Michele Menotta Ph.D.

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1 Michele Menotta Ph.D. Department of Biomolecular Sciences University of Urbino "Carlo Bo" Via Saffi Urbino (PU) ITALY Tel: Fax: EDUCATION Jan Ph.D. in Biochemistry University of Urbino Carlo Bo Dissertation title: Identification of genes expressed during the presymbiotic phase of T. borchii-t Americana: Cell signalling phenomena. Advisor: Prof. Vilberto Stocchi Mar B. Sc. Cum laude in Biology University of Urbino Carlo Bo Thesis Title: Genetic variability study, by ITS analysis, of different Tuber species Advisor: Prof. Vilberto Stocchi RESEARCH EXPERIENCE Sep.2012-present Postdoctoral research Department of Biomolecular Sciences University of Urbino Research project: Dexamethasone treatment of AT (Ataxiatelangiectasia) patients. Dexamethasone action in Ataxia- Telangiectasia: seeking the molecular mechanism of phenotype reversion. Sep sep.2012 Postdoctoral research Department of Biomolecular Sciences University of Urbino.

2 Michele Menotta 2 Research project: detection of ODNs-/transcription factors interaction by AFM. Granted by NACBO EU FP6. Research project: Dexamethasone action in Ataxia-Telangiectasia: seeking the molecular mechanism of phenotype reversion. May 2006-Aug Postdoctoral research Department of Biomolecular Sciences University of Urbino. Research project: NF-kB quantification by AFM. Granted by FIRB 2003 NanoMed NANOtechnology for biomedicine. Research project: Drugs delivery by SWNT. Granted by NACBO and NanoMed. Research project (from 2009): Dexamethasone action in Ataxia- Telangiectasia: seeking the molecular mechanism of phenotype reversion. Feb Mar 2006 Postdoctoral research Institute of biochemistry, University of Urbino. Research project: Molecular characterization of T. borchii CDC42, MAPK1 and Rho-GDI. Nov-2000-Nov 2003 Doctoral research Institute of biochemistry, University of Urbino. Research project: Study of cell signaling pathways between T. borchii and T. Americana during pre-symbiosis life stage. Oct-1996-Mar 1999 Undergraduate research Institute of biochemistry, University of Urbino. Research project: Genetic variability of Tuber species by ITS analysis. TEACHING EXPERIENCE Adjunct Professor Proteomics Faculty of Science and Technology. Biology course, University of Urbino.

3 Michele Menotta Adjunct Professor Genetic engineering Faculty of Science and Technology. Biotechnology course, University of Urbino Adjunct Professor Biotechnology laboratory Faculty of Science and Technology. Biotechnology course, University of Urbino Adjunct Lecturer Faculty of Science and Technology. 2 level Master course University of Urbino Teaching Assistant Biotechnology laboratory course, Faculty of Science and Technology, University of Urbino Teaching Assistant Human Biology course Faculty of Health and Physical Exercise, University of Urbino. OTHER EXPERIENXES 02/2005 to 03/2005. Visiting researcher, Institute for Biochemistry I, Joseph-Stelzmann-Strasse 52, D50931 Cologne Advanced AFM methodologies training PARK SYSTEMS KANC 4F lui-dong Suwon South Korea Advanced AFM modules training PARK SYSTEMS, Gambetti Kenologia, Milan Visiting researcher, Nanomed Lab University of Genoa European Park AFM User Meeting Lille (F). TECHNICAL EXPERTISE DNA-RNA extraction and manipulation, gel electrophoresis, blotting, radio isotope handling and labelling.

