ConRad 2013, 13.–16.5.2013, Global Conference on Radiation ...

CHALLENGECHALLENGE CBRN MEDICAL DEFENSE INTERNATIONAL

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Special Edition April 2013A publication of Beta Publishing Group

German Society for Military Medicine and Pharmacy(GSMMP/DGWMP)

Bundeswehr Institute ofRadiobiology affiliated to the University of Ulm

ConRad 2013, 13.–16.5.2013, Global Conference on Radiation Topics, Munich

Abstracts

www.nmd-conference.org

The German Society for Military Medicine and Pharmacy (GSMMP/DGWMP)

Deutsche Gesellschaft für Wehrmedizin und Wehrpharmazie e.V.

-Vereinigung deutscher Sanitätsoffiziere (VdSO)-

Neckarstraße 2a, D-53175 Bonn, GermanyPhone: +49 228 632420, Fax: +49 228 698533

E-Mail: [email protected], Internet: www.dgwmp.de

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Welcoming address by the Conference Chairperson and Secretary. . . . . . . . . . . . . . . . . . . . . . . . . . . p. 2Welcoming address by the President of the German Society for Military Medicine and Pharmacy (GSMMP/DGWMP) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . p. 3Welcome letter of beta-publishing . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . p. 4

Oral Presentations

11. Fukushima. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . p. 18 12. Radiation risk perception and communication. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . p. 1913. Radiation emergency medical preparedness and response . . . . . . . . . . . . . . . . . . . . . . . . . . . . p. 10 14. National, international and global radiation accident management . . . . . . . . . . . . . . . . . . . . . . p. 12 15. External exposure assessment . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . p. 1316. Decontamination measures and monitoring . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . p. 14 17. Biological dosimetry and EPR. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . p. 14 18. Radiation health effects and medical countermeasures . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . p. 18 19. Effects of low dose ionizing radiation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . p. 25 10. Radiation epidemiology . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . p. 26 11. Radiation protection . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . p. 27 12. Radiation biology/radiation physics . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . p. 29 13. Non-ionizing radiation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . p. 30

Poster Presentations . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . p. 31

Table of Contents

International Editorial Advisory GroupChairmanBrigadier General (ret) MC Dr Torsten Sohns, Bonn/Germany

MembersLtCol (Res) MC Dr Walter Biederbick, Bonn/GermanyCol MC Dr Roman Chlibek, PhD, Hradec Kralove/Czech RepublicCol MC Prof Dr Marek K. Janiak, MD, PhD, Warsaw/PolandCol MC Prof Dr Jiri Kassa, CSc., Hradec Kralove/Czech RepublicProf Dr Simo Nikkari, PhD, Helsinki/FinlandCol MC Prof Dr Viktor Meineke, Munich/GermanyAss Prof Dr Lars Schaade, Berlin/GermanyCol MC Prof Dr Horst Thiermann, Munich/GermanyBrigadier General (ret) MC Dr Christoph Veit, Bonn/GermanyMajor General (ret) MC Dr Prof Victor A. Voicu, Bucuresti/RomaniaCol MC Prof Dr Lothar Zöller, Munich/Germany

Editor-in-chief Col (ret) MC Dr Ernst-Jürgen FinkeE-mail: [email protected]

Project ManagerMr Helmar WinkelTel.: +49 (228) 919 37-22E-mail: [email protected]

Manager Sales InternationalMrs Juliane Schneider, Tel.: +49 (228) 919 37-59E-mail: [email protected]

Production ManagerMrs Renate Stieler, Tel.: +49 (228) 919 37-29E-mail: [email protected]

Published byBeta Verlag & Marketinggesellschaft mbHCelsiusstr. 43, 53125 Bonn / GermanyTel.: +49 (228) 919 37-10, Fax: +49 (228) 919 37-23

E-mail: [email protected], www.mci-forum.com

PublisherHeike Lange, Heinz-Jürgen Witzke

Index of Advertisers:DGWMP 2nd coverHeyl 3rd coverMeta-System p. 7Dr. Westmeier 4th cover

Imprint

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ConRad 2013, 13.–16.5.2013, Conference, Munich

Dear Colleagues,

It is a pleasure for us to personally welcome you to the

Global Conference on Radiation Topics (20th Nuclear Med-

ical Defence Conference) from May 13th to 16th, 2013 in Munich!

