and kidney cancers; several hundred cases of other common neoplasms, and over 19,000 controls. Collection of biological samples has been pursued for new studies on several cancer sites, including laryngeal, bladder and colorectal cancer, which will allow to analyze selected genetic polymorphisms, insulin-like growth factors (IGF), adiponectin and other biomarkers. The results of the main analyses conducted within 2008 are summarized below. Tobacco smoking and upper aero-digestive tract cancers: a reanalysis Although tobacco smoking has long been recognized as a major risk factor for cancer of the upper aero-digestive tract (UADT, i.e., oral cavity, pharynx, larynx, and oesophagus), only a few studies provided estimates of the effect of very low tobacco consumption. We evaluated the dose-response relationship between UADT cancers and tobacco smoking using logistic regression spline models. We included 1,241 UADT male cases and 2,835 male controls pooled from a large series of case-control studies conducted in northern Italy and in the Swiss Canton of Vaud during the last 2 decades. For cancers of the pharynx, larynx and oesophagus the risk steadily increased with number of cigarettes/day and was significantly higher already for smokers of 2 cigarettes/day as compared to nonsmokers. The effect of tobacco smoking at low levels seemed less evident for laryngeal cancer since the excess risk begun with 6 cigarettes/day. In conclusion, for all the examined UADT sites, a dose-response relationship between cancer risk and cigarette smoking emerged. The excess risk among people smoking 2 cigarettes/day highlights the absence of any harmless level for cigarette smoking. Diet diversity and upper aero-digestive tract cancers We analyzed the role of diet diversity (the variety of foods consumed) on the risk of UADT cancers, including oral and pharyngeal, oesophageal and laryngeal cancer. The study of oral and pharyngeal cancer included 805 cases and 2,081 controls. A significant inverse association was observed with total diet diversity: the multivariate odds ratio (OR) was 0.78 for subjects in the highest tertile of diversity. Inverse relations were found also for diversity within vegetables (OR = 0.62) and fruits (OR = 0.67). The study of squamous cell esophageal cancer included 304 cases and 743 controls. ORs in the highest quartile of intake were 0.42 for total diversity, 0.34 for vegetable diversity and 0.51 for fruit diversity. The study of laryngeal cancer included 527 cases and 1297 controls. A significant inverse association was observed for vegetable diversity 203

205 (OR=0.41 for the highest versus the lowest quartile) as well as for fruit diversity (OR=0.40). Conversely, a direct association was found for meat diversity (OR=1.67). Dietary factors and oral and pharyngeal cancer We reviewed data from six cohort studies and approximately 30 case-control studies on the relation between selected aspects of diet and the risk of oral and pharyngeal cancer. The pooled relative risk (RR) for high vegetable consumption were 0.65 from three cohort studies on UADT cancer, and 0.52 from 18 case-control studies of oral and pharyngeal cancer. Corresponding RRs for fruit consumption were 0.78, and Family history and gastric cancer The relation between family history of cancer in first-degree relatives and the risk of stomach cancer has been considered in our study including 230 cases and 547 matched controls. Relative to subjects with no history, those with a family history of gastric cancer had an OR of gastric cancer of 2.5. The OR was higher when the affected relative was a sibling (OR = 5.1) rather than a parent (OR = 2.2), although the heterogeneity test was not significant. Food groups, micronutrients and gastric cancer In the same study of gastric cancer, we examined the relation with selected dietary factors. A direct association was observed for cereals (OR=2.07 for the highest compared to the lowest quintile of intake), soups (OR=1.94), and potatoes (OR=2.04). Conversely, inverse relations were observed with vegetables (OR=0.47) and fruit intake (OR=0.53). No specific constituent of plant foods has been consistently identified to explain the inverse association between nonstarchy vegetables and fruit and gastric cancer. In our study, we estimated micronutrient intake using information from a validated and reproducible food frequency questionnaire, through an Italian food composition database. We found decreased ORs for the highest vs. lowest quartile of vitamin E (OR = 0.50), alpha-carotene (OR = 0.52) and beta-carotene (OR = 0.42) intake. Gastric cancer was directly associated with sodium, with ORs of 2.22 for the second, 2.56 for the third and 2.46 for the fourth quartile of intake. No significant relation emerged with iron, calcium, potassium, zinc, vitamin C, thiamin, riboflavin, niacin, vitamin B6, folate, vitamin D, retinol, beta-cryptoxanthin, lycopene and lutein plus zeaxanthin. Vitamin D and colorectal cancer Epidemiologic evidence indicates that vitamin D is inversely associated with risk of colon or rectal cancer or both. In our analyses on 1953 cases and 4154 controls, we found that adjusted ORs for colon cancer decreased after the fifth decile of vitamin D intake, reaching 0.69 for the ninth and tenth deciles. The inverse association appeared to be somewhat more pronounced for the proximal than the distal colon, and was similar among strata of geographic region and calcium intake. Rectal cancer was unrelated to vitamin D intake in this population. Vitamin D and breast cancer risk We investigated whether vitamin D was associated with risk of breast cancer in our study including 2569 cases of breast cancer and 2588 hospital controls. After allowance for major risk factors for breast cancer and dietary covariates, including calcium and energy intake, there was no association up to the seventh decile between vitamin D intake and breast cancer. Thereafter, the OR declined, so that the overall trend was statistically significantly inverse. The OR for subjects in the three highest deciles of consumption of vitamin D compared with those in the lowest ones combined was 0.79 (95% CI, ). Only intake of vitamin D over 3.57 μg or 143 IU appeared to have a protective effect against breast cancer. 204

206 Indoor air pollution from solid fuel use and lung cancer In some areas of China, indoor air pollution (IAP) originating principally from the combustion of solid fuels has a relevant role in lung cancer. We analyzed the relationship between IAP from solid fuel use and selected chronic lung diseases and lung cancer risk in Harbin, Northeast China, an area with a very high baseline risk of lung cancer for both sexes. Data included 218 patients with incident, histologically confirmed lung cancer and 436 hospital controls. We calculated an index of IAP from solid fuel use exposure using data on heating type, cooking fuel used, and house measurements. Cases reported more frequently than controls an exposure to coal fuel for house heating and/or cooking, and the OR for ever vs. never exposed was The ORs of lung cancer according to subsequent tertiles of IAP exposure index were 1.82 and 1.99 as compared with the lowest tertile. The ORs of lung cancer for participants with a history of chronic bronchitis and tuberculosis were 3.79 and 3.82, respectively. This study gave further support and quantification of the positive association between IAP, history of selected nonmalignant lung diseases, and lung cancer risk for both sexes. Gail model risk factors for breast cancer We evaluated the feasibility and impact of adding an extended family history as a new breast cancer risk factor into the Gail model using data from a hospital-based case-control study conducted by the National Cancer Institute of Naples (southern Italy) between 1997 and We compared the model with a first-degree relative (FDR) (standard Gail model) with a model with information on second-degree relative (SDR); and the FDR with a model with the combination of FDR and SDR. We computed the C-statistic by comparing the risks found in our population to those in the Gail-US population. The concordance for the model with FDR was 0.55; the model with SDR showed a modest but significant discriminatory accuracy (0.56), and the model with the combination of FDR and SDR gave a concordance statistic of 0.57, indicating a good comparison between the two models. The results of the study showed that extended family history information could be useful to improve the discriminatory power of the Gail model risk factors. Alcohol and breast cancer, according to estrogen receptor status Studies suggested that the association between alcohol consumption and breast cancer risk is stronger or limited to tumors expressing estrogen receptors (ER). We considered the issue using data from a subset of subjects (989 cases and 1350 controls) from our breast cancer study, for which biological information was available. Alcohol drinking was significantly associated with ER+ tumors (OR = 2.16 for an intake of 13.8 g/day as compared with nondrinkers), but not with ER- tumors (OR = 1.36). When breast cancers were further classified according to progesterone receptors (PR), the findings for ER+PR+ cancers were similar to those for all ER+ ones, with an OR of 2.34 for an intake of 13.8 g/d. No significant association emerged for ER- PR- tumors (OR = 1.25). Consistently with previous data, in this study alcohol was more strongly related to ER+ than to ER- breast tumors. Diet and endometrial cancer We analyzed data from a case-control study on endometrial cancer, conducted during in three areas of Italy, including 454 cases and 908 controls, to investigate the relationship between various aspects of diet and risk of endometrial cancer. With reference to main food groups, we found a significant direct association with consumption of red meat, and a moderate increase in risk for consumption of sweets and poultry. On the other hand, a high consumption of cereals and vegetables decreased the risk of endometrial cancer by 44% and 41%, respectively, while other food groups investigated were not associated with the risk of this cancer. Another study considered the role of allium vegetables, which were found to be inversely related with risk of various other cancers in an earlier Italian study. For endometrial 205

207 cancer too, we found significant inverse associations with consumption of both onions (OR = 0.40 for the highest vs. lowest level of consumption) and garlic (OR = 0.62). With reference to micronutrients, a selection of carotenoids was inversely related with risk of endometrial cancer. In particular, we found a beneficial effect for high levels of beta-carotene (OR = 0.69), betacryptoxanthin (OR = 0.65) and lutein/zeaxanthin (OR = 0.59), while other carotenoids (such as alpha-carotene and lycopene) and micronutrients were unrelated with endometrial cancer. In conclusion, this study suggested that diet might have an important role on endometrial cancer, and in particular that a diet rich in meat and poor in vegetables and related micronutrients could have an adverse effect. Family history of cancer and endometrial cancer risk With reference to family history of cancer, our questionnaires had detailed information on history of all cancers in first-degree relatives and thus allowed a thoroughly investigation of the relation with endometrial cancer risk. The OR of endometrial cancer for women with at least one first-degree relative diagnosed the same neoplasm was 2.13, while those who had relatives with a diagnosis of uterine and colorectal cancer had OR of endometrial cancer of 1.76 and 1.62, respectively. Risks were also increased in women younger than 55 years at diagnosis (OR = 2.60 for family history of uterine cancer, OR = 2.79 for family history of colorectal cancer). We found direct associations with some other cancer sites, while there was no association with the family history of breast cancer. Thus, our study confirmed that a family history of endometrial, uterine and colorectal cancer increases the risk of endometrial cancer. Flavonoids and ovarian cancer Flavonoids intake has been associated with a decreased risk of various cancers in a number of epidemiological studies. We considered the issue in relation to ovarian cancer, using data from an Italian hospital-based case-control study including a total of 1031 cases and 2411 controls. We found an inverse relation between flavonols (OR = 0.63 for the highest vs. lowest quintile) and isoflavones (OR = 0.51) and ovarian cancer, with significant trend in risk. Adjustment for fruit and vegetable intake did not modify these associations, suggesting that isoflavones and flavonols may have a distinct role in explaining the effect of fruit and vegetable against ovarian cancer. Oral contraceptives and ovarian cancer Data of our case-control study of ovarian cancer were included in a collaborative reanalysis of 45 epidemiological studies from 21 countries, for a total of 23,257 women with ovarian cancer (cases) and 87,303 without ovarian cancer (controls). These data were analysed to assess the relation between oral contraceptives (OC) use and ovarian cancer. Though an inverse association is well known, the public-health effects of this reduction in risk will depend on how long the protection lasts after use ceases. Overall 7308 (31%) cases and 32,717 (37%) controls had ever used OC, for average durations among users of 4.4 and 5.0 years, respectively. The longer that women had used OC, the greater the reduction in ovarian cancer risk (p<0.0001). This reduction in risk persisted for more than 30 years after OC use had ceased but became somewhat attenuated over time. The incidence of mucinous tumours (12% of the total) seemed little affected by OC, but otherwise the proportional risk reduction did not vary much between different histological types. It was estimated that OC already prevented some 200,000 ovarian cancers and 100,000 deaths from the disease, and that over the next few decades the number of cancers prevented will rise to at least 30,000 per year. Macronutrients, fatty acids and cholesterol and renal cell cancer Using data elicited from a validated food frequency questionnaire, including 78 food groups and recipes, we examined the role of selected macronutrients, fatty acids and cholesterol in the 206

208 etiology of renal cell cancer (RCC). We found a direct association with starch intake (OR = 1.9 for highest vs. lowest quintile of intake), and an inverse association with fats from vegetable sources (OR = 0.6), unsaturated fatty acids (OR = 0.5) and polyunsaturated fatty acids (OR = 0.5), particularly linoleic acid (OR = 0.5) and linolenic acid (OR = 0.7). When 6 major macronutrients were included in the same model, the adverse effect of high intake of starch remained statistically significant, together with the protective effect of polyunsaturated fatty acids. Reproductive and hormone-related factors and renal cell cancer Using information from our study of RCC, including 273 female cases and 546 female controls, we evaluated the effect of reproductive, menstrual and other gender-specific variables among women. RCC risk was inversely related to age at first birth (OR = 0.7, for 25 vs. <25 years), whereas hysterectomy increased more than 2-fold the risk of RCC (OR = 2.3). No associations were found with parity, menopausal status, age at menopause and use of hormone replacement therapy or oral contraceptives. Our findings supported a role of hysterectomy in increasing RCC risk without corroborating, however, a major role of female hormone-related factors. Glycaemic index and load and cancer risk We were involved in a meta-analysis of published studies on the factors linked to glucose metabolism, glycemic index (GI) and glycemic load (GL), and cancer risk. Thirty-nine studies were included in the meta-analysis. Overall, both GL and GI were significantly associated with a greater risk of colorectal (summary RR=1.26 and RR=1.18, respectively) and endometrial (RR=1.36 and RR=1.22) cancer than of breast and pancreatic cancer. The association between GL and breast cancer disappeared when publication bias was taken into account. No association was found for pancreatic cancer. Relation between Mediterranean diet and obesity The Mediterranean diet is rich in fat and starch, and hence may be related to overweight. We therefore investigated the relation between adherence to a Mediterranean diet and body mass index (BMI) and waist-to-hip ratio (WHR) using data from the control group of our network of case-control studies on cancer, including 6619 subjects (3090 men, 3529 women). A Mediterranean diet score (MDS) was derived on the basis of eight characteristics of the Mediterranean diet. In multiple linear regression models, the MDS was not related to BMI (beta = 0.05 for men and for women) or WHR (beta = and 0.001, respectively) in both sexes. Thus, in our study adherence to the major characteristics of the Mediterranean diet was unrelated to BMI and WHR, confirming previous data from Greece and Spain. Smoking prevalence in Italy in 2008 A survey was conducted in 2008 on a sample of 3,035 individuals (1,459 men and 1,576 women) aged 15 years or over representative of the Italian population in terms of age, sex, geographic area, and socio-economic characteristics. In 2008, 22.0% of Italians described themselves as current cigarette smokers (26.4% of men, 17.9% of women); ex-smokers were 18.4% (24.1% of men, 13.2% of women). By the year 2012 the number of former could exceed that of current smokers. The smoking prevalence in the young (15-24 years) was around 30% in males, and almost 20% in females. For both sexes, current smoking was less prevalent in higher (22.9% of men, 20.1% of women) than in lower educated participants (34.8% of men, 22.1% of women), and in northern (22.5% of men, 16.1% of women) than southern Italy (31.8% of men, 18.4% of women). In 2008 smoking prevalence was the lowest observed over the last 50 years, in Italy. Subjects with less privileged socio-economic characteristics should be considered target populations for tobacco control. 207

209 HI-WATE project on colorectal cancer and drinking water by-products In 2008, the Department started the collection of data for a case-control study on colorectal cancer within a project of the 6-FP on the Health Impact of long term exposure to disinfection by-products in drinking water (DBPs) (HI-Wate study). The study is conducted in the greater Milan area and in the Provinces of Pordenone and Udine, and includes incident, histologically confirmed cases enrolled in the major general and teaching hospital of the study area, and frequency-matched controls, admitted to the same hospital as cases for acute, nonneoplastic conditions. Cases and controls are interviewed using an extensive questionnaire. Detailed information on water use and water-related habits for etiologically relevant time periods is collected, including not only water consumption, but also water related activities, such as showering, bathing and swimming, that may influence exposure assessment. Moreover, in order to assess the subjects exposure to DBPs and THMs in water, current and historical THM levels, water source and year of starting chlorination in the study area will be collected from local companies, authorities and municipalities. About 250 colorectal cancer cases and 250 corresponding controls have been collected, i.e., about half of the total number of cases and controls which should be enrolled at the end of the study. Giving the large number of people exposed to chlorinated drinking-water and its DBPs, the study has potentially important implications in terms of public health. Even modest excess risk in relation to DBP exposure may in fact have a relevant impact on a population level, and may be responsible of a considerable number of colorectal cancer cases. Health status of a sample of illegal immigrants in the municipality of Milan from 2005 to 2008 We conducted an analysis on a sample of 20,800 immigrants (of which 85% did not have a regular VISA) collected by a Consultorio from Milan over the period From this analysis we observed that the diseases most frequently recorded in this sample were various symptoms, such as asthenia, paresis, headache (ICD-9: ), followed by arthritis and backpain (found in 10-11% of subjects). It was also noted that the subjects who do not have a regular VISA tend to have a higher frequency of acute infections of the upper respiratory tract and diseases of the esophagus, stomach and duodenum (about 10% of the 16,033 irregular immigrants, compared to around 6% of those in possession of a regular VISA). There were no other important differences in the frequencies of disease between these two groups of immigrants. Impact of body mass index and obesity on clinical response to systemic treatment for Psoriasis. Evidence from the PSOCARE project To assess the role of BMI on early clinical response to systemic treatment for psoriasis, we analyzed data from a nationwide registry and cohort study involving compelling registration of patients receiving for the first time in their life a new systemic treatment for psoriasis at reference centres in Italy. Information was gathered by treating physicians with the aid of a webbased electronic form. Patients with a clinical diagnosis of chronic plaque psoriasis and with 8 and 16 weeks of completed follow-up by March 31 th, 2007 were eligible. A relative reduction of Psoriasis Area Severity Index (PASI) of at least 75% at follow-up compared to baseline was evaluated as clinical endpoint (PASI-75). A total of 2,368 patients were analysed at 8 weeks and 2,042 at 16 weeks. PASI-75 was achieved by 819 (34.5%) patients at 8 weeks and 1,034 (50.6%) patients at 16 weeks. The proportion steadily decreased with increased values of BMI, from 41.7% in patients with BMI < 20 to 29.1% in patients with BMI 30 at 8 weeks, and from 59% in patients with BMI <20 to 42.2% in patients with BMI 30, at 16 weeks. Compared to normal-weight (BMI 20-24), obese patients (BMI 30) had a lower chance of improving at 8 and 16 weeks, adjusted ORs for achieving PASI-75 being 0.73 and 0.62, respectively. Thus, BMI affects early clinical response to systemic treatment for psoriasis irrespectively of the administered drug. 208

210 Distribution of congenital melanocytic naevi and congenital naevus-like naevi in a survey of 3406 Italian children To estimate the prevalence of congenital melanocytic naevi (CMN) and congenital naevus-like naevi (CNLN) in Italian schoolchildren, and to assess variations according to potential risk factors for melanoma, we conducted a survey in 13 Italian areas on 3406 schoolchildren aged years. Children were examined by dermatologists who made a count of small (6-15 mm in diameter) and medium/large (> 15 mm) CMN/CNLN on 19 anatomical areas. Overall, 592 children (17.4%) had one or more CMN/CNLN. Prevalence of small CMN/CNLN was 16.1%, and that of medium/large CMN/CNLN was 1.8%. There was no difference between age groups and sexes. CMN/CNLN were more frequent in children with a higher number of common melanocytic naevi (multivariate odds ratio, OR = 7.1 for the highest vs. the lowest quartile). Family history of malignant melanoma (OR = 1.4) and personal history of diabetes (OR = 4.4) appeared to be directly, and sun exposure inversely associated with CMN/CNLN. No relation was evident between CMN/CNLN and pigmentary traits, anthropometric characteristics, dietary habits, freckles, sunburns, sunscreen use or history of selected diseases. Public health prevention and information The major products of our activity have also been published in the lay press, in order to increase the project impact on prevention and public health. LABORATORY OF EPIDEMIOLOGICAL METHODS Strategies and best practices for the reduction of injuries Since 2006, our Department is involved in the Apollo project, an European project aiming at identifying strategies and best practices for the reduction of injuries. Within this project, our Department is coordinator of the Workpackage 4 on the development and implementation of recommendations for the prevention of falls among elderly people in the European Union (EU). The majority of falls in elderly people are due to a combination of several interacting factors. Many studies have investigated the role several factors on the risk of falling. Thus we conducted a systematic review and a quantitative meta-analysis of risk factors for falls in communitydwelling elderly people, including relevant studies published up to Fifty-nine studies were included and 28 risk factors were analyzed, including socio-demographic characteristics, mobility, sensory, psychological and medical factors, and medication use. The strongest associations were found for history of falls, vertigo, gait problems, use of walking aids, and use of antiepileptics. Moreover, we investigated the amount of information on the participation rates reported from studies investigating the effectiveness of selected intervention for the prevention of falls among community-dwelling elderly people. We identified 32 studies implementing interventions based on an exercise program, an exercise program in combination with other measures, or other types of interventions. Fourteen studies did not report information on the refusal rate; most studies reporting the information had a refusal rate between 25 and 50%. We also conducted a survey to investigate the attitudes of elderly people towards selected intervention for the prevention of falls. The study was conducted in Italy, Poland, Greece, Hungary, and Slovenia, countries for which limited data is available on this issue. A total of 1497 subjects aged 65 to 89 years were interviewed to evaluate their beliefs and attitudes towards two evidence-based interventions, i.e. a social activity aimed at improving muscle strength and balance (such as exercise class or dancing), and a home safety assessment and modification program. Among the respondents about 47% would accept to participate to a social activity, and about 38% would accept a 209

211 program of home hazard assessment and modification. However, we found marked differences between countries in the acceptability of the two proposed interventions. Monitoring of cancer mortality in Europe In 2008, we conducted various analyses within a project launched in 1992 for the monitoring of cancer mortality in Europe and other areas of the world. The main objective of the project is to maintain and improve an integrated system for analysis, modeling and interpretation of death statistics in Europe, created by our group. The project is based on mortality data of the WHO, and includes the number of cancer deaths in many countries, sex, cause, period, and age of death in Europe and some other countries outside Europe, together with the estimates of the resident population. The project is not only descriptive, as a specific effort is devoted to the interpretation of the observed data on the basis epidemiological evidence. It offers a unique opportunity for the continued exploitation of mortality data in Europe, with the primary aim of improving the monitoring and prevention of cancer by optimizing the allocation of economic resources in health. Cancer mortality in the European Union We analyzed cancer mortality in the EU over the period Overall, cancer mortality leveled off in men since 1988 and declined in (annual percent change, APC = - 1.3%). In women, a steady decline has been observed since the early 1970s. The decline in male cancer mortality has been driven by lung cancer, which leveled off since the late 1980s and declined thereafter (APC = 2.7% in ). Recent decreases were also observed for other tobacco-related cancers, as oral cavity/pharynx, esophagus, larynx and bladder, as well as for colorectal and prostate cancers. In women, breast cancer mortality leveled off since the early 1990s and declined thereafter (APC = -1.0% in ). Female mortality declined through the period for colorectal and uterine cancer, while it increased over the last three decades for lung cancer (APC = 4.6% in ). In both sexes, mortality declined in for stomach cancer and for a few cancers amenable to treatment. This update analysis of the mortality from cancer in the EU shows favorable patterns over recent years in both sexes. Cancer mortality in Italy We updated a previous work on Italian cancer mortality with data for 2002 and analyzed trends over the period. Total cancer deaths for 2002 in Italy were 163,070 (93,398 men, 69,672 women). Male cancer mortality rose until 1988 and since then has had a 1.4% yearly fall. The first cause of cancer death in males was lung cancer, accounting for 28% of deaths. The decrease in mortality from male lung cancer started in the late 1980 s and was the main reason for the favorable trends in total male cancer mortality, reflecting the change in smoking prevalence in Italian males. Female total cancer mortality trends have also been favorable, with an overall yearly drop of 1.1% since The most frequent causes of cancer deaths in females were breast and colorectal cancers, accounting for 16% and 14% of cancer deaths, and both showed declining trends. Female lung cancer rose over the last decades because of the increase in cigarette smoking since the 1970's in Italian women. Thus, mortality from the most common cancers in Italy showed a favorable trend over recent years. Incidence and mortality from non-hodgkin lymphoma We updated trends in mortality from non-hodgkin lymphomas (NHL) considering data up to 2004 in several European countries, and for comparative purpose in the USA and Japan. We also analyzed patterns in incidence for selected European countries providing national data. In most European countries, NHL mortality rose up to the mid 1990s, and started to level off or decline in the following decade. The rates were, however, still increasing in eastern Europe. Overall, in the EU, mortality from NHL declined from 4.3/100,000 to 4.1 in men and from

212 to 2.5 in women between the late 1990s and the early 2000s. Similarly, NHL mortality rates declined from 6.5/100,000 to 5.5 in US men and from 4.2 to 3.5 in US women. In most countries considered, NHL incidence rates rose up to , while they tended to level off or decline thereafter, with particular favorable patterns in countries from northern Europe. Thus, the epidemic of NHL observed during the second half of the 20th century has now started to level off in Europe as in other developed areas of the world. Trends in mortality from hepatocellular carcinoma in Europe Age-standardized (world standard) mortality rates from hepatocellular carcinoma (HCC) were computed for 23 European countries over the period Male overall mortality from HCC increased in Austria, Germany, Switzerland, and other western countries, while it significantly decreased over recent years in countries such as France and Italy, which had large upward trends until the mid-1990s. In the early 2000s, among countries allowing distinction between HCC and other liver cancers, the highest HCC rates in men were in France (6.8/100,000), Italy (6.7), and Switzerland (5.9), whereas the lowest ones were in Norway (1.0) and Ireland (0.8). In women, a slight increase in overall HCC mortality was observed in Spain and Switzerland, while mortality decreased in several other European countries, particularly since the mid-1990s. In the early 2000s, female HCC mortality rates were highest in Italy (1.9/100,000), Switzerland (1.8), and Spain (1.5) and lowest in Greece and Ireland (0.3). HCC mortality remains largely variable across Europe. Favorable trends were observed in several European countries mainly over the last decade, particularly in women and in young adults. Trends in oesophageal cancer incidence and mortality in Europe We analyzed recent trends in mortality from oesophageal cancer in 33 European countries. For selected European cancer registration areas, we also analyzed incidence rates for different histological types. For men in the EU, mortality rates were stable around 6/100,000 between the early 1980 s and the early 1990 s, and slightly declined in the last decade (5.4/100,000 in the early 2000s, annual percent change, APC = -1.1%). In several western European countries, male rates have started to level off or decline during the last decade (APC = -3.4% in France, and - 3.0% in Italy). Also in Spain and the UK, which showed upward trends in the 1990 s, the rates tended to level off in most recent years. A leveling of rates was observed only more recently in countries of central and eastern Europe, which had had substantial rises up to the late 1990 s. Oesophageal cancer mortality rates remained comparatively low in European women, and overall EU female rates were stable around /100,000 over the last 2 decades (APC = - 0.1%). In northern Europe a clear upward trend was observed in the incidence of oesophageal adenocarcinoma, and in Denmark and Scotland incidence of adenocarcinoma in men is now higher than that of squamous-cell carcinoma. Squamous-cell carcinoma remained the prevalent histological type in southern Europe. Changes in smoking habits and alcohol drinking for men, and perhaps nutrition, diet and physical activity for both sexes, can partly or largely explain these trends. Declining mortality from bladder cancer We updated trends in bladder cancer mortality in 32 European countries and the EU as a whole, over the period In the EU overall, bladder cancer mortality rates (age-standardized, world standard population) were stable up to the early 1990 s at approximately 7/100,000 men and 1.5/100,000 women, and declined thereafter by approximately 16% in men and 12% in women, to reach values of 6 and 1.3/100,000, respectively, in the early years of the present decade. Over recent years, most countries showed decreasing trends. The favorable trends in men are partly or largely due to the recent declines in the prevalence of smoking in European men, together with reduced occupational exposure to occupational carcinogens. The decreases in women are more difficult to explain. Better control of urinary tract infections has probably 211

213 played a role, while the role of diet and other potential urinary tract carcinogens remains undefined. The changing pattern of kidney cancer incidence and mortality in Europe We updated trends in kidney cancer mortality in 32 European countries and the EU as a whole, over the period In men in the EU, mortality rates from kidney cancer peaked at 4.8 per 100,000 in , and declined to 4.1 (-13%) in In women in the EU, the corresponding values were 2.1 in and 1.8 (-17%) in The main decreases were in Scandinavian countries, and other western European countries. In most eastern European countries kidney cancer mortality rates tended to stabilize, even if values remained high, especially in the Czech Republic and Baltic countries. For kidney cancer incidence, there were decreases in rates for both sexes in Sweden throughout the 25-year calendar period considered. In the last 10 years considered, incidence rates decreased or tended to stabilize also in other northern European countries in both sexes, except in the UK. Record-linkage for cohort analyses With reference to our project to create an Italian cohort study based on record-linkage, during 2008 we prepared the archives that will allow to link data between our network of case-control studies (conducted since 1982) and those from the historical archives of the Local Health Unit (ASL) of Milan, that include several health information such as vital status and date of death of the subjects. Several stages of the project have been undertaken and completed during the year. In particular, we finished the identification of subjects recruited in the case-control studies among the historical archives of the ASL, and a database with univocal information on subjects from the two sources is now available, thus allowing a linkage of the archives; at the same time, we completed data collection from all case-control studies in a unique database (using original data and through the preparation of a codebook that allowed to re-codify studies conducted in various periods and on several diseases) that includes information from interviews to approximately 27,000 subjects. During 2008, we searched in various other historical databases individuals that were not linked in historical archives of the ASL. On 18,257 residents in Lombardy, 15,860 (86.9%) subjects, including 7,680 cases (82.2%) and 8,911 controls (91.8%) were linked. LABORATORY OF EPIDEMIOLOGY OF CHRONIC DISEASES Organization for data and biological sample collection for case-control studies Data collection of epidemiological data is going on and it includes: 1) interview and interviewer management and training activity for new interviewers; 2) contacts with hospital department and ethical committee for study approval and conduction; 3) check and codification of patient questionnaires; 4) diagnosis and histological exam check; 5) organization and management of biological sample collection; 6) data input management. Ongoing case-control studies include: cancer of the oral cavity, pharynx, larynx, esophaguscardias, biliary tract, colorectum, lymphomas, myelomas and sarcomas. The overall updated dataset include about: cases of cancers of oral cavity and pharynx, 700 of the esophagus, of the stomach, of the colorectum, 600 of the liver, 120 of the biliary tract, 600 of the pancreas, 850 of the larynx, 500 cutaneous malignant melanoma, of the breast, of the cervix, of the endometrium, 200 of trophoblastic gestational disease, 200 of the vulva, of the ovary, of the prostate, 700 of the bladder, 800 of the kidney and renal pelvis, 600 of the tyroid, 200 of Hodgkin disease, 500 of non-hodgkin 212

214 disease, 150 of sarcomas, 300 of myelomas and about controls. Biological sample collection includes cancers of the oral cavity, pharynx, larynx, bladder and colorectum aimed to study genetic polymorphisms. Definition and management of a cohort of subjects living at Asti for cardiovascular risk assessment This study is conducted in collaboration with the ALMA association and the Ordine dei Medici di Asti and includes data collection and first analyses of follow-up. Case-control studies Physical activity and endometrial cancer In the case-control study of endometrial cancer, we investigated the relation with physical activity at different ages. Taking into account various potential confounding factors, including age, body mass index, diabetes, hormonal/reproductive factors and energy intake, the OR of endometrial cancer in women at high level of occupational physical activity, as compared with those at the lowest level, were 1.69 at 12 years of age, 1.33 between 15 and 19 years, 1.17 between 30 and 39 years and 0.82 between 50 and 59 years. No significant trend in risk with level of occupational physical activity was found at any age. Similarly, no significant association was detected with recreational physical activity in different periods of life, since the OR for the highest vs. lowest level of physical activity were 0.82 at 12 years of age, 0.78 between 15 and 19 years, 1.12 between 30 and 39 years and 0.97 between 50 and 59 years. Therefore, our results ruled out a strong relation between physical activity and endometrial cancer. Coffee consumption and endometrial cancer Some studies found an inverse relation between coffee consumption and endometrial cancer risk. We conducted a meta-analysis of published studies on this issue, including 2 cohort (201 cases) and 7 case-control studies (2409 cases). We observed a summary RR of 0.80 (95% CI: ) for coffee drinkers as compared to nondrinkers. Compared to nondrinkers, the summary RR were 0.87 (95% CI: ) for low-to-moderate coffee drinkers and 0.64 (95% CI: ) for heavy drinkers. Coffee, decaffeinated coffee, tea intake and risk of renal cell cancer The relation between coffee, decaffeinated coffee and tea intake and RCC risk was analyzed in our case-control study on 767 cases with RCC and 1,534 controls. Coffee intake (mostly espresso and mocha) was not associated with RCC (OR = 1.02 in drinkers of 4 or more cups/day compared with drinkers of less than 1 cup/day). No relation was observed with decaffeinated coffee and tea intake, the ORs being 1.38 and 0.78 for drinkers compared with nondrinkers respectively, although these two estimates were based on a low number of drinkers. This study, based on a large dataset, provided further evidence that coffee, decaffeinated coffee and tea consumption is not related to RCC risk. Fish consumption and cancer risk The relation between consumption of fish and n-3 polyunsaturated fatty acid (PUFA) and the risk of selected neoplasms has been analysed in two series of integrated case-control studies conducted in northern Italy during and In the first series of studies, the OR for an increase in fish intake of 1 portion/week were 0.7 for rectal cancer; 0.8 for oral/pharyngeal, esophageal, gastric, colon and laryngeal cancer; 0.90 for pancreatic, endometrial and ovarian cancer and for multiple myelomas. No consistent relation was found for liver, gallbladder, breast, prostate, bladder, kidney and thyroid cancers, Hodgkin disease and 213

215 non-hodgkin lymphomas, although most OR were below unity for these neoplasms, too. In the second series of studies, the OR for the highest quintile of n-3 PUFAs compared with the lowest one were 0.5 for oral/pharyngeal, 0.5 for esophageal, 1.4 for gastric, 0.7 for colon, 0.8 for rectal, 1.5 for laryngeal, 0.8 for breast, 1.0 for endometrial, 0.6 for ovarian and 1.0 for renal-cell cancers; the estimates and the trends in risk were significant for oral/pharyngeal, esophageal, colon and ovarian cancer. Thus, although prospective studies overall do not provide evidence of a significant association between n-3 PUFA and cancer incidence, these Italian studies suggest that consumption of even relatively small amounts of fish decreases the risk of several cancers, especially of cancers of some parts of the digestive tract. LABORATORY OF MEDICAL INFORMATICS Development of the BPCO.CARE website This web based medical index has been developed by the Laboratory of Medical Informatics in order to collect, classify, evaluate, and describe the most useful medical information on the web related to the chronic obstructive pulmonary disease (COPD). The description associated to each web site included in the BPCO.CARE index illustrates the main contents, the services, and the tools offered to the users, including those related to the web 2.0 technology such as podcasts, RSS feeds, blogs, webcasts, and webinars. A section of the BPCO.CARE website also includes a classified collection of the podcast services available on the net in the COPD and pulmonology area and information on how to subscribe to. The BPCO.CARE website (available at has been developed in collaboration with the Department of Cardiovascular Research. Maintenance of the CARDIO.CARE, ONCO.CARE, GASTRO.CARE, NEURO.CARE, PNEUMO.CARE, PAIN.CARE and DERMA.CARE websites These indexes have been developed by the Laboratory of Medical Informatics in order to collect, classify, evaluate, and describe the most useful medical information on the web, and to provide Internet users with an easy means to surf the net. Several medical areas are covered including oncology ( neurology ( gastroenterology ( cardiology ( pulmonology ( the pain care and management ( and dermatology ( The project is in collaboration with intramural departments (Department of Oncology, Laboratory of Neurological Disorders and Department of Cardiovascular Research, Laboratory of General Practice Research, Laboratory of Translational and Outcome Research in Oncology) and extramural research groups (Italian Group for Epidemiologic Research in Dermatology, GISED). Studies on the typology of the web 2.0 applications in medicine The Laboratory of Medical Informatics is involved in studies and surveys which aim is to describe the typology of the web 2.0 applications and tools (including social networks, podcasts, feed RSS, blogs, and wikis) in medicine available on the net, and how these are perceived by the medical community. Internet as a research and formative tool on the chronic pain in cancer patient This research project was born in the framework of the project "Pain in the patient with cancer", in collaboration with the Laboratory of Medical Research and Consumer Involvement and the Laboratory of Translational and Outcome Research in Oncology. Its aim is to make available 214

216 for doctors, patients and families correct information about therapies for cancer pain and to produce greater tests on the effectiveness of therapies based on the use of analgesic drugs. This project is articulated in two main activities: - Paincare, set-up a catalogue of selected web pages dedicated to the cancer pain and relative periodical up-to-date, - adaptation of the methods and instruments of Evidence Based Medicine to the resources available on the Internet about chronic pain in patients with cancer. This is done through a specific questionnaire. We are also considering the opportunity to set up a new Italian cancer pain website. Studies on the quality of health information available on the Internet We are conducting an analysis on the quality and reliability of the information available on the Internet related to the management of cancer pain. Through an ad hoc form using worldwide applied criteria and principles for the evaluation of health websites and the evidence based medicine tools, an interdisciplinary team of reviewers is involved in the evaluation of the quality of information offered by the websites included in the PAIN.CARE medical index. The same study will also allow to compare the level of the quality of health related information among different classes of websites. The study, in collaboration with the Laboratory of Medical Research and Consumer Involvement, is ongoing and the results will be available in Training activities In 2008, the Laboratory of Medical Informatics continued its training activity on issues related to the use of the Internet in medicine, and extended it to the use of the recent web 2.0 technologies and tools in the medicine area. The members of the laboratory staff activated (or attended as invited teachers) a number of training courses, workshops, and master courses. Onsite CME courses for the Italian physicians have also been organized using the training/educational facilities and equipment available at the Mario Negri Institute. 215

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218 DEPARTMENT OF PUBLIC HEALTH STAFF Head Department Maurizio BONATI, MD. "Angelo & Angela Valenti" Centre for Health Economics (CESAV) Head of Laboratory Livio GARATTINI, Econ.D. Laboratory of Clinical Epidemiology Head of Laboratory Guido BERTOLINI, MD. Clinical Knowledge Engineering Unit Head Unit Davide LUCIANI, MD. Laboratory for Mother and Child Health Head Laboratory Maurizio BONATI, MD. 217

219 CURRICULA VITAE Maurizio Bonati has a Medical School degree at the University of Milan. Areas of interest: Monitoring and epidemiological evaluation of drug utilisation and effects of drugs and vaccines in motherhood and childhood. Research methodology in general hospital and paediatric community practice. Transfer of information to the community. Epidemiology of paediatric and perinatal care. Past and present roles both at the Mario Negri Institute and in other institutions: Research Fellow at the IRFMN, within the Neurochemistry Lab.; Research Assistant at the IRFMN, within the Clinical Pharmacology Lab.; Chief of the Perinatal Clinical Pharmacology Unit at the IRFMN; Advisor to WHO for the Drug Utilization Research Group (pregnancy, paediatrics and breastfeeding); coordinator of the International Cooperative Study of Drug Use in Pregnancy, under the auspices of WHO and the support of EEC; co-editor of The Kangaroo; coordinator of the European Cooperative Study: Development of the European register of clinical trials on medicines for children (DEC-net), under the 5 th Framework Programme s Quality of life and Management of Living Resources; since 1989 he has been director of the Centre for Drug Information; since 1993 head of the Lab. for Mother and Child Health; since 1997 teacher for the Lombardy region s professional training courses; since 2000 teacher for the Lombardy region s professional training courses; since 2002 Editor of the Ricerca & Pratica scientific journal; since 2003 professor of the School of Specialisation in Paediatrics - University of Milan Bicocca; teacher at the annual European course Evaluation of Medicinal Products in Children (promoted by ESDPPP and Eudipharm); from May 2008 Head of Department Public Health"Mario Negri" Institute for Pharmacology Research. Selected publications Bonati M, Campi R. What Can We Do to Improve Child Health in Southern Italy? PLos Medicine 2005;2(9):e250. Pandolfini C, Bonati M. A literature review on off-label drug use in children. Eur J Ped 2005;164: Santoro E, Rossi V, Pandolfini C, Bonati M. DEC-net: the development of the European Register of Clinical Trials on Medicines for Children. Clinical Trials 2006;3: Clavenna A, Rossi E, De Rosa M, Bonati M. Use of Psychotropic Medications in Italian Children and Adolescents. Eur J Pediatr 2007;166: Maschi S, Clavenna A, Schiavetti B, Campi R, Bernat M, Bonati M. Neonatal outcome following pregnancy exposure to antidepressants: a prospective controlled cohort study. BJOG 2008;115: Usala T, Clavenna A, Zuddas A, Bonati M. Randomised controlled trials of Selective Serotonin Reuptake Inhibitors in treating depression in children and adolescents: a systematic review and meta-analysis. European Neuropsychopharmacology 2008;18: Livio Garattini: got his degree in Economics in March 1983 at the Bocconi University in Milan. Educational activities: King s Fund College, London: courses of health care management; Centre for Health Economics, York: review of publications on the English NHS; Ecole Nationale de la Santé Publique, Rennes: courses of health policy. Areas of interest: Health Economics and Health Policy Analysis. At present he is the Director of CESAV (Centre of Health Economics A. e A. Valenti - M. Negri Institute); : researcher at M. Negri Institute; : clerk at Banca Commerciale Italiana in Milan; : junior consultant at Sogess srl in Milan; : researcher at Bocconi University in Milan. Selected publications: Garattini L, Ghislandi S (2006) Off-patent drugs in Italy- A short-sighted view? The European Journal of Health Economics 7(1): Garattini L, Ghislandi S (2007) Should we really worry about launch delays of new drugs in OECD countries? (Editoriale) The European Journal of Health Economics 8(1): 1-3. Cornago D, Li Bassi L, De Compadri P, Garattini L (2007) Pharmacoeconomic studies in Italy: a critical review of the literature The European Journal of Health Economics 8(2): Koleva D, Motterlini N, Banfi P, Garattini L (2007) Healthcare costs of COPD in Italian referral centres: A prospective study Respiratory Medicine 101: Garattini L, Motterlini N, Cornago D (2008) Prices and distribution margins of in-patent drugs in pharmacy: A comparison in seven European countries Health Policy 85(3): Garattini L, De Compadri P, Koleva D, Pasina L, Nobili A (2008) A critical review of the full economic evaluations of pharmacological treatments for colorectal cancer JME 11(1):