4 Michele Menotta 4 Phage and yeast library screening. Real time quantitative PCR and microarray analysis. Protein engineering: cdna cloning, mutagenesis, expression in bacteria, yeast, D. discoideum, insects and mammalian cells. Protein purification: column chromatography, FPLC, affinity tag. Protein characterization: SDS PAGE, western blotting, chemical cross-linking, in vitro activity. Protein interaction: Yeast two hybrid screening, pull-down experiments. Mass spectrometry: GC-Ion Trap and ESI-Q-TOF. In gel protein digestion and MS/MS protein identification. De novo MS/MS sequencing. Microscopy: Immunohistochemical / immunofluorescence staining and analysis of cells and tissues. Atomic Force Microscopy: NCM, CM, Tapping and FMM surface imaging, in air and in liquid. Force volume spectroscopy with or without chemically modified tips. Chemical and biological functionalization of AFM tips and surfaces. Data statistical analysis and statistical software skill (i.e. SPSS, PRISM, ORIGIN, R). Computer programming by JAVA, C++, VB PATENTS Paola Fanzio, Giuseppe Firpo, Mauro Magnani, Chiara Manneschi, Michele Menotta, Valentina Mussi, Luca Repetto, Paola Scaruffi, Sara Stigliani, Gian Paolo Tonini, Ugo Valbusa. Device and single-molecule analysis method by means of detection of the collisions of a target molecule on functionalized nanopores. WO A PUBBLICATIONS

5 Michele Menotta 5 1. Paola Fanzio *, Valentina Mussi, Michele Menotta, Giuseppe Firpo, Luca Repetto, Patrizia Guida, Mauro Magnani, and Ugo Valbusa. Protein detection with a dslnafunctionalized nanopore. Biosensors and Bioelectronics. Accepted. 2. Michele Menotta, Sara Biagiotti and Mauro Magnani. Quantitative analysis of FK506 by AFM. Biosensors and Bioelectronics. In revision. 3. Sara Biagiotti, Michele Menotta, Marzia Bianchi, Luciana Chessa and Mauro Magnani. Differential gene expression following dexamethasone administration in ATM-/- lymphoblastoid cell lines. The Journal of Steroid Biochemistry and Molecular Biology Jul;392(1-2): Mauro De Santi, Antonella Antonelli, Michele Menotta, Carla Sfara, Sonja Serafini, Simone Lucarini, Giorgio Brandi and Mauro Magnani. Single-Walled Carbon Nanotubes Functionalization for the Delivery of the Water Insoluble Anticancer Agent Indole-3- carbinol Cyclic Tetrameric Derivative CTet. J. Nanopharmaceutics Drug Delivery 1, (2013). 5. Michele Menotta, Sara Biagiotti, Luciana Chessa and Mauro Magnani. Dexamethasone Partially Rescues Ataxia Telangiectasia-mutated (ATM) Deficiency in Ataxia Telangiectasia by Promoting a Shortened Protein Variant Retaining Kinase Activity. J Biol Chem Oct 10; 287(49): Michele Menotta, Rita Crinelli, Elisa Carloni, Valentina Mussi, Ugo Valbusa and Mauro Magnani. Binding force measurement of NF-kB - ODNs interaction: an AFM based decoy and drug testing tool. Biosensors and Bioelectronics Oct. 15; 28(1): Epub 18 July E. Omiccioli, P. Maltese, M. Menotta, G. Amagliani, M. Magnani. Magnetic capture hybridization technology: a new tool for selective isolation of cell-free circulating DNA from human plasma. JOURNAL OF BIOTECHNOLOGY : Mussi V, Fanzio P, Stigliani S, Scaruffi P, Firpo G, Repetto L, Menotta M, Magnani M, Tonini GP, and Valbusa U. Electrical characterization of DNA-functionalized solid state nanopores for bio-sensing. J Phys Condens Matter Nov 17; 22(45): Epub 2010 Oct 29.

