BiO-dOSimetric Tools for triage to Responders. 14 th International Congress of Radiation Research Warsaw, Poland, August 28 - September 1, 2011

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1 BiO-dOSimetric Tools for triage to Responders 14 th International Congress of Radiation Research Warsaw, Poland, August 28 - September 1, 2011 Theme 10: Security Call FP7-SEC

2 5 countries 7 partners Start date of project: 1 st July 2010 Duration: 36 months Budget: 4.5 M BiO-dOSimetric Tools for triage to Responders Project number: Instrument: Collaborative Project Thematic priority: SECURITY Call identifier: SEC

3 5 countries 7 partners Beneficiary name Short name Country COMMISSARIAT A L ENERGIE ATOMIQUE CEA France NATIONAL UNIVERSITY OF IRELAND, GALWAY KARLSRUHE INSTITUTE OF TECHNOLOGIES IZOTOPKUTATO INTEZET - MAGYAR TUDOMANYOS AKADEMIA NUIG KIT IKI Ireland Germany Hungary CANBERRA CANBERRA France UNIVERSIDAD POLITECNICA DE VALENCIA UPVLC Spain ORSZAGOS ATOMENERGIA HIVATAL HAEA Hungary

4 What issue the project addresses? How quickly sort out victims of a radiological attack? First responders have to know the dose received by each person to optimize the triage A integrated toolbox

5 In case of a radiological attack a fast evaluation of the dose is required to sort out the victims 3 ways to be exposed Irradiation External Contamination Internal Contamination

6 Irradiation External Contamination Internal Contamination Blood Nasal mucus Saliva Mouth cells SAMPLES + Retrospective dose Calculation of the dose Urine Hairs Radioactive nanoparticles Low background, spectrometry IC-PMS

7 Fast Evaluation of the dose by the cell damages New biodosimetric tools measurement Evaluation of Radioactive Sources calculation Dose received A best triage

8 Fast Evaluation of the dose by the cell damages Blood Nasal mucus Saliva Mouth cells SAMPLES Cells damages New Biodosimetric Tools Hairs Urine

9 Biodosimetric tools Biodosimetry using the detection of DNA damage responses Biodosimetry using centrosome quantification Principle Level of -H2AX depends on number of DNA double-strand breaks which is dose-dependent Principle Centrosome amplification is a dose-dependent response to DNA damages Measurement of -H2AX level in the cells Measurement of proteins ( -tubulin, centrin, pericentrin and ninein) related to centrosomes amplification.

10 Real field exercise to test the applicability of the tools and systems in a realistic situation Organizer: Hungarian Atomic Energy Authority (HAEA), Hungary Objective: The objective of the exercise to test the 1. Tools, systems 2. Procedures and 3. Methodologies of the bio-dosimetric toolbox. Location: KFKI campus of the Hungarian Academy of Sciences (Budapest). This site has already been used for similar exercises, the latest being the 2008 Hungarian National Exercise on Combating Illicit Trafficking

11 BOOSTER WP 100 Management WP 110 Management Activities WP 120 Dissemination Activities WP 200 System requirements & Design concept WP 300 Fast evaluation WP 400 New biodosimetric tools WP 500 Software dev. & integration WP 600 System validation & Training WP 210 Needs & requirement WP 310 Miniaturization of gamma camera WP 410 Characterization of the exposure WP 510 Hardware development WP 610 System validation WP 220 System architecture design WP 320 Assessment of other sensors WP 420 Biodosimetry using the detection of DNA damages WP 520 Software dev. (interoperability) WP 620 Training courses WP 230 Exploitation strategy WP 430 Biodosimetry using centrosome detection WP 530 Decision-making software WP 440 Validation of the tools WP 540 Interface SW/HW

12 A scenario persons in a stadium to support their team, waiting for the start of the match 5 explosions in less 15 minutes

13 In case of a crisis management (accident, terrorist attack ), involving radioactive material, different areas are commonly defined around the scene 1,2. 1 Manual for First Responders to a Radiological Emergency (EPR-FIRST RESPONDERS), IAEA, Vienna, TMT Hanbook, Carlos Rojas-Palma et al., NRPA, Norway, 2009.

14 SIMACOP SIMACOP : the backbone of BOOSTER for communication Controled Area Exclusion Area Support Area

15 First Responders radiological equipment In the exclusion area Gampix Colibri UltraRadiac Falcon 5000 Gamma camera Localisation of radioactives sources Dosimeters Map of the radioactivities Gamma spectrometer Identification of radioactives sources Exclusion Area Information sent to the support area

16 Exclusion Area Decontamination Area Blood Nasal mucus Saliva Mouth cells SAMPLES Decontamination Urine Hairs Gamma Probes for body gamma contamination X-Ray Probes for hands and face contamination measurement Alpha Beta Gamma Probes for foot contamination measurement

17 Low background spectrometry Goal : Identification of radioactive material by gamma spectrometry Principle: measurement of the energy of photons emitted by the sample The iron chamber Diameter: 300 mm (diameter) Height: 370 mm Wall thickness: 100 mm. Weight: ~ 200 kg. The sample measurement : 10 cm (diameter) x 17 cm (height). Detector: PGT IGC10 coaxial HpGe crystal Support Area The lead chamber Diameter: 200 mm height: 360 mm weight: ~ 90 kg. The sample chamber: is 56 mm x 56 mm x 100 mm. Detector: GL2020R planar HpGe crystal Skin swabs, saliva, nasal mucus or from the environment samples

18 Retrospective dosimetry Thermoluninescence analysis on SMD materials Principle: some material can emit light when they are heated if they have been irradiated. The amount of light is proportional to the dose received by the material. Idea: use SMDs from the cell phones carried by victims Support Area TL glow curves of 100k type SMD resistor irradiated with different doses

19 Biodosimetric tools H2AX quantification analysis Centrosome quantification analysis Cytogenic analysis Time On field measurement On lab measurement How to do that? Standard technics

20 H2AX quantification analysis Goal : to develop a test assaying the dose received by a person in less than 20 min Standard approach: Immunofluorescent by counting foci X Too long! Global counting by extracting proteins H2AX and H2AX and calculate the ratio H2AX/( H2AX + H2AX)

21 Hairs Blood Nasal mucus Saliva Mouth cells SAMPLES Hairs Urine How to quickly sample hairs?

22 Hairs Extraction of proteins in less than 20 s Microwave Signal proportionnal to the dose

23 Blood

24 Centrosome quantification Centrosome amplification is a dose-dependent response to DNA damage (1) Develop a novel biodosimetry assay using centrosome quantification to measure medium-term exposure to ionizing radiation Support Area Purify centrosome components: immunogens for monoclonal antibodies development Use in immunofluorescence Automated centrosome counting using High Content Microscopy (1) Bourke, 2007, EMBO Reports 8:603-9

25 Centrosome quantification Operetta: High Content Imaging System Fluorescent microscope 96 or 384 well plates Automated image acquisition User-configurable High Content Analysis Harmony software Support Area

26 Centrosome quantification 9 candidate proteins for antibody production Centrosome composed of several hundred proteins Support Area

27 Centrosome quantification Centrosome amplification in chicken lymphocytes Support Area

28 Number of centrsome per cell Centrosome quantification % of cells with > 2 centrosomes Quantification of centrosome amplification in DT40 cells after IR. 3, , , , Dose (Gy) Dose (Gy) R 2 =0.996 R 2 =0.999 Support Area

29 Conclusion Id victim Dose map Biodosimeters Gamma camera Retrospective dose.. Dose BOOSTER web site

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