220 Guido Bertolini got his Medical degree in 1989 at the University of Bologna, and the specialization in Pharmacological Research in 1993 at the Mario Negri Institute and in Gastroenterology in 1994 at the University of Pavia. He founded and chaired from 1997 to 2000 the School of Clinical Methodology and Quality of Care Improvement at the Ospedali Riuniti di Bergamo and the Istituto di Ricerche Farmacologiche Mario Negri. From 1999 to 2003 he has been contract professor at the post-doctoral schools in Anaesthesia and Intensive Care, University of Brescia and Milano; from 2002 to 2005 he has been contract professor of Educational Science at the Faculty of Lettere e Filosofia, University of Bergamo. Current research interests: Clinical Research Methodology, Continuous Quality of Care Assessment and Improvement, Health services research and outcome, Medical decision making, Medical Education. These interests are mainly developed within the fields of Intensive Care Medicine and Rare Diseases. Since 1997 he chairs the GiViTI Coordinating Center for research in intensive care medicine. He has been Head of the Unit of Epidemiology and Education for Clinical Practice at the Mario Negri Institute and since 2001 he is the Head of the Laboratory of Clinical Epidemiology. From 2001 to 2005 he has been Vice-chairman of the Research Group on Cost-effectiveness, Section on Health Services Research and Outcomes European Society of Intensive Care Medicine and, from 2001 to 2005, he has been President of the Scientific Committee of the Ospedale maggiore in Crema. Selected publications Malacarne P, Langer M, Nascimben E, Moro ML, Giudici Daniela, Lampati L, Bertolini G, GiViTI. Building a continuous multicenter infection surveillance system in the intensive care unit: Findings from the initial data set of 9,493 patients from 71 Italian intensive care units. Crit Care Med 2008; 36: Bertolini G, Rossi C, Anghileri A, Livigni S, Addis A, Poole D. Use of drotrecogin alfa (activated) in Italian intensive care units: the results of a nationwide survey. Intensive Care Med 2007; 33: Rossi C, Simini B, Brazzi L, Rossi G, Radrizzani D, Iapichino G, Bertolini G. Variable costs of ICU patients: a multicenter prospective study. Intensive Care Med. 2006;32: Vanoli M, Daina E, Salvarani C, Sabbadini MG, Rossi C, Bacchiani G, Schieppati A, Baldissera E, Bertolini G. Takayasu's arteritis: A study of 104 Italian patients. Arthritis Rheum 2005; 15;53: IF: Galli M, Luciani D, Bertolini G, Barbui T. Anti-beta 2-glycoprotein I, antiprothrombin antibodies, and the risk of thrombosis in the antiphospholipid syndrome. Blood 2003;102: Simini B, Bertolini G. Should same anaethesist do preoperative anaesthetic visit and give subsequent anaeshetic? Questionnaire survey of anaesthetists. BMJ 2003; 327: Davide Luciani got his Medical Degree at the University of Bologna in 1995, and the Diploma in "Tropical Medicine and Hygiene" at the University of Liverpool in In 2001, he spent one year at the Department of Statistical Science (University College London). Bayesian probabilistic applications, decision theory and the graphical approach to pathophysiological modeling represent his main interests. Within his research activity, these skills are meant as the main methodological ingredients in the formalization of clinical reasoning, in order to improve its effectiveness and to exploit its educational value. Since 2005 he is responsible of the Unit of Clinical Knowledge Engineering. Selected publications Luciani D, Cavuto S, Antiga L, Miniati M, Monti S, Pistolesi M, Bertolini G Bayes pulmonary embolism assisted diagnosis: a new expert system for clinical use Emerg Med J 2007 ; 24 : M.Cesana, R.Cerutti, E.Grossi, E.Fagiuoli, M.Stabilini, F.Stella, D Luciani.Bayesian Data Mining Techniques: The Evidence Provided by Signals Detected in Single-Company Spontaneous Reports Databases. Drug Information Journal, vol. 41, pp , 2007 Latronico N, Bertolini G, Guarneri B, Botteri M, Peli E, Andreoletti S, Bera P, Luciani D, Nardella A, Vittorielli E, Simini B, Candiani A Simplified electrophysiological evaluation of peripheral nerves in critically ill patients: the Italian multi-centre CRIMYNE study Crit Care. 2007;11(1):R11. Bertolini G, Luciani D, Biolo G Immunonutrition in septic patients: A philosophical view of the current situation Clin Nutr 2007 ; 26 : Luciani D, Marchesi M, Bertolini G. The role of Bayesian Network in the diagnosis of pulmonary embolism. J Thromb Haemost 2003; 1: Galli M, Luciani D, Bertolini G, Barbui T. Anti-beta 2-glycoprotein I, antiprothrombin antibodies, and the risk of thrombosis in the antiphospholipid syndrome. Blood,2003 Oct 15;102(8): Haemostasis, 2003 Apr;1(4):

221 INTRODUCTION TO THE DIPARTMENT S ACTIVITIES The main aim of the Public Health Department is to understand which factors affect the health of individuals or entire populations and to define effective interventions for responding to their health needs. Special emphasis is therefore placed on prevention, so that the risks of contracting illness are lowered, and on the dissemination of independent, evidence-based information. The department s effort cannot disregard the National Health System, however, which must guarantee access to, and quality of, care that is based on principles of equity and appropriateness and must guarantee it especially to the more vulnerable patient groups. It is in this context that the Public Health Department carries out its activities. In addition to its formal research activity, the department participates in, and organises, initiatives involving information dissemination, training, and debate aimed at healthcare operators and social care workers, but also at the general population. These activities are also supported by the publication of the department s two journals: Ricerca&Pratica and Quaderni di Farmaco Economia. "A. and A. Valenti" Centre for Health Economics (CESAV) The "Angelo e Angela Valenti" Centre for Health Economics (CESAV) was established in 1992 at the "M. Negri Institute" and based at Villa Camozzi- Ranica (Bergamo)-Italy. CESAV is primarily a research centre, but also does educational work. The centre is involved in health economics and health policy research. The main areas of research are: Economic Evaluation of Health Care Programs (i.e. assessment of costs and benefits of alternative health care treatments and services) and Comparative Health Policy Analysis (i.e. study of domestic and foreign health care systems, in particular aimed at identifying possible innovations for European countries). The general aim of the Laboratory of Clinical Epidemiology is to contribute to the improvement of health care in different medical fields. The guiding principles are mainly two: to help physicians use the available knowledge and resources at their best; to play a role in the growth of useful knowledge for clinical practice. The Laboratory operates particularly in the field of Intensive Care Medicine and Rare Diseases. Within the Laboratory, the Unit of Clinical Knowledge Engineering aims to bring the value of clinical reasoning out, through the implementation of probabilistic models for its formalization, thus favoring the evaluation and the continuous improvement of complex clinical activities. Laboratory of Clinical Epidemiology Research, as a multidimensional approach to producing knowledge, characterises the Laboratory s activity. Research provides the basis for planning and carrying out the Laboratory s activity in a critical way and involves the participation of health professionals, social workers, mothers, children, and parents. Special attention is given to activities involving countries in the north and south of the world. Laboratory for Mother and Child Health The main objective of the Laboratory for Mother and Child Health is to ensure a better mother and child well-being by undertaking interdisciplinary and collaborative work in the field. Four broad areas, or spheres, of research have been selected: - monitoring and epidemiological evaluation of utilisation and effects of drugs and vaccines; - research methodology in general hospital and paediatric community practice; 220

222 - public health determinants of children s well-being; - transfer of health information to the community. Special attention is given to activities involving countries in the north and south of the world. In addition to the formal research activities, the Laboratory promotes initiatives in the public health field, in particular those involving mother and child health care. The initiatives involve the participation in, and the organisation of, educational, training, and information-dissemination activities. The critical and active transfer of scientific knowledge is a continuous, daily stimulus to the Laboratory s activity. NATIONAL COLLABORATIONS "A. and A. Valenti" Centre for Health Economics (CESAV) Public and private institutions, other health care organizations (Ministry of Health, Regional and Local Health Authorities, Hospital Trusts). Laboratory of Clinical Epidemiology Dipartimento di Neurologia, Ospedale Regionale S. Maria dei Battuti Cà Foncello, Treviso Dipartimento di Specialità Chirurgiche, Scienze Radiologiche e Medico Forensi, Cattedra di Anestesia dell Università degli Studi di Brescia Servizio Anestesia e Rianimazione, Osp. San Giovanni Bosco, Torino Istituto Mediterraneo per i Trapianti e Terapie ad Alta Specializzazione, Palermo U.O. Anestesia e Rianimazione 1, Ospedale A. Manzoni, Lecco Cattedra di Fisica e Informatica Medica, Facoltà di Medicina e Chirurgia, Firenze Laboratory for Mother and Child Health Centre for Child Health, (CSB) Cultural Paediatric Association, (ACP) Federfarma Lombardia Hospital of Bergamo Ospedali Riuniti, Poison Control Center, Unit Clinical Toxicology Italian Drug Agency, (AIFA) Italian Global Health Watch (OISG) Italian National Institute of Health (ISS) Italian Society of Preparers Pharmacists (SIFAP) Italian Society of Clinical Pharmacy (SIFO) Il Pensiero Scientifico Editore Operating unity of Neuropsychiatry of the childhood and of the adolescence, Fondation Policlinico di Milano, (UONPIA) University of Cagliari, Department of Neuroscience, Clinic of Child and Adolescent Neuropsychiatry University of Milan-Bicocca, Faculty Medicine, Paediatric Clinic University of Milan, Faculty of Political Science 221

223 INTERNATIONAL COLLABORATIONS "A. and A. Valenti" Centre for Health Economics (CESAV) CES (Collège des Economistes de la Santé) of Paris Corvinus University of Budapest Global Fund of Geneva WidO of Bonn University Carlos III of Madrid University of Birmingham University of Hannover University of York University Pompeu Fabra of Barcelona University Erasmus of Rotterdam Laboratory of Clinical Epidemiology Bloomsbury Institute of Intensive Care Medicine, Institute of Biomedical Research, University College London, UK Department of Statistical Science, University College London, UK Klink fur Anaesthesiologie und Intensivtherapie, Friedrich-Schiller-Universitat, Jena, Germany Machine Intelligence Group, University of Aalborg, Denmark American Board of Family Medicine, Kentucky, US Institute of Anaesthesia and Intensive Care, Semmelweis University, Budapest, Hungary Department of Anaesthesiology and Intensive Care, Warsaw Medical University, Poland Department of Intensive Care, General Hospital Novo Mesto, Slovenia Department of Pulmonary and Intensive Care Medicine, Nicosia General Hospital, Cyprus Hospital Das Clínicas, Federal University of Minas Gerais, Belo Horizonte, Brazil Laboratory for Mother and Child Health Catalan Institute of Pharmacology, Barcellona, Spagna Centre for Tropical Diseases (CECOMET), Ecuador Clínica Infantil Colsubsidio, Bogotà, Colombia European Medicines Agency (EMEA) European Society for Developmental, Perinatal and Paediatric Pharmacology. (ESDPPP) European Union (EU) International Society of Drug Bulletins (ISDB) Hospital Robert Debré, Paris, France World Health Organization (WHO) University of Nottingham, Derbyshire Children s Hospital, Derby, United Kingdom 222

224 EDITORIAL BOARD MEMBERSHIP "A. and A. Valenti" Centre for Health Economics (CESAV) INTERNATIONAL: Acta Bio Medica; Applied Health Economics and Health Policy; Biomedical Statistics and Clinical Epidemiology; Health Policy; Journal of Medical Economics; The European Journal of Health Economics. NATIONAL: Economia e Politica del Farmaco; FarmacoEconomia News; Farmeconomia e Percorsi Terapeutici; L'Internista; PharmacoEconomics Italian Research Articles; Quaderni di FarmacoEconomia. Laboratory of Clinical Epidemiology Ricerca & Pratica; Dedalo. Gestire i sistemi complessi in sanità. Laboratory for Mother and Child Health INTERNATIONAL: European Journal Clinical Pharmacology; Journal of Clinical Pharmacology & Pharmacoepidemiology; Paediatric & Perinatal Drug Therapy; Pediatria (São Paulo); Saludarte; NATIONAL: Dialogo sui Farmaci; Disturbi d Attenzione e Iperattività; Quaderni ACP; Quaderni di Farmacoeconomia; Ricerca & Pratica. PEER REVIEW ACTIVITIES "A. and A. Valenti" Centre for Health Economics (CESAV) Applied Health Economics and Health Policy; Health Policy; PharmacoEconomics; The European Journal of Health Economics. Laboratory of Clinical Epidemiology INTERNATIONAL: British Medical Journal; Intensive Care Medicine; Critical Care Medicine; Clinical Journal of the American Society of Nephrology. NATIONAL: Ricerca & Pratica Laboratory for Mother and Child Health INTERNATIONAL: Archives of Diseases Childhood; British Medical Journal; BMC Pregnancy and Childbirth; BMC Public Health; British Medical Journal; Current Drug Metabolism; European Journal of Clinical Pharmacology; Future Medicine; Expert Review of Clinical Pharmacology; Health 223

225 Education Research; Journal of Antimicrobial Chemotherapy; Journal of Clinical Pharmacology & Pharmacoepidemiology; Pharmacoepidemiology and Drug Safety; Prescrire. NATIONAL: Assistenza Infermieristica e Ricerca; Bollettino SIFO; Dialogo sui Farmaci; Epidemiologia e Prevenzione; Giornale Italiano di Farmacia Clinica; Medico e Bambino; Quaderni ACP. NATIONAL AND INTERNATIONAL COMMITTEE MEMBERSHIP "A. and A. Valenti" Centre for Health Economics (CESAV) Commissione Accordi di Programma, AIFA, Roma Comitato di Bioetica, Provincia Autonoma di Trento Laboratory of Clinical Epidemiology Comitato Etico Azienda Ospedaliera Bolognini di Seriate (BG) Commissione per la Ricerca sanitaria finalizzata, Provincia autonoma di Trento Commissione di valutazione per il Bando Ricerca Finalizzata, Ministero della Salute Laboratory for Mother and Child Health Comitato Etico dell'azienda Ospedaliera "Ospedale Maggiore" di Crema Comitato scientifico ADHD, ISS Commissione Nazionale per le Vaccinazioni, Ministero della Salute Commissione tecnico-scientifica per la programmazione e verifica delle vaccinazioni, Regione Lombardia Comitato Scientifico del Gruppo di Lavoro Farmaci e Bambini, AIFA Commissione tecnica per l'elaborazione, gestione e aggiornamento del Prontuario Terapeutico Regionale (P.T.R.), Regione Autonoma Valle d'aosta Gruppo di Lavoro Pediatrico, AIFA Gruppo di Lavoro "Promozione allattamento al seno", Regione Lombardia Paediatric Expert Group (P.E.G.), EMEA EVENT ORGANIZATION "A. and A. Valenti" Centre for Health Economics (CESAV) Congress: Convegno Nazionale di Farmacoeconomia: Economia del farmaco: fra soluzioni tecniche e decisioni politiche. 7-8 May, Ranica (BG) Laboratory of Clinical Epidemiology Congress, GiViTI Annual Meeting, October 29-31, Pesaro. Workshop, Meeting Compact, April15, Ranica (BG). 224

226 Laboratory for Mother and Child Health Stage in Cooperation and Public Health part of the Master s Course in Analysis and Management of Projects for Development. From 25 March at 4 april, Milan. Course La medicina basata sull evidenza (EBM) dalla teoria alla pratica nella neuropsichiatria dell infanzia e dell adolescenza. 11 June, Milano. Seminary Prevenzione della Cervice Uterina: incontro-confronto su conoscenze e prospettive. 17 June, Milan, Italy. Course La sperimentazione clinica in pediatria di famiglia June, Santa Maria Imbaro (CH), Italy. PARTICIPATION IN EVENTS IN WHICH THE DEPARMENT WAS INVOLVED "A. and A. Valenti" Centre for Health Economics (CESAV) May Congress: I Vaccini HPV: Le Esperienze Cliniche e le Strategie di Sanità Pubblica. Le strategie di sanità pubblica. I costi e la sostenibilità. Milan. Course: Le maculopatie e la degenerazione maculare legata all età: gestione clinicoterapeutica ed economia. L impatto socioeconomico delle maculopatie: Farmacoeconomia e Qualità di Vita. Baveno (VB). June Congress: Prevenzione del carcinoma della cervice uterina: incontro-confronto su conoscenze e prospettive. Valutazione costi-benefici. Milan. Seminar: Health Economics Ad Board for Almorexant (panel of experts). Frankfort. Workshop: Strumenti e metodi di trasparenza per il processo decisionale. Le possibili prospettive della prevenzione in funzione della qualità e della sostenibilità. Introduzione alle principali tecniche adottate nelle valutazioni economiche in sanità. Ruolo e sviluppo delle strategie di prevenzione vaccinale nel soggetto adulto. Ragusa. July Congress: 7th European Conference on Health Economics. Rome. November Course: Corso di farmacoeconomia ed economia sanitaria. Rome. Laboratory of Clinical Epidemiology February Congress, Trauma e Subintensive, Bologna Congress, "Riusciremo a morire in pace?", Milano Congress, Emorragia intracerebrale (ICH): quale paziente si giova di un trattamento intensivo in ambiente neurochirurgico? Torino 225

227 Course, Statistica di Base, Torino March Congress, Fine Vita, Padova Course, Statistica di Base, Torino Course, Statistica di Base, Torino Course, Excel e MargheritaDue: applicazione pratica, Torino Congress, GiViTI Bergamo, Zingogna (BG) Congress, La sorveglianza delle infezioni in TI, Zingonia April Course, Statistica di Base, Torino Congress, Le infezioni in TI: Presentazione dei dati regionali del gruppo GiViTI, Bologna Course, Statistica di Base, Torino Course, Statistica di Base, Torino Course, Margherita Due (Course, avanzato), Torino May Congress, Analgesia, anestesia, terapia intensiva in ostetricia, Torino Workshop, Perfezionamento e aggiornamento in medicina intensiva, Mendrisio Course, Statistica di Base, Torino Congress, Appropriatezza dei ricoveri in terapia intensiva, Torino June Congress, Valutazione della qualità della Assistenza sulla base degli indicatori di gravità del paziente, Avezzano September Congress, ESPEN Congress, Firenze Congress, Sepsi nel trauma - Trauma Update, Roma Course, MargheritaTre: stato dell'arte e pianificazione dei lavori, Ivrea October Congress, Incontri GiViTI-Piemonte, Torino Course, Migliorare la compilazione di Margherita Due ed imparare a leggere il report, Torino Course, Migliorare la compilazione di Margherita Due ed imparare a leggere il report, Torino Workshop, Accademia della cura, Rho (MI) Congress, Meeting GiViTI, Pesaro November Congress, Trauma Update, Cesena Course, MargheritaTre: presentazione dei nuovi moduli, Torino December Workshop, PNS: Euronetwork Sindromi Paraneoplastiche, Treviso Congress, Sorveglianza delle infezioni in ti con margherita due: Presentazione dei dati 2007, Torino 226

228 Laboratory for Mother and Child Health January Congress: In Toscana, dai dati alle scelte: l ospedalizzazione pediatrica e neonatale. Regione Toscana; Ospedale Meyer, Agenzia Regionale Sanità Toscana e Servizio Sanitario della Toscana; Firenze. Regional Congress: Seminare e Raccogliere. Guadagnare Salute. Associazione Culturali Pediatri Campagna (ACP) e Università degli Studi di Napoli Federico II, Dipartimento di Pediatria; Napoli. February Course of Formation: Bioetica e Sperimentazione. Centro di bioetica e biodiritto dell Università di Siena e Comitato Etico - AOU Senese; Siena. Meetings in the library. Azienda U.L.SS. n. 6 Vicenza, International Society of Drug Bulletins (ISDB), Biblioteca Biomedica Ospedale S. Bortolo; Vicenza. March III Regional Seminary: Attualità in farmacovigilanza. Università degli Studi di Siena, Sistema di farmacovigilanza Regione Toscana; Siena. IV National Congress: Pediatria On Line. Sirmione (BS). April Congress: Un esperienza sanitaria locale per un confronto globale. Fondazione Ivo de Carneri Onlus e Fondazione Policlinico San Matteo; Pavia. Course: Allestimento dei medicinali in pediatria. Università degli Studi di Milano, Facoltà di Farmacologia; Milano. SIII 2008 Integrated interfaculty, interdisciplinary seminar: Alimentazione: sicurezza, accesso, qualità, culturauniversità degli studi di Milano, Facoltà di Scienze Politiche; Milano. May Forum ISDB Italia: Il ruolo dell informazione indipendente. Dialogo sui farmaci; Verona. Seminary: IX Giornata dell allattamento al seno. La Leche League Italia; Imola (BO). 4th Updating Course: Novità e criticità nell attività regolatoria di farmaci e dispositivi medici. Società Italiana di Farmacia Ospedaliera (SIFO) e Società Italiana Attività Regolatorie (SIAR); Verona. First Conference: Anticipating changes in drug development for children building on paediatric rheumatology. Marie Curie ActionsTRiPR, Istituto Gaslini e Scuola Internazionale di Scienze Pediatriche (SISP), Genova. SEFAP Master (Servizio di Epidemiologia e Farmacologia Preventiva) : Corso di perfezionamento in Farmacovigilanza. Università degli Studi di Milano, Dipartimento di Scienze Farmacologiche; Milano. June Course: 2007 l anno dei farmaci per i bambini?. Associazione Laureati della Facoltà di Farmacia (a.l.far.). Istituto di Ricerche Farmacologiche Mario Negri; Milano. Congress: Prevenzione del carcinoma della cervice uterine: incontro-confronto su conoscenze e prospettive. Regione Lombardia e Istituto Mario Negri; Milano. September 227

229 XII Regional Congress Cure Primarie: Il farmaco tra appropriatezza e sicurezza. Regionale Friuli Venezia Giulia; Grado (GO). First Conference: Un disturbo conosciuto in tutto il mondo!. Regionale Lombardia A.I.F.A. onlus; Milano. Festivaletteratura 2008: Il corpo oltre. Come superare i limiti biologici: la vita dei farmaci. Mantova. Congress: Nuovi orizzonti per l influenza. ASL Provincia di Lodi Servizio Medicina Preventiva nella città; Lodi. October XX National Congress ACP: Venti di ACO. Associazione Culturale Pediatri; Cagliari. XXIX National Congress SIFO: SIFO e istruzioni. Funzioni e competenze del farmacista per un paese ed un SSN in evoluzione. Società Italiana di Farmacia Ospedaliera; Napoli. Seminary: Star bene a scuola. Migliorare la qualità di vita per favorire l apprendimento. Municipality of Milan; Milano. Course: III edizione del percorso di formazione per rappresentanti di associazioni di cittadini & pazienti Orientarsi in salute & sanità per fare scelte consapevoli. Partecipasalute, Istituto di Ricerche Farmacologiche Mario Negri ; Milano. Meetings Beyond a look: Salute: bene per pochi o diritto per tutti?. Coordinamento Comasco per la Pace, Comune di Lurate Caccivio, Encuentro, Biblioteca Comunale e Pro Loco; Lurate Caccivio (CO). Meeting: Premio Ercole Pisello. Regione Umbria, Associazione Giuseppe Corradi, Comune di Bevagna; Bevagna (PG). November Course Formazione alla ricerca: inquinamento ambientale e funzionalità respiratoria in bambini di un area urbana. ASL TA/1, Federazione Italiana Medici Pediatri (FIMP); Taranto. Continuing education Disagio scolastico e patologie neuropsichiatriche. ASL Provincia di Milano 3; Monza. Bioetica Congress : La trent anni di sanità tra bioetica e prassi quotidiana. Commissione Regionale di Bioetica, Regione Toscana, Servizio Sanitario della Toscana; Firenze. 1th International Congress of UENPS: Global Neonatology & Perinatology. Union of European Neonatal and Perinatal Societies (UENPD), Società Italiana di Neonatologia (SIN) e Società Italiana di Medicina Perinatale (SIMP); Roma. Course ECM for Paediatricians and Parents: Dall infanzia all età avanzata: la farmacovigilanza nelle età a maggior rischio. Servizio Farmaceutico AUSL Ferrara; Ferrara. Course: Farmaci generici e note AIFA in pediatria: analisi dei dati prescrittivi. Dipartimento Cure Primarie ASL di Milano; Milano. December XXI National Congress Confronti in Pediatria: Pediatria 2008: attraverso le immagini. IRCCS Burlo Garofolo, Università di Trieste; Trieste. Formation Course: In tema di antibioticoterapia, infezioni e malattie riemergenti. Centro per la Formazione Permanente e l'aggiornamento del Personale del Servizio Sanitario (CEFPAS); Associazione Culturale Pediatri Centro-Sicilia; Caltanisetta. 228

230 Seminary: Prevenire il carcinoma della cervice uterina: serve il vaccino? A che età?. Consorzio Mario Negri Sud; Santa Maria Imbaro (CH) GRANTS AND CONTRACTS "A. and A. Valenti" Centre for Health Economics (CESAV) Abbott EG Grunenthal-Prodotti Formenti Novartis Farma SpA Ratiopharm Sanofi Aventis Sanofi Pasteur MSD Vivisol Laboratory of Clinical Epidemiology ARESS Piemonte ASL TO-2 Piemonte AstraZeneca Azienda ULSS 16, Padova Italia Bellco SpA Draeger Italia Regione Lombardia Regione Toscana Regione Piemonte Sanofi-Aventis Italia Laboratory for Mother and Child Health Boehringer Ingelheim European Union Il Pensiero Scientifico Editore Italian Drug Agency, (AIFA) Provinve of Milan Regional Health Ministry - Lombardy Region Regional Health Ministry - Valle d Aosta Region 229

231 SELECTION OF SCIENTIFIC PUBLICATIONS FROM 2008 "A. and A. Valenti" Centre for Health Economics (CESAV) Garattini L, Motterlini N, Cornago D. Prices and distribution margins of in-patent drugs in pharmacy: A comparison in seven European countries. Health Policy 2008;85(3): De Compadri P, Koleva D, Mangia A, Motterlini N, Garattini L. Cost minimisation analysis of 12 or 24 weeks of peginterferon alfa-2b + ribavirin for hepatitis C virus. JME 2008;11(1): Garattini L, De Compadri P, Koleva D, Pasina L, Nobili A. A critical review of the full economic evaluations of pharmacological treatments for colorectal cancer. JME 2008;11(1): Garattini L, Casadei G. Health technology assessment: for whom the bell tolls? The European Journal of Health Economics 2008; Beghi E, Atzeni L, Garattini L. Economic Analysis of Newer Antiepilectic Drugs. CNS Drugs 2008;22(10): Laboratory of Clinical Epidemiology Bertolini G. From national to global outcome research in the intensive care unit: A challenge to win. Crit Care Med 2008;36: Guarneri B, Bertolini G, Latronico N. Long-term outcome in patients with critical illness myopathy or neuropathy. The Italian multi-centre CRIMYNE study. J Neurol Neurosurg Psychiatry : Malacarne P, Langer M, Nascimben E, Moro ML, Giudici Daniela, Lampati L, Bertolini G, GiViTI. Building a continuous multicenter infection surveillance system in the intensive care unit: Findings from the initial data set of 9,493 patients from 71 Italian intensive care units. Crit Care Med 2008; 36: Di Bartolomeo S, Valent F, Rossi C, Beltrame F, Anghileri A, Barbone F. Geographical differences in mortality of severely injured patients in Italy. Eur J Epidemiol. 2008; 23: Laboratory for Mother and Child Health Bonati M. An independent contribution to the growth of paediatric clinical pharmacology in Italy. Eur J Clin Pharmacol 2008;64: Clavenna A, Bonati M. A missed opportunity. BMJ Font M, Miselli M, Conforti A, Bonati M. An Italian Story with wider implications? BMJ Maschi S, Clavenna A, Schiavetti B, Campi R, Bernat M, Bonati M. Neonatal outcome following pregnancy exposure to antidepressants: a prospective controlled cohort study. BJOG 2008;115: Pandolfini C, Bonati M. European paediatric research and children s therapeutic needs. A trial review. Acta Paediatrica 2008;97: Pandolfini C, Bonati M. Something is moving in European drug research for children, but a more focused effort concerning all therapeutic needs is mandatory. Arch Dis Child 2008;93:715. Pandolfini C, Bonati M, Rossi V, Santoro E, Choonara I, Naylor C, Sammons E, Jacqz-Aigrain E, Zarrabian S, Arnau JM, Castel JM, Danés I, Fuentes I. The DEC-net European register of paediatric drug therapy trials: contents and context. Eur J Clin Pharmacol 2008;64: Santos DB, Clavenna A, Bonati M, Coelho HL. Off-label and unlicensed drug utilization in hospitalized children in Fortaleza, Brazil. Eur J Clin Pharm 2008;64: Usala T, Clavenna A, Zuddas A, Bonati M. Randomised controlled trials of Selective Serotonin Reuptake Inhibitors in treating depression in children and adolescents: a systematic review and meta-analysis. European Neuropsychopharmacology 2008;18:

232 LAY PRESS SELECTION PUBLISHED IN 2008 "A. and A. Valenti" Centre for Health Economics (CESAV) Garattini L, Gritti S. Confronto di prezzi e margini alla distribuzione coperti da brevetto in sette Paesi Europei. Focus On 18 Febbraio Garattini L, Gritti S. Prezzi dei farmaci coperti da brevetto. Ricerca & Pratica 2008;24: Garattini L, Cornago D, De Compadri P, Gritti S. Confronto di prezzi e margini alla distribuzione di farmaci coperti da brevetto in sette Paesi Europei. Quaderni di Farmaco Economia 2008;5:15-21 (1a parte). Gritti S, Casadei G, De Compadri P, Garattini L. ECM: analisi comparativa in sei diversi Paesi europei. Quaderni di Farmaco Economia 2008;6: De Compadri P, Koleva D. I costi della psoriasis vulgaris nei pazienti sottoposti a terapia sistemica: una rassegna della letteratura e una stima preliminare di costo in Italia. Quaderni di Farmaco Economia 2008;6:7-15. Garattini L, Motterlini N, Cornago D. Confronto di prezzi e margini alla distribuzione di farmaci coperti da brevetto in sette Paesi Europei. Quaderni di Farmaco Economia 2008;7:7-16 (2a parte). Garattini L. Finanziaria Dialogo sui farmaci 2008;1: Garattini L. Finanziaria e farmaci. Editoriale Ricerca & Pratica 2008;24(2): Laboratory for Mother and Child Health Biasini G, Bonati M, Corchia C, Marchetti F, Napoleone E, Panei P, Rossi Paolo, Rossi Rossella, Toffol G. Lettera aperta di nove componenti esterni del Gruppo Italiano sui farmaci pediatrici dell'agenzia Italiana del Farmaco. R&P 2008;24: Bonati M. Farmaci essenziali per i bambini. Reazioni 2008;6:1. Bonati M. Farmaci essenziali per i bambini. BIF 2008;XV: Bonati M. Il Pap-test? Certamente! La vaccinazione anti-hpv? Sì, eventualmente. Medico e Bambino 2008;4: Bonati M. I determinanti della salute e le disuguaglianze sociali. Quaderni ACP 2008; 15:138. Bonati M. Indipendenti da chi, da cosa? Dialogo sui Farmaci 2008;3:154. Bonati M. L AIFA al servizio dei bambini. Medico e Bambino 2008;27: Bonati M. L AIFA dalla parte dei bambini. CARE 2008;4: Bonati M. La vicenda dell Agenzia Italiana del Farmaco (AIFA). Quaderni acp 2008;15: 224. Bonati M. Prevenzione non è solo vaccinazione: il caso del carcinoma della cervice uterina. Quaderni di farmacoeconomia 2008;5:5-6. Bonati M. Redattori per passione, indipendenti per scelta. R&P 2008;24: Clavenna A, Fortinguerra F. Contro la tosse è meglio il miele. Quaderni acp 2008;15:85. Clavenna A, Fortinguerra F. Informazione sui farmaci: novità in Europa e in America. Quaderni ACP 2008;15:181. Clavenna A, Fortinguerra F. Molte novità sui farmaci per bambini. Quaderni ACP 2008;15:131. Clavenna A, Fortinguerra F. Paracetamolo e felilefrina: aggiornamenti sulla sicurezza di impiego. Quaderni acp 2008;15:259. Font M, Miselli M, Conforti A, Bonati M. An Italian story with wider implications? R&P 2008;24: Gaillard P, Bullio P, Martinod D, Fortinguerra F, Bonati M. Valutazione dell attività della Commissione tecnicoscientifica della Valle d Aosta. R&P 2008;24: Maschi S, Bonati M. Additivi nei farmaci per bambini, effetti indesiderati e appropriatezza. Dialogo sui farmaci 2008;6:

233 Pandolfini C, Clavenna A, Bonati M. La qualità dell informazione sulla fibrosi cistica nei siti internet italiani. R&P 2008;24: Pirola ME, Bettinelli ME, Fortinguerra F. Prevalenza, esclusività e durata dell allattamento al seno in Lombardia. R&P 2008;24: Taddia A, Zeni B, Campomori A, Campi R. Mifepristone e misoprostolo: efficacia e sicurezza nell'aborto medico. R&P 2008;24:3-16. OTHER PRODUCTS PUBLISHED IN 2008 "A. and A. Valenti" Centre for Health Economics (CESAV) Boffelli S, Rossi C, Bertolini G. Progetto Margherita. Promuovere la ricerca e la valutazione in Terapia Intensiva. Rapporto Bergamo: Ed. Sestante, [Report] Laboratory for Mother and Child Health Bonati M, Zuddas A. Metilfenidato, atomoxetina. In: Enciclopedia Medica Italiana. II tomo; III aggiornamento. 2008; [Chapter Book] Bonati M, Campi R e Gruppo di Lavoro per la Convenzione sui Diritti dell Infanzia e dell Adolescenza. I diritti dell infanzia e dell adolescenza in Italia. 4 rapporto di aggiornamento sul monitoraggio della Convenzione sui diritti dell infanzia e dell adolescenza in Italia Gruppo di lavoro per la CRC c/o Save the Children, Roma. 2008; [Chapter Book] Bonati M. Cuba. In: Salute globale e aiuti allo sviluppo. Diritti, ideologie, inganni. 3 Rapporto dell Osservatorio Italiano sulla Salute Globale. Edizioni ETS, Pisa. 2008; [Chapter Book] Bonati M. Neonatal Clinical Pharmacology In: 1st International Congress of UENPS. Rome, Italy, November 17-19, [abstract] Clavenna A, Sequi M, Bortolotti A, Merlino L, Foretino I, Bonati M. Drug utilisation profile in the paediatric population of Lombardy Region, Italy. In: Safe and Effective Medicines for Children. The 11th biennal ESDP Congress. Rotterdam 2008;105. [abstract] Fortinguerra F, Clavenna A, Bonati M. Drug related problems in children: the role of a drug information centre. In: Safe and Effective Medicines for Children. The 11th biennal ESDP Congress. Rotterdam 2008; 106. [abstract] Fortinguerra F, Clavenna A, Labate L, Maschi S, Bonati M, Advertisement Reviewer Group. Pharmaceutical advertisements in Italian paediatric general practitioner journals. In: Safe and Effective Medicines for Children. The 11th biennal ESDP Congress. Rotterdam 2008;107. [abstract] Pandolfini C, Bonati M. Lesson from DEC-net. Development and application of the European register of clinical trials on medicines for children. In: First Conference of the Marie Curie Actions TRiPR. Genoa, Italy, May 29 th -June 1 st, [abstract] Racca F, Bonati M, Berta G, Testa R, Benini F, Benedetti M, Bignamini E, Maspoli M, Salvo I, Ranieri MV. Long term ventilation of children in Italy: preliminary data from questionnaire survey. In: 21st Annual Congress European Society of Intensive Care Medicine; Lisbon, Portugal September [abstract] Racca F, Bonati M, Ottonello G, Berta G, Pavone M, Morandi F, Wolfler A, Tardivo I, Testa R, Sequi M, Maspoli M, Bignamini E, Salvo I, Ranieri VM. Home discharge and monitoring program for home mechanically ventilated children in Italy. In: 2nd Congress of the European Academy of Paediatrics. Nice, France, October 24-28, [abstract] Usala T, Rocchi F, Knellwolf AL, Bonati M, Masi G, Zuddas A, Arcieri R, Panei P. Long term safety of psychotropic drugs used for Attention-deficit/hyperactivity disorder (ADHD) in Italian children and adolescent population. In: Safe and Effective Medicines for Children. The 11th biennal ESDP Congress. Rotterdam 2008;119. [abstract] During 2008 the laboratory s activities were covered by the national media 96 times 232

234 RESEARCH ACTIVITIES "A. and A. Valenti" Centre for Health Economics (CESAV) Educational activity Educational activities are developed only if related to research studies, in order to offer original contributions which naturally reinforce the research aims. Economic Evaluation of Health Care Programs The aim of this research area is to assess the costs of pathologies and the cost-effectiveness ratios of the diagnostic/therapeutic existing alternatives. In general, analyses can be classified into two groups: partial economic evaluations (e.g. cost of illness analysis) and full economic evaluations (e.g. cost-effectiveness analyses). Comparative Health Policy Analysis The aim of this research area is to study the organization of health care systems, in order to draw lessons from international comparisons. This is particularly important in a "market" like health care where economic competition lacks by definition and therefore public regulation plays a crucial role. Laboratory of Clinical Epidemiology Appropriateness in Intensive Care Units The main purpose of these research activities is the assessment and improvement of the quality of care in Italian Intensive Care Units (ICUs). It is a multi-annual project promoted on behalf of GiViTI, a collaborative network composed by half of the Italian ICUs and coordinated by the Laboratory. The main focus is the Margherita project. Its aim is the continuous evaluation of the quality of care and it is based on a free software developed by the Laboratory and distributed to all the ICUs adhering to the GiViTI group. The software has been realized on a modular structure, which enables to easily integrate the basic data collection (the core of Margherita) with the data collection of specific research projects (the petals of Margherita). Additionally, in the current year, a software based on a probabilistic model (bayesian network) covering a large set of clinical variables, has been further implemented. Such an activity, lead by the Unit of Clinical Knowledge Engineering, and also involving the Department of Statistical Science (University College London) and the Machine Intelligence Group of the University of Aalborg (Denmark), pursues the realization of tools for the retrospective evaluation of specific treatments in ICU. By means of this model, it is meant to overcome the current limits of an overall evaluation of ICU, likewise it happens when the traditional models for mortality prediction are adopted. Within this activity, a bayesian system to monitor the Standardised Mortality Ratio of the single ICU. This system, which resembles the bayesian pharamcovigilance system currently in use at the WHO and FDA, will identify periods during which some problems occurred, allowing the physicians of the Center to promptly and efficacy react. Studies on Multiple Organ Failure pathological processes The Laboratory of Clinical Epidemiology has lead several investigations to clarify which pathophysiological mechanisms induce multiple organ failure, a condition still burdened with high mortality. Among these, the investigation on the neuromuscular impairment in critical patients (the observational study 'Crimyne'), the study on the impact of enteral 233