6 Michele Menotta 6 9. Raffaella Pierleoni ; Michele Menotta, Antonella Antonelli ; Carla Sfara; Giordano Serafini; Sabrina Dominici; Maria Elena Laguardia ; Annalisa Salis ; Gianluca Damonte ; Lucia Banci ; Marco Porcu ; Paolo Monini ; Barbara Ensoli ; Mauro Magnani. Effect of the redox state on hiv-1 TAT protein multimerization and cell internalization and trafficking. Mol Cell Biochem Dec;345(1-2): Epub 2010 Aug Antonelli, S Serafini, M Menotta, C Sfara, F Pierigè, L Giorgi, G Ambrosi, L Rossi and M Magnani. Improved cellular uptake of functionalized single-walled carbon Nanotubes. Nanotechnology Oct 22;21(42): Epub 2010 Sep Menotta M, Crinelli R, Carloni E, Bianchi M, Giacomini E, Valbusa U, Magnani M. Labelfree quantification of activated NF-kappaB in biological samples by atomic force microscopy. Biosens Bioelectron Jul 15;25(11): Epub 2010 Apr Crinelli R, Carloni E, Menotta M, Giacomini E, Bianchi M, Ambrosi G, Giorgi L, Magnani M.Oxidized ultrashort nanotubes as carbon scaffolds for the construction of cellpenetrating NF-kappaB decoy molecules. ACS Nano May 25;4(5): Amicucci A, Zambonelli A, Iotti M, Polidori E, Menotta M, Saltarelli R, Potenza L, Stocchi V. Morphological and molecular modifications induced by different carbohydrate sources in Tuber borchii. J Mol Microbiol Biotechnol. 2010;18(2): Epub 2010 Mar Maltese P, Canestrari E, Palma L, Ruzzo A, Corini F, Menotta M, Andreoni F, Latiano A, Annese V, and Magnani M. (2009). High resolution melting (HRM) analysis for the detection of ER22/23EK, BclI, and N363S polymorphisms of the glucocorticoid receptor gene. The Journal of steroid biochemistry and molecular biology 113(3-5): Menotta M., Amicucci A., Basili G., Polidori E., Stocchi V., and Rivero F. (2008). Molecular and functional characterization of a Rho GDP dissociation inhibitor in the filamentous fungus Tuber borchii. BMC. Microbiol. 8: Crinelli R., Bianchi M., Menotta M., Carloni E., Giacomini E., Pennati M., and Magnani M. (2008). Ubiquitin over-expression promotes E6AP autodegradation and reactivation of the p53/mdm2 pathway in HeLa cells. Mol. Cell Biochem. 17. Gioacchini A.M., Menotta M., Guescini M., Saltarelli R., Ceccaroli P., Amicucci A., Barbieri E., Giomaro G., and Stocchi V. (2008). Geographical traceability of Italian white truffle

7 Michele Menotta 7 (Tuber magnatum Pico) by the analysis of volatile organic compounds. Rapid Commun. Mass Spectrom. 22: Polidori E., Ceccaroli P., Saltarelli R., Guescini M., Menotta M., Agostini D., Palma F., and Stocchi V. (2007). Hexose uptake in the plant symbiotic ascomycete Tuber borchii Vittadini: biochemical features and expression pattern of the transporter TBHXT1. Fungal. Genet. Biol. 44: Ceccaroli P., Saltarelli R., Guescini M., Polidori E., Buffalini M., Menotta M., Pierleoni R., Barbieri E., and Stocchi V. (2007). Identification and characterization of the Tuber borchii D- mannitol dehydrogenase which defines a new subfamily within the polyol-specific medium chain dehydrogenases. Fungal. Genet. Biol. 44: Menotta M., Amicucci A., Basili G., Rivero F., Polidori E., Sisti D., and Stocchi V. (2007). Molecular characterisation of the small GTPase CDC42 in the ectomycorrhizal fungus Tuber borchii Vittad. Protoplasma 231: Menotta M., Pierleoni R., Amicucci A., Sisti D., Cerasi A., Millo E., Chiarantini L., and Stocchi V. (2006). Characterization and complementation of a Fus3/Kss1 type MAPK from Tuber borchii, TBMK. Mol. Genet. Genomics 276: Cervasi B., Paiardini M., Serafini S., Fraternale A., Menotta M., Engram J., Lawson B., Staprans S.I., Piedimonte G., Perno C.F. et al. (2006). Administration of fludarabine-loaded autologous red blood cells in simian immunodeficiency virus-infected sooty mangabeys depletes pstat-1-expressing macrophages and delays the rebound of viremia after suspension of antiretroviral therapy. J. Virol. 80: Amicucci A., Guidi C., Menotta M., Pierleoni R., Polidori E., Zeppa S., and Stocchi V. Towards a better understending of the ectomycorrhizal fungus Tuber borchii Vittad. Life cycle. Recent Res. Devel. Microbiology, (9)(2005): ISBN: G. Giomaro, D. Sisti, A. Amicucci, C. Guidi, M. Menotta, E. Polidori, R.. Pierleoni, S. Zeppa, A. M. Gioacchini, M. Guescini, V. Stocchi."Sintesi micorrizica in vitro fra piante forestali e funghi del genere Tuber: un modello per lo studio dei meccanismi morfologici e molecolari coinvolti nel processo di micorrizazione". Informatore Botanico Italiano, 37(x, sup.1 parte 1) 2005.