For the first time the conference will be presented under this new name, ConRad - Global Conference

on Radiation Topics - Preparedness, Response, Protection and Research, taking into account the unique

opportunity given at this event to bring together such a broad spectrum of international experts, civil-

ian and military, from all over the world as a forum for professional and multidisciplinary exchange of

experience and expertise in this particular domain. New insights in the fields of radiation accident pre-

paredness, management and response and radiation protection as well as new research findings in radi-

ation medicine and radiobiology will be the key topics.

This timeframe of three days, launched in 2011, will be maintained for the upcoming conference to

provide adequate time to the different subjects. Special attendance is addressed to the fact that these

aspects will be presented in a comprehensive form for a broader audience in relation to the idea of con-

tinuous education. An exciting and diversified scientific program has been scheduled covering the top-

ics “Fukushima”, “Radiation risk perception and communication”, “Radiation emergency medical pre-

paredness and response”, “National, international and global radiation accident management”,

“External exposure assessment”, “Decontamination measures and monitoring”, “Biological dosimetry

and EPR”, “Radiation health effects and medical countermeasures”, “Effects of low dose ionizing radia-

tion”, “Radiation epidemiology”, “Radiation protection”, “Radiation biology/radiation physics” and

“Non-ionizing radiation”.

We hope you also enjoy the accompanying industrial exhibition arranged by the German Society for

Military Medicine. There, companies will present their latest products and technologies in the field of

radiation protection and laboratory equipment for radiation research.

About 250 participants from 40 nations are registered to attend this year`s conference. This event will

offer to you plenty of opportunities for extensive discussions, making of new contacts and strengthen-

ing of existing relationships after the oral presentations, during the poster sessions, while visiting the

exhibition or at the social events. The conference dinner will take place at “Schlossgut Odelzhausen” a

former seat of nobility with a private brewery.

We are sure that you will enjoy being in Munich, the capitol city of the Free State of Bavaria, for this

exciting conference. We look forward to you being part of this further key event dealing with radiation

topics affecting all of us.

Sincerely,

Colonel Prof. Dr. Viktor Meineke Dr. Christina Beinke

Conference Chair Conference Secretary

Director Bundeswehr Institute of Radiobiology

affiliated to the University of Ulm

Welcoming address by the Conference Chairperson andSecretary

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Greetings by the President of the GSMMP/DGWMP

Dear participants of the 20th Nuclear Medical Defence Conference 2013,

It is again a great honour and pleasure for me to welcome you in Munich. The German Society

for Military Medicine and Pharmacy (GSMMP/DGWMP) supports many important confer-

ences and scientific topics out of the wide catalogue of military medicine. Nuclear Medical

Defence in particular is an even more important field of research than others. The reason for

this is that the emotionalized public interest is almost bare of facts and even basic scientific

knowledge. In addition technical progress in detection of radiation and radioactive elements

has become so sensitive that most of the non-insiders are unable to validate results correctly. It

is not a surprise that as a consequence of the described mismatched relation of facts and emo-

tion, not enough resources exist worldwide for a development of diagnostic and therapeutic

tools in the field of specific Nuclear Medical Research. Military research institutes and organi-

sations are still constantly involved in this area and contribute successfully more and more in

various fields and models of civil military cooperation. The Fukushima radiation accident

again has proved that an enhancement of national and international collaboration is needed

in case of nuclear accidents and incidents, especially if medical treatment would be needed.

The spectrum of the 20th Nuclear Medical Defence Conference is of a wide range. For me this

corresponds to the omnipresence of nuclear medical topics in our modern world, civil and

military. It includes protection against ionizing radiation in the medical and the industrial

field, diagnostic and therapeutic measures, the effects of low dose ionizing radiation and many

other subjects.