235 feeding, and the new treatments proposed for severe sepsis (Xigris and the removal of inflammation mediators through specific filter applied to circuit of plasma-filtration). The value of a strict glycemic control of critical patients has been recently emphasized, given its connection to a drug like insulin that, in spite of its large availability and low cost, induces a relevant reduction of mortality in ICU. The Unit of Clinical Knowledge Engineering has developed a model based on a differential equations system whose aim is to support the physician in dosing both insulin and glucose infusions, in order to extend the possibility of a strict control even to patients with a high risk of hypoglycaemia. This model represents a precious opportunity to investigate the pathophysiological mechanisms behind the benefits of insulin already demonstrated at an empirical level, allowing the explanation of the dynamic behaviour of glycemic fluctuations on the basis of the patient's metabolic profile. The reconstruction of clinical reasoning in the medical practice and education This area represents the main concern of the Unit of Clinical Knowledge Engineering, whose objective is the valorization of clinical reasoning in solving complex clinical problems. The diagnosis of pulmonary embolism still represents a relevant clinical challenge, due to the complexity of the patient's clinical presentation and the variability of diagnostic resources among Centres. In this regards, we are conducting an Italian multicenter study, involving mainly Emergency Units, with the aim of prospectively validating the diagnostic software BayPAD (Bayes Pulmonary embolism Assisted Diagnosis). Such a tool, relying on a probabilistic model covering 72 clinical variables and doing without the need to input all the contemplated observations, would overcome the main reasons which prevented ordinary clinical guidelines to be largely accepted. Moreover, the results of the retrospective validation of the system have been obtained. The Unit started a project for the realization of a software assisting the physician in tracing back the basis of his clinical decisions before the description provided by clinical reports, among those that are typical of particular medical specialty. The software has the double target to create specific applications based on probabilistic models representing complex clinical decision problems, and to involve physicians in their construction. The last target is achievable given the strong analogy between the causal structure of the exploited models (bayesian networks) and the pathophysiological structure of medical knowledge. By this, it will be given the chance to adopt this system within medical training projects, with a special attention to e-learning programs. Clinical Epidemiology of rare diseases and orphan medicine Our purpose is to find out and describe clinical and research problems related either to rare diseases or to neglected aspects of well-known diseases. We also focus on the needs of the patients with rare diseases. A specific project is connected with this research activity: PNS Euronetwork. It is a European project on paraneoplastic neurological syndromes that is financed by the Fifth and Sixth Framework Program of the European Community. Its purposes are various: to develop a network of reference centers for these pathologies all sharing a common database; to organize a sample bank of biological fluids and cerebrospinal liquid to point out the best antibody indicators for the diagnosis and prognosis of these patients; to realize some research projects on the treatment of this syndrome. Laboratory for Mother and Child Health Pharmacoepidemiology in the Lombardy Region During 2005, 747,790 children and adolescents < 18 years (48% of the population) received at least one drug prescription. A total of 2,177,469 prescriptions were dispensed, corresponding to 3,122,745 medication packages. Each treated child received an average of 3 prescriptions and 4 packages (median 2). The highest prevalence was observed in the

236 year old age range (average value 65%), and then decreased to 38% in the year range. The prevalence was slightly higher in boys than girls (62% vs 59%). Antibiotics were the most prescribed therapeutic class (41% of the population), followed by anti-asthmatics (15%) and anti-histamines (5%). Altogether, these three therapeutic classes comprised 81% of the prescribed packages. In all, 757 drugs were prescribed. Amoxicillin+clavulanic acid was the most prescribed drug (18% of children), followed by amoxicillin (13%) and inhaled beclometasone (9%). The 10 most prescribed drugs, 7 of which were antibiotics, represented 64% of the prescribed packages. Large differences were found in prevalence rates between different LHUs; these ranged between 38 in Milan and 55% in Brescia. No correlation was found between rank distribution at LHU level of the prevalence rate and hospitalisation rates in the paediatric population. The age and residence of the child were the main determinants of drug exposure. In particular, being 1-5 years old (OR 4.51; 95%CI ) and living in Brescia (OR 2.08; 95% CI ) were the factors associated with the highest risk of drug exposure. The general prescribing profile is similar to that observed in other Italian studies, in particular in Northern Italy. Despite the fact that the Lombardy region is a quite homogeneous context, quantitative and qualitative geographical differences were found. The differences observed between local health units suggest that educational interventions for health care professionals and parents could be effective in improving the rational drug use. However, more efforts are needed also in certain contexts characterised by a low prevalence rate, but also by a not always appropriate drug use. Prescription, efficacy, and safety of psychotropic drugs in the Italian paediatric population The safety and effectiveness of psychotropic drug use in the paediatric population is widely debite, in particolar because of the lack of data concerning the long term effects. In Italy the prevalence of psychotropic drug prescriptions increased in the period and decreased afterwards. In such a context, the systematic continuous monitoring of psychopharmacological treatments is essential. The Laboratory coordinates an independent research project supported by the Italian Drug Agency (AIFA) entitled Prescription, efficacy, and safety of psychotropic drugs in the Italian paediatric population, aimed to: estimate the incidence and prevalence of psychotropic drug prescriptions in the paediatric population, to evaluate the prescribing patterns, and to monitor the safety of these drugs. The study population was composed by more than 76,000 children and adolescent < 18 years old living in the local health unit of Verona. Between 1 January 2005 and 31 December 2006, 111 youths (0.8 per 1,000 of the study population) received at least one psychotropic drug prescription. Only 29 patients were in charge to the child and adolescent outpatient psychiatric services. Information concerning diagnostic and therapeutic approaches, and care strategies were collected with the support of the treating physicians for 52 patients (47%). Anxiety-depression syndrome and attention disorders were the most commonly diseases for which psychotropic drugs were prescribed. In all, 20% of youths were chronically treated; 85% received also psychological support; child psychiatrists performed the first diagnosis in 50% of the cases; in 1/3 of the cases parents attended 2 or more different specialists. Workgroup on the Convention for child and adolescent rights The Laboratory for Mother and Child Health is part of the Workgroup on the Convention for child and adolescent rights (CRC) in Italy. The 4 th update report was published this year and, as with the previous reports, came out on May 27 th, anniversary of the CRC s ratification in Italy. The report testifies not only the perseverance and effort of the participating associations in monitoring and keeping a spotlight on child and adolescent rights throughout the years, but also the progress and strengthening of the CRC group. The 4 th CRC report offers an update of the issues addressed in the previous reports, adds more in depth analyses, and enriches the analysis with the inclusion of new topics thanks to the active contribution of an ever increasing number of associations. 235

237 The objective is to expand the workgroup s area of observation so that it addresses all eight groupings into which the United Nations Committee has divided the CRC s articles, corresponding to the report s chapters, in the next United Nations Supplementary Report expected in 2009 (year in which the United Nation s Convention s 20 th anniversary will be celebrated. The 4 th report provides an updated view of the putting into effect, or the violation, of Italian children and adolescent s rights. The CRC Group s associations ask the government to ponder this issue with the hope that such an effort, and the recommendations made, may serve as a useful tool for those who in the new government will be responsible for the welfare of children and adolescents in Italy. In 2009 the Italian government will have to stand before the United Nations Committee and describe the efforts made in the last few years to improve children s rights and to put into place the UN Committee s conclusive observations concerning the extent of the CRC s fulfilment in Italy and of the two CRC Optional Protocols (from June 2006). All this is carried out with the hope that it will stimulate, and contribute to, the development of standard procedures and legislative reforms that will bring about a tangible improvement in the condition of all youths in Italy. National ADHD Register The marketing authorisation for methylphenidate and atomoxetine in Italy has made monitoring the use of these drugs in children with attention deficit hyperactivity disorder (ADHD) necessary. In order to meet this need, a national registry coordinated by the Istituto Superiore di Sanità s drug department, in collaboration with the Italian Drug Agency and the Conferenza permanente degli Assessori alla Sanità delle Regioni and the Province autonome di Trento e Bolzano, was set up and activated on June 18, The Laboratory for Mother and Child Health is part of the scientific committee and participated in writing the protocol, which defines the structure and activities of the national ADHD register and its monitoring. The registry s aims are : to monitor the use of methylphenidate and atomoxetine; to evaluate the safety and compliance of therapies with methylphenidate and atomoxetine, alone or in combination with other therapeutic interventions (pharmacological or not) in the medium and long term; to define the probability of developing ADHD requiring pharmacological treatment in the school-aged population; to define the optimal management strategy for ADHD through the standardisation of the most appropriate diagnostic and therapeutic strategies; to evaluate the effectiveness of psychotropic drugs and/or behavioural therapy in the evolution of ADHD in school-aged children; to calculate the long term risk of persistence of the disorder, of being left behind in school, and of the development of psychosis or other psychiatric disorders. During the register s first year of activity, a total of 851 patients were registered by 83 referral centres, 369 of whom were receiving methylphenidate and 482 atomoxetine. Adverse reactions were reported in 23 patients, 7 of which were classified as severe. In all, 52% of patients concluded the follow-up after 6 months of treatment and 3% did so one year after registration. The Laboratory set up ADHDNews, a newsletter aimed at providing interested health care operators a monthly bibliographic update of the recent scientific literature via . This initiative is part of the Italian Drug Agency s (AIFA) independent research project implemented in collaboration with the Istituto Superiore di Sanità and called Long term safety of drugs used to treat school-aged children with Attention Deficit Hyperactivity Disorder and epidemiology of the disorder in the Italian population. A total of 12 issues of ADHDNews have been produced so far, identifying 440 scientific articles. Over 200 people have registered to receive the update. The newsletter is also viewable on the Mario Negri s website under the section The Mario Negri Institute for the physician (L Istituto Mario Negri per il Medico): 236

238 Ricerca & Pratica Ricerca & Pratica was born in January, 1985, as a manifestation of the Mario Negri Institute for Pharmacological Research. Today, the journal is part of the International Society of Drug Bulletins (ISDB), which represents independent journals. For more than twenty years, the journal has represented an arena for all those professionals who collect data and carry out studies in general practice with the aim to increase their knowledge and to improve their practice. Ricerca & Pratica is also appreciated for its ability to go beyond the merely clinical aspect of medicine, without, however, forgetting that it is to this aspect that the readers dedicate most of their time and effort. Through its activity, Ricerca & Pratica can therefore represent an exclusive, independent observation point. It is also an area that promotes contemplation, evaluation, and information by applying tools such as data trustworthiness and importance, the balance between benefits and risks and between benefits and costs, independence from conflicts of interest, and the realistic objective to contribute to a progressive, equally distributed improvement in the population s health. VII Master s Course An inter-university course aimed at specialising students in the planning and management of co-operation projects for development. The seventh edition of a 10-day Stage in Cooperation and Public Health, part of the Master s Course in Analysis and Management of Projects for Development ( was organised by the Laboratory and held at the Mario Negri Institute in Milan. The Stage involved a in-depth examination of the problems associated with public health in developing countries, taking into consideration their primary importance to the entire world s population in this new millennium. The course was based on both theory and practice. The lessons involved issues such as coping with emergency situations, providing assistance for extreme needs, addressing the problem of access to drugs, health and well-being indexes, analyses of living conditions, and intervention programs, and were complemented by hands-on practice sessions that also involved the basics of epidemiology. OPINING of the Stage: health is a universal right and an essential need for all. Whether a society is rich or poor, its development is judged on the basis of how justly distributed its quality of care is to the population and to what extent it is guaranteed. However, the right to health is often, and in many countries, not reached because economic and social inequalities lead to wide health inequalities. This was the topic that the WHO s Commission on Social Determinants of Health addressed. The final report was sent to all health and finance ministers and to healthcare and social work operators, and was presented by a member of the international commission, Prof. Giovanni Berlinguer, on the stage s opening day. CLOSING Stage: Gherardo Colombo, an ex magistrate ( ) and author of numerous publications who has been involved for years in explaining the meaning of justice to the young, met with the students of the Master s Course in Analysis and Management of Development Projects during the Stage in Cooperation and Public Health. The topic of the discussion was the rules in contemporary Italy and in the globalised world, sixty years after the Universal Declaration on Human Rights (whose principles remain largely unmet worldwide). Social, economic, cultural, and health-related inequalities exist in all countries and characterise the conditions and life expectancies of many populations. The justice that should control and guarantee the distribution of rights and duties is often non-existent or distracted. The rules (of justice) and adherence to them imply the participation in, and respect of, the inalienable rights of mankind on the part of all: a utopia? 237

239 FARMACI E BAMBINI workgroup The huge ordeal with the Italian Drug Agency (AIFA) and the rapid reorganisation of its structure caused the workgroup to suspend its work on drugs for children, after two years of activity. The laboratory was one of the group s members. The group s activity in those two years represented a new methodological approach that increased the AIFA s attention on the paediatric population and that included the organisation of educational and training interventions aimed at health care operators. Given the group s vast experience, the technical-scientific, methodological, and managerial heritage that the AIFA has acquired should not go lost, and the same is true for the approval and credibility gained by the national regulatory agencies in Europe. NASCERE IN CASA The Associazione Nazionale Culturale Ostetriche Parto a Domicilio (National Cultural Association of Home Birth Obstetricians) aims to increase home births, guaranteeing their quality and safety, verifying and comparing the national and international experiences of care outside the hospital, and defining professional selection and assistance criteria based on scientific evidence from the World Health Organization s recommendations and on women s wishes. The Laboratory for Mother and Child Health collaborates with the association, analysing data collected by the obstetricians in order to improve the quality of care offered to women and children through experience, but also through continuous study, continuing professional training, and debate. These improvements will help protect out-of-hospital births and safeguard the obstetricians specific professional role. The prevention of cervical carcinoma The Regional Council approved, with decree n.6683 dated 27 th February 2008, the regional programme on the prevention of cervical carcinoma. The programme involves actions aimed at incrementing adhesion to pap-test screening, especially in disadvantaged populations, and to HPV vaccination in girls born in 2007.The complexity of the interventions to put into place require the specific training of health professionals and the organisation of encounters during which health professionals can discuss the different aspects of the disease, from its epidemiology to its prevention and management. In this context, the laboratory, along with the Lombardy Region, organised the first regional meeting entitled Prevention of cervical cancer: meeting and debate on knowledge and perspectives at the Mario Negri Institute. Co-operation with countries with limited resources As an expression, test, and original method of expression of the choice to make the Laboratory s research transferable and accessible to all populations, the Laboratory promoted and provided assistance to projects in, and for, the South of the world, also in collaboration with the World Health Organization. The technical and organisational support for local groups and international non-governmental organisations for carrying out sociosanitary projects in countries with limited resources, especially Colombia, Ecuador, Brazil, Bolivia, Cuba, Vietnam e the Balkans, continues. ECUADOR - The laboratory supported the campaign Adopt a mom, promoted by the MLAL World-Project, and aimed at the population of Borbón, a town in northern Ecuador. About people, from both the black and indigenous races, live in this health district, which is made up of 170 small communities dispersed along the rivers and from which it can take up to 15 hours by motor-powered canoe to reach the Borbón hospital. Objectives: improve maternal health conditions identify risk factors and obstetric complications in pregnancy, birth, and the perinatal period early on and provide adequate treatment reduce cases of maternal death fight risks of foetal and neonatal death 238

240 train local personnel in the mother and child health field: the communities midwives (parteras), health promoters, and nurses along the rivers that normally assist with 70% of births in the area. COLOMBIA - The Colsubsidio Paediatric Investigation in the Americans 2008 Prize was initiated in Colombia in 1992 with the aim to develop and promote paediatric research. The prize, which has social and community connotations, is aimed at health care workers in Central and South America and was designed to form a network though which knowledge, analysis, study, and equity involving child health can be exchanged. This network joins ethical solidarity with social and professional responsibility. The prize is set up by Colsubsidio (Caja Colombiana Subsidio Familiar), a private, mutual society with more than members. Colsubsidio is a non-profit organisation that has been working in the social, health, and economic areas in different sectors of society since 1957 and is known for its high quality, professional work. Over 90 projects from 10 countries were submitted for the 2004 edition. The projects represented over 240 authors and were evaluated by an international jury (Maurizio Bonati, Italy; Imti Choonara, United Kingdom; Celia Beatriz Gianotti, Brazil; Raúl Mercer, Argentina; Carlos Bernal Parra, Gloria Arias Nieto, Jorge Mauricio Palau Colombia). The abstracts and integrated texts of the projects that received an award are published on the SALUDARTE journal Vol. 6 n.2 November February

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242 LABORATORY OF REGULATORY POLICIES STAFF Head Vittorio BERTELE, M.D. 241

243 CURRICULUM VITAE Vittorio Bertele is a clinical pharmacologist. He got his MD degree in 1977 and the specialization in Internal Medicine in 1982, both at the Milan University Medical School. He was research fellow at the Harvard Medical School and then worked at the Milan University and the Mario Negri Institute. His main areas of interest have been clinical pharmacology of drugs active on the haemostatic and vascular system 1,2, epidemiology of interventions in the cardiovascular area, and clinical trials and drug utilization studies in the cardiovascular area 3,4. He was CPMP expert at the EMEA, and member of the Committee for Drug Price Negotiation at the Italian Ministry of Health 5,6. At present he is Head of the Regulatory Policies Laboratory at the "Mario Negri" Institute, and member of the Technical-Scientific Committee at the Italian Drug Agency. Selected publications Bertele' V., Falanga A., Tomasiak M., Dejana E., Cerletti C., De Gaetano G. Platelet thromboxane synthetase inhibitors with low doses of aspirin: Possible resolution of the "aspirin dilemma". Science 1983; 220: Bertele' V., Falanga A., Tomasiak M., Chiabrando C., Cerletti C., De Gaetano G. Pharmacological inhibition of thromboxane synthetase and platelet aggregation: Modulatory role of cyclooxygenase products. Blood 1984; 63: (1984). Bertele' V, Mussoni L, Pintucci G, Del Rosso G, Romano G, de Gaetano G, Libretti A. The inhibitory effect of aspirin on fibrinolysis is reversed by iloprost, a prostacyclin analogue. Thromb Haemost 1989; 61: The i.c.a.i. Group (Gruppo di studio dell'ischemia cronica Critica degli Arti Inferiori). Prostanoids for chronic critical leg ischemia: A randomized, controlled, open-label trial with prostaglandin E 1. Ann Int Med 1999; 130: Collaborative Group of the Primary Prevention Project (PPP). Low-dose aspirin and vitamin E in people at cardiovascular risk: a randomised trial in general practice. Lancet 2001; 357: Garattini S, Bertele V. Adjusting regulatory rules to public health needs. Lancet 2001; 358: Garattini S, Bertele' V. Efficacy, safety, and cost of new anticancer drugs. BMJ 2002; 325: Garattini S, Bertele V, Li Bassi L. How can research ethics committees protect patients better? BMJ 2003; 326: Joppi R, Bertele' V, Garattini S. Disappointing biotech. BMJ 2005; 331: Garattini S, Bertele' V. Non-inferiority trials are unethical because they disregard patients' interests. Lancet 2007; 370 : INTRODUCTION TO THE LABORATORY'S ACTIVITIES Critical appraisal of clinical methodology Evaluation of the appropriateness of drug legislation, institutions, and regulatory procedures with respect to public health needs. Cooperation to the design and conduct of pharmacovigilance and pharmacoepidemiology studies in Europe Cooperation to the development and functioning of the pan-european Infrastructure for clinical trials (ECRIN, European Clinical Research Infrastructure Network) Optimisation of drug use and healthcare fund stewardship including potential reforms and initiatives to achieve this Critical appraisal and recommendations for European Pricing and Reimbursement systems including generics, interchangeable products within a class and new innovative medicines Assessment of emerging technologies Evaluation of marketing authorization applications submitted to the European regulatory agency (EMEA) and of subsequent variations 242

244 Cooperation to the development and solution of regulatory issues in developing countries FINDINGS/MAIN RESULTS Critical appraisal of clinical research methodological aspects as the adoption of non-inferiority design in clinical trials, the choice of inadequate comparators or outcome measures, etc Raising awareness among interested parties about the deficiencies of the present EU pharmaceutical legislation and about our proposals to improve it in the public health interest Evaluation of pan-european clinical pilot studies to be run with the support of ECRIN (European Clinical Research Infrastructure Network) to assess the validity of the overall organisation and of the quality assurance system, and to refine cost evaluation Participation in a consortium of public and private European pharmacovigilance and pharmacoepidemiology centres aimed to plan and conduct a pharmacovigilance project in Europe under the coordination of the European Medicines Agency (EMEA) Development of Pan-European strategies for the rational use of new and existing drugs: establishment of the Piperska Group Recommendations for Pan-European pricing policies for generics as well as interchangeable brands in a class once generics are available Assessment of emerging technologies in the frame of the Italian Horizon Scanning Project which provides decision makers with timely information on the potential clinical impact and cost-effectiveness of new health technologies Critical review of drug documentation at the basis of marketing authorizations Critical review of the criteria to assess pharmaceutical innovation and include new drugs in the national reimbursement schemes. NATIONAL COLLABORATIONS Italian Drug Agency (AIFA) Istituto Superiore di Sanità Department of Health Lombardy Region Italian Horizon Scanning Project 243

245 European Medicine Agency (EMEA) INTERNATIONAL COLLABORATIONS European Clinical Research Infrastructures Network (ECRIN) European Network of Centres in Pharmacoepidemiology and Pharmacovigilance (ENCePP) Karolinska Institutet, Division of Clinical Pharmacology, Department of Laboratory Medicine, and Department of Drug Management and Informatics, SE University of Liverpool Management School, Prescribing Research Group, UK International Information Network on New and Emerging Health Technologies (EuroScan) World Health Organisation (Department of Essential Drugs and Medicines Policy) Association of South East Asian Nations (ASEAN) EDITORIAL BOARD MEMBERSHIP Ricerca & Pratica Dialogo sui Farmaci NATIONAL AND INTERNATIONAL COMMITTEE MEMBERSHIP Technical Scientific Committee at the Italian Drug Agency (AIFA) Subcommittee of the Centralised Procedure at the Italian Drug Agency (AIFA) Scientific Committee of the Italian Horizon Scanning Project SELECTION OF SCIENTIFIC PUBLICATIONS FROM 2008 Araszkiewicz AA, Szabert K, Godman B, Wladysiuk M, Barbui C, Haycox A. Generic olanzapine: health authority opportunity or nightmare? Expert Rev Pharmacoeconomics Outcomes Res 2008; 8: Bertele' V, Angelici L, Barlera S, Garattini S. Thrombolysis or nothing for acute myocardial infarction? It's all the same! Br J Clin Pharmacol 2008; 65: Garattini S, Bertele' V. Do we learn the right things from clinical trials? Eur J Clin Pharmacol 2008; 64: Garattini S, Bertele' V. The mandate of the European Medicines Evaluation Agency. In : Pharmacoepidemiology and therapeutic risk management Harvey Whitney Books, Cincinnati, 2008; Garattini S, Bertele' V, Godman B, Haycox A, Wettermark B, Gustafsson L L, Piperska Group. Enhancing the rational use of new medicines across European healthcare systems. Eur J Clin Pharmacol 2008; 64:

246 Godman B, Bucsics A, Burkhardt T, et al. Insight into recent reforms and initiatives in Austria; implications for key stakeholders. Expert Rev. Pharmacoeconomcis Outcomes Res 2008; 8: Godman B, Haycox A, Schwabe U, Joppi R, Garattini S. Having your cake and eating it: Office of Fair Trading proposal for funding new drugs to benefit patients and innovative companies. Pharmacoeconomics 2008; 26: 91-8 Vitry A, Lexchin J, Sasich L, Dupin-Spriet T, Reed T, Bertele' V, Garattini S, Toop L, Hurley E. Provision of information on regulatory authorities websites. Intern Med J 2008; 38: Wettermark B, Godman B, Andersson K, Gustaffson L L, Haycox A, Bertele' V. Recent national and regional drug reforms in Sweden. Implications for pharmaceutical companies in Europe. Pharmacoeconomics : Wettermark B, Godman B, Andersson K, Gustafsson L, et al. Recent national and regional drug reforms in Sweden implications for pharmaceutical companies in Europe. Pharmacoeconomics 2008; 26: RESEARCH ACTIVITIES Critical appraisal of clinical methodology Raising awareness about potential biases in clinical research Critical evaluation of the EU pharmaceutical legislation Raising awareness among interested parties about the deficiencies of the present EU pharmaceutical legislation and about our proposals to improve it in the public health interest. Critical appraisal of ongoing reforms including pricing reforms in major European countries Evaluation of ongoing reforms across Europe to enhance generic prescribing rates, drive down generic prices and corresponding originator brands, as well as potential prices of interchangeable brands once standards become available as generics, and the potential for cross cultural learning to release valuable resources to fund increased volumes and new innovative drugs in the future without prohibitive increases in general taxation or health insurances to continue to provide equitable and comprehensive healthcare in Europe. Development of a Pan-European Infrastructure for clinical trials Participation in a distributed infrastructure linking national networks of clinical research centres and clinical trials units (ECRIN, European Clinical Research Infrastructure Network) which provides integrated one-stop shop services to investigators and sponsors in multinational studies. Development of Pan-European strategies for pharmacovigilance Developing and testing innovative methods to integrate and present information on benefits and risks in order to provide all stakeholders (patients, prescribers, regulators and pharmaceutical companies) with accurate and useful information on drug-related risks and benefits. Development of Pan-European strategies for the rational use of drugs Enhancing rational use in line with an approach that has become known as the five Es, namely: evaluation; economics; enforcement; education and engineering to further fund increased volumes and new valuable innovative drugs. 245

247 Assessment of emerging technologies Collecting information on emerging medicines with respect to their potential clinical impact and their cost effectiveness and ranking the new products according to their possible marketing authorization date, their potential innovation grade, therapeutic and economic impact, possible price and NHS sustainability with the aim to provide decision makers with timely information on the potential clinical impact and cost effectiveness of new health technologies. Assessment of drug dossiers for regulatory approvals Expert support to the Rapporteurship for marketing authorisation applications and variations to the conditions of marketing authorisation Activities for the Technical Scientific Committee at the AIFA Consultative activities for the Italian Drug Agency regarding regulatory duties with respect to drug quality, safety, efficacy, and cost. Activities for the sub-committee for the European Procedures Assessment of the dossiers for marketing authorisation applications through centralized, decentralized or mutual recognition procedures. 246

248 CENTRE OF COMPUTER SCIENCE ENGINEERING STAFF Research and Communication Informatics Head of Division ROSSI Lorenzo Marco Division of I.C.T. Services and Management Head of Division BAZZI Davide 247

249 CURRICULA VITAE Lorenzo Marco Rossi graduated in Biomedical Engineering with specialization in Hospital Clinical Instrumentation at Politecnico of Milan. He has been working with the Institute Mario Negri since Main areas of interest: 1. Planning and realization of software for clinical research 2. Planning and realization of software system for in-plant automatization 3. Planning and realization of products for multimedial divulgation Davide Bazzi graduated in Informatics with specialization in ABACUS at Istituto Tecnico Industriale Statale of Corsico. He has been working with the Institute of Mario Negro since Main areas of interest: 1. Planning, realization and management of communication Network and Data Center 2. Definement and management of quality levels in ICT services distribution 3. Planning and realization of technological innovation for ICT systems 4. Definement and application of organization s methodologies and processes for the informatics security management INTRODUCTION TO THE CENTER'S ACTIVITIES In order to fulfill even more specialization needs in informatics development, the Centre of Computer Science Engineering during 2008 has been reorganized itself considering the acquired skills and created three distinct division bound each other by a strong collaborative relationship. The Centre of Computer Science Engineering gathering informatics multidisciplinary aspects promotes and propose itself to coordinate and harmonize the development of the tools for the management information, improving the integration between informative procedures making more efficacious communication process and management of scientific and administrative datas, in order to support and fasten decisional, management, clinical trials and scientific processes. RESEARCH ACTIVITIES Implementation e of Clinical Trials gathering forms (E-CRF) Lab. Clinical Trials o Trial COMETS o Trial TAILOR o Trial HEAD & NECK Outer o Trial CIPOMO Maintenance and management of datas gathering forms for the following clinical trials Lab. Neurological Disorders (Dep. Neuroscience): o Estensione Registro Europeo SLA o Trial L-ACETYLCARNITINE o Trial ANTIEPILETTICI o Trial EPILESSIA E STROKE 248

250 o o Trial EPO VS MP IN SPINAL SHOCK Trial VALPROATO Lab. Clinical Trials (Dep. Oncology) o Trial FOLFOX o Trial TOP Lab. New Drug Development Strategies (Dep. Oncology) o Trial MAPS o Trial STARPAN Dep. Epidemiology o Trial CADASIL Lab. Quality Assessment of Geriatric Therapies and Services (Dep. Neuroscience) o Trial GISAS o Patients registry for Polipathologies and Politherapies SIMI web Web based applications connected to the projects - Utility system for the current trials gathering data program (realized) - Analysis software for interaction between drugs and integrations between interaction database and drugs reference book (in collaboration with Lab. Quality Assessment of Geriatric Therapies and Services) (developed). - Reusable electronic poll system for the Mango group - Registration form to the INSILICO workshop (in collaboration with the Lab. Environmental Chemistry and Toxicology) (developed) - Forum "Under 40" (created) - Electronic board "Negri Community" (developed and realized) - Database concerning hospitalizations, recipes and medical visitation provided from Regione Lombardia for covenant data analysis Other projects Web based application for in-plant automation New database for the Institute Orders New database for the Institute Distributed Publications Management of the database of the Institute Staff Multimedial communication - Planning videoconferences and web connection between quarters - Presentation of the new Institute: interviews and videos - DVD video editing and realization of new Institute presentation (in collaboration with Photographer Office) 249

251 Websites - Management of the Institute website - "Adamant" project website - "Fondazione Mattioli" website Informatics infrastructures - Realization of the MPLS communication data infrastructures among the Milan, Bergamo and Ranica locations - Planning and realization of the accesses and management and supporting processes control system 250

252 ITALIAN COCHRANE CENTRE STAFF Head Alessandro LIBERATI, M.D. 251

253 CURRICULUM VITAE Alessandro Liberati obtained his MD Degree in 1978 at the University of Milano and his post doctoral degree in Hygiene and Preventive Medicine in 1981 at the same university. Teaching activities: Primary responsibility of several academic and non academic training courses on the methodology of clinical research and systematic reviews/metanalyses. He is Director of the advanced Master Evidence based medicine e metodologia della ricerca sanitaria, at the Università degli Studi di Modena e Reggio Emilia. Areas of scientific expertise: methodology of clinical research with particular reference to controlled clinical trials, epidemiological methods for research synthesis (systematic reviews and metanalyses); methods of practice guidelines production and implementation, evaluation of ethical implications of clinical research. Past and current roles at the Mario Negri Institute: Junior researcher at the Laboratory of Clinical pharmacology; Head, Unit of Clinical Epidemiology and Health Services Research; Head Laboratory of Clinical Epidemiology; since 1994 he is Director of the Italian Cochrane Centre; since 1998 he is Associate Professor of Clinical Biostatistics and Epidemiology at the University of Modena and Reggio Emilia; since 1997 he is President of the Associazione per la Ricerca sulla Efficacia dell Assistenza Sanitaria - Centro Cochrane Italiano (AREAS-CCI); since 2005 he is President of the Local Ethics Committee of the Local Health Unit of Bologna; since 2004 he is Member of the Commissione Nazionale Ricerca Sanitaria; since 2005 he is Member of the Commissione Ricerca e Sviluppo dell Agenzia Italiana del Farmaco (AIFA). Selected publications Filippini G, Moja L, Liberati A, Gensini GF, Gusinu R, Conti AA. When drug companies select what they want to publish patients are denied relevant therapeutic information. Intern Emerg Med Sep;3(3):255-7 Moja L, Moschetti I, Cinquini M, Sala V, Compagnoni A, Duca P, Deligant C, Manfrini R, Clivio L, Satolli R, Addis A, Grimshaw JM, Dri P, Liberati A. Clinical evidence continuous medical education: a randomised educational trial of an open access e-learning program for transferring evidence-based information - ICEKUBE (Italian Clinical Evidence Knowledge Utilization Behaviour Evaluation) - study protocol. Implement Sci Jul 17;3:37 Iorio A, Moja L, Liberati A, Gensini GF, Gusinu R, Conti AA. Selecting references that match constructs: the difficult job of citing the parachute hyperbole. Intern Emerg Med Jun;3(2): Guyatt GH, Oxman AD, Kunz R, Jaeschke R, Helfand M, Liberati A, Vist GE, Schünemann HJ; GRADE Working Group. Incorporating considerations of resources use into grading recommendations. BMJ May 24;336(7654): Guyatt GH, Oxman AD, Kunz R, Falck-Ytter Y, Vist GE, Liberati A, Schünemann HJ; GRADE Working Group. Going from evidence to recommendations. BMJ May 10;336(7652): Banzi R, Moja L, Moschetti I, Liberati A, Gensini GF, Gusinu R, Conti AA. Rimonabant for overweight and "metabolic syndrome": the attempt to supersize disease and risk by pharmaceutical marketing. Intern Emerg Med Mar;3(1):53-6. De Palma R, Liberati A, Ciccone G, Bandieri E, Belfiglio M, Ceccarelli M, Leoni M, Longo G, Magrini N, Marangolo M, Roila F; Programma Ricerca e Innovazione Emilia Romagna Oncology Research Group. Developing clinical recommendations for breast, colorectal, and lung cancer adjuvant treatments using the GRADE system: a study from the Programma Ricerca e Innovazione Emilia Romagna Oncology Research Group. J Clin Oncol Mar 1;26(7):

254 INTRODUCTION TO THE CENTRE'S ACTIVITIES The Italian Cochrane Centre (ICC) ( was founded in 1994 and is affiliated to the Cochrane Collaboration (CC). The CC is an international non profit organization that prepares, maintains and promotes systematic reviews of the effects of health care interventions. The main product of the Cochrane Collaboration is the Cochrane Library, a quarterly publication containing Cochrane systematic reviews and other relevant databases of other siblings international organizations. The objectives of the ICC are centered around supporting various activities of the Cochrane Collaboration within Italy. In particular: a) to disseminate the knowledge of CC and CC activities throughout Italy; b) to provide methodological and practical support to all individuals and groups who are interested in collaborating with the CC; c) to contribute to the adoption and dissemination of Evidence-based Medicine in Italy. FINDINGS/MAIN RESULTS The ICC has created a national network with researchers and health care providers who are producing systematic reviews for the Cochrane Collaboration and are actively involved in other activities related to the dissemination of evidence based medicine. NATIONAL COLLABORATIONS Agenzia Italiana del Farmaco (AIFA), Roma Istituto Superiore di Sanità, Roma Ministero della Salute, Roma Centro Valutazione Efficacia Assistenza Sanitaria (CeVEAS), Modena Dipartimento di Epidemiologia della ASL Roma E Istituto Neurologico "Carlo Besta", Milano Agenzia Sanitaria Regionale, Regione Emilia Romagna, Bologna Università degli Studi di Milano Università di Modena e Reggio Emilia Azienda Sanitaria Locale 20, Alessandria INTERNATIONAL COLLABORATIONS The Cochrane Collaboration, Oxford, UK Centre for Reviews and Dissemination, University of York, York, UK British Medical Journal Publishing Group, London, UK Centre for Statistics in Medicine, Oxford, UK Thomas Chalmers Centre for Systematic Reviews, Ottawa, Canada The Campbell Collaboration, Philadelphia, USA 253

255 EDITORIAL BOARD MEMBERSHIP Clinical Evidence (Alessandro Liberati) Evidence Based Health Policy and Research (Alessandro Liberati) Journal of Clinical Epidemiology (Alessandro Liberati) Journal of Health Services Research (Alessandro Liberati) PEER REVIEW ACTIVITIES Annals of Internal Medicine (Alessandro Liberati) British Medical Journal (Alessandro Liberati) JAMA (Alessandro Liberati) Evidence Based Health Policy and Research (Alessandro Liberati) Canadian Medical Association Journal (Alessandro Liberati) EVENT ORGANIZATION V Edition Master on Evidence Based Medicine e metodologia della ricerca sanitaria, March April 2008, Università degli Studi di Modena e Reggio Emilia 1-day Cochrane Workshop in Evidence for health care decision-making Focus in: Orthopedy and physiatry - 16 May 2008, Milan Focus in: General Medicine - 12 June 2008, Milan Workshop for physiotherapists - 26 June 2008, Milan Focus in: Neurology and Internal Medicine - 10 July 2008, Milan Annual Continental European Cochrane Entities Meeting (CECEM), 8-9 May 2008, Milan Istituto Ortopedico Galeazzi Raccomandazioni cliniche : come la ricerca potrebbe orientare la pratica clinica - 16 May 2008, Milan Workshop on the 4th November 2008, Naples: -The EBM in rehabilitation: the contribution of the Cochrane Collaboration -The method GRADE to produce recommendations: how the research can guide clinical practice -Meeting of the Working Group on "New biotech drugs to treat breast cancer XIII Annual Meeting of the Italian Cochrane Le conoscenze e le innovazioni in medicina: il ruolo delle revisioni sistematiche 3 November 2008, Naples 254

256 PARTICIPATION IN EVENTS IN WHICH THE CENTRE WAS INVOLVED V Edition Master on Evidence Based Medicine e metodologia della ricerca sanitaria, March April 2008, Università degli Studi di Modena e Reggio Emilia Prove di efficacia e pratica terapeutica: EBM e Cochrane Collaboration, Master per Coordinatori di Unità Operativa e/o di Dipartimento per le professioni infermieristiche e ostetriche, Università degli Studi di Milano, anno (October 2008). Annual Continental European Cochrane Entities Meeting (CECEM) 8 9 May 2008, Milan XVI Cochrane Colloquium, 3-7 October 2008, Fribug, Germany GRANTS AND CONTRACTS Istituto di Ricerche Farmacologiche Mario Negri, Milano AIFA - Agenzia Italiana del Farmaco, Roma Ministero della Salute 255

257 SELECTION OF SCIENTIFIC PUBLICATIONS FROM 2008 De Andrea S, Montanari M, Moja L, Apolone G. Prevalence of undertreatment in cancer pain. A review of published literature. Ann Oncol Dec;19(12): Filippini G, Moja L, Liberati A, Gensini GF, Gusinu R, Conti AA. When drug companies select what they want to publish patients are denied relevant therapeutic information. Intern Emerg Med Sep;3(3): Moja L, Moschetti I, Cinquini M, Sala V, Compagnoni A, Duca P, Deligant C, Manfrini R, Clivio L, Satolli R, Addis A, Grimshaw JM, Dri P, Liberati A. Clinical evidence continuous medical education: a randomised educational trial of an open access e-learning program for transferring evidence-based information - ICEKUBE (Italian Clinical Evidence Knowledge Utilization Behaviour Evaluation) - study protocol. Implement Sci Jul 17;3:37. Iorio A, Moja L, Liberati A, Gensini GF, Gusinu R, Conti AA. Selecting references that match constructs: the difficult job of citing the parachute hyperbole. Intern Emerg Med Jun;3(2): Guyatt GH, Oxman AD, Kunz R, Jaeschke R, Helfand M, Liberati A, Vist GE, Schünemann HJ; GRADE Working Group. Incorporating considerations of resources use into grading recommendations. BMJ May 24;336(7654): Guyatt GH, Oxman AD, Kunz R, Falck-Ytter Y, Vist GE, Liberati A, Schünemann HJ; GRADE Working Group. Going from evidence to recommendations. BMJ May 10;336(7652): Banzi R, Moja L, Moschetti I, Liberati A, Gensini GF, Gusinu R, Conti AA. Rimonabant for overweight and "metabolic syndrome": the attempt to supersize disease and risk by pharmaceutical marketing. Intern Emerg Med Mar;3(1):53-6. De Palma R, Liberati A, Ciccone G, Bandieri E, Belfiglio M, Ceccarelli M, Leoni M, Longo G, Magrini N, Marangolo M, Roila F; Programma Ricerca e Innovazione Emilia Romagna Oncology Research Group. Developing clinical recommendations for breast, colorectal, and lung cancer adjuvant treatments using the GRADE system: a study from the Programma Ricerca e Innovazione Emilia Romagna Oncology Research Group. J Clin Oncol Mar 1;26(7):

258 RESEARCH ACTIVITIES Educational and dissemination activities In 2008 the Italian Cochrane Centre (ICC) participated in the organization of fifth year of the Master s program in Evidence Based Medicine (EBM) and health research methodology in collaboration with the University of Modena and Reggio Emilia in Italy. ICC organized and conducted a series of workshops about basic EBM concepts for healthcare decision-making directed toward general practitioners, specialists and doctors in training. These workshops were held at the Mario Negri Institute in Milan. In collaboration with AIFA and Zadig (a scientific publishing group), ICC has made a significant contributions toward the ECCE project (continuing education centred on evidence based medicine). The ECCE project was established in 2005 and promoted by AIFA as a part of a large educational project for continuing medical education (CME) of all Italian doctors. Within the first year of the project, ECCE attracted 17,000 doctors (2005); by the end of 2007 that number rose to AIFA and Zadig have also been partners of the ICC in the translation, adaptation, distribution and evaluation of Clinical Evidence - an international source of the best available evidence for effective healthcare (published by BMJ publishing group). This past year we have been working on the 6 th Italian edition of Clinical Evidence which corresponds to the 17th English edition. In collaboration with PartecipaSalute project, ICC began to translate and disseminate the Cochrane Collaborations' press releases (plain language summaries of Cochrane Systematic Reviews) to scientific journalists, doctors and consumers Research activities ICC researchers have collaborated in the production of 7 Cochrane Systematic Reviews and 7 protocols currently available in the Cochrane Library. ICC researchers are involved in methodological projects focused on the study of the quality of Systematic Reviews. In 2008 the ICC has collaborated with PartecipaSalute project. PartecipaSalute aims to involve consumers and their disease-related associations with follow objectives: PartecipaSalute ("Participate in Health Care") is a project initiated at September 2003 to foster a strategic alliance between patients groups and professional societies with the common goal of promoting better health and shared decision-making. The project s main aims are: - to increase patients associations involvement in healthcare assistance and in decision making processes; - to promote a partnership between patients associations and medical societies soliciting medical societies attention to patients need and perspectives - to develop a partnership between citizens, patients and healthcare system. The project developed a website in order to offer critical tools to read and understand medical and healthcare information, for better healthcare decisions. 257