8 Michele Menotta Gioacchini A.M., Menotta M., Bertini L., Rossi I., Zeppa S., Zambonelli A., Piccoli G., and Stocchi V. (2005). Solid-phase microextraction gas chromatography/mass spectrometry: a new method for species identification of truffles. Rapid Commun. Mass Spectrom. 19: Menotta M., Gioacchini A.M., Amicucci A., Buffalini M., Sisti D., and Stocchi V. (2004). Headspace solid-phase microextraction with gas chromatography and mass spectrometry in the investigation of volatile organic compounds in an ectomycorrhizae synthesis system. Rapid Commun. Mass Spectrom. 18: Menotta M., Amicucci A., Sisti D., Gioacchini A.M., and Stocchi V. (2004). Differential gene expression during pre-symbiotic interaction between Tuber borchii Vittad. and Tilia americana L. Curr. Genet. 46: Gioacchini A.M., Menotta M., Polidori E., Giomaro G., and Stocchi V. (2002). Solid-phase microextraction gas chromatography/ion trap mass spectrometry and multistage mass spectrometry experiments in the characterization of germacrene D. J. Mass Spectrom. 37: Amicucci A., Guidi C., Bertini L., Menotta M., Potenza L., Rossi I., Stocchi V.. Identification of white Tuber species by multiplex PCR. Libro degli Atti. Vth International Congres: Science and cultivation of truffle and other edible hypogeous mushrooms. Aix-en- Provence, France 4-6 Marzo Proceedings INTERNATIONAL MEETING PRESENTATIONS 1. Michele Menotta, Sara Biagiotti, Marzia Bianchi, Luciana Chessa and Mauro Magnani. Dexamethasone action in Ataxia-Telangiectasia: seeking the molecular mechanism of phenotype reversion. ATW th - 31st July 2013, Birmingham UK. 2. Sara Biagiotti, Michele Menotta, Marzia Bianchi, Luigia Rossi, Luciana Chessa and Mauro Magnani. Gene expression profiling of Dexamethasone treatment on Ataxia- Telangiectasia cells. ATW th - 31st July 2013, Birmingham UK.