Ladies and gentlemen, this conference would not take place under such comfortable and

favourable circumstances without the substantial support of the industrial exhibition and the

sponsoring companies. Please, make use of the opportunity to inform you about technical

innovations and to discuss solutions with the representatives of the industry. And at the end

of the day don’t forget to tighten your personal network, to make new friends and to renew

old friendships.

I wish you a very successful conference.

Sincerely

Yours

Christoph Veit (MD)

Brig.Gen. (ret.)

President of the GSMMP/DGWMP

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ConRad 2013, 13.–16.5.2013, Conference, Munich

Dear participants,

CHALLENGE CBRN MEDICAL DEFENSE INTERNATIONAL would also like to warmly welcome

you to the Global Conference on Radiation Topics (20th Nuclear Medical Defence Conference)

in Munich.

Many Nations have improved their preparedness for the protection against nuclear threads

after the latest sad developments in Japan, although there still is a gap between possible

threads, resources and preparedness to these, even though international and military –civil

cooperation have become more intense in the past. CHALLENGE Magazine contributes a plat-

form for specialists and those involved in operational, scientific, medical, socio-ethical as well

as public health and overall security to strengthen this exchange. CHALLENGE was first pub-

lished by Beta Publishing in 2011 and currently appears quarterly, in addition to our well

known MEDICAL CORPS INTERNATIONAL FORUM Magazine. On the occasion of the 20th

Nuclear Medical Defence Conference, the present special issue of CHALLENGE is dedicated to

selected topics in radiation protection and research for a mutual international exchange. We

encourage you to contribute to our magazine and welcome your feedback. From the publisher,

we wish a pleasant stay in Munich, and we are sure that the outstanding scientific program

will give you an interesting and fruitful conference with many networking opportunities and

hope that you will enjoy your stay in Munich.

Sincerely

Heike Lange Helmar Winkel

Publisher Executive Director

Beta Publishing Group Beta Publishing Group

Welcome letter

1. Fukushima

1 Institute of Development, Aging and Cancer, 2Graduate School of Sci-ence, 3Graduate School of Agricultural Sciences, 4Center for theAdvancement of Higher Education, 5Graduate School of Dentistry,Tohoku University, 6Graduate School of Science and Engineering, Yama-gata University, 7Faculty of Agriculture, Niigata University, 7NationalInstitute of Public Health, 8RIKEN Bioresource Center (BRC), 9NationalInstitute of Radiological Sciences of Japan.

The Fukushima Daiichi Nuclear Power Plant (FNPP) accidentreleased large amounts of radioactive substances into the environ-ment. We collected organ specimens from over 200 cows, 50 pigsand 4 monkeys within and adjacent to the 20-km radius evacuationzone of FNPP. We are performing determination of the concentrationof radionuclides in the organs of animals. In all the specimens exam-ined, photopeaks of both Cesium-134 (134Cs, half-life: 2.065 y) and137Cs (30.07 y) was observed in a gamma-ray spectrometer and thelevel of their radioactivity concentrations in each organ was quite sim-ilar after the decay correction from the day of specimen collection toMarch 15, 2011, the day of major release of radionuclides fromFNPP. Linear correlation was found between radioactivity concentra-tion of 137Cs in whole peripheral blood (PB) and that in each organ.The slope of the regression line was organ dependent and was thehighest in the skeletal muscles. Thus, radioactivity concentrations of134Cs and 137Cs in the organ could be estimated from their radioac-tivity concentrations in PB. The level of radioactive Cs in an organ ofboth fetus and infants was higher than that of the correspondingmaternal organ. Over all levels of radioactivity concentration of 137Cswere dependent on the geological location and environment of thecattle examined. Radioactivity concentration of 137Cs in PB was sig-nificantly higher in pigs than in cows but the concentration ratio of PBto skeletal muscles was significantly lower in pigs than in cows.Organ-specific deposition of radionuclides with relatively short half-lifewas detected, such as silver-110m (110mAg, 249.8 d) in the liverand tellurium-129m (129mTe, 33.6 d) in the kidney of adult cattlewithin the evacuation zone. However, these were not detectable inany organ of fetus and infants of cows, pigs or monkeys. This study isthe first to reveal in detail the distribution of radionuclides attributedto the FNPP accident through the entire body of animals.