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260 THE CATULLO AND DANIELA BORGOMAINERIO CENTER One of the buildings on the Mario Negri Institute campus is The Catullo and Daniela Borgomainerio Center built in 1987 thanks to a donation from Mrs. Angela Marchegiano Borgomainerio. This is a Center for the study of rare childhood diseases and even today some of the laboratories housed in the building still conduct this research. For example, the study of new therapies used to treat a very rare form of acute myeloid leukemia, know as acute promyelocytic leukemia. A number of new studies are being done to identify new drugs having different mechanisms able to synergize with trans retinoic acid. Research on epidemiological childhood leukemia is also done at the Borgomainerio and a similar line of research involves testicular cancer in adolescents and young adults. We also do research aimed at finding evidence based therapies for children. Paediatric research activities done at the Borgomainerio Center are also performed in collaboration with groups located at other Institute locations including, The Aldo and Cele Daccò Center for Clinical Research on Rare Diseases at Ranica in Bergamo, the Regional Centre for Drug Information (CRIF) and the Laboratory for Mother and Child Health (Department of Public Health) which are both located in Milan. 259

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262 THE LIBRARY STAFF Head Librarian Vanna Pistotti 261

263 The Library, specialised in pharmacology and clinical epidemiology, was founded in 1963 thanks to a generous donation from the Gustavus and Louise Pfeiffer Research Foundation, in Denville, New Jersey, USA. Numerous public and private organisations help keep it operative, through donations in money or books, and subscriptions to periodicals. STAFF One Head and two Assistants plus a clerical worker WHAT THE LIBRARY OFFERS The library has a collection of about 5000 textbooks, monographs and congressional proceedings, and 200 periodicals of which a major part are in an electronic format. The books are classified according to the US National Library of Medicine Classification and the Medical Subject headings of Medline (MeSH). Besides the internal collection, the Library has access to the National Periodical Catalogue and to other Library systems (SBBL, GIDIF-RBM). DATABESES AND ELECTRONIC JOURNALS From every computer in the Institute it is now possible to have access to more than 2000 electronic journals and to three of the most important databases, PubMed, the Cochrane Library and Embase. SPECIAL PROJECTS The Library cooperates to the realisation of the Italian Information Specialists (GIDIF, RBM) journal catalog which is updated annually and to the catalog of the Lombardy Biomedical Library Consortium, a network that serves, through Internet, the scientific community in this District. It collaborates to the maintenance of the Institute web site, particularly taking care of the Publications section, both scientific and lay press. TRAINING Every year courses on the use of the database and electronic journals are organised. These courses are designed for use by those working at the Institute but outsiders who are interested may attend. 262

264 Pistotti V Conoscere e usare Google Ricerca & Pratica 2008 n.142 : PUBLICATIONS Pistotti V, Santoro E Navigare sulla rete alla ricerca di informazioni di salute In :La dispensa di Partecipasalute: orientarsi in salute e sanità per fare scelte consapevoli IRFMN, Milano, 2008; CONTRACTS Since 1994 the library has been part of the Lombard Biomedical Library System. 14 university and research organisation libraries in Lombardy take part in this project, which allows easy, free access to scientific information to over 140 centres and institutions the Lombardy Region. 263

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266 Negri Bergamo Laboratories ANNUAL REPORT 2008 departments and laboratories 265

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268 DEPARTMENT OF MOLECULAR MEDICINE STAFF Head Ariela BENIGNI, Biol.Sci.D., Ph.D. Unit of Gene Therapy Head Susanna TOMASONI, Biol.Sci.D., Ph.D. Laboratory of Cell Biology and Xenotransplantation Head Marina MORIGI, Biol.Sci.D., Ph.D. Unit of Platelet-Endothelial Cell Interaction Head Miriam GALBUSERA, Biol.Sci.D. Laboratory of Immunology and Genetics of Organ Transplantation and Rare Diseases Head Marina NORIS, Chem.Farm.D., Ph.D. Unit of Cellular biology of Autoimmunity and Transplant Rejection Head Sistiana AIELLO, Biol.Sci.D Unit of Cellular and Molecular Biology of Transplantation Tolerance Head Federica CASIRAGHI, Chemist Laboratory of Experimental Models of Kidney Diseases Head Carla ZOJA, Biol.Sci.D., Ph.D. Unit of Pathology and Immunophatology Head Mauro ABBATE, M.D. 267

269 CURRICULA VITAE Ariela Benigni got the Biol.Sci. degree in 1979 at the University of Milano, Italy, and the Ph.D. at Maastricht University, Netherlands, in Educational training: in 1979 Post Doctoral Fellow, Istituto di Ricerche Farmacologiche Mario Negri (IRFMN), Laboratory of Cancer Chemotherapy, Milan, Italy; in Post Doctoral Fellow, Associazione Bergamasca per lo Studio delle Malattie Renali, Laboratory of the Division of Nephrology and Dialysis, Ospedali Riuniti di Bergamo, Italy; in 1982 Post Doctoral Fellow, Centre Regional de Transfusion Sanguigne de Strasbourg, France. Areas of interest: vasoactive and inflammatory mediators of progressive renal injury with a particular emphasis on endothelin-1; combined treatment of antipertensive and renoprotective drugs to halt progressive renal injury; use of stem cells for tissue regeneration in acute renal failure in vivo e in vitro gene transfer; prevention of acute graft rejection through gene therapy; induction of kidney transplant tolerance by gene therapy; correction of genetic deficiency in rare diseases. Employement: in 1983 Scientist, IRFMN, Laboratory of Kidney Disease, Bergamo, Italy; in Head Laboratory of Prostaglandin and Leukotriene Metabolism, IRFMN, Bergamo, Italy; from January 1991 Scientific Secretary, IRFMN, Bergamo, Italy; in Head Laboratory of Vasoactive and Inflammatory Mediators of Tissue damage, IRFMN, Bergamo, Italy; from January 2000 Head, Department of Molecular Medicine, IRFMN, Bergamo, Italy; from March 2007 Consultant World Health Organization (WHO) for the multicentre observational study Screening for Pre-eclampsia: evaluation of the predictive ability of angiogenic factors for Pre-eclampsia ; during 2007 Senior Fellow at the University of Oxford, Nuffield Department of Obstetrics & Gynaecology. Selected publications: Macconi D, Chiabrando C, Schiarea S, Aiello S, Cassis L, Gagliardini E, Noris M, Buelli S, Zoja C, Corna D, Mele C, Fanelli R, Remuzzi G, Benigni A. Proteasomal processing of albumin by renal dendritic cells generates antigenic peptides. J Am Soc Nephrol Jan;20(1): Epub 2008 Dec 17. Vlahou A, Charonis A, Benigni A. Report on the first combined working group and management committee meeting of EuroKUP (Urine and Kidney Proteomics cost action). J Proteomics Jan 30;71(6): Epub 2008 Nov 17. Perico N, Benigni A, Remuzzi G. Present and future drug treatments for chronic kidney diseases: evolving targets in renoprotection. Nat Rev Drug Discov Nov;7(11): Epub 2008 Oct 10. Review. Morigi M, Introna M, Imberti B, Corna D, Abbate M, Rota C, Rottoli D, Benigni A, Perico N, Zoja C, Rambaldi A, Remuzzi A, Remuzzi G. Human bone marrow mesenchymal stem cells accelerate recovery of acute renal injury and prolong survival in mice. Stem Cells Aug;26(8): Epub 2008 May 22. Pezzotta A, Mister M, Monteferrante G, Cassis L, Azzollini N, Aiello S, Satta M, Benigni A, Remuzzi G, Noris M. Effect of seliciclib (CYC202, R-roscovitine) on lymphocyte alloreactivity and acute kidney allograft rejection in rat. Transplantation May 27;85(10): de Borst MH, Benigni A, Remuzzi G. Primer: strategies for identifying genes involved in renal disease. Nat Clin Pract Nephrol May;4(5): Epub 2008 Mar 25. Review. Marina Morigi got her Biol.Sci. degree in 1987 at the University of Milano, Milano, Italy and the Ph.D. at Maastricht University, Netherlands, in Educational training: in Research training, IRFMN, Bergamo, Italy; in Post Doctoral Fellow, IRFMN, Bergamo, Italy; in 1991 Stage at Brigham and Women s Hospital, Laboratory of Dr. P. Marsden, Boston, USA. Employement: since 1995 Scientist, IRFMN, Bergamo, Italy; in Head, Unit of Renal and Endothelial Cell Biology; since 2000 Head, Laboratory of Cell Biology and Xenotransplantation, IRFMN, Bergamo, Italy. Areas of interest: role of Shigatoxin in the pathogenesis of endothelial dysfunction and microvascular thrombosis in Hemolytic Uremic Syndrome; in vitro model of hyperacute xenograft rejection (porcine endothelium exposed to human serum as a source of xenoreactive natural antibodies and complement); renal toxicity of the proteins filtered through the capillary barrier. In vitro model to study intracellular signals, gene expression and production of inflammatory mediators in cultured proximal tubular cells and glomerular epithelial cells; cell therapy and tissue regeneration: Capability of adult stem cells to differentiate and to regenerate renal tissue in acute and chronic experimental models of renal disease. 268

270 Murine embryonic stem cell therapy to correct the genetic defect characteristic of Fabry disease in an experimental mouse model. Selected publications Pretransplant infusion of mesenchymal stem cells prolongs the survival of a semiallogeneic heart transplant through the generation of regulatory T cells. Casiraghi F, Azzollini N, Cassis P, Imberti B, Morigi M, Cugini D, Cavinato RA, Todeschini M, Solini S, Sonzogni A, Perico N, Remuzzi G, Noris M. J Immunol Sep 15;181(6): Human bone marrow mesenchymal stem cells accelerate recovery of acute renal injury and prolong survival in mice. Morigi M, Introna M, Imberti B, Corna D, Abbate M, Rota C, Rottoli D, Benigni A, Perico N, Zoja C, Rambaldi A, Remuzzi A, Remuzzi G. Stem Cells Aug;26(8): Epub 2008 May 22. The regenerative potential of stem cells in acute renal failure. Morigi M, Benigni A, Remuzzi G, Imberti B. Cell Transplant. 2006;15 Suppl 1:S Review. M. Morigi, B. Imberti, C. Zoja, D. Corna, S. Tomasoni, M. Abbate, D. Rottoli, S. Angioletti, A. Benigni, N. Perico, M. Alison, G. Remuzzi. Mesenchymal Stem Cells Are Renotropic, Helping to Repair the Kidney and Improve Function in Acute Renal Failure. J Am Soc Nephrol 2004;15: Morigi M, Buelli S, Zanchi C, Longaretti L, Macconi D, Benigni A, Moioli D, Remuzzi G, Zoja C. Shigatoxin-induced endothelin-1 expression in cultured podocytes autocrinally mediates actin remodeling. Am J Pathol Dec; 169(6): Imberti B, Morigi M, Tomasoni S, Rota C, Corna D, Longaretti L, Rottoli D, Valsecchi F, Benigni A, Wang J, Abbate M, Zoja C, Remuzzi G. Insulin-like growth factor-1 sustains stem cell mediated renal repair. J Am Soc Nephrol Nov;18(11): Marina Noris got her degree in Pharmaceutical Chemistry and Technologies in 1986 at the University of Rome La Sapienza) and the Ph.D. at Maastricht University, Netherlands, in Educational training: in Fellow, Istituto di Chimica Farmaceutica e Tossicologica, University of Rome, Italy; in Post Doctoral Fellow, Istituto di Chimica Farmaceutica e Tossicologica, University of Rome, Italy; in September 1987-March 1994 Post Doctoral Fellow, IRFMN, Unit of Mediators of Inflammation and Tissue Damage, Laboratory of Kidney Disease, Bergamo, Italy. Areas of interest: immunology of transplantation, tolerance induction; genetics of hemolytic uremic syndrome, thrombotic thrombocytopenic purpura, focal segmental glomerulosclerosis, diabetic nephropathy, role of nitric oxide and arginine dysfunctions in uremia and in pre-eclampsia. Employment: in Head, Unit of Endothelial Cell Pathophysiology, IRFMN, Bergamo, Italy; Head, Laboratory of Cellular and Molecular Biology of the immune response and autoimmunity, IRFMN, Italy; from January 2000: Head, Laboratory of Immunology and Genetics of Rare Diseases and Organ Transplantation, Department of Molecular Medicine, IRFMN, Bergamo, Italy. Selected publications Castelletti F, Donadelli R, Banterla F, Hildebrandt F, Zipfel PF, Bresin E, Otto E, Skerka C, Renieri A, Todeschini M, Caprioli J, Caruso RM, Artuso R, Remuzzi G, Noris M. Mutations in FN1 cause glomerulopathy with fibronectin deposits. Proc Natl Acad Sci U S A. 2008;105(7): Fang CJ, Fremeaux-Bacchi V, Liszewski MK, Pianetti G, Noris M, Goodship TH, Atkinson JP. Membrane cofactor protein mutations in atypical hemolytic uremic syndrome (ahus), fatal Stx-HUS, C3 glomerulonephritis, and the HELLP syndrome. Blood;111(2): Casiraghi F, Azzollini N, Cassis P, Imberti B, Morigi M, Cugini D, Cavinato RA, Todeschini M, Solini S, Sonzogni A, Perico N, Remuzzi G, Noris M. Pretransplant infusion of mesenchymal stem cells prolongs the survival of a semiallogeneic heart transplant through the generation of regulatory T cells. J Immunol. 2008;181(6): Caprioli J, Noris M, Brioschi S, Pianetti G, Castelletti F, Bettinaglio P, Mele C, Bresin E, Cassis L, Gamba S, Porrati F, Bucchioni S, Monteferrante G, Fang CJ, Liszewski MK, Kavanagh D, Atkinson JP, Remuzzi G. Genetics of HUS: the impact of MCP, CFH and IF mutations on clinical presentation, response to treatment, and outcome. Blood, 2006; 108(4): Noris M, Casiraghi F, Todeschini M, Cravedi P, Cugini D, Monteferrante G, Aiello S, Cassis L, Gotti E, Gaspari F, Cattaneo D, Perico N, Remuzzi G. Regulatory T Cells and T Cell Depletion: Role of Immunosuppressive Drugs. J Am Soc Nephrol. 2007; 18 (3): Noris M, Bucchioni s, Galbusera M, Donadelli R, Bresin E, Castelletti F, Caprioli J, Brioschi S, Scheiflinger F, Remuzzi G and the International Registry of Recurrent and Familial HUS/TTP. Complement factor H mutation in familial thrombotic thrombocytopenic purpura with ADAMTS13 deficency and renal involvement. J Am Soc Nephrol 2005; 16: Noris M, Brioschi S, Caprioli J, Todeschini M, Bresin E, Porrati F, Gamba S, Remuzzi G, on behalf of the International Registry of Familial and Recurrent HUS/TTP. Familial haemolytic uraemic syndrome and an MCP mutation. Lancet 2003; 362: Carlamaria Zoja got her Biol.Sci. degree at the University of Milano, Italy, in 1979 and the Ph.D. at the University of Maastricht, The Netherlands in Educational Training: in Post Doctoral Fellow, Associazione Bergamasca per lo studio delle Malattie Renali, Laboratory of the Division of Nephrology and Dialysis, Ospedali Riuniti di Bergamo, 269

271 Italy; in Post Doctoral Fellow, Center for Thrombosis and Vascular Research, Department of Research Katholieke Universiteit, Leuven, Belgium; in : Post Doctoral Fellow, IRFMN, Laboratory of Kidney Disease, Bergamo, Italy. Areas of interest: experimental models of kidney diseases of immunological and non immunological origin; vasoactive and inflammatory mediators of renal disease progression; role of proteinuria in progressive kidney damage; protection of renal disease progression by a multidrug approach; novel immunosuppressive and anti-inflammatory strategies for the treatment of lupus nephritis; role of Shigatoxin in the pathogenesis of endothelial dysfunction in Hemolytic Uremic Syndrome. Employement: since 1985 Scientist, IRFMN, Bergamo, Italy; in : Head, Unit of Experimental Modelling for Human Renal Diseases, Laboratory of Kidney Diseases, IRFMN, Bergamo, Italy; since 1995: Head, Laboratory of Experimental Models of Kidney Diseases, IRFMN, Bergamo, Italy. Selected publications C.Zoja, S. Angioletti, R. Donadelli, C. Zanchi, S. Tomasoni, E. Binda, B. Imberti, M. te Loo, L. Monnens, G.Remuzzi, M. Morigi. Shiga toxin-2 triggers endothelial leukocyte adhesion and transmigration via NF-kB dependent up-regulation of IL-8 and MCP-1. Kidney Int 2002;62: C. Zoja, D. Corna, D. Camozzi, D. Cattaneo, D. Rottoli, C. Batani, C. Zanchi, M. Abbate, G. Remuzzi. How to fully protect the kidney in a severe model of progressive nephropathy: a multidrug approach. J Am Soc Nephrol 2002;13: R. Donadelli, C. Zanchi, M. Morigi, S. Buelli, C. Batani, S. Tomasoni, D. Corna, D. Rottoli, A. Benigni, M. Abbate, G. Remuzzi, C. Zoja. Protein overload induces fractalkine upregulation in proximal tubular cells through NF-kB and p38 MAPK dependent pathways. J Am Soc Nephrol 2003; 14: C.Zoja, F.Casiraghi, S.Conti, D.Corna, D.Rottoli, R.A.Cavinato, G.Remuzzi, A.Benigni. Cyclin-Dependent kinase inhibition limits glomerulonephritis and extends lifespan of mice with systemic lupus. Arthritis & Rheumatism 2007; 56; C.Zanchi, C.Zoja, M. Morigi, F.Valsecchi, XY Liu, D. Rottoli, M. Locatelli, S. Buelli, A.Pezzotta, P.Mapelli, J. Geelen, G.Remuzzi, J.Hawiger J. Fractalkine and CX3CR1 mediate leukocyte capture by endothelium in response to Shiga toxin. J Immunol. 2008; 181: M.Abbate, C.Zoja, D.Corna, D.Rottoli, C.Zanchi, N.Azzollini, S. Tomasoni, S.Berlingeri, M.Noris, M.Morigi, G.Remuzzi. Complement-mediated dysfunction of glomerular filtration barrier accelerates progressive renal injury. J Am Soc Nephrol. 2008; 19: Mauro Abbate obtained his M.D. degree in 1988 at the University of Brescia, Italy. Educational training: in Graduate Student, IRFMN, Bergamo, Italy; in Post Doctoral Fellow, IRFMN, Bergamo, Italy; in Research Fellow, The Renal Unit, Massachusetts General Hospital, HMS, Boston, USA. Areas of interest: renal disease progression: the role of proteinuria, complement, and mediators of injury in progressive kidney damage; mechanisms of glomerular injury; anti-gbm glomerulonephritis; mechanisms of tubular injury; kidney fibrosis; the renal biopsy; membranous nephropathy. Employement: in : Scientist, IRFMN, Bergamo, Italy; from 2000 Head, Unit of Renal Pathology and Immunopathology, IRFMN, Bergamo, Italy. Principali pubblicazioni Abbate M, Zoja C, Morigi M, Rottoli D, Angioletti S, Tomasoni S, Zanchi C, Longaretti L, Donadelli R, Remuzzi G: Transforming Growth Factor-beta 1 Is Up-Regulated by Podocytes in Response to Excess Intraglomerular Passage of Proteins: A Central Pathway in Progressive Glomerulosclerosis. Am J Pathol.2002;161, Macconi D, Abbate M, Morigi M, Angioletti S, Mister M, Buelli S, Bonomelli M, Mundel P, Endlich K, Remuzzi A, Remuzzi G. Permselective dysfunction of podocyte-podocyte contact upon angiotensin II unravels the molecular target for renoprotective intervention. Am J Pathol Apr;168(4): Abbate M, Zoja C, Remuzzi G. How does proteinuria cause progressive renal damage? J Am Soc Nephrol Nov;17(11): Review Ruggenenti P, Cravedi P, Sghirlanzoni MC, Gagliardini E, Conti S, Gaspari F, Marchetti G, Abbate M, Remuzzi G. Effects of rituximab on morphofunctional abnormalities of membranous glomerulopathy. Clin J Am Soc Nephrol Nov;3(6): Morigi M, Introna M, Imberti B, Corna D, Abbate M, Rota C, Rottoli D, Benigni A, Perico N, Zoja C, Rambaldi A, Remuzzi A, Remuzzi G. Human bone marrow mesenchymal stem cells accelerate recovery of acute renal injury and prolong survival in mice. Stem Cells Aug;26(8): Abbate M, Zoja C, Corna D, Rottoli D, Zanchi C, Azzollini N, Tomasoni S, Berlingeri S, Noris M, Morigi M, Remuzzi G. Complement-mediated dysfunction of glomerular filtration barrier accelerates progressive renal injury. J Am Soc Nephrol Jun;19(6):

272 Sistiana Aiello got the Biol.Sci. degree in 1993 at the University of Milano, Italy, and the Specialization in Pharmacology Research in 1996, at IRFMN, Bergamo, Italy. Educational training: in research training, IRFMN, Bergamo; in post doctoral fellow, IRFMN, Bergamo. Areas of interest: transplant immunology with a particular interest on dendritic cell biology and mechanisms by which T regulatory cells arise and work; in vitro and in vivo studies on new compounds with immunosuppressive capacity or capable to prevent ischemia/reperfusion tissue injury; vasoactive and inflammatory mediators of progressive renal injury with a particular emphasis on platelet activating factor (PAF) and nitric oxide (NO). Employement: since 2000 Scientist within Laboratory of Immunology and Genetics of Rare disease and Organ Transplantation; IRFMN, Bergamo; since 2006 Head, Unit of Cellular Biology of Autoimmunity and Transplant Rejection, IRFMN, Transplant Research Center Chiara Cucchi de Alessandri e Gilberto Crespi, Ranica. Selected publications: Pezzotta A, Mister M, Monteferrante G, Cassis L, Azzollini N, Aiello S, Satta M, Benigni A, Remuzzi G, Noris M. Effect of seliciclib (CYC202, R-roscovitine) on lymphocyte alloreactivity and acute kidney allograft rejection in rat. Transplantation. 2008; 85(10): S. Aiello, P. Cassis, L. Cassis, S. Tomasoni, A. Benigni, A. Pezzotta, R.A. Cavinato, D. Cugini, N. Azzollini, M. Mister, L. Longaretti, A.W. Thomson, G. Remuzzi, M. Noris. DnIKK2-transfected dendritic cells induce a novel population of inos-expressing CD4 + CD25 - cells with tolerogenic properties. Transplantation 2007; 83: S. Tomasoni, S. Aiello, L. Cassis, M. Noris, L. Longaretti, R.A. Cavinato, N. Azzollini, A. Pezzotta, G. Remuzzi, A. Benigni. Dendritic cells genetically engineered with adenoviral vector encoding dnikk2 induce the formation of potent CD4 + T regulatory cells. Transplantation 2005;79: L. Cassis, S. Aiello, M. Noris. Natural versus adaptive regulatory T cells. Contrib Nephrol 2005; 146: M. Mister, M. Noris, J. Szymczuk, N. Azzollini, S. Aiello, A. Arduini, L. Trochimowicz, E. Gagliardini, M. Abbate, N. Perico, G. Remuzzi. Propionyl-L-carnitine prevents renal function deterioration due to ischemia/reperfusion in isolated perfused rat kidney and in kidney graft. Kidney Int 2002; 61: S. Aiello, M. Noris, G. Piccinini, S. Tomasoni, F. Casiraghi, S. Bonazzola, M. Mister, M.H. Sayegh, G. Remuzzi. Thymic dendritic cells express inducible nitric oxide synthase and generate nitric oxide in response to self- and alloantigens. J Immunol 2000;164: Federica Casiraghi has obtained his degree in Industrial Chemistry in 1988, and the degree in Clinical Monitoring and in Biochemical Research in at IRFMN, Bergamo, Italy. Educational Training: research fellow, IRFMN, Bergamo. Areas of interest: Transplant immunology with particular focus on pharmacological and cellular therapies for induction and maintenance of transplantation tolerance. Characterization of regulatory T cells in renal transplant patients and in experimental models of allograft tolerance. Impact of different immunosuppressive drugs on T cell function in renal transplant patients. Vasoactive and inflammatory mediators of progressive renal injury with a particular emphasis on arachidonic acid metabolites. Employment: since 1994 Scientist within Laboratory of Immunology and Genetics of Rare Disease and Organ Transplantation, IRFM, Bergamo; since 2006 Head, Unit of Cellular and Molecolar Biology of Transplantation Tolerance, Transplant Research Center Chiara Cucchi de Alessandri e Gilberto Crespi, Ranica. Selected publications: Casiraghi F, Azzollini N, Cassis P, Imberti B, Morigi M, Cugini D, Cavinato RA, Todeschini M, Solini S, Sonzogni A, Perico N, Remuzzi G, Noris M. Pretransplant infusion of mesenchymal stem cells prolongs the survival of a semiallogeneic heart transplant through the generation of regulatory T cells. J Immunol. 2008;181(6): Noris M, Casiraghi F, Todeschini M, Cravedi P, Cugini D, Monteferrante G, Aiello S, Cassis L, Gotti E, Gaspari F, Cattaneo D, Perico N, Remuzzi G. Regulatory T Cells and T Cell Depletion: Role of Immunosuppressive Drugs. J Am Soc Nephrol. 2007; 18 (3): Cavinato RA, Casiraghi F, Azzollini N, Cassis P, Cugini D, Mister M, Pezzotta A, Aiello S, Remuzzi G, Noris M. Pretransplant donor peripheral blood mononuclear cells infusion induces transplantation tolerance by generating regulatory T cells. Transplantation, 2005;79(9): Zoja C, Benigni A, Noris M, Corna D, Casiraghi F, Pagnoncelli M, Rottoli D, Abbate M, Remuzzi G. Mycophenolate mofetil combined with a cyclooxygenase-2 inhibitor ameliorates murine lupus nephritis. Kidney Int. 2001; 60(2): Tomasoni S, Noris M, Zappella S, Gotti E, Casiraghi F, Bonazzola S, Benigni A, Remuzzi G. Upregulation of renal and systemic cyclooxygenase-2 in patients with active lupus nephritis. J Am Soc Nephrol.1998; 9(7):

273 Casiraghi F, Ruggenenti P, Noris M, Locatelli G, Perico N, Perna A, Remuzzi G. Sequential monitoring of urine-soluble interleukin 2 receptor and interleukin 6 predicts acute rejection of human renal allografts before clinical or laboratory signs of renal dysfunction. Transplantation 1997; 63(10): Miriam Galbusera got her Biol.Sci. degree in 1981 at the Università degli Studi di Milano. Educational training: in Post Doctoral Fellow, Istituto di Patologia Speciale Medica dell'università degli Studi di Milano, Italy; in Post Doctoral Fellow, IRFMN, Bergamo, Italy; in Post Doctoral Fellow at Scripps Clinic and Research Foundation, Laboratory of Thrombosis and Hemostasis, La Jolla, CA, USA; in Post Doctoral Fellow, IRFMN, Bergamo, Italy. Areas of interest: ADAMTS-13 and VWF in thrombotic microangiopathies, VWF biochemistry, xenotransplantation, platelet-endothelial cell interaction under flow condition, platelet pathophysiology in uremia, receptor studies in kidney and platelets. Employement: : Scientist, IRFMN, Bergamo, Italy; from 2000 Head, Unit of Platelet- Endothelial Cell Interaction, IRFMN, Bergamo, Italy. Selected publications Tripodi, A., Chantarangkul, V., Bohm, M., Budde, U., Dong, J.-F., Friedman, K.D., Galbusera, M., Girma, J.-P., Moake, J., Rick, M.E., Studt, J.-D., Turecek, P.L., Mannucci, P.M.: Measurement of von Willebrand factor cleaving protease (ADAMTS-13): results of an international collaborative study involving 11 methods testing the same set of coded plasmas. J Thromb Haemost 2004; 2: Galbusera, M., Morigi, M., Buelli, S., Gastoldi, S., Macconi, D., Testa, C., Angioletti, S., Remuzzi, G.: Xenogeneic serum-induced thrombus formation on porcine endothelium is mediated by vitronectin receptor and P-selectin: role of reactive oxygen species. Xenotransplantation 2005;12: Ruiz-Torres, M.P., Casiraghi, F., Galbusera, M., Macconi, D., Gastoldi, S., Todeschini, M., Porrati, F., Belotti, D., Pogliani, E.M., Noris, M., Remuzzi, G.: Complement activation: the missing link between ADAMTS13 deficiency and microvascular thrombosis of thrombotic microangiopathies. Thromb Haemost 2005; 93: Noris, M., Bucchioni, S., Galbusera, M., Donadelli, R., Bresin, E., Castelletti, F., Caprioli, J., Brioschi, S., Scheiflinger, F., Remuzzi, G.: Complement factor H mutation in familial thrombotic thrombocytopenic purpura with ADAMTS13 deficiency and renal involvement. J Am Soc Nephrol 2005; 16: Galbusera, M., Bresin, E., Noris, M., Gastoldi, S., Belotti, D., Capoferri, C., Daina, E., Perseghin, P., Scheiflinger, F., Fakhouri, F., Grunfeld, J-P., Pogliani, E., Remuzzi, G.: Rituximab prevents recurrence of thrombotic thrombocytopenic purpura: a case report. Blood 2005;106: Rieger, M., Mannucci, P.M., Kremer Hovinga, J.A., Herzog, A., Gerstenbauer, G., Konetschny, C., Zimmermann, K., Scharrer, I., Peyvandi, F., Galbusera, M., Remuzzi, G., Böhm, M., Plaimauer, B., Lämmle, B., Scheiflinger, F.: ADAMTS13 autoantibodies in patients with thrombotic microangiopathies and other immunomediated diseases. Blood 2005;106: Susanna Tomasoni got her Biological Science degree in 1991 at the University of Milan. Educational training: in Graduate student, University of Milan; in PhD student, University of Milan; in 1994 Research Fellow, Renal Division, Brigham & Women s Hospital, Harvard Medical School, Boston, USA; in 1995 PhD degree in Physiological Science, University of Bologna; : Post Doctoral Fellow, IRFMN, Bergamo, Italy. Areas of interest: construction of adenoviral vectors for gene therapy; gene transfer to the kidney in the context of transplantation; transfection of dendritic cell for cell therapy; progression of renal disease. Employement: in Scientist, IRFMN, Bergamo, Italy; from 2000 Head, Unit of Gene Therapy, IRFMN, Bergamo, Italy Selected publications Allograft rejection: acute and chronic studies. Tomasoni S, Remuzzi G, Benigni A. Contrib Nephrol. 2008;159: Review. Tomasoni S, Azzollini N, Casiraghi F, Capogrossi M C, Remuzzi G, Benigni A. CTLA4Ig gene transfer prolongs survival and induces donor-specific tolerance in a rat renal allograft. J Am Soc Nephrol 2000; 11: Tomasoni S, Benigni A. Gene therapy: How to target the kidney. Promises and pitfalls. Curr Gene Ther 2004; 4: Tomasoni S, Longaretti L, Azzollini N, Gagliardini E, Mister M, Buehler T, Remuzzi G, Benigni A. Favorable effect of cotransfection with TGF-beta and CTLA4Ig of the donor kidney on allograft survival. Am J Nephrol 2004; 24: Tomasoni S, Aiello S, Cassis L, Noris M, Longaretti L, Cavinato R, Azzollini N, Pezzotta A, Remuzzi G, Benigni A. Dendritic cells genetically engineered with adenoviral vector encoding dnikk2 induce the formation of potent CD4+ T- regulatory cells. Transplantation 2005; 79: Benigni A, Tomasoni S, Turka LA, Longaretti L, Zentilin L, Mister M, Pezzotta A, Azzollini N, Noris M, Conti S, Abbate M, Giacca M, Remuzzi G, Adeno-associated virus-mediated CTLA4Ig gene transfer protects MHC-mismatched renal allografts from chronic rejection. J Am Soc Nephrol, 2006, 17:

274 INTRODUCTION TO THE DEPARTMENT'S ACTIVITIES The Department of Molecular Medicine was established in 1999 at the Negri Bergamo laboratories to coordinate the work of three laboratories and three units. The activities of the Department of Molecular Medicine are strictly interrelated with those of the Department of Renal Medicine of the Clinical Research Center for Rare Diseases Aldo e Cele Daccò. The following major objectives have been pursued: 1) identification of mediators and mechanisms responsible for the relentless decline of renal function in kidney diseases and development of therapeutic interventions to slow or even halt the disease progression to end-stage renal failure; 2) understanding the mechanisms underlying endothelial cell dysfunction in thrombotic microangiopathies and hyperacute rejection of xenograft 3) finding new strategies for modulating the immune response and preventing acute and chronic rejection of kidney allograft as well as exploration of immunological pathways leading to donor specific unresponsiveness and tolerance of the graft; 4) investigation of the molecular and genetic basis of rare diseases such as hemolytic uremic syndrome/thrombotic thrombocytopenic purpura and pre-eclampsia and search for diseasesusceptibility genes or gene polymorphisms predicting the patient's response to drug therapy in more common and complex polygenic disorders. Such goals have been pursued using various approaches: 1) experimental models of kidney diseases of immunological and non-immunological origin mimicking human renal diseases to study vasoactive and inflammatory mediators and to test novel antiproteinuric and renoprotective drugs; 2) in vitro cultures of renal cells to address the toxicity of protein overload reproducing the condition of exaggerated protein traffic of proteinuric progressive nephropathies; 3) in vitro models to assess the interaction of vascular endothelial cells with leukocytes and platelets under controlled flow conditions; 4) experimental models of kidney allotransplant to study immunological processes responsible for acute and chronic rejection, the nephrotoxicity of immunosuppressor drugs as well as to explore pathways responsible for accomodation; 5) gene transfer of viral constructs carrying genes encoding immunomodulatory molecules to overcome acute rejection of allotransplantation avoiding immunosuppression; 6) identification of candidate genes with linkage analysis and search for mutations as well as assessment of gene polymorphisms. FINDINGS/MAIN RESULTS Cord blood mesenchymal stem cells prolong survival of mice with acute kidney injury. Pre-transplant infusion of adult or embryonic mesenchymal stem cells prolongs survival of heart transplant in mice through the generation of regulatory T cells. C3 filtered in the urine induces renal damage. Fractalkine induces micro vascular lesions typical of HUS because it induces leucocyte adhesion to the endothelium. 273

275 Self proteins become antigenic in disease conditions and promote renal injury. A cell cycle inhibitor is a potent immunosuppressor in transplant setting. First description of fibronectin 1 mutations as a cause of a rare disease, the glomerulopathy with fibronectin deposit. Abnormal function of mutated MCP which predisposes to atypical HUS Proof of concept that gene therapy with adenoviral vectors corrects ADAMTS13 deficiency in Thrombotic Thrombocytopenic Purpura NATIONAL COLLABORATIONS Laboratorio di Terapia genica e cellulare, G. Lanzani, Divisione di Ematologia, Ospedali Riuniti di Bergamo Laboratorio di Tecnologie riproduttive, Istituto Sperimentale Lazzaro Spallanzani, Cremona Centro Trasfusionale e di Immunologia dei Trapianti, IRCCS Ospedale Maggiore, Milano Dipartimento di Istologia Microbiologia e Biotecnologie Mediche, Università di Padova International Centre for Genetic Engineering and Biotechnology, Molecular Medicine Group, Trieste U.O. di Ostetricia e Ginecologia, Ospedale San Gerardo di Monza U.O. di Ostetricia e Ginecologia, Azienda Ospedaliera Ospedali Riuniti di Bergamo U.O. di Ostetricia e Ginecologia, Azienda Ospedaliera Spedali Civili di Brescia I.R.C.C.S. Policlinico San Matteo, Pavia INTERNATIONAL COLLABORATIONS Academisch Ziekenhuis Maastricht, Interne Geneeskunde, Maastricht, Olanda Beth Israel Deaconess Medical Center and Harvard Medical School, Boston, USA Children's Hospital and Regional Medical Center, University of Washington, Seattle, USA Departements of Pediatrics and Human Genetics, University of Michigan, Ann Arbor, USA Deparment of Medicine, Division of Rheumatology, Washington University School of Medicine, St. Louis, USA Erasmus University of Rotterdam, Olanda Hans-Knoll Institute for Natural Products Research, Jena, Germania INSERM, Paris, Francia Klinikum der Ludwig Maximillians Universitat Munchen, Germania Max Delbruck Center for Molecular Medicine, Berlin, Germania Max-Plank Gesellschaft zur Forderung der Wissenshaften, Hpi of experimental endocrinology, Hannover, Germania National Institute of Health, Bethesda, USA Necker Hospital, Paris, Francia Osaka University School of Medicine, Osaka, Giappone Pediatric Nephrology and Hypertension, University of Utah, USA Renal Transplant Unit, Hospital de Bellvitge, Barcelona, Spagna Rosalind Franklin University of Medicine and Science, Chicago, USA The Scripps Research Institute, La Jolla, USA Universitaet Hamburg, Institut fur Molekulare Neuropathobiologie, Hamburg, Germania 274

276 University of Aarthus, Danimarca University of Colorado Cardiovascular Institute, Denver, USA University of Groningen, Olanda University of Iowa, Department of Internal Medicine and Pediatrics, Iowa City, USA University of Liverpool, School of Biological Sciences, UK University of Oxford, UK University of Oulu, Danimarca University of Pittsburgh School of Medicine, Pittsburgh, USA University of Zurich, Svizzera Weizman Institute of Science, Rehovot, Israele World Health Organization, Geneva, Svizzera EDITORIAL BOARD MEMBERSHIP Journal of American Society of Nephrology (Marina Noris) PEER REVIEW ACTIVITIES American Journal of Pathology American Journal of Physiology Cell Proliferation Diabetologia Hypertension Immunology Kidney International Journal of Clinical Investigation Journal of the Renin Angiotensin Aldosterone System Journal of the American Society of Nephrology Journal of Thrombosis and Haemostasis Nature Medicine Nephrology, Dialysis and Transplantation Nephron Clinical Practice New England Journal of Medicine Pediatric Nephrology Plos Medicine The American Journal of Pathology The Canadian Journal of Physiology and Pharmacology The Journal of Experimental Medicine The Lancet The Open Urology & Nephrology Journal Transplant Immunology Trends in molecular medicine 275

277 PARTICIPATION IN EVENTS IN WHICH THE DEPARTMENT WAS INVOLVED Meeting per progetto GENECURE: Berlino, gennaio 2008; Ranica (BG), April 2008; Bruxelles, 15 Dicembre 2008 Nefropatie primitive e secondarie, Firenze, November 2008 III Convegno Internazionale sulle cellule Staminali: le cellule staminali tra terapia e formazioni dei tumori, Agrigento-Favara, November 2008 Workshop Mesenchymal stem cells, 18 November 2008, Milano American Society of Nephrology, Philadelphia, 5-10 November 2008 Annual GRIP Investigator Meeting, 9-10 November 2008, Philadelphia "Rare diseases and Orphan Drugs" Istituto Superiore di Sanità, Roma dal October 2008 Focus on Rare Diseases: the Genetic and Molecular basis of Rare kidney Disorders, Bergamo, October XXII International complement workshop, Basilea, 28 settembre-2 October 2008 European Society for Pediatric Nephrology, Lione, 14 September 2008 Corso di aggiornamento "Renal Biopsy in Medical Diseases of the Kidneys", Columbia University, New York, September 2008 Meeting of the Dense Deposit Disease Focus group, Hinxton (London), August 2008 Workshop Kidstem Network, 3-4 July 2008, Dresden American Transplant Congress, Toronto, 31 May-4 June 2008 ERA-EDTA course on Complement in renal disease and transplantation, Leiden, April 2008 Podonet Steering Committee meeting, Heidelberg, 20 April 2008 Advanced Workshop of Nephoprotection, April 2008, Centro Daccò, Ranica (BG) EuReGene Yearly Meeting- Symposium on Complex (Renal) Genetics, Nizza, 31 gennaio 2008 EuroKUP First Meeting, Bruxelles, May 2008 Annual European Congress of Rheumatology EULAR 2008, Parigi, June nd Annual Conference of the Council for High Blood Pressure Research, Atlanta, September