9 Michele Menotta 9 3. Michele Menotta, Sara Biagiotti, Marzia Bianchi, Luigia Rossi, Laura Streppa and Mauro Magnani. Label-free quantification of Tacrolimus by Atomic Force Microscopy. CNBXI 27th-29th June 2012, Varese. 4. Biagiotti S, Menotta M, Bianchi M, Giacomini E, Crinelli R, Savio C, Rossi L, Chessa L, Magnani M. New applications for old drugs: molecular mechanism involved in glucocorticoid analogue action in the treatment of Ataxia Telangiectasia FEBS th-25th June Torino. 5. Michele Menotta, Rita Crinelli, Elisa Carloni, Valentina Mussi, Ugo Valbusa and Mauro Magnani. Binding force measurement of NF-kB - ODNs interaction: an AFM based decoy and drug testing tool. FEBS th-25th June Torino. 6. Elisa Giacomini, Marzia Bianchi, Rita Crinelli, Elisa Carloni, Michele Menotta, Mauro Magnani. Different intron-mediated enhancement mechanisms for the 5'- UTR intron of the human UbC gene. SIB th-17th September Milano 7. E. Carloni, R. Crinelli, M. Bianchi, E. Giacomini, M. Menotta, E. Gabucci and M. Magnani. Role of heat shock transcription factors in the induction of UbC gene expression in MG132-treated cells. SIB th-17th September Milano 8. Michele Menotta, Rita Crinellia, Elisa Carloni, Valentina Mussi, Ugo Valbusa and Mauro Magnani. Binding force measurement of NF-kB - ODNs interaction: an AFM based decoy and drug testing tool. SIB th-17th September Milano 9. Mauro De Santi, Sonja Serafini +, Simone Lucarini ^, Gianluca Ambrosi*, Michele Menotta +, Giorgio Brandi +, Mauro Magnani. Use of nanotubes (SWNTs) to increase bioavailability of Indole-3-Carbinol tetrameric derivative. 2nd NACBO International Nanobiotechnology Conference 7th- 9th July 2009 University of Rome 'Tor Vergata. 10. E. Carloni, *M. Menotta, *E. Giacomini, *M. Bianchi, *S. Dominici, #G. Ambrosi, *M. Magnani and *R. Crinelli Covalent functionalization of SWCNT with ODNs targeting Nuclear Factor - B: toward the construction of cell-penetrating decoy molecules. 2nd NACBO International Nanobiotechnology Conference 7th- 9th July 2009 University of Rome 'Tor Vergata. 11. Michele Menotta, Rita Crinelli, Elisa Carloni, Marzia Bianchi, Elisa Giacomini and Mauro Magnani. Analysis of transcription factor-dna interaction by Atomic Force

10 Michele Menotta 10 Microscopy. 2nd NACBO International Nanobiotechnology Conference 7th- 9th July 2009 University of Rome 'Tor Vergata. 12. Michele Menotta, Rita Crinelli, Marzia Bianchi, Elisa Carloni, Elisa Giacomini and Mauro Magnani. AFM imaging could bring NF-kB to the clinic. National Nanomedicine Congress Auditorium Advanced Biotechnology Center (CBA) Genova November 28-29, Antonella Amicucci1, Michele Menotta1, Giorgio Basili1, Emanuela Polidori2, Paola Ceccaroli1, Vilberto Stocchi1, Francisco Rivero. Opening a window for understanding the cell signaling processes in the filamentous fungus Tuber borchii: RhoGdi, another element in the molecular network th European conference on fungal genetics, Edinburgh 14. Zambonelli A., Amicucci A., Menotta M., Polidori E., Iotti M., Saltarelli R., Potenza L., Stocchi V. - Influence of host plant and different supplies on morphology and gene expression in Tuber borchii. Spoleto Tuber Amicucci A., Zambonelli A., Iotti M., Menotta M., Saltarelli R., Zeppa S., Stocchi V., Morphological and molecular modifications induced by different carbohydrate sources in Tuber borchii Vittad. mycelium, in:, book of Abstracts, SORRENTO, s.n, 2007, pp. 193 (atti di: XIII international Congress on molecular plant-microbe interactions, Sorrento, Italy, luglio R. Crinelli, M. Bianchi, M Menotta, E. Carloni, E. Giacomini, L. Gentilini, M Magnani. Ubiquitin Overexpression leads to stabilization of p53 and increased p21 (cip/waf1) transcription by promoting e6ap autodegradation in HeLa cells. SIB 2006, Riccione, 28-30/09/ M. Bianchi, R. Crinelli, E. Giacomini, L. Palma, M. Menotta, E. Carloni e M. Magnani. Overexpression of wild-type ubiquitin downregulates transcription of endogenous polyubiquitin genes in HeLa cells. SIB 2006, Riccione, 28-30/09/ Zeppa S, Barbieri E, Rossi I, Guescini M, Curci R, Falcieri E, Menotta M e Stocchi V. Analisi dell'espressione genica durante il processo di differenziazione miogenica. II Convegno Internazionale "Il ruolo dell'esercizio fisico nella prevenzione delle malattie e nel miglioramento della qualità della vita", Rimini, maggio 2005.