Comparison of performance of three different dosimetricsystems used by US Armed Forces in aftermath of FukushimaDaiichi reactor accident

A. RomanyukhaNaval Dosimetry Center, US Navy, Bethesda, MD, USA

On 11 March 2011, the 9.0 magnitude earthquake and subse-quent massive tsunami caused severe damage of several reactors atthe Fukushima Daiichi Nuclear Power Plant. Three of the six reactorsexperienced a partial meltdown which resulted in the release ofradioactive materials into the environment. The U.S. Department ofDefense (DoD) support operation in response to this disaster wasnamed Operation Tomodachi (“friend” in English) and consisted ofmany activities including humanitarian aid and disaster relief toJapan. There was immediate participation of U.S. Navy vessels, air-planes, and helicopters that were already in the area at the time ofthe disaster or arrived shortly afterward. The Naval Dosimetry Center

Oral Presentations

An accident at the Fukushima Daiichi Nuclear Power Plant in2011Makoto Akashi, Executive DirectorNational Institution of Radiological Sciences (NIRS), Chiba-city, CHIBA,JAPAN

An earthquake measuring 9.0 struck the northeast coast of HonshuIsland of Japan at 14:46 on March 11, 2011, triggering a hugetsunami. The earthquake left 15,881 people dead and 2,668 missing(as of March 8, 2013). In the northeast coast area, all nuclear powerplants (NPPs) in operation automatically shut down immediately afterthe earthquake. Since the connection to off-site power for units faileddue to the earthquake, off-site power was lost immediately at theFukushima Daiichi NPPs of Tokyo Electric Power Company (TEPCO).Therefore, the diesel generators, designed to start up after losing off-site power, began providing electricity to the cooling system of thereactors. Unfortunately, a huge tsunami disabled the emergency dieselgenerators. Thus, a station blackout was caused and the cooling sys-tems lost their function. As consequence, large amounts of radioactivematerials were released into the environment and the trouble with thecooling systems also led to hydrogen explosions and core melt-down.The major nuclides released were I-131, Cs-134 and Cs-137. Thedeposition of these radioactive materials on land resulted in a highambient dose of radiation around the NPPs. Therefore, almost170,000 people had to be evacuated or stay indoors. This earthquakealso affected infrastructures such as the monitoring system for radia-tion and the telecommunications system. Since community lifelinessuch as the water supply and electricity were severely damaged, theoff-site command center located 5 km from the NPPs could not func-tion. Moreover, even simple countermeasures for decontaminationsuch as removing clothes and wiping the skin with wet towels couldnot be performed for evacuees at the shelters. Furthermore, hospitalsthat had been designated as radiation emergency hospitals lost theirfunction because of the damage to their facilities, and they were alsolocated in the evacuation areas. Personnel of the local fire depart-ments and hospitals were also asked to evacuate and the staff of localambulance service refused to transport work ers contaminated to ahospital, the reason being no hospitals could be found to receivethem. Thus, a lack of knowledge prevented the se personnel frombeing able to transport or accept contaminated work ers from the NPPsbecause of concerns about the health effects of radia tion from work-ers. In this accident, no responders including on-site workers from theNPPs or residents around the site required treatment from the view-point of radiation exposure. Radiation accidents can cause medical, environmental, psychological, and economic problems. We lear nedagain that basic knowledge on radiation and its effects is extremelyimportant for health care providers.