278 GRANTS AND CONTRACTS Comitato Telethon Fondazione ONLUS Commissione Europea FP6 e FP7 Fondazione Aiuti per la Ricerca sulle Malattie Rare (ARMR) Fondazione ART per la Ricerca sui Trapianti ONLUS Fondazione Cariplo Fondazione La Nuova Speranza Lotta alla Sclerosi focale ONLUS Fondazione ROTRF/JDRF Istituto Superiore di Sanità ACRAF (Aziende Chimiche Riunite Angelini Francesco Spa) Farmaceutici Damor Spa Genzyme Corporation Giuliani Spa Pfizer Ltd Sanofi-Aventis Spa Speedel Pharma Ltd SELECTION OF SCIENTIFIC PUBLICATIONS FROM 2008 M. Abbate, G. Remuzzi, C. Zoja. Role of proteinuria in progression. In: The Kidney: Physiology and Pathophysiology, 4 th ed., chapter n. 89. Edited by R.J. Alpern, S.C. Hebert. Elsevier, London 2008, pp M. Noris, G. Remuzzi, T.H.J. Goodship. Hemolytic uremic syndrome/thrombotic thrombocytopenic purpura. In: Handbook of Systemic Autoimmune Diseases, Vol. 7, chapter n. 14. Edited by J.C. Mason, C.D. Pusey. Elsevier, London 2008, pp F. Castelletti, R. Donadelli, F. Banterla, F. Hildebrandt. P.F. Zipfel, E. Bresin, E. Otto, C. Skerka, A. Renieri, M. Todeschini, J. Caprioli, M.R. Caruso, R. Artuso, G. Remuzzi, M. Noris. Mutations in FN1 cause glomerulopathy with fibronectin deposits. Proc Natl Acad Sci USA 2008;105: M. Noris, G. Remuzzi. Translational Mini-Review Series on Complment Factor H: Therapies of renal diseases associated with complement factor H abnormalities: atypical haemolytic uraemic syndrome and membranoproliferative glomerulonephritis. Clin Exp Immunol 2008;151: J. Caprioli, G. Remuzzi. A mouse model of non-shiga toxin-associated haemolytic uraemic syndrome. (Editorial) Nephrol Dial Transplant 2008;23: R. Martinez-Barricarte, G. Pianetti, R. Gautard, J. Misselwitz, L. Strain, V. Fremeaux-Bacchi, C. Skerka, P.F. Zipfel, T. Goodship, M. Noris, G. Remuzzi, S.R. de Cordoba on behalf of the European Working Party on the genetics of HUS. The complement factor H R1210C mutation is associated with atypical hemolytic uremic syndrome. J Am Soc Nephrol 2008;19: A. Pezzotta, M. Mister, G. Monteferrante, L. Cassis, N. Azzollini, S. Aiello, M. Satta, A. Benigni, G. Remuzzi, M. Noris. Effect of seliciclib (CYC202, R-roscovitine) on lymphocyte alloreactivity and acute kidney allograft rejection in rat. Transplantation 2008;85: C. Zanchi, C. Zoja, M. Morigi, F. Valsecchi, X.Y. Liu, D. Rottoli, M. Locatelli, S. Buelli, A. Pezzotta, P. Mapelli, J. Geelen, G. Remuzzi, J. Hawiger. Fractalkine and CX3CR1 mediate leukocyte capture by endothelium in response to Shiga toxin. J Immunol 2008;181: M.H. de Borst, A. Benigni, G. Remuzzi. Primer: strategies for identifying genes involved in renal disease. Nat Clin Pract Nephrol 2008;4:

279 M. Abbate, C. Zoja, D. Corna, D. Rottoli, C. Zanchi, N. Azzollini, S. Tomasoni, S. Berlingeri, M. Noris, M. Morigi, G. Remuzzi. Complement-mediated dysfunction of glomerular filtration barrier accelerates progressive renal injury. J Am Soc Nephrol 2008;19: S. Tomasoni, G. Remuzzi, A. Benigni. Allograft rejection: acute and chronic studies. In: Contribution to Nephrology: Gene therapy for renal diseases and transplantation. Edited by C. Ronco, A. Benigni, G. Remuzzi. Karger, Basel 2008, Vol. 159, pp N. Perico, A. Benigni, G. Remuzzi. Present and future drug treatments for chronic kidney diseases: evolving targets in renoprotection. Nature Reviews Drug Discovery 2008;7: M. Morigi, M. Introna, B. Imberti, D. Corna, M. Abbate, C. Rota, D. Rottoli, A. Benigni, N. Perico, C. Zoja, A. Rambaldi, A. Remuzzi, G. Remuzzi. Human bone marrow mesenchymal stem cells accelerate recovery of acute renal injury and prolong survival in mice. Stem Cells 2008;26: P. Ruggenenti, P. Bettinaglio, F. Pinares, G. Remuzzi. Angiotensin converting enzyme insertion/deletion polymorphism and renoprotection in diabetic and nondiabetic nephropathies. Clin J Am Soc Nephrol 2008;3: F. Casiraghi, N. Azzollini, P. Cassis, B. Imberti, M. Morigi, D. Cugini, R.A. Cavinato, M. Todeschini, S. Solini, A. Sonzogni, N. Perico, G. Remuzzi, M. Noris. Pretransplant infusion of mesenchymal stem cells prolongs the survival of a semiallogeneic heart transplant through the generation of regulatory T cells. J Immunol 2008;181: P. Ruggenenti, M. Noris, G. Remuzzi. Thrombotic Microangiopathies. In: Therapy in Nephrology and Hypertension A companion to Brenner and Rector s The Kidney (3 rd Edition), chapter n. 26. Edited by C.S. Wilcox. Saunders Elsevier, Philadelphia 2008, pp Azzollini N, Cugini D, Cassis P, Pezzotta A, Gagliardini E, Abbate M, Arduini A, Peschechera A, Remuzzi G, Noris M. Propionyl-L-carnitine prevents early graft dysfunction in allogeneic rat kidney transplantation. Kidney Int Dec;74(11): Epub 2008 Aug 13. Figliuzzi M, Adobati F, Cornolti R, Cassis P, Remuzzi G, Remuzzi A. Assessment of in vitro differentiation of bovine pancreatic tissue in insulin-expressing cells. JOP Sep 2;9(5): Caprioli J, Mele C, Mossali C, Gallizioli L, Giacchetti G, Noris M, Remuzzi G, Benigni A. Polymorphisms of EDNRB, ATG, and ACE genes in salt-sensitive hypertension. Can J Physiol Pharmacol Aug;86(8): Ruggenenti P, Cravedi P, Sghirlanzoni MC, Gagliardini E, Conti S, Gaspari F, Marchetti G, Abbate M, Remuzzi G. Effects of rituximab on morphofunctional abnormalities of membranous glomerulopathy. Clin J Am Soc Nephrol Nov;3(6): Epub 2008 Aug 6. Remuzzi G, Cattaneo D, Perico N. The aggravating mechanisms of aldosterone on kidney fibrosis. J Am Soc Nephrol Aug;19(8): Epub 2008 Jun 11. Tomasoni S, Remuzzi G, Benigni A. Allograft rejection: acute and chronic studies. Contrib Nephrol. 2008;159: Review. 278

280 RESEARCH ACTIVITIES Laboratory of Cell Biology and Xenotransplantation Life-sparing effect of human cord-blood mesenchymal stem cells in mice with acute kidney injury In collaboration with the Laboratory of Experimental Models of Kidney Diseases New approaches for the cure of acute kidney injury (AKI), a highly life-threatening clinical setting, have identified in the transplantation of bone marrow-derived mesenchymal stem cells (MSCs) a new tool for renal repair in experimental models. To further explore innovative interventions for AKI, we investigated the potential of human cord blood MSCs (CB-MSCs) in preventing cisplatin-induced AKI and prolonging survival in an immunodeficient NOD/SCID mouse model. NOD/SCID mice were subcutaneously injected with the nephrotoxic drug cisplatin (12.7 mg/kg) and one day later were intra vein injected with saline or human CB-MSCs (5x10 5 cells) stained with the fluorescent dye PKH26. Renal function was measured at different time points as blood urea nitrogen. Mice were followed during time for survival studies or sacrificed at 4 days after cisplatin. Kidneys were taken for ultrastructural analysis and for studying proliferation and apoptosis. Intracellular pathways, involved in the regenerative process induced by human CB-MSCs, were also investigated. Infusion of human CB-MSCs in cisplatin mice with AKI ameliorated renal function and markedly decreased proximal tubular epithelial cell injury and mortality (cisplatin mice: human CB-MSCs 14% vs saline 100% mortality at 9 days p<0.001). Transplanted human CB- MSCs localized predominantly in peritubular areas and acted to significantly increase tubular cell proliferation, to reduce apoptosis by virtue of their intrinsic capacity to produce high amount of growth and pro-survival factors. Peritubular capillary ultrastructural changes were less severe, with less polymorphonuclear infiltration in cisplatin-mice treated with human CB-MSCs. Here, an important step forward in the understanding human CB-MSC protection attributes to these cells the capacity to inhibit tubular oxidative stress, by reducing nitrotyrosine expression, and to induce the phosphorylation of the pro-survival factor Akt in tubular epithelium. These findings indicate that human MSC of cord blood origin hold potential to prolong survival in human AKI and should be considered for testing shortly in a clinical trial. Embryonic stem cells prolong the survival of semiallogeneic transplants In collaboration with the Laboratory of Immunology and Genetic of Rare Diseases and Organ Transplantation Transplantation is the only therapeutic choice for end-stage failure of several organs, and is often associated to immunosuppressive drug administration and sometimes to graft failure. In animal models, an innovative strategy to prolong survival of transplanted organ, consists in the administration, prior to transplantation, of hematopoietic or mesenchymal stem cells. Our group has studied the use of embryonic stem cells (ESC) to induce transplant tolerance. By in vitro fertilization, we have obtained an ESC line (from B6C3 mice) and we have verified the typical expression of ESC markers such as stage specific antigen-1 (SSEA-1), alkaline phosphatase the transcription factor Oct-4 and we have confirmed that they could form teratoma in syngeneic animals. To study the immunomodulatory effect of ESC we have evaluated by mixed lymphocyte reaction the proliferative response of splenocytes from semiallogeneic animals (B6) which have been injected, 7 or 45 days before, with ESC. Results showed that, ESC administration, after 45 days, induce T-cell hyporesponsiveness towards donor, possibly setting 279

281 the basis for graft tolerance. Therefore, we have studied whether the pre-treatment with ESC could modulate tolerogenic properties in semiallogeneic ectopic heart transplantation (B6C3 into B6 recipients). Transplanted mice, pre-infused with ESC, 7 or 45 days before transplantation, exhibited a prolongation of graft survival as compared to mice receiving fibroblasts (P<0.05 and 0.01 respectively). More than 44% of the animals infused with ESC 45 days before transplantation showed graft survival longer than 100 days versus 10% of animals injected with ESC 7 days before. In addition to ESC derived by in vitro fertilization, we have also obtained an ESC line by nuclear transfer (NT-ESC). In a semiallogeneic model, NT-ESC induced prolongation of graft survival similar to that obtained by ESC (> 100 days in 40% of the animals). The study of the mechanism possibly involved in graft tolerance, by adoptive transfer studies, showed that regulatory T cells are not involved. Laboratory of Experimental Models of Kidney Diseases Complement-mediated dysfunction of glomerular filtering barrier accelerates progressive renal injury Intrarenal complement activation leads to chronic tubulointerstitial injury in animal models of proteinuric nephropathies and is candidate target of renoprotective therapy. The present study investigates the role of abnormally filtered plasma proteins in this injury and mechanisms responsible for C3 accumulation in kidney. We find that mice with C3 deficiency are protected to a significant degree against the interstitial inflammatory response and tissue damage elicited by protein overload. These effects are partly related to less severe podocyte injury and lower proteinuria. Both C3 protein deposition and C3 mrna upregulation in proximal tubule are features in common with other progressive models. Here, antiproteinuric treatment with angiotensin-converting enzyme inhibitor, lisinopril, limits filtered plasma protein and C3 accumulation by proximal tubular cells and lessens interstitial inflammation and damage in wild-type mice. C3 deficient kidneys transplanted before induction of disease into wild type mice develop glomerular injury, C3 accumulation in podocytes and proximal tubules, and tubulointerstitial changes in response to protein overload; in contrast, wild type kidneys given to C3-deficient mice reveal no abnormal C3 deposition and milder disease. These data show that 1. the presence of C3 enhances susceptibility to injury of glomerular filtering barrier, 2. ultrafiltered C3 has greater influence than locally synthesized C3 on tubulointerstitial damage induced by protein overload, and 3. local C3 synthesis is irrelevant to the development of proteinuria. Our results suggest that complement-targeted approaches should be combined with interventions to minimize proteinuria as a way to more effectively prevent progression of renal disease and associated cardiovascular events. Fractalkine and CX3CR1 mediate leukocyte captare by endothelium in response to Shiga toxin In collaboration with the Laboratory of Cell Biology and Xenotransplantation Shiga toxins (Stx) are the virulence factors of enterohemorrhagic E.coli O157:H7, a world-wide emerging diarrheal pathogen, which precipitates post-diarrheal hemolytic uremic syndrome, the leading cause of acute renal failure in children. Here we show that Stx2 triggered expression of fractalkine, a CX3C transmembrane chemokine, acting as both adhesion counterreceptor on endothelial cells and soluble chemoattractant. Stx2 caused in HUVEC expression of fractalkine mrna and protein, which promoted leukocyte capture, ablated by antibodies to either endothelial fractalkine or leukocyte CX3CR1 receptor. Exposure of human glomerular endothelial cells to Stx2 recapitulated its fractalkine-inducing activity and fractalkine-mediated leukocyte adhesion. Both processes required phosphorylation of Src-family protein tyrosine kinase and p38 MAPK in endothelial cells. Furthermore, they depended on nuclear import of NF-kB and other stress-responsive transcription factors. Inhibition of their nuclear import with 280

282 the cell-penetrating SN50 peptide reduced fractalkine mrna levels and fractalkine-mediated leukocyte capture by endothelial cells. Adenoviral overexpression of IkB inhibited fractalkine mrna upregulation. The fractalkine-mediated responses to Stx2 were also dependent on AP-1. In mice both virulence factors of Stx-producing E.coli Stx and LPS- are required to elicit hemolytic uremic syndrome. Here, fractalkine was detected within glomeruli of C57BL/6 mice injected with Stx2, and further increased after Stx2 plus LPS co-administration. This was associated with recruitment of CX3CR1-positive cells. Thus, in response to Stx2, fractalkine is induced playing essential role in the promotion of leukocyte-endothelial cell interaction thereby potentially contributing to the renal microvascular dysfunction and thrombotic microangiopathy that underlie hemolytic uremic syndrome due to enterohemorrhagic E.coli O157:H7 infection. Upon renal injury self-proteins generate antigenic peptides through a proteasome-dependent pathway In collaboration with the Laboratory of Renal Biophysics (Department of Bioengineering) and the Laboratory of Analytic Biochemistry (Department of Environmental Health Sciences) In proteinuric nephropathies loss of the permselective properties of the glomerular membrane leads to abnormal filtration of proteins, mainly albumin, that cause distress of the proximal tubular cell inducing the release into the interstitium of chemokines responsible for the recruitment and activation of inflammatory cells. The interstitial infiltrates are also characterized by the presence of immune-competent cells including dendritic cells (DCs) and T lymphocytes that accumulate within renal parenchyma even in absence of an immune insult. As antigen presenting cells, DCs can initiate immune response or tolerance. The outcome depends on context. The current view is that immunity is controlled by internal communication between the tissue, where DCs are resident, and the immune-competent cells, so that tolerance is maintained by the healthy organ, whereas immunity can be stimulated by a distressed organ. Within the kidney, DCs are in close contact with the tubular epithelium and work as immune sentinels to probe renal interstitium in search for foreign antigens. It is conceivable that upon injury, the inflammatory stimuli released from the tubular cell represent danger signals that alert DCs making them immunogenic instead of tolerogenic. This might favor the presentation of normally ignored self-antigens triggering an immune response. The evidence that the antiproteinuric effect of the angiotensin converting enzyme inhibitors is associated with a significant reduction of interstitial infiltrates and renal damage further supports this hypothesis and the idea that abnormally filtered plasmatic proteins represent self-antigens capable to trigger an immune response. To verify our hypothesis, we investigated how albumin interacts with the immune system. Rat proximal tubular cells exposed in vitro to autologous albumin concentrations, mimicking those present in the ultrafiltrate in proteinuric conditions, release into the supernatant peptides identified, by mass spectrometry, as N-terminal 24, 27, or 30 amino acid albumin fragments. Given the abundance of the 24 amino acid peptide, we focused later studies on this peptide we named Alb1-24. This fragment is not formed in the presence of pepstatin A, suggesting that an acid protease with pepsin-like activity is responsible for the selective N-terminal truncation of albumin at position 24. Alb1-24 is taken up by bone marrowderived rat dendritic cells and processed, through a proteasome dependent pathway, into shorter peptides that are loaded on the major histocompatibility complex class I (MHC-I) and presented to syngeneic CD8 + T cells. The first encounter between Alb1-24-pulsed DCs primes naive CD8 + T cells, so that when exposed a second time, CD8 + T cells become activated and release interferon γ. In a cell-free system, purified proteasome cleaves albumin into peptides some of them have a size compatible with MHC-I binding (8 to 10 amino acid long). Within the products of in vitro proteasomal degradation of Alb1-24, the 15 to 22 amino acid peptide was identified by bioinformatic tools, used to search for potential binders to MHC-I, as a top-ranked 8-mer ligand for H2Kb allele. The functional role of DC proteasome in generating potentially antigenic peptides from albumin was next demonstrated in vivo in a model of non-immune 281

283 proteinuric nephropathy. Four weeks after the 5/6 nephrectomy of renal mass, a decrease in the percentage of DCs in the kidney was paralleled by an enhancement of these cells in the renal lymph nodes, suggesting migration of DCs from the renal interstitium. The CD8 + T cells isolated from renal lymph nodes produced interferon γ, at the first encounter, when incubated with Alb1-24- pulsed DCs, suggesting that they were educated in vivo during the course of the disease. By contrast, the in vivo treatment with the proteasome inhibitor bortezomib, that prevent the formation of peptides for recognition by CD8 + T cells, made these cells resistant to activation by Alb1-24-pulsed DCs either in a primary or secondary culture. Our findings indicate that albumin can become the source of potentially antigenic peptides upon renal injury and outline the relationship between tubular cell biology and the role of the DC in processing self-proteins through a proteasome-dependent pathway. Laboratory of Immunology and Genetic of Rare Diseases and Organ Transplantation Effect of seliciclib (CYC202, R-roscovitine) on lymphocyte alloreactivity and acute kidney allograft rejection in rat T cell stimulation by alloantigens is followed by cell cycle progression, an event that is critically dependent on cyclin-dependent kinases (CDKs). We conducted a study to evaluate whether the CDK inhibitor seliciclib affected rat lymph node cell (LNc) activation and proliferation induced by either concanavalin A or allogeneic splenocytes in vitro and studied the mechanisms underlying the suppressive effect. We also investigated the immunosuppressive properties of seliciclib in vivo. Seliciclib completely inhibited in vitro proliferation of LNc and CD8 + Tcells, in response to either concanavalin A or allogeneic splenocytes. The percentage of activated LNc was lower in MLR added with seliciclib than in MLR added with vehicle. The percentages of viable and apoptotic cells at the end of MLR with seliciclib were comparable to those of MLR with vehicle. LNc pre-exposed in MLR to seliciclib did not respond to further stimulation with alloantigens, and neither IL-2 nor IL-15 restored proliferation. These data indicate that the inhibitory effect of seliciclib on T cell alloreactivity is not due to cytotoxic effect but is associated with induction of profound T cell anergy. LNc harvested at the end of the primary MLR with seliciclib did not suppress the proliferation of syngeneic LNc cells toward allogeneic splenocytes, thus excluding that seliciclib induced the formation of regulatory cells. Finally, seliciclib partially prolonged grafted animal survival in a rat model of fully MHC-mismatched kidney transplantation. Altogether these results document that seliciclib regulates lymphocyte reactivity and may exert an immunosuppressive effect in vivo in the setting of transplantation. Pretransplant infusion of mesenchymal stem cells prolongs the survival of a semiallogeneic heart transplant through the generation of regulatory T cells. In this study, we investigated whether mesenchymal stem cells (MSC) had immunomodulatory properties in solid organ allotransplantation, using a semiallogeneic heart transplant mouse model, and studied the mechanism(s) underlying MSC tolerogenic effects. Either single (portal vein, day -7) or double (portal vein, day -7 and tail vein, day -1) pretransplant infusions of donor-derived B6C3 MSC in B6 recipients induced a profound T cell hyporesponsiveness and prolonged B6C3 cardiac allograft survival. The protolerogenic effect was abrogated when donor-derived MSC were injected together with B6C3 hematopoietic stem cells (HSC), suggesting that HSC negatively impact MSC immunomodulatory properties. Both the induction 282

284 (pretransplant) and the maintenance phase (>100 days posttransplant) of donor-derived MSCinduced tolerance were associated with CD4 + CD25 + Foxp3 + Treg expansion and impaired antidonor Th1 activity. MSC-induced regulatory T cells (Treg) were donor-specific since adoptive transfer of splenocytes from tolerant mice prevented the rejection of fully MHC-mismatched donor-specific secondary allografts but not of third-party grafts. In addition, infusion of recipient-derived B6 MSC tolerized a semiallogeneic B6C3 cardiac allograft, but not a fully MHC-mismatched BALB/c graft, and expanded Treg. A double i.v. pretransplant infusion of recipient-derived MSC had the same tolerogenic effect as the combined intraportal/i.v. MSC infusions, which makes the tolerogenic protocol applicable in a clinical setting. In contrast, single MSC infusions given either peritransplant or 1 day after transplant were less effective. Altogether these findings indicate that MSC immunomodulatory properties require HSC removal, partial sharing of MHC Ags between the donor and the recipient and pretransplant infusion, and are associated with expansion of donor-specific Treg. Mutations in FN1 cause glomerulopathy with fibronectin deposits. Glomerulopathy with fibronectin (FN) deposits (GFND) is an autosomal dominant disease with age-related penetrance, characterized by proteinuria, microscopic hematuria, hypertension, and massive glomerular deposits of FN that lead to end-stage renal failure. The genetic abnormality underlying GFND was still unknown. We hypothesized that mutations in FN1, which encodes FN, were the cause of GFND. In a large Italian pedigree with eight affected subjects, we found linkage with GFND at the FN1 locus at 2q32. We sequenced the FN1 in 15 unrelated pedigrees and found three heterozygous missense mutations, the W1925R, L1974R, and Y973C, that cosegregated with the disease in six pedigrees. The mutations affected two domains of FN (Hep-II domain for the W1925R and the L1974R, and Hep-III domain for the Y973C) that play key roles in FN-cell interaction and in FN fibrillogenesis. Mutant recombinant Hep-II fragments were expressed, and functional studies revealed a lower binding to heparin and to endothelial cells and podocytes compared with wild-type Hep-II and an impaired capability to induce endothelial cell spreading and cytoskeletal reorganization. Overall dominant mutations in FN1 accounted for 40% of cases of GFND in our study group. These findings may help understanding the pathogenesis of proteinuria and glomerular FN deposits in GFND and possibly in more common renal diseases such as diabetic nephropathy, IgA nephropathy, and lupus nephritis. To our knowledge no FN1 mutation causing a human disease was previously reported. Membrane cofactor protein mutations in atypical hemolytic uremic syndrome (ahus), fatal Stx-HUS, C3 glomerulonephritis, and the HELLP syndrome. The hemolytic uremic syndrome (HUS) is a triad of microangiopathic hemolytic anemia, thrombocytopenia, and renal impairment. Genetic studies demonstrate that heterozygous mutations of membrane cofactor protein (MCP;CD46) predispose to atypical HUS (ahus), which is not associated with exposure to Shiga toxin (Stx). Among the initial 25 MCP mutations in patients with ahus were 2, R69W and A304V, that were expressed normally and for which no dysfunction was found. The R69W mutation is in complement control protein module 2, while A304V is in the hydrophobic transmembrane domain. In addition to 3 patients with ahus, the A304V mutation was identified in 1 patient each with fatal Stx-HUS, the HELLP (hemolysis, elevated liver enzymes, and low platelets) syndrome, and glomerulonephritis with C3 deposits. A major goal was to assess if these putative mutations lead to defective complement regulation. Permanent cell lines expressing the mutated proteins were complement "challenged," and membrane control of C3 fragment deposition was monitored. Both the R69W and A304V MCP mutations were deficient in their ability to control 283

285 the alternative pathway of complement activation on a cell surface, illustrating the importance of modelling transmembrane proteins in situ. Unit of Gene Therapy Gene therapy to correct Thrombotic Thrombocytopenic Purpura Thrombotic thrombocytopoenic purpura (TTP) is a thrombotic microangiopathy, characterized by anemia, thrombocytopenia and formation of microvascular platelet rich thrombi. TTP patients show prevalent, but not exclusive, neurological symptoms and renal dysfunction due to accumulation in the plasma of ultralarge von Willebrand multimers. In healthy subjects the VWF-cleaving protease ADAMTS13 (a disintegrin-like and metalloprotease with thrombospondin type I repeats) reduces the ultra large VWF multimers into smaller forms soon after their secretion. Acquired and familial deficiency of ADAMTS13 causes TTP in humans. Plasma infusion is the treatment of choice for TTP patients, however it exposes patients to a high risk of infections, fluid volume overload and allergic responses. Only 5-6% of ADAMTS13 activity is sufficient to avoid relapses of the disease. In light of these findings gene therapy could be an alternative therapeutic strategy for patients with familial TTP. In order to evaluate the efficacy of gene therapy we constructed two different viral vectors containing human ADAMTS13 cdna, an adeno-associated and an adenoviral vector. Both vectors were able to infect human fibrosarcoma cells and to induce the synthesis of the recombinant protein. The availability of Adamts13-/- knock out mice (Adamts13 -/- ) were instrumental to study if systemic injection of the vectors was able to restore protein levels. Adamts13 -/- mice were given 1x10 11 vgu AAV-ADAMTS13 resulting in appreciable expression of radamts13 mrna into the liver, the constitutive site of ADAMTS13 production. Time course experiments showed that ADAMTS13 mrna was expressed at 4, 8 and at much lower levels at 16 weeks from injection. Antigen levels were measured in plasma of treated mice by ELISA but no protein was detected, probably due to the low infectivity of the virus. It is well known that large cdnas impair correct packaging of AAV. The AAV serotype used in this experiment (AAV2) has an insertional capacity similar to ADAMTS13 size. We are now going to test different seroptypes with a larger insertional capacity. We also tested the adenoviral vector in vivo. Systemic injection of 1x10 9 Ad-ADAMTS13 pfu induced ADAMTS13 mrna expression in the liver, kidneys and lung in Adamts13 -/- mice sacrified 1 week after the treatment, yielding the secretion in the plasma of large amount of protein. The released protein was functionally active even after 2 months from injection. These results would indicate that gene therapy represents a promising therapeutic strategy to correct ADAMTS13 deficiency in patients with familial TTP. 284

286 DEPARTMENT OF BIOMEDICAL ENGINEERING STAFF Head Andrea REMUZZI, Eng. D. Laboratory of Renal Biophysics Head Daniela MACCONI, Biol.Sci.D. Laboratory of Biomedical Technologies Head Bogdan ENE-IORDACHE, Eng.D. Unit of Tissue Engineering Head Marina FIGLIUZZI, Biol.Sci.D. Unit of Medical Imaging Head Luca ANTIGA, Ph.D. 285

287 CURRICULA VITAE Andrea Remuzzi got his degree in Mechanical (Biomedical) Engineering in 1979, Politecnico di Milano. Research experience: 1980 Politecnico di Milano, Dipartimento di Ingegneria Biomedica; 1981 Istituto Mario Negri (Milano), Laboratorio di Farmacologia Cardiovascolare; Massachusetts Institute of Technology, Mechanical Engineering Department, Cambridge, USA. Areas of interest: biological transport phenomena, mathematical models, renal pathophysiology, cellular response to mechanical stimulation, tissue engineering, pancreatic islet transplantation, clinical databases, computational fluid dynamics. Chronology of appointment: From 1984 to 1986 Ricercatore Istituto Mario Negri (Bergamo), Laboratorio di malattie renali, Head, Unità di Bioingegneria, Istituto Mario Negri, Head, Laboratorio di Bioingegneria, Istituto Mario Negri, Head, Dipartimento di Ricerca Renale, Istituto Mario Negri, from 2000 Head, Dipartimento di Bioingegneria, Istituto Mario Negri. From 1998 to 2007 contract professor of Bioengineering, Politecnico di Milano. For 2007 Professor of Bioengineering, University of Bergamo. Selected publications Davies PF, Remuzzi A, Gordon EJ, Dewey CF Jr, Gimbrone MA Jr. Turbulent fluid shear stress induces vascular endothelial cell turnover in vitro. Proc Natl Acad Sci U S A Apr;83(7): PMID: Remuzzi A, Puntorieri S, Battaglia C, Bertani T, Remuzzi G. Angiotensin converting enzyme inhibition ameliorates glomerular filtration of macromolecules and water and lessens glomerular injury in the rat. J Clin Invest Feb;85(2): PMID: Noris M, Morigi M, Donadelli R, Aiello S, Foppolo M, Todeschini M, Orisio S, Remuzzi G, Remuzzi A. Nitric oxide synthesis by cultured endothelial cells is modulated by flow conditions. Circ Res Apr;76(4): PMID: Giavazzi R, Foppolo M, Dossi R, Remuzzi A. Rolling and adhesion of human tumor cells on vascular endothelium under physiological flow conditions. J Clin Invest Dec;92(6): PMID: Antiga L, Ene-Iordache B, Remuzzi A. Computational geometry for patient-specific reconstruction and meshing of blood vessels from MR and CT angiography. IEEE Trans Med Imaging May;22(5): PMID: Remuzzi A, Gagliardini E, Sangalli F, Bonomelli M, Piccinelli M, Benigni A, Remuzzi G. ACE inhibition reduces glomerulosclerosis and regenerates glomerular tissue in a model of progressive renal disease. Kidney Int Apr;69(7): Ruggenenti P, Remuzzi A, Remuzzi G. Decision time for pancreatic islet-cell transplantation. Lancet : Antiga L, Piccinelli M, Fasolini G, Ene-Iordache B, Ondei P, Bruno S, Remuzzi G, Remuzzi A. Computed tomography evaluation of autosomal dominant polycystic kidney disease progression: a progress report. CJASN : Daniela Macconi got her Biol.Sci.D. degree in Milan in the Research experience: CNR Institute of Neuroscience - Cell Mol Pharmacology - and Department of Medical Pharmacology, University of Milan, Milan, Italy; Laboratory of the Division of Nephrology and Dialysis, Ospedali Riuniti di Bergamo, Bergamo, Italy; University of Michigan, Medical School, Department of Pathology, Medical Science I, Ann Arbor Michigan, USA; Mario Negri Institute for Pharmacological Research, Laboratory of Kidney Disease, Bergamo, Italy. Areas of interest: glomerular permeability, renal disease progression, podocytes, angiotensin II, reactive oxygen species Chronology of appointment: From 2000 Head Laboratory of Renal Biophisics, Department of Biomedical Engineering; Head, Unit of Inflammatory Mediator of Leucocyte Origin; Scientist, post-doctoral fellow Mario Negri Institute for Pharmacological Research, Bergamo, Italy; fellow Laboratory of the Division of Nefrology e Dialysis, Ospedali Riuniti di Bergamo, Bergamo, Italy Selected publications Macconi D, Remuzzi G. Candesartan and renal protection: more than blocking angiotensin type 1 receptor? Kidney Int. 74:1112-4, Macconi D, Abbate M, Morigi M, Angioletti S, Mister M, Buelli S, Bonomelli M, Mundel P, Endlich K, Remuzzi A, Remuzzi G: Permselective dysfunction of podocyte-podocyte contact upon angiotensin II unravels the molecular target for renoprotective intervention. Am J Pathol 168: , Macconi D, Bonomelli M, Benigni A, Plati T, Sangalli F, Longaretti L, Conti S, Kawachi H, Hill P, Remuzzi G, Remuzzi A. Pathophysiologic implications of reduced podocyte number in a rat model of progressive glomerular injury. Am J Pathol 68:42-54, Ruiz-Torres MP, Casiraghi F, Galbusera M, Macconi D, Gastoldi S, Todeschini M, Porrati F, Belotti D, Pogliani EM, Noris M, Remuzzi G: Complement activation: the missing link between ADAMTS-13 deficiency and microvascular thrombosis of thrombotic microangiopathies. Thromb Haemost 93:443-52,

288 Morigi M, Macconi D, Zoja C, Donadelli R, Buelli S, Zanchi C, Ghilardi M, Remuzzi G: Protein overload-induced NFkappaB activation in proximal tubular cells requires H(2)O(2) through a PKC-dependent pathway. J Am Soc Nephrol 13: , 2002 Macconi D, Ghilardi M, Bonassi ME, Mohamed EI, Abbate M, Colombi F, Remuzzi G, Remuzzi A: Effect of angiotensinconverting enzyme inhibition on glomerular basement membrane permeability and distribution of zonula occludens-1 in MWF rats. J Am Soc Nephrol 11:477-89, Bogdan Ene-Iordache got MSc in Mechanical Engineering in 1990 at the Oil & Gas Institute in Ploiesti (Romania). In 1992 he joined the Bioengineering Laboratory at NegriBERGAMO Laboratories. Main interests: renal research (hemodynamics and remodeling of arteriovenous fistula for vascular access, morfometrical analysis of glomerular capillaries) and controlled clinical trials (data management and data analysis for controlled clinical studies); biomedical informatics, web sites development, coordination of IT activity in the Clinical Research Center for Rare Diseases Aldo e Cele Daccò ; applied clinical informatics (Nephrology, Diabetology and Hematology Units, Bergamo Hospital). Roles: since January 2000 is head of the Biomedical Technologies Laboratory, Department of Biomedical Engineering. Selected publications Ene-Iordache B, Imberti O, Foglieni O, Remuzzi G, Bertani T and Remuzzi A. Effects of angiotensin-converting enzyme inhibition on glomerular capillary wall ultrastructure in MWF/Ztm rats. J Am Soc Nephrol 5: , Ene-Iordache B and Remuzzi A. Numerical analysis of blood flow in reconstructed glomerular capillary segments. Microvasc Res 49: 1-11, Remuzzi A and Ene-Iordache B. Capillary network structure does not affect theoretical analysis of glomerular size selectivity. Am J Physiol 268: F972-F979, Ene-Iordache B, Mosconi L, Remuzzi G, Remuzzi A. Computational fluid dynamics of a vascular access case for hemodialysis. J Biomech Eng 123(3): , Ene-Iordache B, Mosconi L, Antiga L, Bruno S, Anghileri A, Remuzzi G, Remuzzi A. Radial artery remodeling in response to shear stress increase within arteriovenous fistula for hemodialysis access. Endothelium 10(2): , Ruggenenti P, Fassi A, Ilieva AP, Bruno S, Iliev IP, Brusegan V, Rubis N, Gherardi G, Arnoldi F, Ganeva M, Ene-Iordache B, Gaspari F, Perna A, Bossi A, Trevisan R, Dodesini AR, Remuzzi G for the Bergamo Nephrologic Diabetes Complications Trial (BENEDICT) Investigators. Preventing microalbuminuria in type 2 diabetes. NEJM 351(19): , Ene-Iordache B, Carminati S, Antiga L, Rubis N, Ruggenenti P, Remuzzi G and Remuzzi A. Developing regulatorycompliant electronic case report forms for clinical trials: experience with the DEMAND trial. JAMIA 15(2), Marina Figliuzzi got her Biol.Sci.D. degree in Milan in the Research experience : Mario Negri Institute for Pharmacological Research, Bergamo, Italy. Areas of interest: isolation of pancreatic islets from human, bovine, pig and rat pancreas, cell culture, immunoisolation devices for pancreatic islets, differentiation of progenitor pancreatic cells in insulin containing cells, immunhistochemistry. Chronology of appointment: From 2000 Head Unit of Tissue Engineering, Department of Biomedical Engineering; fellow laboratory of Renal research, Mario Negri Institute for Pharmacological Research, Bergamo, Italy. Selected publications Figliuzzi M, Adobati F, Cornolti R, Cassis P, Remuzzi G, Remuzzi A.Assessment of in vitro differentiation of bovine pancreatic tissue in insulin-expressing cells. JOP 9(5):601-11, Figliuzzi M, Plati T, Cornolti R, Adobati F, Fagiani A, Rossi L, Remuzzi G, Remuzzi A. Biocompatibility and function of microencapsulated pancreatic islets. Acta Biomater Mar;2(2): Figliuzzi M, Cornolti R, Plati T, Rajan N, Adobati F, Remuzzi G, Remuzzi A: Subcutaneous xenotransplantation of bovine pancreatic islets. Biomaterials. 26: , Figliuzzi M, Zappella S, Morigi M, Rossi P, Marchetti P, Remuzzi A: Influence of donor age on bovine pancreatic islet isolation. Transplantation. 70: , 2000 Morigi M, Micheletti G, Figliuzzi M, Imberti B, Karmali MA, Remuzzi A, Remuzzi G, Zoja C. Verotoxin-1 promotes leukocyte adhesion to cultured endothelial cells under physiologic flow conditions. Blood.: , Zoja C, Morigi M, Figliuzzi M, Bruzzi I, Oldroyd S, Benigni A, Ronco P, Remuzzi G. Proximal tubular cell synthesis and secretion of endothelin-1 on challenge with albumin and other proteins. Am J Kidney Dis.26: , Morigi M, Zoja C, Figliuzzi M, Foppolo M, Micheletti G, Bontempelli M, Saronni M, Remuzzi G, Remuzzi A. Fluid shear stress modulates surface expression of adhesion molecules by endothelial cells. Blood. 85: ,

289 Luke Antiga graduated in 1999 in Biomedical Engineering and got his PhD in Bioengineering in 2003, Politecnico di Milano, having worked at the research laboratory of Biomedical Technology, Department of Bioengineering Institute Mario Negri. Training activities: 2003 Post-doctoral fellow at Imaging Research Laboratories, Robarts Research Institute, London, Ontario. Areas of interest: acquisition and image processing for medical and microscopy applications, numerical modeling of transport phenomena. Roles: from 2000 to 2002 Doctoral Student at the Laboratory of Biomedical Technology, Department of Bioengineering, from 2002 to 2003 visiting scientist Robarts Institute for medical Imaging, London, Ontario, Canada, from 2004 to 2006 resercher at the Laboratory of Biomedical Technology, Department of Bioengineering, from 2007 Head Unit of Medical Imaging, Department of Bioengineering. Selected publications Lee SW, Antiga L, Spence JD and Steinman DA. Geometry of the carotid bifurcation predicts its exposure to disturbed flow. Stroke, in press, Antiga L, Piccinelli M, Fasolini G, Ene-Iordache B, Ondei P, Ruggenenti P, Remuzzi G and Remuzzi A. Computed tomography evaluation of ADPKD progression: a progress report. Clinical Journal of the American Society of Nephrology (CJASN), 1(4): , Jul Thomas JB, Antiga L, Che S, Milner JS, Hangan Steinman DA, Spence JD, Rutt BK and Steinman DA. Variation in the carotid bifurcation geometry of young vs. older adults: Implications for "geometric risk" of atherosclerosis. Stroke, 36(11): , Nov Antiga L, Steinman DA. Robust and objective decomposition and mapping of bifurcating vessels. IEEE Transactions on Medical Imaging, 23(6): , June Antiga L, Ene-Iordache B and Remuzzi A. Computational geometry for patient-specific reconstruction and meshing of blood vessels from MR and CT angiography. IEEE Transactions on Medical Imaging, 22(5): , May Antiga L, Ene-Iordache B, Remuzzi G and Remuzzi A. Automatic generation of glomerular capillary topological organization. Microvascular Research, 62: , June

290 INTRODUCTION TO THE DEPARTMENT'S ACTIVITIES The Department of Bioengineering conducts research and development in biomedicine, both at experimental and clinical level. Physiopathological processes are studied through the use of engineering techniques and to develop innovative treatment strategies. Several lines of research in basic, applied research and clinical are currently active within the Department. The main instruments used for this research consist of: theoretical models; diagnostic imaging; histological measures; physical and chemical parameters in both experimental and clinical studies; cell culture techniques; biomaterials; technologies for archiving and processing of clinical data. The ongoing studies relate to four main areas: 1) study of the mechanisms involved in the progression of chronic nephropathy; 2) studies on the role hemodynamics in the development of vascular diseases; 3) development of laboratory techniques for tissue engineering, 4 ) developement of information systems to manage clinical data and images generated in the context of controlled clinical trials and in routine clinical practice. FINDINGS/MAIN RESULTS We produced evidence demonstrating a possible relationship between the geometry of the cerebral arterial vessels, hemodynamic conditions and location of cerebral aneurysms. These findings are opening new perspectives in the understanding of the mechanisms responsible for this disease. Demonstration of a significant relationship between morphometric parameters of the renal parenchyma, quantified through elaboration of CT images, and the loss of renal function in patients with polycystic kidney disease. Demonstration that the technique of islet immunoisolation does not preclude oxygenation and consequently that this technique can be used for pancreatic islet transplantion. Demonstration that the mechanism responsible for regeneration of the glomerular capillary, induced by drugs that inhibit the angiotensin II, depends on the proliferation of parietal cells of Bowman capsule and their migration on the glomerular capillary loops. Implementation of a web-based system for the management, in compliance with GCP, of clinical data generated in controlled clinical trials. Set up a new network of specialists in Nephrology for data collection aimed at monitoring the quality of treatment of chronic progressive nephropathy in current clinical practice. NATIONAL COLLABORATIONS Dipartimento di Bioingegneria, Politecnico di Milano, Milano. Fidia Advanced Biopolymers, Abano Terme, Padova. Istituto di Fisiologia Clinica CNR, Pisa. Unità di Diabetologia, Ospedali Riuniti, Bergamo. 289