11 Michele Menotta A. Amicucci, M. Menotta, G. Basili, E. Polidori, D. Sisti, F. Rivero, F. Palma and V. Stocchi"Characterisation of the small GTPase CDC42 in the Ascomycetous fungus Tuber borchii Vittad.: its putative role in the hyphal apical growth".1st International Conference on Environmental, Industrial and Applied Microbiology, BioMicroWorld-2005, Badajoz (Spain), 15-18Marzo Menotta M, Pierleoni R, Polidori E, Palma F, Sisti D, Amicucci A, Stocchi V. MAPK signaling in the ectomycorrhizal system Tuber borchii- Tilia Americana. ECFG7 Copenhagen A.Amicucci, M. Menotta, E. Polidori, F. Palma and V. Stocchi. Molecular cloning of Cdc42 and Rho-GDI genes involved in the actin cytoskeleton development in the ectomycorrhizal fungus T. borchii Vittad. SIB Ferrara M. Menotta, R. Pierleoni, E. Polidori, D. Sisti, A. Amicucci and V. Stocchi. MAPK signaling in the ectomycorrhizal system Tuber borchii- Tilia americana. SIB Ferrara A.M. Gioacchini, M. Menotta, R. pierleoni, D. Agostini, V. Stocchi. Characterization by ESI-Q-TOF tandem Mass spectrometry of phosphorylated proteins involved in ectomycorrhizal symbiosis. Torino XXI SCI giugno A.M. Gioacchini, D. Agostani, G. Piccoli, M. Menotta, L. Vallorani, R. pierleoni, V. Stocchi. Identification of terpenes in Calendula flowers extract by solid-phase microextraction gas chromatography ion trap mass spectrometry. Torino XXI SCI giugno D. Agostini, A.M. Gioacchini, G. Piccoli, M. Buffalini, L. Vallorani, M. Menotta, V. Stocchi. Electrospray Quadruple Time of Flight characterization of Aloe arborescens unfractionated extracts. Torino XXI SCI giugno Menotta M.Early molecular communication between plant and fungus: detection of specifically expressed genes during pre-symbiotic interaction. 6a Riunione Nazionale "A. Castellani" dei Dottorandi di Ricerca in Discipline Biochimiche, Brallo di Pregola (PV), giugno Menotta M., Gioacchini A. M., Buffalini M., Amicucci A., Sisti D. and Stocchi V. Volatile compounds: a role in signalling interaction? 6 ECFG Pisa 6-9 April 2002.

12 Michele Menotta M Menotta, A. Amicucci, A.M. Gioacchini., D. Sisti & V. Stocchi. Early molecular communication between plant and fungus: detection of specifically expressed genes during pre-symbiotic interaction. IMC7 Oslo august Gioacchini A., Menotta M., Zeppa S., Bertini L., Zambonelli A., Stocchi V.. SPME- Gas Chromatography/ion trap mass spectrometry, a new and rapid approach to truffle characterization. 20th Informal Meeting on Mass Spectrometry, May 12-16, M.Menotta, A.M. Gioacchini, A. Amicucci, M. Buffalini, D. Sisti, V. Stocchi"Volatile compounds: a role in mycorrhizal symbiosis signalling interaction?" 20th Informal Meeting on Mass Spectrometry, May 12-16, Gioacchini A.M., Menotta M., Polidori E., Giomaro G., and Stocchi V Solid-phase microextraction gas chromatography/ion trap mass spectrometry and multistage mass spectrometry experiments in the characterization of germacrene D. 20th Informal Meeting on Mass Spectrometry, May 12-16, M. Menotta, A. Amicucci, C. Guidi, E. Barbieri, V. Stocchi. Hypotesis on truffles speciation. TICS Third International Congress on Symbiosis Marburg Germany August Guidi C., Amicucci A., Menotta M., Zambonelli A., Potenza L. and Stocchi V. "Molecular identification of different Tuber species " SIB-BIB Biochimica e Biotecnologie, Villa Olmo (CO) giugno 2000:

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