291 STMicroelectronics, Agrate Brianza, Milano Università di Bergamo Dipartimento di scienze Neurologiche e della visione, Università di Verona. Dipartimento di Ingegneria Industriale e Dipartimento di Ingegneria dell informazione e metodi Matematici Facoltà di Medicina e Chirurgia, Università degli studi di Milano INTERNATIONAL COLLABORATIONS Massachussetts Institute of Technology, Cambridge MA, USA. National Alliance for Medical Imaging Computing, USA. Harvard Medical School, Cambridge MA, USA. Department of Mathematics and Computer Science, Emory University, Atlanta, Georgia, US. Simula Laboratories, Oslo, Norway Academisch Medisch Centrum, Amsterdam, the Netherlands University of Toronto, Ontario, Canada. Ghent University, Ghent, Belgium. Technical University, Eindhoven, The Netherlands. University Hospital, Maastricht, The Netherlands. Universzitetni Klinikni Center Lubjana, Ljubljana, Slovenia. The University of Sheffield, Sheffield, United Kingdom. EDITORIAL BOARD MEMBERSHIP International Journal of Artificial Organs, (Andrea Remuzzi) PEER REVIEW ACTIVITIES Annals of Biomedical Engineering ASME Journal of Biomechanical Engineering Journal of Vascular Research Magnetic Resonance in Medicine Stroke Journal of the American Society of Nephrology Kidney International American Journal of Kidney Diseases American Journal of Pathology American Journal of Physiology Medical & Biological Engineering & Computing New England Journal of Medecine IEEE Transactions on Medical Imaging IEEE Transactions on Biomedical Ingeneering Medical Physics Journal of Biomechanics Medical Engineering and Physics Artificial Organs International Journal of Artificial Organs Biomaterials Contemporary Clinical Trials 290

292 Journal of Biomaterial Application Journal of Endocrinological Investigation EVENT ORGANIZATION Seminar: The Definition of human adult stem cells by substrate interactions John A. Hunt, University of Liverpool, December, Ranica, Bergamo. Seminar: Developments in neuroimaging with applications to Alzheimer's disease Anna Caroli, October, Ranica, Bergamo. Seminar: Hemodynamics in Atherogenesis and Thrombosis, David Ku, Georgia Institute of Technology, Luglio, Ranica, Bergamo. Seminar: MSC in pancreatic islets, Thijs Molder, Maastricht University, July, Bergamo. Seminar: Model Based Drug Development and the FDA Critical Path Initiative, Dr. Robert Powell, Office of Translational Sciences, FDA, Silver Spring, MD, June, Ranica, Bergamo. Seminario: Tecniche avanzate di simulazioni numeriche in vasi sanguinei Christian Vergara, Università di Bergamo, April, Ranica, Bergamo. Seminar: Rotating versus perfusion bioreactor for the culture of engineered vascular constructs based on hyaluronic acid, Chiara Arrigoni, February, Ranica, Bergamo. PARTICIPATION IN EVENTS IN WHICH THE DEPARTMENT WAS INVOLVED 11th Asian Pacific Congress of Nephrology, Kuala Lumpur, Malaysia, May Results of the program for early detection of chronic kidney disease and their risk factors in Mongolia. 29th Annual Meeting of the Society for Clinical Trials. St. Louis, May Randomized and Non-Randomized patients in the Bergamo Nephrologic Diabetes Complications Trial (BENEDICT). 14th International Symposium on Brain Edema and Brain Tissue Injury, June 2008 Warsaw, Poland. Auto regulation of cerebral blood flow and autonomic drive. ASN Renal Week 2008, November, Philadelphia, PA. GFR slopes underestimate renal function decline in type 2 diabetic patients without overt nephropathy. ASN Renal Week 2008, November, Philadelphia, PA. Community Screening and Intervention for Hypertension, Diabetes and Chronic Kidney Disease in Poor Resource Setting in Eastern Nepal. 291

293 ASN Renal week 2008, November, Philadelphia, PA. Structure and Function in ADPKD: Correlation between GFR and Relative Volume of Hypo-Enhanced Parenchyma from Contrast- Enhanced CT Open Symposium agenda, September 2008, Barcelona, Spain. Translation of Virtual Physiological Human concepts into clinical systems and services for disease understanding and interventional planning: ARCH project. Polycystic Kidney Disease Database Consortium Meeting, March 2008, Chicago, USA. Discussion of PKD Clinical Database. 16th Congress of the European Society of Biomechanics, July 2008, Lucerne, Switzerland. Link Between Vortex Structures And Voronoi Diagram In Cerebral Aneurysms. 16th Congress of the European Society of Biomechanics, July 2008, Lucerne, Switzerland. A Open Source Parallel AMR FE Solver For Biological Flows Based On The Libmesh C++ Library. National Alliance for Medical Image Computing, Fourth All Hands Meeting, Salt Lake City, January 2008 National Alliance for Medical Image Computing, Summer Project Week, MIT, Boston, June 2008 ASME Summer Bioengineering Conference, Marco Island, FL, June Hands-on tutorial: the Vascular Modeling Toolkit. Fifth International Bio-Fluid Symposium and Workshop, Pasadena, CA, March CBC Workshop on High-Performance Computing and Biomedical Flows, Oslo, Norway. May GRANTS AND CONTRACTS Research grant AIFA - controlled clinical trials (VARIETY, VALID, ATHENA, ARCADIA). Research grant CHIESI Farmaceutici S.p.A - DEMAND trial A multicenter, randomized, prospective, double-blind study to evaluate the nephroprotective effect of delapril alone or combined with manidipine in patients with type 2 diabetes. Research grant PKD foundation - ALADIN trial Effect of long-acting somatostatin on disease progression in ADPKD: a long-term three year follow-up study. Research grant Baxter ASAP trial Acute Start Access Programme. Research grant ISN per il Kidney Disease Data Center (KDDC ) del COMGAN. Contributo Regione Lombardia per data management del Centro di Coordinamento della Rete Regionale per le Malattie Rare. 292

294 Project - FP6 UE-STEPS "A system approach to tissue engineering products and processes" FP Project - FP7 UE - ARCH "Patient specific image-based computational modeling for improvement of acute and long-term outcomes of vascular access for hemodialysis FP Project Coordination. Research grant PKD foundation - Effect of Long-acting Somatostatin on Disease Progression in ADPKD: A Long-term Three Year Follow-up Study. Project VASCOSILK, Fondazione Cariplo - N.536/ Protesi vascolari in fibroina elettrofilata per la rigenerazione in vivo di arterie di piccolo calibro. Project LIGASILK, Regione Lombardia - N.534/ Bioingegnerizzazione di tendini e legamenti: impiego combinato di supporti tessili in seta e cellule staminali adulte. SELECTION OF SCIENTIFIC PUBLICATIONS FROM 2008 Cravedi P, Mannon RB, Ruggenenti P, Remuzzi A, Remuzzi G. Islet transplantation: need for a time-out? Nat Clin Pract Nephrol Sep 23. Figliuzzi M, Adobati F, Cornolti R, Cassis P, Remuzzi G, Remuzzi A. Assessment of in vitro differentiation of bovine pancreatic tissue in insulin-expressing cells. JOP Sep; 9: Morigi M, Introna M, Imberti B, Corna D, Abbate M, Rota C, Rottoli D, Benigni A, Perico N, Zoja C, Rambaldi A, Remuzzi A, Remuzzi G. Human bone marrow mesenchymal stem cells accelerate recovery of acute renal injury and prolong survival in mice. Stem Cells Aug;26: Arrigoni C, Chittò A, Mantero S, Remuzzi A. Rotating versus perfusion bioreactor for the culture of engineered vascular constructs based on hyaluronic acid. Biotechnol Bioeng Aug;100: Ruggenenti P, Remuzzi A, Remuzzi G. Decision time for pancreatic islet-cell transplantation. Lancet Mar ;371: Lee SW, Antiga L, Spence JD, Steinman DA. Geometry of the carotid bifurcation predicts its exposure to disturbed flow. Stroke Aug;39: Antiga L, Wasserman BA, Steinman DA. On the overestimation of early wall thickening at the carotid bulb by black blood MRI, with implications for coronary and vulnerable plaque imaging. Magn Reson Med Nov;60: Macconi D, Remuzzi G. Candesartan and renal protection: more than blocking angiotensin type 1 receptor? Kidney Int Nov;74: Ruggenenti P, Iliev I, Costa GM, Parvanova A, Perna A, Giuliano GA, Motterlini N, Ene-Iordache B, Remuzzi G; Bergamo Nephrologic Diabetes Complications Trial Study Group. Preventing left ventricular hypertrophy by ACE inhibition in hypertensive patients with type 2 diabetes: a prespecified analysis of the Bergamo Nephrologic Diabetes Complications Trial (BENEDICT). Diabetes Care Aug;31: Ruggenenti P, Perticucci E, Cravedi P, Gambara V, Costantini M, Sharma SK, Perna A, and Remuzzi G. Role of remission clinics in the longitudinal treatment of CKD. J Am Soc Nephrol Jun;19:

295 Antiga L, Piccinelli M, Botti L, Ene-Iordache B, Remuzzi A, Steinman DA. An image-based modeling framework for patient-specific computational hemodynamics. Med Biol Eng Comput Nov;46: RESEARCH ACTIVITIES Laboratory of Renal Biophysics Three dimensional reconstruction of the glomerular capillary network We have recently documented that angiotensin II blockade not only retards the progression of renal diseases, but also induces regression of the glomerular lesions. To quantify the extent of the regression of sclerotic lesions and the potential regeneration of the glomerular capillary induced by a therapy with angiotensin converting enzyme (ACE) inhibitors we are developing three dimensional (3D) reconstruction of tissues by several approaches: one of them is the vascular corrosion casting technique, using a polyurethane resin, that allows to obtain casts of the vascular architecture of the glomerular capillary to be analyzed by scanning electron microscopy. We are also setting up a new technique to study the morphology of the glomerular capillary and its wall by electron microscopy, based on a combination of an electron beam with an ionic one, in order to achieve sectioning of samples and acquisition of serial images for 3D reconstruction of the ultrastructure by mathematical algorithms. Study of the regression of sclerotic lesions and kidney regeneration in response to a therapeutic treatment targeting angiotensin II Clinical and experimental studies have documented that lowering proteinuria by pharmacological interventions retards renal disease progression and even induces remission. We have recently shown in MWF rats, a genetic model of spontaneous glomerular damage, that the ACE inhibitor, lisinopril, induces regression of proteinuria and of existing glomerulosclerosis and stabilizes renal function even when given in advanced stages of nephropathy. Reabsortion of glomerular lesions is associated with remodeling of glomerular cell components which results in a selective increase in podocyte number. These highly differentiated cells are key determinants of perm-selective properties of the glomerular capillary barrier and their loss in the disease correlates with the development of proteinuria and later on of glomerulosclerosis. Beside podocyte restoration, enhanced endothelial cell volume density and reduced mesangial hyperplasia were observed in response to treatment. The mechanism(s) underlying increased podocyte number per glomerulus induced by ACE inhibition has not been established. Podocytes have a limited capability to proliferate and this may be the cause of podocyte depletion we observed in aged MWF rats and generally present in experimental models of progressive nephropathies. However, in untreated MWF rats about 5% of WT1 positive cells (WT1 is a podocyte-specific nuclear antigen) are also positive for Ki-67, a nuclear antigen expressed during all active phases of the cell cycle (G1 to M), and this percentage increase in response to ACE inhibition. At least two possibilities can be taken into account to explain this finding. One is that some podocyte may re-engage and progress throughout the cell cycle. The increase in WT1/Ki-67 positive cells in lisinopril treated MWF rats is associated with a reduced expression of the cyclin-dependent kinase inhibitor p27, a negative modulator of the cell cycle that induces cell cycle arrest. Thus, it is conceivable that reduced p27 may favor proliferation of podocytes that have re-engaged the cell cycle. The second possibility is that WT1/Ki-67 positive cells may derive from podocyte precursor. Ultrastructural features characteristic of podocytes (i.e. foot processes) have been documented in some parietal epithelial cells of the 294

296 Bowman s capsule that express epitopes of mature visceral podocytes and retain the ability to divide. In MWF rats the percentage of parietal podocytes over the total parietal epithelial cells increase in response to ACE inhibition. Electron microscopy examination reveals a continuity between the inner surface of the Bowman s capsule and the outer glomerular capillary membrane supporting the possibility for parietal cells to migrate from Bowman s capsule to the capillary tuft even in normal physiological condition. Moreover, occasional finding of parietal epithelial cells protruding toward the tuft suggests that these cells can also migrate to the tuft through cellular bridges in different areas than the vascular pole. Overall our results suggest that remodeling of parietal epithelial cells of the Bowman s capsule contributes to podocyte restoration in response to ACE inhibition (in collaboration with the Department of Molecular Medicine). Application of confocal microscopy to the study of cells and tissues Fluorescent probes to investigate the distribution and co-localization of proteins and other specific antigens in cell monolayers and tissue specimens are commonly used in the majority of experimental applications. Bidimensional images are not of high resolution to provide correct information, thus the use of 3D reconstruction of fluorescent signal becomes mandatory. These problems can not be solved by the classic fluorescent microscopy that has some limitations. For these reasons we have optimized the laser confocal microscopy by using the LSM510 META microscope from Zeiss (Germany). The microscope is equipped with four laser lines that excite the sample within the whole visible spectrum plus the characteristic META scan head that allow to analyze the emission spectrum of different fluorophores to optimally separate them. The instrument is characterized by and used for its high specificity and for the excellence of the revealed and elaborated signal. Laboratory of Biomedical Technologies Development of computerized systems for controlled clinical trials Numerous clinical trials are conducted in the Clinical Research Center for Rare Diseases Aldo e Celè Daccò. These studies must be carried out with respect to the GCP (Good Clinical Practice) guidelines and require high quality of data. Every clinical study requires a paper case report form (CRF) for collection of patients clinical observations. These data must be checked for inconsistency by dedicated monitoring staff, and then recorded electronically. In our Lab we have developed applications tailored for data management of clinical studies using relational databases systems (RDBMS) and specific programs aimed to data elaboration, validation and extraction for subsequent statistic analyses. REIN, BENEDICT MYSS REIN2, DKG are just few of the trials concluded successfully and published in prestigious medical journals. This accomplishment is, in part, due to the contribution of our Lab. For the DEMAND study we have developed an innovative electronic CRFs based on laptop computers. For the future studies, we are implementing also a web-based framework for electronic data capture and clinical data management for clinical trials. Data Management for the Registry for Rare Disease - Lombardy There is in act a collaboration between our Lab and the Unit of Information Center for Rare Diseases for the management of the Regional Network for Rare Diseases of Lombardy, Italy. We are directly involved for the development and maintenance of the web site of the center and management of a regional Registry for Rare Diseases. We have set-up the databases and developed related web-pages for centers, disease archive, associations of patients and congenital rare diseases. These are publicized on the homepage of the web-site ( 295

297 The Registry for Rare Diseases was born in 2007 as collaboration between Mario Negri Institute, Lombardia Informatica (LI) and Regione Lombardia. The aim was to create a regional registry for rare diseases where all medical staff from Lombardy could register information regarding rare diseases. The application (Sistema Malattie Rare - SMR) was developed by LI in collaboration with our group. SMR application is using web-based technology and can be used jointly with the health patient card. It is actually in use in almost all centers dedicated for rare diseases in Lombardy. Our laboratory is involved in the maintenance of SMR, statistical analyses of registry data and creation of a minimal set of data that are shared at national level by the Istituto Superiore di Sanità (ISS). File download from LI is secured because data are encrypted using an asymmetrical algorithm with public and private key at 2048 bit. Also data transfer at ISS is performed securely by using the htpps Internet protocol. KDDC a coordinating center for data collection and surveillance of prevention programs on non-communicable chronic diseases in emerging countries Chronic kidney diseases are emerging as a global threat to human health. Prevalence and incidence of renal diseases in developing countries are not known, and this is an obstacle to the adoption of preventive measures. Prevention is the only hope for these countries where treatment options for end stage renal failure are simply not available to the vast majority of the population because of their costs. The International Society of Nephrology (ISN), through the Commission for Global Advancement of Nephrology (COMGAN), has established a research committee in order to face the problems about prevention of kidney diseases in developing countries. The coordination of the team and intervention programs was committed to the Mario Negri Institute for Pharmacological Research at the Clinical Research Center Aldo e Cele Daccò. The general aim of the project is to define programs in developing countries to identify those subjects who are at risk of developing a renal disease later in life, in order to design a prevention strategy on national basis by means of interventions of the local ministries of health to governmental and financial level. The Kidney Disease Data Center (KDDC), headquartered in our Laboratory, is dedicated to data management for the prevention programs underway in emerging countries. We have set up an instrument to collect clinical data from different centres located world-wide. Data are stored in a dedicated server in our Laboratory. Results of our epidemiological analyses, shared also with medical staff of the center, allow us to have a general overview on the health of population under study. The prevention program is underway and we have already collected data from participating centers from Nepal, Mongolia, India, China, Moldova, Bolivia and Mozambique. First results on screening on several thousands of subjects, confirm the need to proceed with prevention programs in theses countries. Other countries are willing to start their programs, as for example Mexico, Argentina, Philippines. Through the activity of KDDC it is possible to monitor the course of the prevention programs and to tailor them to fit the needs of each participating country. Hemodynamics and vascular pathology During the last ten years the presence of a close relationship between hemodynamics and vascular pathology has been pointed out both in the biological and in the clinical field. Two typical examples of such relationship are atherosclerosis and intracranial aneurysm disease. In atherosclerosis, a pathology leading to lipid-rich and fibrotic plaque formation in artery walls, lesions tend to form mainly in correspondence to bifurcations (e.g. carotid bifurcation) and great curvature tracts (e.g. coronary arteries). In intracranial aneurysm disease, characterized by the formation of one (or more) artery wall protrusions in the cerebral arterial circulation, cerebral 296

298 aneurysms tend to locate at the distal wall of main bifurcations and at the exteral wall of segments characterized by large curvature. Besides being involved in pathogenesis, hemodynamics also plays a role at a mechanical level for the determination of atherosclerotic plaque rupture, responsible for heart attacks and stroke, or cerebral aneurysm rupture with consequent intracranial hemorrhage. It has been experimentally shown that endothelial cells (lining vessel walls) and smooth muscle cells (the vasoactive component of the wall) exposed to flow exhibit changes in gene expression, cytoskeleton reorganization and permeability according to the characteristics of the imposed flow (e.g. laminar vs turbulent). In spite of this data supporting the hypothesis of a close relationship between hemodynamics and vascular pathology, the nature of such relationship is still under scrutiny, mainly due to the difficulty of measuring in-vivo hemodynamics-related quantities. Thanks to innovative technologies developed during the last few years in the medical imaging and mathematical modeling fields, also within the Biomedical Engineering Department, it is now possible to accurately reproduce patient-specific hemodynamic force distribution from computed tomography (CT) or magnetic resonance (MR) acquisitions. Within the Department of Biomedical Engineering such technologies have three main applications: atherosclerosis (carotid bifurcation and renal artery level), intracranial aneurysm disease and complications of vascular access in hemodialysis. In particular, the Department is currently involved in two international collaborative projects: ARCH, 3-year project funded by the European Union within the Seventh Framework Programme, starting from 2008, aimed at improving the functionality of vascular access in hemodialysis patients, for which the Department is the coordinator centre; ANEURISK, project funded by Siemens Medical and Fondazione Politecnico, aimed at studying the role of hemodynamics in the pathogenesis of intracranial aneurysms and in influencing their rupture risk. Imaging and quantification in renal physiopathology The use of imaging techniques such as CT, MR, and echography, and the application of advanced image processing tools make it possible to perform non-invasive in-vivo quantitative analysis of biological phenomena. Within the Department of Biomedical Engineering, this approach is applied to the investigation of renal physiopathology. Through CT and MR imagebased quantification, new therapies for autosomal dominant polycystic kidney disease (ADPKD) are currently being evaluated. To this purpose, three ongoing clinical trials aimed at reducing the overall kidney cyst volume, one of which funded by the Polycystic Kidney Foundation, are currently relying on image-based quantification. CT image quantification has recently led to the discovery of a high correlation between a specific portion of polycystic kidney tissue and renal function, showing for the first time a likely direct relationship between structure and function, thus opening the way to new therapeutic targets. Beyond ADPKD studies, new methodologies for noninvasive characterization of renal functionality from diffusion-weighted MR images are currently under study. Development of biomedical image analysis and computational modelling tools The employment of quantitative imaging and mathematical modeling techniques aimed at the study of physiopathological processes is directly linked to medical image management and processing methodologies. Within the Department, research activity related to the development of new image processing algorithms and mathematical models for the numerical simulation of biological phenomena, both theoretical and in terms of software development, is actively performed. The department contributes to the development of three main open-source projects in the biomedical imaging field: Vascular Modeling Toolkit ( as main developer, 297

299 Insight Toolkit ( a state-of-the-art library for medical image analysis, and Slicer 3D ( one of the main medical imaging applications. The Department is directly involved in the development activities carried out within the National Alliance for Medical Image Computing, an inter-university consortium gathering the main academic institutions of the United States. Development of devices for the transplantation of immunoisolated islets The project s main objective is to develop an immunisolation device for pancreatic islets that can be implanted in diabetic patients permitting allo-islet transplantation without the use of pharmacological immunosuppression and avoiding allosensitization of the patient. The study started from the design and characterization of the semi-permeable membrane used for the device construction. The device that we are developing can be implanted with minimally invasive surgical procedures and easy to retry. This system is a device made using polysulfone hollow fibers or a tubular membrane in polivinil-alcool. The aims of our studies in the next months will be to improve functionality using nanotechnologies for materials characterization. Moreover we will develop new kinds of device and we will test new implantation sites. Effect of immunoisolation on oxygen consumption by pancreatic islets Immunoisolated pancreatic islet transplantation open new perspectives in the treatment of type 1 diabetes without the use of immunosoppressive drugs. In the last years, our group has been interested in the development of two immunoisolation devices for the transplantation of bovine pancreatic islets in streptozotocin induced diabetic rats. The first device is formed by alginate capsules produced with techniques pointed in our laboratory. The second system is constituted from a device of polysulfone hollow fibers. Both devices must allow adequate diffusion of oxygen to maintain islet viability and function. In the attempt to understand whether immunoisolated islets have adequate oxygen supply in these devices, we have measured their consumption of oxygen. To this purpose, we have set up an experimental technique to measure oxygen consumption rate using a Clark s electrode inserted in a glass thermostated chamber connected to a data recorder and acquisation system. We have then estimated oxygen consumption rate of free and encapsulated islets in alginate capsules and in polysulfone hollow fibers. Both immunoisolation devices allowed oxygen consumption values comparable to that measured in free islets. The data suggests that islet encapsulation in alginate gel and in polysulfone hollow fibers allows an adequate oxygenation to maintain islet function and viability. Mesenchymal stem cells improve vascolarization in pancreatic islet transplantation Type I diabetes is a chronic metabolic disorder that results from the progressive destruction of the ß cells leading to insulin insufficiency and hyperglycemia which is the main determinant of chronic vascular complications. Pancreatic islet transplantation represents a cure for type I diabetes. Recently developed protocols enhanced the success rate of islet transplantation but there are still big limitations including the lack of metabolic capacity of transplanted islets in the long-run. This phenomenon must be mainly attributed to delayed and insufficient islet revascularization that can deprive newly transplanted islets of oxygen, resulting in permanent cell death. Promotion of islet revascularization through locally increased expression of growth factors, such as vascular endothelial growth factor (VEGF), could improve the efficiency of islet transplantation. On this basis, we elected to investigate whether rat MSCs co-transplanted with pancreatic islets may serve as cell therapy to promote therapeutic angiogenesis ultimately leading to an effective metabolic activity of islet grafts. 2,000 syngenic islets alone or in combination with MSCs were transplanted under the kidney capsules of diabetic Lewis rats. Animals transplanted with islets alone never reached normoglycemia. In contrast, in rats 298

300 transplanted with islets and MSCs, glycemia gradually fell after transplantation and normoglycemia was maintained for a long time. In transplanted animals, islet vascularization was quantified by morphometrical analysis. Mean capillare density was significantly increased in islet co-transplanted with MSCs indicating that co-transplantation of MSCs with pancreatic islets improves islet graft function by promoting graft vascularization. (In collaboration with Laboratory of Cellular Biology and Xenotransplantation). Vascular tissue engineering Alternative strategies have been conceived in recent years to develop artificial vascular prothesis to restore the correct transport of blood in the circulation. Currently available synthetic vascular grafts are limited to large internal diameter grafts because of frequent thrombosis and occlusion. The alternative is represented from the use of autologous vascular graft, but they are not always available in patients affected from vascular pathology. Vascular tissue engineering has the objective to generate cellularized vascular prosthesis made of biodegradable materials that can be colonized by endothelial cells. For this purpose (in collaboration with Stazione Sperimentale della Seta in Milano and Politecnico di Milano), we have studied an innovative strategy for the vascular prosthesis realization using biodegradable material, the fibroina of the silk. Tubular structures have been produced, through elettrospinning of the fibrin of the silk, and they will be implanted in small animals (rats) to replace abdominal aorta and, therefore, to evaluate the functionality of the prosthesis. The effect of flow on renal tubular cells ADPKD (Autosomal Dominant Policystic Kidney Disease) is one of the major congenital renal pathologies and results in the development of cysts in the tubular segment, that leads to renal failure. This pathology is due to the modification of the gene encoding for policystin 1, which is contained in the cilia of tubular cells. In the literature it has been shown that cilia work as mechanosensory organs. The bending of the cilium, caused by tubular flow, leads to the activation of the policystin 1 and to the generation of a peak of intracellular calcium concentration. This increase in intracellular calcium activates signalling pathways that modify cellular proliferation and other cellular functions. In pathologic conditions, policystin modification impairs the mechanosensitive function of cilia, altering tubular cellular functions. The aim of this project is the in vitro study of renal tubular cells (MDCK2) in laminar flow conditions, to investigate the role of mechanical stimulation in the pathology development. Preliminary results showed that the application of a constant laminar flow to MDCK2 causes a different tridimensional organization of the cellular layer, as compared to static control. Furthermore, the inhibition of intracellular calcium increase impairs this reorganization when flow is applied. 299

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302 LABORATORY OF BIOLOGY AND THERAPY OF METASTASIS* STAFF Head Raffaella GIAVAZZI, Biol.Sci.D., Ph.D. Head Giulia TARABOLETTI, Biol.Sci.D. * Research activities of this Laboratory are listed in the Department of Oncology section (pag. 7) 301

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304 Aldo and Cele Daccò Center Ranica (Bg) ANNUAL REPORT 2008 departments and laboratories 303

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306 DEPARTMENT OF RENAL MEDICINE STAFF Head Piero RUGGENENTI, M.D. Laboratory of Biostatistics Head Annalisa PERNA, Stat.Sci.D. Unit of Drug Monitoring Head Giulia GHERARDI Laboratory of Clinical Chemistry Head Flavio GASPARI, Chem.D. Laboratory of Advanced Development of Drugs Head Norberto PERICO, M.D. Unit of Pharmacology and Pharmacogenetics Head Dario CATTANEO, Chem.Pharm. D., Ph.D Unit of Early Clinical Evaluation of Drugs Head Aneliya PARVANOVA, M.D. 305

307 CURRICULA VITAE Piero Ruggenenti got his Medicine degree in 1983 at the University of Milan, Italy; he got his specialization in Cardiology in 1985 and in Clinical Nephrology in 1989 at the same University; he specialized in Pharmacological Research in 1988 at IRFMN. Educational training: in researcher at "Centro di Fisiologia Clinica e Ipertensione, Clinica Medica IV", Università degli Studi di Milano; in 1984 Researcher at IRFMN, Bergamo, Italy in Honorary Registrar of the Unit for Metabolic Medicine, Division of Medicine (University of London) of Guy's and St. Thomas's Hospitals, London; in Assistant Professor of the Division of Nephrology and Dialysis of the Ospedali Riuniti di Bergamo. Areas of interest: mechanisms of chronic renal disease progression, diabetes and diabetic complications, clinical transplantation, thrombotic microangiopathies, cardiovascular complications of chronic renal disease, clinical trials, clinical pharmacology. Employment: from 1990 Assistant Professor of the Division of Nephrology and Dialysis of the Ospedali Riuniti di Bergamo; in Head, Unit of Advanced Development of Drugs, Daccò Center, Ranica, Bergamo, Italy; since 2000 Head, Department of Renal Medicine, Daccò Center, Bergamo, Italy. Selected publications P. Ruggenenti, A. Perna, L. Mosconi, M. Matalone, G. Garini, M. Salvadori, C. Zoccali, F. Scolari, Q. Maggiore, G. Tognoni, G. Remuzzi (for The GISEN Group). Randomised placebo-controlled trial of effect of ramipril on decline in glomerular filtration rate and risk of terminal renal failure in proteinuric, non-diabetic nephropathy. Lancet 1997;349: P. Ruggenenti, A. Perna, G. Gherardi, F. Gaspari, R. Benini, G. Remuzzi, on behalf of GISEN. Renal function and requirement for dialysis in chronic nephropathy patients on long-term ramipril: REIN follow-up trial. Lancet 1998;352: P. Ruggenenti, A. Perna, G. Gherardi, G. Garini, C. Zoccali, M. Salvadori, F. Scolari, F.P. Schena, G. Remuzzi. Renoprotective properties of ACE-inhibition in non-diabetic nephropathies with non-nephrotic proteinuria. Lancet 1999;354: G. Remuzzi, C. Chiurchiu, M. Abbate, V. Brusegan, M. Bontempelli, P. Ruggenenti. Rituximab for idiopathic membranous nephropathy. Research Letter. Lancet 2002;360: P. Ruggenenti, A. Fassi, A. Parvanova, S. Bruno, I. Iliev, V. Brusegan, N. Rubis, G. Gherardi, F. Arnoldi, M. Ganeva, B. Ene-Iordache, F. Gaspari, A. Perna, A. Bossi, R. Trevisan, A.R. Dodesini, G. Remuzzi for the Bergamo Nephrologic Diabetes Complications Trial (BENEDICT) Investigators. Preventing microalbuminuria in type 2 diabetes. N Engl J Med 2004;351: G. Remuzzi, P. Cravedi, A. Perna, B.D. Dimitrov, M. Turturro, G. Locatelli, P. Rigotti, N. Baldan, M. Beatini, U. Valente, M. Scalamogna, P. Ruggenenti Dual Kidney Transplant Group. Long-term outcome of renal transplantation from older donors. N Engl J Med 2006;354: Flavio Gaspari got his Chemistry degree in 1977 at the University of Milano, Italy, and the specialization in the same University in Educational training: in Fellow and Researcher at IRFMN, Milan; in at IRFMN, Bergamo, Italy. Areas of interest: pharmacokinetics and the metabolism of xanthines in different animal species; drug pharmacokinetics in uremic patients and in subjects with different degrees of renal function; analytical methods to measure the most important immunosuppressive drugs to determine their pharmacokinetics in kidney, heart, and liver transplant recipients; evaluation of the renal function by using different approaches, in the study of renal disease progression, and in the comparison of different methods for albuminuria determination. Employement: He is Chief of Laboratory of Pharmacokinetics and Clinical Chemistry since January 2000 and he was Chief of this Unit since Selected publications Gotti E, Perico N, Gaspari F, Cattaneo D, Lesti MD, Ruggenenti P, Segoloni G, Salvadori M, Rigotti P, Valente U, Donati D, Sandrini S, Federico S, Sparacino V, Mourad G, Bosmans JL, Dimitrov BD, Iordache BE, Remuzzi G. Blood cyclosporine level soon after kidney transplantation is a major determinant of rejection: insights from the Mycophenolate Steroid-Sparing Trial. Transplant Proc Jun;37(5): Perico N, Gaspari F, Remuzzi G. Assessing renal function by GFR prediction equations in kidney transplantation. Am J Transplant Jun;5(6):

308 D. Cattaneo, F. Gaspari, S. Zanoni, S. Baldelli, E.Gotti, A. Perna, N. Perico, G. Remuzzi. Two-hour post-dose cyclosporine monitoring does not fit all in kidney transplantation. Therapy 2005;2: Ruggenenti P, Fassi A, Ilieva AP, Bruno S, Iliev IP, Brusegan V, Rubis N, Gherardi G, Arnoldi F, Ganeva M, Ene- Iordache B, Gaspari F, Perna A, Bossi A, Trevisan R, Dodesini AR, Remuzzi G; Bergamo Nephrologic Diabetes Complications Trial (BENEDICT) Investigators. Preventing microalbuminuria in type 2 diabetes. N Engl J Med Nov 4;351(19): Epub 2004 Oct 31. Gaspari F, Ferrari S, Stucchi N, Centemeri E, Carrara F, Pellegrino M, Gherardi G, Gotti E, Segoloni G, Salvadori M, Rigotti P, Valente U, Donati D, Sandrini S, Sparacino V, Remuzzi G, Perico N; MY.S.S. Study Investigators. Performance of different prediction equations for estimating renal function in kidney transplantation. Am J Transplant Nov;4(11): Cattaneo D, Merlini S, Pellegrino M, Carrara F, Zenoni S, Murgia S, Baldelli S, Gaspari F, Remuzzi G, Perico N. Related Articles, Links Therapeutic drug monitoring of sirolimus: effect of concomitant immunosuppressive therapy and optimization of drug dosing. Am J Transplant Aug;4(8): Norberto Perico got his Medicine degree in 1983 at the University of Milano, Italy. He got his specialization in Pharmacological Research in 1986 at IRFMN, Bergamo and in Clinical Nephrology in 1989 at the University of Verona, Italy. Educational training: in 1982 Fellow, Department of Pharmacology, New York Medical College, Valhalla, New York, USA; in Post Doctoral Fellow, Laboratory of Kidney Diseases, IRFMN, Bergamo, Italy; in Researcher in the same laboratory. Areas of interest: pathophysiology and pharmacology of cyclosporine nephrotoxicity; new immunosuppressive strategies to prevent renal graft rejection; innovative approach to induce tolerance to organ transplantation; mechanism(s) and management of progression of chronic renal diseases. Employment: in Head, Renal Physiology Unit, Laboratory of Kidney Diseases, IRFMN, Bergamo, Italy; in Assistant Professor, Division of Nephrology and Dialysis, Ospedali Riuniti di Bergamo, Italy; in Head, Laboratory of Transplant Immunology, IRFMN, Bergamo, Italy; from January 2000 Head, Laboratory of Drug Development, Department of Renal Medicine, IRFMN, Bergamo, Italy; from September 2000 Health Director, Daccò Center, IRFMN, Bergamo, Italy. From October 2002 he s Member, ISN-COMGAN Research Committee of the International Society of Nephrology. Selected publications: E. Gotti, N. Perico, A. Perna, F. Gaspari, D. Cattaneo, R. Caruso, S. Ferrari, N. Stucchi, G. Marchetti, M. Abbate, G. Remuzzi. Renal transplantation: can we reduce calcineurin inhibitor/stop steroids? Evidence based on protocol biopsy findings. J Am Soc Nephrol 2003;14: N. Perico, P. Ruggenenti, G. Remuzzi. Losartan in diabetic nephropathy. Expert Rev. Cardiovasc. Ther. 2004; 2(4): Remuzzi G, Lesti M, Gotti E, Ganeva M, Dimitrov BD, Ene-Iordache B, Gherardi G, Donati D, Salvadori M, Sandrini S, Valente U, Segoloni G, Mourad G, Federico S, Rigotti P, Sparacino V, Bosmans JL, Perico N, Ruggenenti P. mofetil versus azathioprine for prevention of acute rejection in renal transplantation (MYSS): a randomised trial. Lancet Aug 7;364(9433): Gaspari F, Ferrari S, Stucchi N, Centemeri E, Carrara F, Pellegrino M, Gherardi G, Gotti E, Segoloni G, Salvadori M, Rigotti P, Valente U, Donati D, Sandrini S, Sparacino V, Remuzzi G, Perico N; MY.S.S. Study Investigators. of different prediction equations for estimating renal function in kidney transplantation. Am J Transplant Nov;4(11): Perico N, Cattaneo D, Sayegh MH, Remuzzi G. Delayed graft function in kidney transplantation. Lancet Nov 13;364(9447): Cattaneo D, Merlini S, Zenoni S, Baldelli S, Gotti E, Remuzzi G, Perico N. Influence of co-medication with sirolimus or cyclosporine on mycophenolic acid pharmacokinetics in kidney transplantation. Am J Transplant Dec;5(12): Annalisa Perna got her Statistical Sciences degree in 1984 at the University of Bologna, Italy. Educational training: She completed her research training at IRFMN, Bergamo Labs. and at the Daccò Center. Areas of interest: statistical methodology of long-term randomised clinical trials in nephrology, statistical methods for calculating sample size and for meta-analytic techniques. She is also involved in performing systematic reviews for the Cochrane Collaboration Renal Review Group. Employment: she is Head of the Laboratory of Biostatistics - Department of Renal Medicine at Daccò Center, Ranica (Bergamo). 307

309 Principal publications: G. Remuzzi, P. Cravedi, A. Perna, B.D. Dimitrov, M. Turturro, G. Locatelli, P. Rigotti, N. Baldan, M. Beatini, U. Valente, M. Scalamogna, P. Ruggenenti. Dual Kidney Transplant Group. Long-term outcome of renal transplantation from older donors. N Engl J Med 2006;354: Ruggenenti P, Perna A, Loriga G, Ganeva M, Ene-Iordache B, Turturro M, Lesti M, Perticucci E, Chakarski IN, Leonardis D, Garini G, Sessa A, Basile C, Alpa M, Scanziani R, Sorba G, Zoccali C, Remuzzi G for the REIN-2 Study group. Blood-pressure control for renoprotection in patients with non-diabetic chronic renal disease (REIN-2): multicentre, randomised controlled trial. Lancet 365: , Schieppati A, Perna A, Zamora J, Giuliano AG, Braun N, Remuzzi G. Immunosuppressive treatment for idiopathic membranous nephropathy in adults with nephrotic syndrome. Cochrane Database Syst Rev. Oct 18(4):CD004293, 2004 Ruggenenti P, Fassi A, Ilieva AP, Bruno S, Iliev IP, Brusegan V, Rubis N, Gherardi G, Arnoldi F, Ganeva M, Ene- Iordache B, Gaspari F, Perna A, Bossi A, Trevisan R, Dodesini AR, Remuzzi G. Preventing microalbuminuria in type 2 diabetes. N Engl J Med 351(19) , The BENEDICT Group. The Bergamo NEphrologic Diabetes Complications Trial (BENEDICT): design and baseline characteristics. Control Clin Trials 24(4): , Plata R, Cornejo A, Arratia C, Anabaja A, Perna A, Dimitrov BD, Remuzzi G, Ruggenenti P for the Commission on Global Advancement of Nephrology (COMGAN), Research Subcommittee of the International Society of Nephrology. Angiotensin-converting-enzyme inhibition therapy in altitude polycythaemia: a prospective randomised trial. Lancet 359: , Dario Cattaneo got the Pharmacy degree in 1996 at the University of Milan, and the specialisation in Pharmacology (2001) awarded by the same University. In 2000 he got the specialization in Pharmacological Research at the Mario Negri Institute for Pharmacological Research (IRFMN) and in 2005 he has been awarded the PhD degree by the Open University of London, UK. Educational Training: in 1997 Post Doctoral Fellow, IRFMN, Laboratory of Pharmacokinetics and Clinical Chemistry; in 2000 beneficiary of the Fellowship Girola and from 2001 to 2005 recipient of the Monzino Fellowship for his research activity done at the IRFMN. Areas of interest: pharmacology (pharmacokinetics, pharmacodynamics and pharmacogenetics) of immunosuppressants, antiviral agents and hypolipidemic drugs; study of secondary forms of dyslipidemia; polypharmacological approaches for the treatment of chronic kidney diseases; assessment of humoral response as predictor of acute or chronic rejection after organ transplantation. Employement: in 1997 researcher, IRFMN, Laboratory of Pharmacokinetics and Clinical Chemistry,. Since 2006, Head of the Unit of Pharmacology and Pharmacogenetics, IRFMN, Ranica Bergamo. Component of the Ethical Committee (2003) of the Hospital Bolognini (Seriate, Italy) and the Hospital E.Medea (Bosisio Parini, Italy) since Member of the editorial board of Current Clinical Pharmacology and affiliate of the International Association of Therapeutic Drug Monitoring and Clinical Toxicology (IATDMCT) since Selected publications Cattaneo D, Merlini S, Zenoni S, Baldelli S, Gotti E, Remuzzi G, Perico N. Influence of co-medication with sirolimus or cyclosporine on mycophenolic acid pharmacokinetics in kidney transplantation. Am J Transplant Dec;5(12): Cattaneo D, Gotti E, Perico N, Bertolini G, Kainer G, Remuzzi G. Cyclosporine formulation and Kaposi's sarcoma after renal transplantation. Transplantation Sep 27;80(6): Perico N, Cattaneo D, Sayegh MH, Remuzzi G. Delayed graft function in kidney transplantation. Lancet Nov 13-19;364(9447): Cattaneo D, Merlini S, Pellegrino M, Carrara F, Zenoni S, Murgia S, Baldelli S, Gaspari F, Remuzzi G, Perico N. Therapeutic drug monitoring of sirolimus: effect of concomitant immunosuppressive therapy and optimization of drug dosing. Am J Transplant Aug;4(8): Cattaneo D, Perico N, Gaspari F, Gotti E, Remuzzi G. Glucocorticoids interfere with mycophenolate mofetil bioavailability in kidney transplantation. Kidney Int Sep;62(3): Giulia Gherardi got her Scientific High School Diploma on 1989 at the Liceo Scientifico Marie Curie in Zogno (Bergamo), the Nurse Diploma on 1995 at the Scuola per Infermieri Professionali, Ospedali Riuniti, Bergamo. Educational training: Clinical Research Nurse Diploma on 1997 at IRFMN Daccò Center. Areas of interest: statistical methodology of long-term randomised clinical trials in nephrology, diabetology; the organisation and the monitoring of clinical trials. Emplyment: In involved as co-organazing, speaker, co-speaker and tutor for the Clinical Research Course for Nurse at IRFMN Daccò Center (Ranica Bergamo). Several training activities for Nurses in Clinical Research area. In , Clinical Research Monitor at IRFMN Daccò Center; since 2000 Monitoring Unit Chief. 308

310 Selected pubblications Gaspari F, Ferrari S, Stucchi N, Centemeri E, Carrara F, Pellegrino M, Gherardi G, Gotti E, Segoloni G, Salvadori M, Rigotti P, Valente U, Donati D, Sandrini S, Sparacino V, Remuzzi G and Perico N on the behalf of the MY.S.S. study investigators. Performance of different prediction equations for estimating renal function in kidney transplantation. American Journal of Transplantation. 2004; 4: Ruggenenti P, Fassi A, Parvanova Ilieva A, Bruno S, Petro Iliev I, Brusegan V, Rubis N, Gherardi G, Arnoldi A, Ganeva M, Ene-Iordache B, Gaspari F, Perna A, Bossi A, Trevisan R, Dodesini AR, Remuzzi G, for the Bergamo Nephrologic Diabetes Complications Trial (BENEDICT) Investigators. Preventing microalbuminuria in type 2 diabetes. N Engl J Med. 2004; 351: Remuzzi G, Lesti M, Gotti E, Ganeva M, Dimitrov BD, Ene-Iordache B, Gherardi G, Donati D, Salvadori M, Sandrini S, Valente U, Segoloni G, Mourad G, Federico S, Rigotti P, Sparacino V, Bosmans JL, Perico N, Ruggenenti P, for the MY.S.S. Group. Mycophenolate mofetil versus azathioprine for prevention of acute rejection in renal transplantation (MYSS): a randomized trial. Lancet Aug 7; 364: Ruggenenti P, Perna A, Gherardi G, Benini R, Remuzzi G. Chronic proteinuric nephropathies: outcomes and response to treatment in a prospective cohort of 352 patients with different patterns of renal injury. Am J Kidney Dis Jan; 35 (6): Ruggenenti P, Perna A, Zoccali C, Gherardi G, Benini R, Testa A, Remuzzi G. Chronic proteinuric nephropathies. II. Outcomes and response to treatment in a prospective cohort of 352 patients: differences between women and men in relation to the ACE polymorphism. Gruppo Italiano di Studi Epidemiologici in Nefrologia. J Am Soc Nephrol Jan; 11 (1): Ruggenenti P, Perna A, Gherardi G, Garini G, Zoccali C, Salvadori M, Scolari F, Schena FP, Remuzzi G. Renoprotective properties of ACE-inhibition in non-diabetic nephropathies with non-nephrotic proteinuria. Lancet Jul 31; 354 (9176): Aneliya Parvanova Ilieva got her Medical Doctor degree at the Faculty of Medicine, Thracian University (former Higher Medical Institute), Stara Zagora, Bulgaria in 1988, and the specialisation in Pharmacology in Department of Pharmacology, Faculty of Medicine, of the same university in Educational training: : Teaching of 3 rd, 4 th and 5 th -year medical students and 2 nd and 3 rd -year clinical nurses in a general pharmacology and clinical pharmacology, Thracian University, Stara Zagora, Bulgaria. Examiner of these students in theoretical and practical, oral and written exams and tests. 1993: Course on investigation of isolated organs Bulgarian Academy of Sciences, Sofia. 1998: Visiting scientist, IRFMN, Ranica, Bergamo, Italy. 1998: Proficiency in the methods for insulin sensitivity evaluation (hyperinsulinemic euglicaemic clamp technique) and glomerular filtration rate evaluation (plasma clearance of iohexol). Areas of interest: primary and secondary prevention of the chronic microvascular diabetic complications (diabetic nephropathy, diabetic retinopathy and diabetic neuropathy); role of insulin resistance and hyperhomocysteinemia in these pathologies. Employment: She participated as investigator in several clinical studies. She is Chief Unit of Early Clinical Evaluation of Drugs at IRFMN since She is a member of the Union of Bulgarian Doctors (since 1989), of the Union of Pharmacologists in Bulgaria (since 1990), and member of the Union of Scientists in Bulgaria (since 1991). Selected publications Parvanova A, Chiurchiu C, Ruggenenti P, Remuzzi G. Inhibition of the renin-angiotensin system and cardio-renal protection: focus on losartan and angiotensin receptor blockade. Expert Opinion on Pharmacotherapy 2005 Sep; 6 (11): Ruggenenti P, Fassi A, Parvanova A, Bruno S, Iliev I, Brusegan V, Rubis N, Gherardi G, Arnoldi F, Ganeva M, Ene- Iordache, Gaspari F, Perna A, Bossi A, Trevisan R, Dodesini A, Remuzzi G. Preventing Microalbuminuria in Type 2 Diabetes. NEJM 2004;351(19): Parvanova A, Iliev I, Filipponi M, Dimitrov BD, Vedovato M, Tiengo A, Trevisan R, Remuzzi G, Ruggenenti P. Insulin resistance and proliferative retinopathy: a cross-sectional, case-control study in 115 patients with type 2 diabetes. J Clin Endocrinol Metab 2004 Sep; 89(9): The BENEDICT Group. The Bergamo Nephrologic DIabetes Complications Trial (BENEDICT): design and baseline characteristics. Controlled Clinical Trials 2003; 24: Parvanova A, Iliev I, Dimitrov BD, Arnoldi F, Zaletel J, Remuzzi G, Ruggenenti P. Hyperhomocysteinemia and increased risk of retinopathy: a cross-sectional, case-control study in patients with type 2 diabetes. Diabetes Care 2002; 25 (12):

311 INTRODUCTION TO THE DEPARTMENT'S ACTIVITIES The Department of Renal Medicine was established on 1999 at the Clinical Research Center for Rare Diseases Aldo e Cele Daccò Villa Camozzi, Ranica to coordinate the activities of three Laboratories and two Units. The activities of the Department are mainly focused on the study of the mechanisms of progression of chronic nephropathies, of new prevention and intervention strategies for diabetic nephropathy, non diabetic chronic nephropathies, chronic allograft dysfunction, of cardiovascular complications of diabetes, chronic renal disease, dialysis and transplantation and of thrombotic microangiopathies. The main aims of these activities are: 1. To identify screening and intervention strategies aimed to prevent the onset of nephropathy and of other chronic complications of diabetes and/or hypertension. 2. To define intervention strategies to prevent or slow the progression of chronic nephropathies and eventually obtain remission/regression of renal dysfunction. 3. To optimize immunosuppressive protocols in kidney transplantation and to define new donor selection criteria in order to expand the pool of available organs. These aims will be pursued through the following modalities: 1. Pilot pathophysiology and clinical pharmacology studies fully finalized at the Clinical Research Center to test new pathogenetic hypotheses and new treatment modalities. 2. National and international networks and multicenter trials aimed to verify the efficacy of treatments of potential interest identified as described at point Meta-analyses and probabilistic models to test new risk factors and treatments in large samples of patients and to transfer this information at individual level. Many of these activities rest on the possibility of a tight cooperation with the Department of Molecolar Medicine, the Department of Bioengineering and the Public-Private Department of Specialist and Transplant Medicine. This cooperation allows to plan the research activities of the Department on the basis of new information derived from basic research and of problems of major clinical relevance emerging from routine clinical activities. FINDINGS/MAIN RESULTS Definition and validation of specific treatments aimed to prevent the development and progression of nephropathy and related micro and macrovascular complications in subjects with type 2 diabetes Definition and validation of new integrated treatment protocols aimed to slow the progression and/or to achieve remission/regression of diabetic and non-diabetic chronic nephropathies Institution of a standardized protocol on line (The Remission Clinics ) finalized to achieve regression/remission of chronic nephropathies and limit overall renal and radiovascular risk in hospital practice in the setting of a multicenter Network Characterization of the antiproteinuric, nephroprotective and cardioprotective effect of maximized and polypharmacologic renin-angiotensin system inhibition, intensified blood pressure and lipid control and identification of novel treatments to reduce the blood pressure and ameliorate insulin sensitivity in subjects at increased cardiovascular risk 310

312 Identification of acquired or congenital risk factors for chronic complications of diabetes and cardiovascular morbidity and mortality Identification and validation of early markers of acute kidney failure and of methods for direct and indirect measurements of kidney function and GFR decline Definition and validation of new, specific treatments for idiopathic membranous nephropathy and for HUS forms associated with genetic defect of complement factors including the standardization of combined liver and kidney transplantation to prevent post transplant recurrence of genetic associated HUS. Definition and validation of new laboratory procedures and predictive models to help monitoring and optimizing immunosuppressive therapy in clinical transplantation with particular focus on pharmacokynetic markers of drug exposure and genetic predictors of drug tolerability and efficacy Definition and validation of selection and allocation criteria of kidneys from marginal and oldvery old donors to increase the donor pool and the transplant activity Finalization and activation of multicenter clinical trials aimed to prevent onset and progression of diabetic and non-diabetic chronic nephropathies, to achieve remission of the nephrotic syndrome in primary glomerular diseases, minimize maintenance immunosuppression in kidney transplantation and prevent cardiovascular morbidity and mortality in chronic hemodialysis. Computerization of data acquisition and monitoring procedures for the conduction of controlled clinical trials NATIONAL COLLABORATIONS 2008 Lombardia - Ospedale C. Cantù, Abbiategrasso (MI) - Ospedale Civile di Asola, Asola (MN) - Ospedale Fenaroli, Alzano Lombardo (BG) - Azienda Ospedaliera OO.RR., Bergamo - Ospedale Caduti Bollatesi, Bollate (MI) - Azienda Ospedaliera Spedali Civili, Brescia - Ospedale San Biagio, Clusone (BG) - Ospedale S. Anna, Como - Azienda Ospedaliera Istituti Ospedalieri, Cremona - Ospedale di Desio (MI) - Ospedale Briolini, Gazzaniga (BG) - Azienda Ospedaliera di Melegnano, Melegnano Vizzolo Predabissi (MI) - Ospedale San Leopoldo Mandic, Merate (LC) - Ospedale Maggiore Policlinico, Milano - Ospedale Provinciale San Carlo Borromeo, Milano - Azienda Ospedaliera - Polo Universitario L. Sacco, Milano - Ospedale Fatebenefratelli, Milano - Ospedale Niguarda Ca' Granda, Milano - Clinica Pediatrica G. e D. De Marchi, Milano - Ospedale San Raffaele, Milano - Ospedale Pediatrico di Montichiari, Montichiari (BS) 311

313 - Ospedale San Gerardo, Monza (MI) - Istituti Clinici Zucchi, Monza (MI) - Università degli Studi di Pavia, Dipartimento di Medicina Interna e Terapia Medica, Pavia - Centro Antidiabetico, Ponte San Pietro (BG) - Azienda Ospedaliera Ospedale Treviglio Caravaggio, Romano di Lombardia (BG) - Istituto Clinico Humanitas, Rozzano (MI) - Ospedale Bolognini, Seriate (BG) - Ospedale Civile, Ivrea - Azienda Ospedaliera Ospedale Treviglio Caravaggio, Treviglio (BG) - A.O. della Valtellina e della Valchiavenna, Sondrio - Azienda Ospedaliera Santa Croce e Carli, Cuneo - Ospedale Regionale di Circolo Fondazione Macchi, Varese - USL 60, Unità Operativa di Nefrologia e Dialisi, Vimercate (LC) - Ospedale Bassini, Cinisello Balsamo, Milano - IRCCS Multimedia, Sesto San Giovanni, Milano Piemonte - A.S.O. Maggiore della Carità, Novara - Azienda Ospedaliera San Giovanni Battista, Torino - Ospedale Mauriziano Umberto I, Torino - Ospedale Regina Margherita, Torino - Ospedale Martini, Torino - Ospedale Luigi Einaudi, Torino Veneto, Trentino Alto-Adige e Friuli Venezia Giulia - Casa di Cura Abano Terme, Abano Terme (PD) - Ospedale Civile, Belluno - Ospedale S. Giacomo Apostolo, Castelfranco Veneto, Treviso - Ospedale Provinciale Umberto I, Mestre (VE) - Ospedale Giustinianeo, Padova - Università degli Studi di Padova, Istituto di Anatomia Patologica, Padova - Ospedale Civile, Padova - Ospedale S. Camillo dé Lellis, Schio (VI) - Ospedale Regionale Santa Maria dei Battuti, Treviso - Ospedale Ca Fondello, Divisione Nefrologia e Dialisi, Treviso - Ospedale Civile Maggiore Borgo Trento, Verona - Ospedale Policlinico Borgo Roma, Verona - Ospedale Civile San Bortolo, Vicenza - Ospedale Santa Chiara, Trento - Istituto Scientifico per l'infanzia Burlo Garofalo, Trieste - Ospedale S. Antonio, S. Daniele del Friuli, Udine - Università degli Studi di Udine, Centro Trapianti Fegato-Rene-Pancreas, Udine Liguria, Emilia Romagna e Toscana - Azienda Ospedaliera San Martino, Genova - Istituto G. Gaslini, Genova - Ospedale S. Orsola Malpighi, Bologna - Ospedale Policlinico, Modena - Istituto di Clinica Medica e Nefrologia, Parma - Ospedale Santa Maria delle Croci, Ravenna - Arcispedale Santa Maria Nuova, Reggio Emilia 312

314 - Ospedale Santa Maria Annunziata, Bagno a Ripoli, Firenze - Azienda Ospedaliera Careggi-Monna Tessa, Firenze - Ospedale Nuovo S. Giovanni di Dio, Firenze - Azienda Ospedaliera Meyer, Firenze - Ospedale di S. Miniato, S. Miniato (FI) - Azienda Ospedaliera Cisanello, Pisa - Ospedale di Pistoia, Pistoia - AUSL Rimini, Rimini - ASL Ravenna, Ravenna - Ospedale G.B. Morgagni, Forlì - Ospedale Civile, Imola Bologna - Università di Pisa, Ospedale S. Chiara, Pisa Marche - Ospedale Regionale Torrette, Torrette di Ancona, Ancona - Ospedale I.N.R.C.A., Ancona - Azienda Ospedaliera S. Salvatore, Pesaro Lazio, Basilicata e Campania - Ospedale Polispecializzato, Anzio, Roma - Ospedale Fatebenefratelli, Roma - Ospedale Pediatrico Bambino Gesù, Roma - Policlinico Gemelli, Roma - Ospedale Policlinico Umberto I, Roma - Ospedale San Camillo Forlanini, Roma - Università Cattolica del Sacro Cuore, Roma - Dipartimento di Biopatologia Umana, Università La Sapienza, Roma - Ospedale Grande degli Infermi, Viterbo - Ospedale Riuniti, Matera - Azienda Ospedaliera Ospedale Civile, Caserta (NA) - Università Federico II di Napoli, Cattedra di Nefrologia, Napoli - Università di Napoli, Policlinico Nuovo, Napoli - Azienda Ospedaliera S. G. di Dio e Ruggi d Aragona, Salerno Abruzzo - Ospedale G. Bernabeo, Ortona, Chieti - Presidio Ospedaliero San Massimo, Penne (PE) - Presidio Ospedaliero "G.Mazzini", Teramo Puglia, Calabria, Sicilia e Sardegna - Ospedale Regionale Miulli, Acquaviva delle Fonti, Bari - Ospedale Pediatrico Giovanni XXIII, Bari - Ospedale Policlinico, Bari - Azienda Ospedaliera V.Fazzi, Lecce - Ospedale Casa Sollievo dalla Sofferenza, S.Giovanni Rotondo (FG) - Presidio Ospedaliero di Martina Franca, Martina Franca, Taranto - A.U.S.L. TA/1 - Presidio Ospedaliero, Taranto - Azienda Ospedaliera Ospedale Pugliese Ciaccio, Catanzaro - Ospedale dell'annunziata, Cosenza - Centro di Fisiologia Clinica del CNR, Divisione di Nefrologia, Reggio Calabria - Azienda Ospedaliera Bianchi-Melacrino-Morelli, Reggio Calabria - Ospedale N. Giannettasio, Rossano Calabro, Cosenza 313

315 - Nuovo Presidio Ospedaliero, Acireale, Catania - Azienda Ospedaliera "Ferrarotto", Catania - Ospedale Zonale Maggiore, Modica (RG) - Ospedale Civico, Palermo - Ospedale V. Cervello, Palermo - Azienda Ospedaliera "Umberto I", Siracusa - Azienda Sanitaria G. Brotzu, Ospedale San Michele, Cagliari - Istituto di Clinica e Biologia dell Età Evolutiva, Cagliari - Ospedale A. Segni, Ozieri, Sassari - Ospedale SS. Annunziata, Sassari - Istituto di Patologia Speciale Medica dell'università degli Studi di Sassari - Ospedale Policlinico, Sassari - Istituto Mediterraneo per i Trapianti e Terapie ad Alta Specializzazione (IsMeTT), Palermo - A.O. Universitaria Ospedali Riuniti, Foggia Umbria - Azienda Ospedaliera di Perugia, Perugia INTERNATIONAL COLLABORATIONS University Medical Center, Ljubljana Slovenia - University Hospital Ziekenhuius, Edegem Antwerpen, Belgium - Clinique de Nephrologie-Dialyse Chu Brugmann, Bruxelles, Belgium - University Ziekenhuius Gent, Gent, Belgium - U.Z. Gasthuisberg, Leuven, Belgium - General Hospital Maria Middelares, Sint Niklaas, Belgium - University of Groningen, AV Groningen, The Netherlands - Academisch Ziekenhuis, Maastricht, The Netherlands - Thracian University, Stara Zagora, Bulgaria - The Birmingham Children's Hospital, Birmingham, UK - Guy s Hospital, London, UK - Manchester Children's Hospital, Manchester, UK - Nottingham City Hospital, Nottingham, UK - Aalborg Hospital, Aalborg, Denmark - Nephrological Department, University of Copenaghen, Copenaghen, Denmark - Steno Diabetes Center, Gentofte, Denmark - Department of Nephrology, Odense University Hospital, Odense, Denmark - Department of Nephrology, Sahlgrenska University Hospital, Goteborg, Sweden - Ospedale San Giovanni, Bellinzona, Switzerland - Department of Nephrology, University of Wien, Wien, Austria - Carl Thiem Klinikum, Cottbus, Germany - Klinikum der Johann Wolfgang, Frankfurt am Main, Germany - Arbeitsgruppe fyr Biomolekulare Medizin, Hamburg, Germany - Univeristatklinik Heidelberg, Heidelberg, Germany - Medizinische Klinik, Mannheim, Germany - Luitpold Krankenhaus Med. Universitatklinik/Dialyse, Wurzburg, Germany 314

316 - Hospital Ntra Sra. de Sonsoles, Avila, Spain - Hospitalet de Llobregat, Institut Català de la Salut, Barcellona, Spain - Fundacion Jimenez Diaz, Madrid, Spain - Hospital Clinico Martin Logas, Madrid, Spain - Hospital 12 de Octubre, Madrid, Spain - Hospital Gregorio Maranon, Madrid, Spain - Hospital La Paz, Madrid, Spain - Hospital Puerta de Hierro, Madrid, Spain - Hospital Ramon y Cajal, Madrid, Spain - Hospital Severo Ochoa, Leganes, Madrid, Spain - Hospital Universitario de Tarragona Joan XXIII, Tarragona, Spain - Hospital Garcia de Orta, Almada, Portugal - Brigham & Women's Hospital, Boston, USA - Hennepin County Medical Center, Minneapolis, USA - SIU School of Medicine, Springfield, USA - The Toronto Hospital, Toronto, Canada - INCUCAI, Buenos Aires, Argentina - Hospital Italiano de Buenos Aires, Buenos Aires, Argentina - Hospital Regional de Valdivia, Valdivia, Cile - Soroka Medical Center, Beer Sheva, Israel EDITORIAL BOARD MEMBERSHIP 2008 Journal of Nephrology (Piero Ruggenenti) Current Diabetes Reviews (Piero Ruggenenti) Clinical Journal of the American Society of Nephrology (Piero Ruggenenti) Current Pharmacology Reviews (Dario Cattaneo) PEER REVIEW ACTIVITIES American Journal of Kidney Diseases American Journal of Physiology American Journal of Transplantation Biomed Central Edition Brazilian Journal of Medical and Biological Research Circulation Clinical Journal of the American Society of Nephrology (CJASN) Clinical Nephrology Clinical Transplantation 315

317 Current Diabetes Reviews Diabetic Nephropaty Diabetologia Dove Press Expert Opinion on Emerging Drugs Informa Healthcare Journal of Acta Diabetologica Journal of Clinical Investigation Journal of Nephrology Journal of the American Society of Nephrology (JASN) Kidney International Nature Clinical Practice Nephrology Nephrology Dialysis and Transplantation Nephron Clinical Practice New England Journal of Medicine Pediatric Nephrology Plos Medicine Talanta The Brazilian Journal The Lancet The Open urology &Nephrology Journal Translational Research Transplantation Vascular Pharmacology PARTICIPATION IN EVENTS IN WHICH THE DEPARTMENT WAS INVOLVED Pharmacokinetics of mycophenolate sodium and comparison with the mofeil formulation in stable kidney transplant recipients. In oportunidades para maximizar o sucesso do doente, Lisboa, Portugal, February 23, Pharmacogenetics of immunosuppressants: progress and promises. In GENECURE Agenda, Ranica, Italy, April Statins, dyslipidemia and novel hypolipidemic agents. Advanced Workshop for Turkish Nephrologists, Ranica, April 18-19, La farmacogenetica. Forum annuale del Comitato di Bioetica, organizzato dall Azienda Ospedaliera Ospedali Riuniti di Bergamo, 12 May Polymorphisms in ABCB1, the gene encoding for p-glycoprotein, predicts recovery of graft function early after kidney transplantation. Oral presentation for the American Transplant Congress, Toronto, May 31 June 4, La ricerca in campo genetico. Azienda Ospedaliero-Universitaria di Parma, Parma 11 July

318 Strategie farmacologiche di renoprotezione. All interno del Simposio Terapia conservativa, dietetica e farmacologica della malattia renale cronica, Pisa 26 September Valutazione del filtrato glomerulare e utilizzo dell esame delle urine nella pratica clinica, 2 Convegno sulla funzione renale, Centro Congressi Villa Gualino, Torino 27 June 2008 Society for Clinical Trials, St. Louis, Missouri, USA, May Institute for Health Metrics and Evaluation, Global Health Metrics and Evaluation: current state and future directions. Research Conference, Seattle, Washington, USA, April Geni, proteine e malattie. 20 November 2008, Borsa Merci Bergamo Prima Giornata Europea delle Malattie rare, 29 February 2008, Centro Congressi Giovanni XXIII, Bergamo XLV ERA-EDTA Congress, Stoccolma, May 2008 Convegno La ricerca indipendente sui farmaci promossa dall AIFA. Roma, 30 September 2008 Course in Renal Endocrinology. Stoccolma, October Congresso Nazionale della Società Italiana di Nefrologia. Rimini, October2008 Convegno Nefropatie Primitive e Secondarie, Firenze, November Corso di Aggiornamento in Nefrologia e Metodiche Dialitiche, Milano, December 2008 Corso di Aggiornamento La Remission Clinic nella Pratica Clinica, Ranica (BG), 16 December 2008 GRANTS AND CONTRACTS AIFA (Agenzia Italiana del Farmaco) PKD Foundation Abbott GmbH & Co. Astrazeneca SPA ACRAF Spa (Aziende Chimiche Riunite Angelini Francesco) BOHERINGER INGELHEIM Italia Spa Chiesi Farmaceutici Dompè Spa Genzyme Europe BV Roche SpA Sanofi-Aventis Spa Sigma Tau Spa Solvay Pharmaceuticals 317

319 SELECTION OF SCIENTIFIC PUBLICATIONS FROM 2008 G. Remuzzi, N. Perico, M.E. De Broe. Tubulointerstitial Diseases. In: Brenner and Rector's The Kidney, 8 th ed., Vol. II, chapter n. 33. Edited by B.M. Brenner. Saunders Elsevier, Philadelphia 2008, pp S.K. Sharma, N. Perico, P. Ruggenenti, G. Remuzzi. Prevention of chronic renal diseases in the elderly. In: The Aging Kidney in Health and Disease, chapter n. 13. Edited by J.F. Macias Nunez, J.S. Cameron, D.G. Oreopoulos. Springer, New York 2008, pp P. Ruggenenti, I. Iliev, G.M. Costa, A. Parvanova, A. Perna, G.A. Giuliano, N. Motterlini, B. Ene-Iordache, G. Remuzzi, and the BENEDICT Study Group. Preventing left ventricular hypertrophy by ACE inhibition in hypertensive patients with type 2 diabetes. Diabetes Care 2008;31: P. Ruggenenti, E. Perticucci, P. Cravedi, V. Gambara, M. Costantini, S.K. Sharma, A. Perna, G. Remuzzi. Role of remission clinics in the longitudinal treatment of CKD. J Am Soc Nephrol 2008;19: P. Ruggenenti, A. Remuzzi, G. Remuzzi. Decision time for pancreatic islet-cell transplantation. (Comment) Lancet 2008;371: M. Cortinovis, D. Cattaneo, N. Perico, G. Remuzzi. Investigational drugs for diabetic nephropathy. Expert Opin Investig Drugs 2008;17: N. Perico, A. Benigni, G. Remuzzi. Present and future drug treatments for chronic kidney diseases: evolving targets in renoprotection. Nature Reviews Drug Discovery 2008;7: P. Ruggenenti, P. Bettinaglio, F. Pinares, G. Remuzzi. Angiotensin converting enzyme insertion/deletion polymorphism and renoprotection in diabetic and nondiabetic nephropathies. Clin J Am Soc Nephrol 2008;3: F. Casiraghi, N. Azzollini, P. Cassis, B. Imberti, M. Morigi, D. Cugini, R.A. Cavinato, M. Todeschini, S. Solini, A. Sonzogni, N. Perico, G. Remuzzi, M. Noris. Pretransplant infusion of mesenchymal stem cells prolongs the survival of a semiallogeneic heart transplant through the generation of regulatory T cells. J Immunol 2008;181: P. Ruggenenti, M. Noris, G. Remuzzi. Thrombotic Microangiopathies. In: Therapy in Nephrology and Hypertension A companion to Brenner and Rector s The Kidney (3 rd Edition), chapter n. 26. Edited by C.S. Wilcox. Saunders Elsevier, Philadelphia 2008, pp Saland JM, Ruggenenti P, Remuzzi G; Consensus Study Group. Liver-Kidney Transplantation to Cure Atypical Hemolytic Uremic Syndrome. J Am Soc Nephrol Dec 17. Ruggenenti P, Cravedi P, Remuzzi G. Rituximab for membranous nephropathy and immune disease: less might be enough. Nat Clin Pract Nephrol Dec 2 Cravedi P, Mannon RB, Ruggenenti P, Remuzzi A, Remuzzi G. Islet transplantation: need for a time-out? Nat Clin Pract Nephrol Dec;4(12): Epub 2008 Sep 23. Cravedi P, Mannon RB, Remuzzi G. Lymphocyte depletion for kidney transplantation: back to the past? Nat Clin Pract Nephrol Oct;4(10): Epub 2008 Aug 26. Braun N, Schmutzler F, Lange C, Perna A, Remuzzi G, Risler T, Willis NS. Immunosuppressive treatment for focal segmental glomerulosclerosis in adults. Cochrane Database Syst Rev Jul 16;(3): Ruggenenti P, Cravedi P, Sghirlanzoni MC, Gagliardini E, Conti S, Gaspari F, Marchetti G, Abbate M, Remuzzi G. Effects of rituximab on morphofunctional abnormalities of membranous glomerulopathy. Clin J Am Soc Nephrol Nov;3(6): Epub 2008 Aug 6. Remuzzi G, Cattaneo D, Perico N. The aggravating mechanisms of aldosterone on kidney fibrosis. J Am Soc Nephrol Aug;19(8): Epub 2008 Jun

320 Cattaneo D, Bitto A, Baldelli S, Cortinovis M, Gotti E, Perico N, Remuzzi G. Pharmacokinetic/pharmacodynamic drug interaction between rosiglitazone and mycophenolate mofetil in kidney transplantation: a case report. Transplantation Mar 27;85(6): Fassi A, Rubis N, Parvanova A, Iliev I, Zamora J, Giuliano GA, Ene-Iordache B, Perna A, Anabaya A, Motterlini N, Ruggenenti P, Remuzzi G for the BENEDICT Study Investigators. Randomized and non-randomized patients in the Bergamo Nephrologic Diabetes Complications Trial (BENEDICT). Abstract. 29 th Annual Meeting of the Society for Clinical Trials St Louis, Missouri, May 18-21, 2008 RESEARCH ACTIVITIES Laboratory of Biostatistics European Study for Preventing by Lipid-lowering agents ANd Aceinhibition Dialysis Endpoints (ESPLANADE) clinical trial: final analyses. ESPLANADE is a randomised, multicenter trial aimed to test the antiproteinuric effects of statins added to combined ACE-inhibitor (ACEi) and angiotensin II receptor antagonist (ATA) therapy in proteinuric, chronic nephropathies. The primary aim of the study is to assess whether combined therapy of ACEi, ATA and statins is more effective than ACEi and ATA alone in reducing proteinuria. During 2008 we completed data collection and statistical analysis of final data. Acetyl-carnitine in insulin resistance clinical trial: final analyses. Acetyl-carnitine in insulin resistance clinical trial is a pilot, longitudinal, prospective cohort study. It is aimed to evaluate the short-term effects of acetyl-carnitine on insulin resistance and on metabolic sindrome in subjects with increased risk of type II diabetes. During 2008 we completed data collection and statistical analysis of final data. BErgamo NEphrologic DIabetes Complications Trial (BENEDICT) Phase A and B: comparison of baseline characteristics of randomised and non randomised subjects. BENEDICT is a randomised, double-blind, multicentre clinical trial, aimed to assess the effects of an ACE inhibitor, alone or in combination with a non diidropiridinic calcium channel blocker, on progression to microalbuminuria in hypertensive, normoalbuminuric, type II diabetic subjects (Phase A) and on progression to macroalbuminuria in hypertensive, microalbuminuric, type II diabetic subjects (Phase B). During 2008 we completed statistical analysis of baseline characteristics in randomised and non randomised subjects. BErgamo NEphrologic DIabetes Complications Trial (BENEDICT) Phase A and PPARγ2 P12A polymorphism: final analyses. BENEDICT is a randomised, double-blind, multicentre clinical trial performed in 1204 subjects with normoalbuminuria, hypertension and type II diabetes. Primary efficacy analyses have shown that Trandolapril and Verapamil in combination and Trandolapril alone significantly slowed the progression to microalbuminuria as compared to placebo. On the contrary Verapamil alone did not significantly differ from placebo (Phase A). Data on PPARγ2 P12A polymorphism were collected and it was explored whether the above effects on progression to microalbuminuria were regulated by the PPARγ2 P12A. During 2008 we performed statistical analysis of final data. 319

321 MYcophenolate Steroid Sparing Study (MYSS) clinical trial and ABCB1 - exons 21 and 26 genotype: final analyses. MYSS is a randomised, multicentre, clinical trial which showed that in cadaveric renal transplanted patients with immunosuppressive therapy with ciclosporine Neoral, Mychophenolate Mofetil was not more effective than Azathioprine in preventing acute rejections at 21 months after transplant. It was also evaluated whether adverse events associated to the use of cyclosporine during the MYSS trial were regulated by ABCB1 exons 21 and 26 genotype. During 2008 we performed statistical analysis of final data. Long-term renal survival in recipients of grafts from old (aged years) and very old donors (aged >70 years): final analyses. Aim of this prospective, longitudinal, controlled study is to compare long-term renal survival in a cohort of recipients of one or two kidneys from donors aged 60 to 69 years (N=67) with that of kidney recipients from donors aged 70 years or older (N=71). During 2008 we completed data collection and statistical analysis of final data. Laboratory of Clinical Chemistry Increase of urinary excretion of NGAL (neutrophil gelatinase-associated lipocalin) as an early marker in predicting acute renal dysfunction in patients with solid organ cancer given cisplatin as chemotherapeutic agent Cisplatin has a major impact in cancer medicine and is widely used for therapeutic management of several tumors, such as those of ovary, testes, and the head and neck. However, while several anti-neoplastic agents frequently exhibit nephrotoxicity, the platinum derivatives are among the most frequent compounds leading to renal injury. Indeed, approximately 25-35% of patients develop evidence of nephrotoxicity following an initial dose of cisplatin and these findings indicate that there is a pressing need for way to protect the kidney while administering effective chemotherapeutic agents such as cisplatin. Unfortunately, creatinine is an unreliable indicator during acute changes in kidney function and among compounds that might serve as a novel biomarkers for the initiation phase of acute renal failure, neutrophil gelatinase-associated lipocalin (NGAL) has been identified as one of the most strikingly upregulated genes and overexpressed proteins in the kidney after ischemia. Markedly increased NGAL concentrations were easily detected in urine early after renal ischemia in mouse and rat models. Recent studies have demonstrated that NGAL is a useful early predictor of acute renal failure also in humans. Taken altogether these findings indicate that urinary and/or serum NGAL concentration monitoring may represent a sensitive, specific, and highly predictive early biomarker for acute renal failure. So far, however, no data are available on the reliability of NGAL to predict the development of acute renal dysfunction in cancer patients receiving cisplatin treatment. To this purpose, we set up a study in 46 patients with solid tumors aimed to evaluate whether serum and/or urinary NGAL concentrations soon after cisplatin infusion (determined by ELISA assay) were suitable to predict development of subsequent acute renal dysfunction. Our findings indicated that urinary NGAL is an early and reliable marker of development of acute renal dysfunction in cancer patients receiving cisplatin treatment. Among the 46 patients receiving cisplatin, acute renal failure developed in 12, as documented by the increase in serum creatinine concentration that occurred between day 3 to 7 after cisplatin infusion. In all patients developing acute renal failure a mild increase in serum NGAL was observed at day 1 post-cisplatin treatment. Thereafter the concentration of the marker progressively declined 320

322 below baseline values. However, the serum profile of NGAL was comparable in both patients with and without acute renal injury. At variance, the concentration of NGAL in the urine progressively increased from day 1 to day 7 post-cisplatin administration only in patients who had the increase in serum creatinine and thus developed acute renal failure. Thus, urinary but not serum NGAL concentrations were proven to be a useful surrogate marker to predict patients who may develop acute kidney injury after single cisplatin infusion. Increase of urinary NGAL concentration more than 3500% over baseline value at day 2 post-drug infusion allows to identify a subgroup of patients with persistent renal dysfunction. Evaluation of renal function decline determined by 13 different GFR estimating formulas in type 2 diabetic patients Evaluation of glomerular filtration rate (GFR) is of critical importance in the clinical management of both clinic and outclinic diabetic patients since diabetic nephropathy affects 25-40% of the patients, and diabetes is the leading cause of end-stage renal disease. However, the measurement of the true GFR either by the renal clearance of the gold standard inulin or by plasma clearance of contrast agents is time consuming, difficult to perform and cannot be easily implemented in clinical daily practice. Furthermore, creatinine clearance is inconvenient and sometimes inaccurate due to variable creatinine metabolism and tubular secretion. To circumvent these drawbacks, a number of equation has been developed to provide an estimate of renal function from serum creatinine and demographic characteristics. Recently we demonstrated in both transplant and diabetic patients that prediction formulas were inaccurate in predicting true GFR especially in subjects with renal function values higher than 60 ml/min/1.73m 2. However, their performance and suitability to monitor renal function decline in patients with type 2 diabetes have not been assessed so far. Thus, we evaluated the decline of renal function by calculating the regression line vs. measured GFR (by iohexol plasma clearance) or GFR estimated by 13 different prediction equations in 337 normo- and microalbuminuric subjects, enrolled in the BENEDICT and in the DEMAND study, who had at least 4 serial renal function determinations. Each patient had 4-18 GFR determinations performed in a 0.8 to 8 years period. The measured renal function declined by ml/min/1.73m 2 /year. All the prediction formulas markedly underestimated the GFR decline and neither baseline albuminuria (normo vs. micro) nor follow-up duration (more or less than 3.5 years) affected the results. Estimated GFR decline ranged from (Ibrahim equation) to ml/min/1.73m 2 /year (Hull formula). In diabetics patients with baseline GFR >120 (hyperfiltering) or between 80 and 120 ml/min/1.73m 2 GFR declined by and ml/min/1.73m 2 /y, respectively. In the two cohorts estimated slopes under-estimated GFR decline by 6.14 and by 3.33 folds, respectively. These findings showed that none of the 13 GFR models do not reliably predict GFR decline in diabetics with normo or microalbuminuria, in particular in those at increased risk because of hyperfiltration. Failure to early detect progressive kidney dysfunction may delay the institution of renoprotective therapies in this population. This may have major clinical implications since glomerular hyperfiltration has a pathogenic role in onset and progression of diabetic renal disease and early intervention to ameliorate hyperfiltration may prove renoprotective in the long term. Development of a new equation to predict creatinine clearance in Cuban population Determination of glomerular filtration rate (GFR) provides the measure of the filtering capacity of the kidneys and is considered the best overall index of renal function currently used to determine the effectiveness of therapies designed to slow the progression of chronic renal 321

323 diseases. However, the measurement of the true GFR either by the renal clearance of the gold standard inulin or by plasma clearance of contrast agents is time consuming, difficult to perform and cannot be easily implemented in clinical daily practice and particularly in poor or third world countries. Urinary creatinine clearance is widely used to measure GFR, but it is inconvenient and sometimes inaccurate due to variable creatinine metabolism and tubular secretion. To overcome these limitations, the National Kidney Foundation-Kidney Disease Outcomes Quality Initiative recommends the estimation of GFR using prediction equations based on serum creatinine, demographic and anthropometric data. A relevant number of prediction formulas developed mainly in caucasic subjects have been published so far, but none of them correctly estimated renal function in Cuban population. Thus, together with the National Institute of Nephrology of Havana City, we aimed to develop a new equation to predict creatinine clearance in Cuban population. The database consisted of 9014 subjects who attended the Cuban Institute of Nephrology during the period % of the subjects were male and 6408 were older than 18 years. For all the subjects body weight, height, body surface area age, and gender were collected together with the measurement of serum creatinine and creatinine clearance. The new developed equation, suitable to estimate creatinine clearance in adult population, included age, weight, height, gender and serum creatinine as independent variables. The correlation between measured creatinine clearance and predicted values by the new equation was good, (Spearman correlation coefficient r = 0.794) and better than with both Cockcroft-Gault and MDRD formula (r = and 0.667, respectively). The accuracy in creatinine clearance estimation was markedly higher using the new equation: the number of values predicted with an error within 10% were 50% more than with Cockcroft-Gault and MDRD formulas. In particular, at variance with the results obtained with Cockcroft-Gault and MDRD equations, the new developed formula was proven to be more accurate in predicting creatinine clearance in both male and female subjects with renal function higher than 60 ml/min/1.73m 2 GFR. Laboratory of Drug Development Blood cell count and lipid lowering therapy affect sirolimus exposure in kidney transplant recipients on calcineurin inhibitor-free regimen Sirolimus (SRL) is an immunosuppressive agent characterized by a narrow therapeutic index. Studies in organ transplant recipients given this drug in combination with cyclosporine (CsA) have shown that SRL exposure is extremely variable. However, no data are available on the pharmacokinetics of SRL in calcineurin inhibitor-free protocols. Fifty-five pharmacokinetics profiles were collected from 20 kidney transplant patients given SRL over a 36 months follow-up. None of them were on CsA or tacrolimus. Large interindividual variability in the daily distribution of SRL concentrations was documented. Patients with low blood cell count showed a significant decrease in daily SRL exposure, requiring 35% increments in the daily dose to reach drug concentration targets. At variance, concomitant administration of omega-3 polyunsaturated fatty acids increased SRL AUC 0-24 by 25% compared to values found in normolipidemic patients. Altogether, these information can be used to optimize SRL dose-adjustments in the routine clinical practice. These results have been presented at the meeting of the International Association of Therapeutic Drug Monitoring and Clinical Toxicology (Nice, September 2007) and are now submitted for publication in the Journal of Clinical Pharmacology. 322

324 Abcb1 genotypes predict cyclosporine-related adverse events and graft outcome in kidney transplantation: a pre-specified analysis of the mycophenolate steroids sparing (Myss) study Cyclosporine A (CsA) is a substrate of P-glycoprotein, an efflux transporter encoded by the ABCB1 gene. As compared to carriers of the wild-type gene, carriers of T allelic variants in exons 21 or 26 have reduced P-glycoprotein activity and secondarily increased CsA intracellular concentration. Thus, carriers of T variants might be at increased risk of CsA-related events. We evaluated the associations between ABCB1 genotypes in exon 12, 21 and 26 and CsA-related outcomes in 147 renal transplant recipients on CsA-based immunosuppression included in the Mycophenolate Steroids Sparing study and followed prospectively for a median of 65.5 months. As compared to carriers of the wild genotype, carriers of T allelic variants in exons 21 or 26 had a three-fold excess risk of delayed graft function (p=0.011 and p=0.019, respectively), a trend to slower renal function recovery and lower glomerular filtration rate at study end, significantly higher incidence of new-onset diabetes and cytomegalovirus reactivation. T variants in both exon 21 (p=0.022) and 26 (p=0.034) were independently associated with 3.8 and 3.5 folds excess of DGF. Incidence of acute rejections and mean CsA dose and blood levels were comparable in genotype groups. Renal transplant recipients with T allelic variants in exon 21 or 26 of the ABCB1 gene are at increased risk of CsA-related adverse events. Genetic evaluation may help identify patients at risk and modulate CsA therapy to optimize graft and patient outcomes. These results have been presented at the meeting of the American Society of Transplantation (Toronto, June 2008) and are now submitted for publication in the Journal of the American Society of Nephrology. 323

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326 LABORATORY OF COORDINATION OF DIAGNOSIS AND INFORMATION ON RARE DISEASES STAFF Head Arrigo SCHIEPPATI, M.D. Information Centre for Rare Diseases Head Erica DAINA, M.D. 325

327 CURRICULA VITAE Arrigo Schieppati got his degree in Medicine at the University of Milan in 1978 and the specialisation in Medical Nephrology in 1984 at the same University. He performed his training at the Mario Negri Bergamo Laboratories with Dr. Remuzzi, and completed it with stages at the laboratories of prof. Patrono (Catholic University in Rome), prof. John Gordon (Cambridge, GB), and at the Division of Renal Diseases - University of Colorado Medical School, directed by Dr. Schrier (Denver, USA). Since 1982 he works at the Division of Nephrology and Dialysis Riuniti Hospital Bergamo, where he is in charge of Outpatients Clinic and Day Hospital. From 1992 to 1995 he was Head of the Information Center for Rare Diseases and since 1996 he is Head of the Laboratory for Coordination of Information and Diagnosis of Rare Disease at the Clinical Research Center for Rare Diseases Aldo e Cele Daccò of the Mario Negri Institute. Areas of interest: diagnosis and therapy of chronic renal diseases, hypertension and rare kidney diseases. Affiliations: ethical committee Riuniti Hospital - Bergamo; member of the working group of the regional network for rare diseases in Lombardy; scientific committee Bolognini Hospital Seriate (BG); member of the Task Force on Rare Diseases (DG Health and Consumer Protection); International Society of Nephrology; American Society of Nephrology; Editorial Board Journal of Nephrology. Selected publications Schieppati A, Henter J I, Daina E, Aperia A. Why rare diseases are an important medical and social issue. Lancet June 14; 371: Schieppati A, Remuzzi G. Chronic renal diseases as a public health problem: epidemiology, social, and economic implications. Kidney Int Suppl Sep;(98):S7-S10. Remuzzi G, Schieppati A, Boissel JP, Garattini S, Horton R. Independent clinical research in Europe. Lancet Nov 6-12;364(9446): Schieppati A, Perna A, Zamora J, Giuliano GA, Braun N, Remuzzi G. Immunosuppressive treatment for idiopathic membranous nephropathy in adults with nephrotic syndrome. Cochrane Database Syst Rev Oct 18;(4):CD Remuzzi G, Schieppati A, Ruggenenti P. Clinical practice. Nephropathy in patients with type 2 diabetes. N Engl J Med Apr 11;346(15): Schieppati A, Remuzzi G, Garattini S. Modulating the profit motive to meet needs of the less-developed world. Lancet Nov 10;358(9293): Erica Daina got his degree in Medicine at the University of Milan in 1987 and the specialisation in Medical Nephrology in 1990 at the same University. She performed her training at the II Medical Division - San Raffaele Hospital - Milan, and at the Division of Nephrology and Dialysis - Riuniti Hospital - Bergamo. In March 1988 she started her collaboration with the Mario Negri Institute and since June 1993 she works as full-time clinical researcher at the Clinical Research Center for Rare Diseases Aldo e Cele Daccò. Since 1996 she is Unit Head Information Center for Rare Diseases. Areas of interest: rare diseases, Takayasu arteritis, Hemolytic Uremic Syndrome/Thrombotic Thrombocytopenic Purpura, Fabry s disease, Alport s syndrome. Since January 2002 she is representative of Coordinating Centre - Regional Network for Rare Diseases - and collaborates to didactics activity at the University of Turin (School of Clinical Pathology Specialization - 2 nd Level Master in Rare Diseases). Selected publications Bresin E, Daina E, Noris M, Castelletti F, Stefanov R, Hill P, Goodship T H J, Remuzzi G, International Registry Recurrent Familial HUS/TTP. Outcome of renal transplantation in patients with non-shiga toxin-associated Hemolytic Uremic Syndrome: Prognostic significance of genetic background. Clinical Journal American Society Nephrology 2006; 1: Galbusera M, Bresin E, Noris M, Gastoldi S, Belotti D, Capoferri C, Daina E, Perseghin P, Scheiflinger F, Fakhouri F, Grünfeld JP, Pogliani E, Remuzzi G. Rituximab prevents recurrence of thrombotic thrombocytopenic purpura: a case report. Blood Aug 1;106(3): Vanoli M, Daina E, Salvarani C, Sabbadini MG, Rossi C, Bacchiani G, Schieppati A, Baldissera E, Bertolini G; Itaka Study Group. Takayasu's arteritis: A study of 104 Italian patients. Arthritis Rheum Feb 15;53(1): Remuzzi G, Galbusera M, Noris M, Canciani MT, Daina E, Bresin E, Contaretti S, Caprioli J, Gamba S, Ruggenenti P, Perico N, Mannucci PM; Italian Registry of Recurrent and Familial HUS/TTP. von Willebrand factor cleaving protease (ADAMTS13) is deficient in recurrent and familial thrombotic thrombocytopenic purpura and hemolytic uremic syndrome. Blood Aug 1;100(3): Daina E, Schieppati A, Remuzzi G. Mycophenolate mofetil for the treatment of Takayasu arteritis: report of three cases. Ann Intern Med Mar 2;130(5):

328 INTRODUCTION TO THE LABORATORY S ACTIVITIES Rare Diseases (RD) represent about ten percent of all human medical illnesses and infirmities. It is difficult to define what exactly is intended as a RD. The US Congress in the Orphan Drug Act has given the first definition in Under this law it is considered rare a disease that affects less than Americans (prevalence 0.75 per 1 000). Recently, the European Parliament adopted a more strict definition; they consider rare a condition that affects not more than five individuals per in the European Community (prevalence 0.5 per 1 000). Besides the epidemiological parameter, RD have certain characteristics in common: 1) most of them are of genetic origin; 2) rarity often brings a difficult and/or late diagnosis; 3) generally, RD are heavy social burdens both to the family and community and cause early mortality. The greatest barrier to prevention, diagnosis and treatment of RD is inadequate knowledge. Once a diagnosis of RD is put, a major complaint of patients and those involved in their care is the difficulty to obtain pertinent information about causes, symptoms and either established or experimental treatments. Often, patients with RD are willing to participate in clinical studies, but they do not know where and how, and physicians or health authorities are seldom able to help them. RD is not a very attracting field for basic and clinical investigators for several reasons: it is difficult to find adequate animal models for many rare disorders; clinical trials may require more patients than available; financial support is insufficient. Few countries have a central body or system to disseminate information on RD. Accurate information on the incidence and prevalence of RD is extremely important for both basic and clinical investigators. Invaluable help to research advances in RD would come from the availability of registries and databases containing diagnostic, clinical and biological data of patients with rare disorders. A pilot experience has been established at the Clinical Research Centre for Rare Diseases Aldo and Cele Daccò since This Centre is unique with its integration of an Information Centre for Rare Diseases, clinical facilities for the implementation and development of clinical studies, educational activities for physicians, nurses and patients. In 2001 it has been nominated Coordinating Centre of the Regional Network for Rare Diseases in the Lombardy Region, an area of 9 million people in Northern Italy. As Coordinating Centre is also working with the National Centre of Rare Diseases at Istituto Superiore di Sanità. All the up-to-date information regarding the activities of the Coordinating Centre are available at the web site: FINDINGS/MAIN RESULTS The database of the Information Centre for Rare Diseases contains data about patients affected by 882 different rare disorders. In the Bank of biological materials, samples from 1394 patients with rare conditions and their families have been collected. The Centre has established contacts with 327 Italian Associations for rare diseases. It was even possible that patients with 88 different rare diseases - for which no Associations have been established in Italy yet - to meet among themselves. 327

329 In July 2000, the European Commission recognized the Laboratory as a site for "Postgraduate training on rare diseases" (contract No. QLK ). In December 2001 (Delibera della Giunta Lombarda N. 7328), the Centre was identified as "Coordinating Centre of the Regional Network for Rare Diseases". The Laboratory coordinates the International Registry of Recurrent and Familial Hemolytic Uremic Syndrome (HUS) and Thrombotic Thrombocytopenic Purpura (TTP), since The research projects developed in collaboration with the Laboratory of Immunology and Genetic of Rare Diseases and Organ Transplantation have allowed to better comprehend the pathogenesis of these diseases and to identify some genetically determined forms of HUS, associated to defects of complement regulatory factors, and of TTP, associated to congenital deficiency of ADAMTS13 enzyme. NATIONAL COLLABORATIONS Italian National Institute of Health G. Bosco Hospital, Turin, Regional Coordinating Center for Rare Diseases Riuniti Hospital, Bergamo Italian Takayasu's Arteritis Study Group - ITAKA Group Italian Registry of the Membranoproliferative Glomerulonephritis Italian Registry of the Familial Focal Segmental Glomerulosclerosis Biotechnology Laboratory, IRCCS Policlinico San Matteo, Pavia Assessorato alla Sanità, Lombardia Region University of Torino, School of Clinical Pathology, Faculty of Medicine and Surgery Italian Network for Promotion of Folic Acid to Prevent Birth Defects University of Turin, Department of Experimental Medicine and Oncology, 2 nd Level Master in Rare Diseases Italian Society of Neonatology (Lombardy section), Rare Congenital Respiratory Diseases Study Group University of Milan, 1 st Level Master in Clinical Research Department of Molecular Biology, Unit of Medical Genetics, Policlinico Le Scotte, Siena BergamoScienza Association INTERNATIONAL COLLABORATIONS International Registry of Recurrent and Familial Hemolytic Uremic Syndrome and Thrombotic Thrombocytopenic Purpura Information Centre for Rare Diseases and Orphan Drugs ICRDOD, Bulgaria Podonet: Consortium for Clinical, Genetic and Experimental Research into Hereditary Diseases of the Podocyte Coordinator: Heidelberg University Hospital, Germany International Network and Registry for Thrombotic Microangiopathy (TMA) Coordinator: Schneider Children s Hospital, Albert Einstein College of Medicine, USA EDITORIAL COMMITTEE MEMBERSHIP Journal of Nephrology (Arrigo Schieppati) Quaderni di Farmacoeconomia (Erica Daina) 328

330 NATIONAL AND INTERNATIONAL COMMITTEE MEMBERSHIP Network for Rare Diseases Lombardy Region (Delibera Regione Lombardia N 7328, 11/12/2001) Task Force on Rare Diseases (established by DG Health and Consumer Protection on 21 January 2004) Scientific Committee A.O. Bolognini di Seriate Ethical Committe, A.O. Ospedali Riuniti di Bergamo Working group Classification and coding of rare diseases coordinated by Italian National Institute of Health EVENT ORGANIZATION La Fondazione IRCCS Ospedale Maggiore Policlinico, Mangiagalli e Regina Elena incontra il Centro di Ricerche Cliniche per le Malattie Rare Aldo e Cele Dacco Clinical Research Centre for Rare Diseases Aldo and Cele Daccò Mario Negri Institute Ranica (Bergamo), January 24, 2008 Una Speranza per le malattie rare, La Pre-Eclampsia Clinical Research Centre for Rare Diseases Aldo and Cele Daccò Mario Negri Institute Ranica (Bergamo), May 16, 2008 Focus on Rare Diseases The Genetic and Molecular Basis of Rare Kidney Disorders Bergamo, October 9-11, 2008 PARTICIPATION IN EVENTS IN WHICH THE LABORATORY WAS INVOLVED 11 Convegno Patologia Immune e Malattie Orfane Torino, January 24-26, 2008 Prima Giornata Europea delle Malattie Rare Un giorno raro per persone molto speciali Bergamo, February 29, 2008 Prima Giornata Europea delle Malattie Rare Sviluppi della Rete Regionale per le Malattie Rare Milan, February 29, 2008 XXI Convegno Nazionale Associazione Italiana per le Mucopolisaccaridosi e Malattie Affini - AIMPS San Donato Milanese (Milan), march 28-30, 2008 II Convegno Nazionale Associazione Italiana Sindrome di Shwachman-Diamond Dal difetto genetico alla terapia: venti anni nella conoscenza di una malattia rara Rimini, April 18-19,

331 Convegno : Le malattie Croniche in età pediatrica Brescia, April 19, 2008 Malattie Rare Impegno Comune Rome, June 24, 2008 XXII International Complement Workshop Basilea, September 28 October 2, 2008 Corso di aggiornamento Malattie Polmonari: percorsi diagnostici e terapeutici Milan, October 16, 2008 III Congresso Nazionale Associazione Pediatri in Gruppo - APeG Florence, October 17-18, 2008 Congresso Internazionale Rare Diseases and Orphan Drugs Rome, October 27-31, 2008 Workshop Consorzio MIA Lupus Eritematoso Sistemico Milan, November 14, 2008 Progetto PARTECIPASALUTE: Le Associazioni definiscono le priorità della ricerca: uno scenario possibile in Italia? Milan, December 2, 2008 Progetto di formazione sul campo: registro Regionale Malattie Rare Presidi Rete Regionale, 2008 European Commission "Fondazione Ricerca Malattie Rare", Bergamo Lombardy Region Fondazione Telethon GRANTS AND CONTRACTS SELECTION OF SCIENTIFIC PUBLICATIONS FROM 2008 A. Schieppati, J.I. Henter, E. Daina, A. Aperia Why rare diseases are an important medical and social issue Lancet; june , vol. 371, pp F. Castelletti, R. Donadelli, F. Banterla, F. Hildebrandt, P. Zipfel, E. Bresin, E. Otto, C. Skerka, A. Renieri, M. Todeschini, J. Caprioli, M.R. Caruso, R. Artuso, G. Remuzzi, M. Noris Mutation in fn1 cause glomerulopathy with fibronectin deposits Proc Natl Acad Sci Usa. 2008, vol. 105, pp A. Schieppati G. Remuzzi Novel Therapies of Lupus Nephritis 330

332 Curr. Opin. Nephrol. Hypertens. 2008, Vol. 17, pp OTHER PRODUCTS PUBLISHED IN 2008 Schieppati A, Daina E, Gamba S Parla anche con me. L esperienza di 18 anni del Centro di Informazione per le Malattie Rare In: Dire, fare, curare Parole tra medici e malati Collana Salute e Società, FrancoAngeli Editore, novembre 2008 Cattaneo D, Daina E Malattie Autoimmuni In: Antagonisti recettoriali dell Endotelina: quali composti e quali patologie Il Pensiero Scientifico Editore, dicembre 2008 RESEARCH ACTIVITIES Information Centre for Rare Diseases In 1992, in the frame of the Clinical Research Centre for Rare Diseases Aldo e Cele Daccò was established an Information Centre for Rare Diseases. The Information Service is available to patients with rare diseases, their families and doctors. It offers information, update and useful addresses free of charge with particular emphasis on etiology, pathogenesis, genetics, treatments and availability of referral research centres. One of the most difficult issues to address when studying rare diseases are those, related to the recruitment of a sufficient number of patients with a given disease. The database of the Information Centre for Rare Diseases has become a useful tool for identification of potentially eligible patients for clinical studies. Till now, the Information Centre for Rare Diseases has collected patients clinical data for about 200 different rare conditions with 10 or more cases. Furthermore, intensive collaboration with groups with the same interest from Italy and abroad provides many possibilities for starting of clinical projects with a multicentral design. Bank of biological samples and description of hereditary nephropathies The aim of this project is to collect clinical data and biological samples from patients and their families with rare genetic conditions. A database with clinical data and a Bank for biological samples collection and preservation has been created. The availability of clinical data and biological samples is useful to perform new biochemical and genetic tests within specific research projects aimed to better reveal the mechanisms of the diseases, their manifestation and therapeutic opportunities. In particular, the attention is focused on rare genetic disorders of the kidney. A thorough clinical evaluation, including clinical data collection, medical physical examination, renal ultrasonography, laboratory tests of blood and urine is offered to patients, affected by hereditary nephropathies (Alport syndrome, Fabry disease, Familial Focal Glomerulosclerosis, Glomerulopathy with Fibronectin deposits, Membranoproliferative glomerulonephritis, Medullary Cystic Kidney disease, Cystinuria), who are addressing our Centre. After obtaining a written informed consent, biological samples from patients and their relatives are collected, labelled with specific codes to assure the anonymity and conserved in the Bank for biological samples. In case the responsible gene for a hereditary nephropathy is known (Alport syndrome, Fabry disease, Medullary Cystic Kidney disease, Cystinuria), the blood samples are redirected to the relevant Laboratory of reference. For other nephropathies, where the identification of the 331

333 gene mutation is unknown or still in course, the blood samples are conserved with the aim to be used in specific future research projects. Evaluation of the long term efficacy of enzyme replacement therapy in Fabry disease Fabry disease is an X-linked disorder of the glycosphingolipid catabolism caused by the deficient activity of the lysosomal hydrolase alfa-galactosidase A (A-gal A) in tissues and fluids of affected hemizygous males. Most heterozygous females have an intermediate level of enzymatic activity. The enzymatic defect leads to a systemic deposition of glycosphingolipid in the heart, kidneys, eyes and other organs and tissues. Preliminary studies of enzyme replacement therapy demonstrate that periodic infusions of recombinant human Alfa-galactosidase A are safe (in terms of infusion reactions) and effective in patients with Fabry disease. The purpose of this study is to evaluate the long term efficacy of enzyme replacement therapy in patients with Fabry disease and renal involvement. International Registry of Recurrent and Familial Hemolytic Uremic Syndrome and Thrombotic Thrombocytopenic Purpura Hemolytic Uremic Syndrome (HUS) and Thrombotic Thrombocytopenic Purpura (TTP) are rare diseases of microangiopathic haemolytic anemia and thrombocytopenia with signs of renal (most prevalent in HUS) and cerebral (most prevalent in TTP) damage. There are extremely rare forms of HUS and TTP, often occurring in families, in which the patients relapse even after complete recovery of the first episode with permanent renal and neurological sequelae. In the familial and recurrent forms of HUS and TTP, the attention is concentrated mainly on the genetic predisposition to the disease. The Laboratory coordinates the International Registry of Recurrent and Familial HUS/TTP, since The Registry has collected more than 540 cases of HUS/TTP referred from 100 Italian and 80 European and extra-european Centres. Clinical and laboratory data of all patients referred to the Registry are collected by a uniform data extraction form. The family history and also the personal data of the unaffected relatives are collected, when possible. Biological samples are collected from all patients and available relatives, for the biochemical and genetic analyses. Many research projects in collaboration with the Laboratory of Immunology and Genetic of Rare Diseases and Organ Transplantation and the Laboratory of Cellular Biology Negri Bergamo are developed and have still allowed to identify some genetically determined forms of HUS and TTP. The maintenance of a centralised bank of biological samples ensure the availability of clinical material for new investigative approaches as they will be developed. Rituximab in relapsing and chronic thrombotic thrombocytopenic purpura In most cases, TTP is related to an acquired deficiency of ADAMTS13 activity, due to the presence of autoantibodies anti-adamts13. The absence of ADAMTS13 activity leads to the accumulation of von Willebrand Factor ultralarge multimers (normally cleaved to smaller molecular forms by ADAMTS13 enzyme), which probably induces platelets aggregation and the ensuing process of thrombotic microangiopathy. The purpose of the project is to evaluate the use of a monoclonal anti-cd20 antibody (Rituximab) in patients with relapsing and chronic thrombotic thrombocytopenic purpura, related to the presence of anti-adamts13 antibodies. Rituximab is a humanized monoclonal antibody that targets the CD20 molecule on B cells (white cells producing antibodies). Administration of Rituximab leads to a selective B cell depletion that usually lasts 6 to 9 months. Inhibition of B cell activation or growth by rituximab treatment limit the uncontrolled synthesis of autoantibodies, that may predispose to chronic relapsing TTP. 332

334 Rituximab is infused intravenously once weekly for a total of four infusions. After Rituximab treatment, the patients are periodically controlled with medical examination, blood and urine exams, and with ADAMTS13 plasma activity assay and research of anti-adamts13 autoantibodies, in collaboration with the Laboratory of Cellular Biology Negri Bergamo. Observational period lasts 12 months. Evaluation of urinary podocyte excretion Recent evidence recognize an important role for podocytes in the progression of renal diseases. Podocytes contribute to glomerular permeability and are important target cell for renal disease progression. Glomerular injury is usually associated with the leakage of protein across the filter into the urine and with the disappearance of podocyte foot process. Injuried podocytes either undergo apoptosis or detach from the glomerular basement membrane, with subsequent appearance in the urine. Urinary excretion of podocytes has been reported as possible predictor of disease progression in a number of progressive glomerular diseases, such as IgA nephropathy, focal segmental glomerulosclerosis, diabetic nephropathy. During the past years the Clinical Research Center for Rare Diseases, in collaboration with the Department of Molecular Medicine at Mario Negri Institute, has established an indirect immunohistochemistry method for the detection of podocytes in urinary sediments. This method has allowed to identify and quantitate the extent of urinary podocyte loss in patients with glomerular disease. The present study has the aim to evaluate excretion of urinary podocytes in rare genetic renal diseases. This methodology, applied on a larger number of patients, could provide a non invasive way of indexing the extent of glomerular damage and a useful instrument to evaluate the response to treatment. Effects of an intensified treatment with ACE-inhibitors, Angiotensin II receptor antagonists and Statins in Alport syndrome Alport syndrome (AS) represents a form of progressive hereditary nephritis in which the genetic defect resides in the synthesis of one of several subunits of type IV collagen, the predominant constituent of basement membranes in renal glomeruli. Renal impairment occurs with time and severe renal failure with hypertension and uremia represent the end stage of the disease, even if a high variability in the rate of progression is described. The prognosis is variable. Males are usually affected by a progressive form of the disease. Affected females with X-linked syndrome usually have a good prognosis with a mild renal impairment. Other clinical manifestations are anterior lenticonus which occur in about one third of patients. Other findings are myopia, lens opacities and retinal pigment abnormalities and corneal vesicles or erosions. The disease is also associated to a sensor neural deafness which can occur in approximately half of the patient affected and usually correlates with renal impairment. No definite treatment exists in order to delay the time of dialysis or a kidney transplant. Many studies showed that Angiotensin converting enzyme (ACE) inhibitors slow glomerular filtration rate (GFR) decline and limit progression to end stage renal disease (ERDS) and dialysis in several chronic nephropathies associated with proteinuria. The combination of ACE-I with Angiotensin II receptor antagonists may reduce proteinuria more effectively than the two drugs alone. Moreover the addition of Statins may synergize the antiproteinuric effects of ACE-I and ATAII antagonists in experimental models of chronic renal diseases. The purpose of this study is to evaluate the effect of a standardized multimodal nephroprotection intervention (Remission Clinic) in Alport patients with renal involvement. Nine patients with Alport syndrome and macroalbuminuria will be enrolled in this study. Recruitment of normo or microalbuminuric patients will be stopped and the analysis will be performed on the patients that will be available when the recruitment phase of macroalbuminuric patients will be completed. 333

335 Update of the Congenital Respiratory Malformations database This Project originated from the collaboration between the Study Group on Respiratory Disease of the newborn and the Coordinating Centre for Rare Diseases. The Congenital Respiratory malformations database was conceived to facilitate the diagnostic and assistance procedures for paediatricians facing with more frequent respiratory malformations. The update consisted of a review of the lists of malformation categories and of all syndromes previously included; more syndromes have been added. Furthermore, in the brief description, more detailed than the previous version, the Code for Rare Disease was added, allowing to address the patient to the Referral Centres for Lombardy. A link with the OMIM database is provided for each syndrome for a better description of the disease. Identification of new genes associated to the Autosomal Dominant Familial form of Focal Segmental Glomerulosclerosis Focal segmental glomerulosclerosis (FSGS) is a pathological entity, and is a significant cause of end-stage renal disease (ESRD). Glomerular disease is the third leading cause of ESRD, and FSGS comprises a significant proportion of this subgroup: up to 5% of adults and 20% of children with ESRD. The diagnosis of FSGS is based on renal pathology. The clinical hallmarks include proteinuria, nephrotic syndrome, occasional hematuria and frequent progression to ESRD. Hypertension is also a common finding. At present, there are no consistently reliable treatments for FSGS and response rates to available treatments have been estimated at <30-50%. Because FSGS has become an important cause of ESRD, it is essential to understand the molecular basis and pathogenesis of this disease process. There are various subtypes of FSGS, which include primary (idiopathic or sporadic), secondary, familial and FSGS associated with congenital syndromes. Among familial forms of FSGS, both autosomal recessive and dominant inheritance patterns have been reported. The study, in collaboration with the Laboratory of Immunology and Genetic of Rare Diseases and Organ Transplantation, is aimed at identifying the genetic causes of the autosomal dominant familial forms of FSGS, in order to help in the clinical management. Patients with an autosomal dominant familial form of FSGS (at least two affected subjects in two different generations within a family) and their available relatives, will be enrolled. Clinical data and blood samples will be collected and used to perform routine laboratory exams and to obtain DNA for the genetic analyses. Genetic counselling will be provided to all patients and to their relatives. Rare diseases regional register In 2001 the Italian government approved legislation to establish the National Registry for Rare Diseases coordinated by the Istituto Superiore di Sanità in Rome (DM 279/2001). The National Registry receives epidemiological data from regional reference centres. Lombardy Region, an area of 9 million people in Northern Italy, identified a Coordinating Centre and a network of 29 regional centres that contribute to the Regional Registry for Rare Diseases collecting data in a specific computerized system. Dedicated and on-site training have been implemented to optimize data collection. The Coordinating Center is in charge of Registry management as application maintenance process, data extraction, data sent to Istituto Superiore di Sanità (national minimum data set), Periodic Review Reports. 334

336 Complement abnormalities in primary Membranoproliferative glomerulonephritis Primary membranoproliferative glomerulonephritis (MPGN) is an uncommon cause of chronic renal disease that occurs principally in children and young adults, and that often has an unfavourable prognosis. Data on incidence and prevalence of MPGN are scanty, thus a first aim of this project is to collect through a Registry clinical data and biological samples from well characterized patients with primary MPGN. Thus, a case report form has been designed in order to obtain homogeneous clinical data for all recruited patients. The family history will be also recorded and biological samples will be collected from the patients to perform biochemical and genetic analyses. Genetic counselling will be provided to all patients and relatives. The pathogenesis of primary MPGN is ill defined. The available data indicate that excessive activation of complement due to autoimmune and genetic abnormalities plays a role in inducing damage to the kidney and other organs. The complement is a complex system that mediates the immune response against bacteria and viruses. In normal conditions human cells are protected from complement attack by several inhibitors that restrict complement activation to the surface of pathogens. Such regulatory system is defective in MPGN. Thus a major goal of this project, in collaboration with the Laboratory of Immunology and Genetic of Rare Diseases and Organ Transplantation, will be the identification of the genetic and acquired defects underlying complement activation in MPGN and to establish whether specific complement abnormalities are associated with disease progression, response to therapy and recurrence on a transplanted kidney. 335

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338 The Transplant Research Center Chiara Cucchi De Alessandri e Gilberto Crespi The Transplant Research Center (CRT) was set up in 2002 to support and promote the work of outstanding research scientists throughout the world and to carry out major organ transplant research programs. The Center is housed in the Villa Camozzi, at Ranica, under the same roof as the Mario Negri Institute in Bergamo and is managed in collaboration with the Institute. The Center s staff is mainly made up of senior and junior researchers that were trained in the laboratories of the Mario Negri Institute in Bergamo, focusing on transplant immunology, research for less toxic immunosuppressant drugs, and new gene therapy techniques to prevent acute rejection of transplanted organs. Information on the Center s activities can be found in the sections addressed to the Department of Molecular Medicine (Laboratory of Immunology and Genetics of Organ Transplantation and Rare Diseases) and the Department of Renal Medicine (Laboratory of Pharmacokinetics and Clinical Chemistry). 337

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340 EDUCATIONAL ACTIVITIES Dean, Mario Salmona, PH.D. The Institute's training programs fall under the heading of biomedical science, and are part of the Lombardy Region's professional training schemes. There are courses for specialized laboratory technicians, and for graduates intending to do research. In 1999 the Institute has set up a Ph.D. course, in collaboration with the Open University of London. This degree is recognised throughout Europe and in the USA. In 2006 the Institute also set up a First Level master Course in Clinical Research, in collaboration with the Milan University and a Second Level master Course in rare Diseases, in collaboration with the University of Torino. In 2008 the Institute started a two-years Advanced School in Applied Pharmacology (SAFA). The course provides advanced cultural education and practical experience with experimental work in the laboratories. Students enrolled in formal courses receive one month preparatory training, after which they are assigned study grants. Between 1963 and 2007 the Mario Negri Institute awarded 6,578 grants, 701 of them to foreign researchers who came to the Institute for special training. Everything possible is done to help students find work once they finish the course. The main feature of these courses is that technicians and researchers receive their training "on site". They work full-time in research programs of a high scientific standard, using advanced equipment and learning the latest methods, in regular contact with colleagues in different countries. Besides its scientific value, this approach provides an excellent preparation on the human and personal scale. Students are usually assigned to one of the Institute s Laboratories, where they gradually gain specialized skills by working on specific research projects. They are expected to attend lessons, seminars, courses and congresses and learn to make full use of the Institute s well-stocked library. Students all have access to the internet and to biomedical database and can print out copies of articles they need to consult, from major international journals. Should the opportunity arise, students are expected to be available for trips abroad, to participate in conferences or courses. The Pharmacological Research Specialists and Biochemical Research Technicians will receive diplomas issued officially by the Lombardy Region and the Mario Negri Institute for Pharmacological Research. These have legal value throughout Italy, and are recognised in competitions for public posts, where they are worth a certain number of points. Mario Negri Institute diplomas are widely considered a guarantee of an excellent theoretical and practical training. The Open University of London Ph.D. degree earned at the Institute has legal value throughout Europe and in the USA. Once they have their diploma or Phd., graduates who want to continue doing research at the Institute may be offered a chance to spend a year or two abroad. 339

341 The SAFA course is aimed at preparing young researchers and enabling them to specialize and work for the pharmaceutical industry, for other research institutes and for public institutions, such as the Regions, for instance. Some of our students, after the SAFA course, decide tu enroll the PhD programs offered by our Institute, while others decide to study abroad. At the moment these courses are available: Three-year course for graduates, in Milan or Bergamo, leading to a diploma as Pharmacological Research Specialist. Three-year course for diploma-holders, in Milan or Bergamo, leading to a diploma as Biochemical Research Technician. Research doctorates (Ph.D.), run under an agreement with the Open University of London. And the Universities of Maastricht and Groningen (NL). First Level Master in Clinical Research, in collaboration with the University of Milano. Second Level Master Course in Rare Diseases, in collaboration with the University of Torino. Two-years Advanced School in Applied Pharmacology for graduate students, in Milano and Bergamo. Other training opportunities PREPARING A THESIS FOR A DEGREE: Students can prepare their thesis in scientific subjects at the Institute, with the approval of their university faculty. These students must work at the Institute for at least two years. SUMMER STUDENTS In June and July each year the Institute accepts a certain number of students in their last two years at high school, to give them experience as part of school/work programs. 340

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344 Prof. Silvio Garattini SIlvio Garattini was born in Bergamo (Italy) on 12 November He earned a diploma in Chemistry, then a degree in Medicine and was appointed lecturer in Chemotherapy and Pharmacology. He held the post of Assistant then Deputy Professor at the Milan University Institute of Pharmacology, until A founder of the Mario Negri Institute for Pharmacological Research, when it opened in 1963 he was its director. The Institute now has four locations (Milan, Bergamo, Ranica (Bg), S. Maria Imbaro (Ch)) and more than 950 people work there. Professor Garattini is a member of the Gruppo 2003 (a group of the most cited Italian scientists in international scientific literature) and has hundreds of publications in Italian and English in international scientific journals, and texts on pharmacology. He was a founder of the European Organisation for Research and Treatment of Cancer (EORTC). In the last ten years Professor Garattini has acted in various organizations, including the Italian National Research Council (CNR) - Committee on Biology and Medicine; the National Health Council, the Committee for Italian Research Policy, set up by the Presidency of the Council of Ministers; the Ministry of Health Commissione Unica del Farmaco (CUF). He has held the following posts: President of the UICC Committee on Antitumoral Chemotherapy, President of the European Organisation for Research and Treatment of Cancer (EORTC), Consultant to the World Health Organisation; President of the European Society of Biochemical Pharmacology; member of the Committee for Proprietary Medicinal Products (CPMP) of the European Agency for the Evaluation of Medicinal Products (EMEA); member of the Committee of Experts of Research Policy CEPR - at the Ministry for University and Scientific and Technological Research; member of the Scientific Committee of the Lega Italiana per la Lotta Contro i Tumori; member of the Board of the Istituto Superiore di Sanità; Chairman of the Committee for Research of the Italian Agency for Drugs (AIFA). Other appointments include: Vice-president of the Consiglio Superiore di Sanità; President of the Research and Development Commission of the Agenzia Italiana del Farmaco (AIFA); President of the Angelo and Angela Valenti Foundation and the "Via di Natale" Foundation; President of the Technical Commission for Pharmaceutical Assistance of the Regione Autonoma of Sardinia. He is a member of the Strategic Committee for Welfare, Regione Lombardia, the Consiglio Superiore di Sanità and the Comitato Nazionale di Bioetica, the International Scientific Committee, Centro di Riferimento Oncologico, Aviano, and the Scientific Committee of AISLA, Member Comitato Scientifico, Dipartimento Politiche Antidroga Presidenza del Consiglio dei Ministri, Member, Advisory Board ADAMO Onlus, Milan, Member of Committee of the Recommendation for the Call, Wemos Foundation, Amsterdam,, Member, Development Advisory Board of the International Center for Biomedical Sciences (ICBMS), Luxu Town, Wujiang City, China. Silvio Garattini is a Fellow of the New York Academy of Sciences, the American Association for the Advancement of Science, Honorary Fellow of the Royal College of Physicians (Pharmaceutical Medicine), London, Honorary Fellow of the Italian Society of Pharmacology and a member of numerous other Italian and international scientific societies. He has received many awards for his work, including the French Legion d'honneur for scientific merit, and the Grand Ufficiale della Repubblica Italiana, and holds honorary degrees from the Universities of Bialystok in Poland, and Barcelona in Spain. Recent awards include the Ippocrate Prize, 2003; Mens Sana in Corpore Sano; Nuova Spoleto, 2003; Angelo dell anno; Alkmeon International Prize; International Prize Sant Agostino Città di Bergamo; Il Campione della Scienza; Natta Medal; Coppola Prize, Scienza e Società in the framework of the Premio Città di Firenze and Premio Rana d Oro, Casalbeltrame (Novara). In its 40-plus years, the Mario Negri Institute for Pharmacological Research, under Professor Garattini's leadership, has published more than 11,000 scientific papers and more than 250 books, on topics ranging from cancer and its treatment to tumour immunology, neuropsychopharmacology, and cardiovascular and renal pharmacology. More than 4000 young Italian and 600 foreign graduates and technicians have obtained specialist qualifications at the Institute. 343

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346 Prof. Giuseppe Remuzzi Giuseppe Remuzzi was born in Bergamo, Italy in Upon completion of his medical training at the University of Pavia in 1974, he received specialty training in Hematology and Nephrology at the University of Milan. Since 1975, he has pursued his academic career at the Ospedali Riuniti of Bergamo, where he was appointed Professor of Nephrology and Director of the Department of Medicine and Transplantation in Since 1999, he is Director of the Department of Nephrology and Dialysis of the same hospital. The Negri Bergamo Laboratories, which he has directed since 1984, is a group of basic scientists, physiologists, pharmacologists, molecular and cellular biologists, pathologists and clinicians devoted to the study of renal disease. As Research Coordinator of the Negri Bergamo Laboratories and the affiliated Clinical Research Center for Rare Diseases "Aldo e Cele Dacco", both divisions of the Mario Negri Institute for Pharmacological Research, he conduct a diverse team of researchers studying human renal diseases and their corresponding experimental models from the perspective of pathophysiology and therapeutic intervention. He touched major advances in many areas of nephrology, with particular focus on platelet endothelial interactions, vascular prostaglandin biology, coagulation and renal disease, progression of renal disease, experimental models of glomerular damage, and transplant immunology and tolerance. Particularly his contributions to the understanding of the pathophysiology of hemolytic uremic syndrome, prostaglandin metabolism in pregnancy, renal vascular biology in uremia, the role of protein trafficking in renal disease progression, the induction of graft tolerance by intrathymic injection of donor antigens, the role of the co-stimulatory CD28-B67 pathway in transplant rejection and the prevention of renal and cardiovascular damage in diabetes. Prof. Remuzzi serves on editorial boards of numerous journals including the prestigious New England Journal of Medicine and is member of the International Advisory Board of The Lancet. In recognition of his achievements, he has been awarded in 1998 honorary memberships of the Association of American Physicians and the British Royal College of Physicians. In 2001 he was nominated Chairman of the Research Committee of the COMGAN (Commission on Global Advancement of Nephrology) of the International Society of Nephrology (ISN). He received an Honorary Professorship at the University of Maastricht in In 2005 during the World Congress of Nephrology in Singapore he received the ISN Jean Hamburger Award and in November 2007 he received during the annual congress of the American Society of Nephrology the precious John P. Peters Award. On August 2008, he was appointed "Honoris Causa Professor" by the Catholic University of Cordoba (Argentina). Prof. Remuzzi has authored and co-authored more than 979 scientific articles, reviews and monographs. 345

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348 Mario Negri Institute Milan Executive Office Director Prof. Silvio GARATTINI, M.D. Administrative Office Maria Grazia PEZZONI, Chief Technical Office Fabio BRIGHENTI, D. Arch. General Maintenance Emanuele SINELLI Studies Office Armanda JORI, Pharm.D. Press Office Isabella BORDOGNA, Phil. D. Public Relations Office Claudio PANTAROTTO, Comm. Prevention and Safety Office Emilio BENFENATI, Chem.D. Annamaria SEGALINI, Phys.D. English Style Editor Judi BAGGOTT Photography and Audio-Visual Service Felice DE CEGLIE Purchasing Office Eufrasia COVIELLO Director s Office Rosanna MAPELLI, Chief General Secretariat Elena POZZOLI 347

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350 Negri Bergamo Laboratories Executive Offices Director Research Coordinator Scientific Secretariat Prof. Silvio GARATTINI, M.D. Giuseppe REMUZZI, M.D. Ariela BENIGNI, Biol.Sci.D., Ph.D. Administration of Research Projects/Admn. Assistance Daniela MELACINI, Biol.Sci.D. Press and Communication Office Francesca Di Fronzo, Mod. Lit. D. Audio-Visual Service Antonella PICCINELLI, Biol.Sci.D. Prevention and Safety Office Chief Annamaria SEGALINI, Phys.D. Library Chief Anna BOZZALE Valeria MIGLIOLI General Maintenance Giancarlo GASPARI Director s Office Antoinette van ENGELEN, Chief 349

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352 Centro Aldo e Cele Daccò Ranica (Bg) Executive Offices Director Research Coordinator Scientific Secretariat Health Director Prof. Silvio GARATTINI, M.D. Giuseppe REMUZZI, M.D. Ariela BENIGNI, Biol.Sci.D., Ph.D. Norberto PERICO,M.D. Press and Communication Office Francesca Di Fronzo, Mod. Lit. D. Prevention and Protection Office Chief Annamaria SEGALINI, Phys.D. Paola BOCCARDO, Biol.Sci.D. Library Mansueto Astori Chief Anna BOZZALE Monica MINALI Director s Office Barbara REMONTI, Oriental Languages D. Clinical Trials Office Paola BOCCARDO, Biol.Sci.D. Custodian/general maintenace Giampiero CUGUSI 351