Distribution of Artificial Radionuclides in Animals As s oc i a t edwith the Fukushima Daiichi Nuclear Power Plant Accident

Manabu Fukumoto1, Yasushi Kino2, Tomokazu Fukuda3, Emiko Isogai3, Tsutomu Sekine4, Hisashi Shinoda5, Yasuyuki Abe6, HideakiYamashiro7, Masatoshi Suzuki1, Yoshikazu Kuwahara1, Motoi Fukumoto1, Hidekazu Nihei2, Yosuke Sano2, Ayumi Irisawa2, Tsut omuShimura7, Yuichi Obata8, Shin Saigusa9

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(NDC) responded to the accident by calibrating and delivering over 11,000additional dosimeters to monitor radiation doses of the Navy personnel inJapan. In this presentation we will provide details on the measured dosesand validation of standard procedures used under the conditions of thisnuclear accident. Two other branches of the US Armed Forces (Army andAir Force) also took active participation in Operation Tomodachi. Eachbranch of the US Armed Forces has its own Dosimetry Center, except theMarines which use Navy dosimetry, and use different dosimetric systems tomonitor radiation doses. The Navy uses copper-doped LiF thermolumines-cent dosimeters (TLD) produced by Thermo Fisher Scientific under thename of Harshaw model 8840/8841. The US Air Force Radiation Dosime-try Laboratory employs Thermo Electron EPD-2 (electronic pocket dosime-ter) devices. The U.S. Army Dosimetry Center monitors its personnel withcarbon doped aluminum oxide optically-stimulated luminescence dosime-ters (OSL) produced by Landauer under the name InLight. In some loca-tions during Operation Tomodachi Navy personnel were monitored withdosimetry issued by the Army or Air Force. In order to provide a consis-tency in dose records for Navy personnel the NDC conducted proficiencytesting of all three dosimetric systems performance accordingly to ISO/IEC17011 and ISO/IEC 17025. Here we also report analysis and comparison ofall three dosimetry systems performance which are currently in use by theU.S. Armed Forces.

Disclaimer. The views expressed in this abstract are those of the authorand do not necessarily reflect the official policy or position of the Depart-ment of the Navy, Department of Defense, nor the U. S. Government.

Recovery and Resilience after a Nuclear Power Plant orRadiological/Nuclear Disaster: A Medical-Decision Model forManaging an Effective, Timely, and Balanced Response**

C. Norman Coleman, MD*

Office of the Assistant Secretary for Preparedness and Response,Dept. of Health and Human Services and Radiation Research Pro-gram, National Institute of Health (NIH), USA

The multiple disaster incident in Japan in 2011 provided real-timeexperience with decision-making in a complex mass casualty inci-dent. The evolving situation at the Fukushima Daiichi Nuclear PowerPlant over the first few weeks required decisions to be made whenthere was a limited amount of data and much potential uncertainty asto the outcome. Resilience is a key goal that requires response andrecovery

activities that are appropriately safe, timely, effective, and wellorganized. Informed decisions must be made, and the logic behindthem communicated during the evolution of the incident before thefinal outcome is known. We* propose a medical decision model bywhich to make key health-related decisions that are central drivers tothe overall incident management. In this model, on-site decision mak-ers are empowered to make interim decisions without undue delayusing readily available and high-level scientific, medical, communica-tion, and policy expertise. Ongoing assessment, consultation, andadaption to the changing conditions are done as in medical carewhen competing risks must be continuously considered as the clini-cal course evolves. Indeed, to not make a decision is a decision.Given the central role of health and medical issues in all disasters, wesuggest that this medical decision model be considered for effectivemanagement of complex, large-scale, and large consequence incidents.

*Coleman CN et al Disaster Med Public Health Preparedness. 2013,in press (This abstract is an edited version from this paper)**This abstract is the opinion of the author and does not representopinion or policy of DHHS or any agency within the US Government.

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2. Radiation risk perception and communication

Ionizing radiation: risk perception, risk communicationW. Rüegg (Consultant)

ObjectivesIn the public there is a widespread and often extremely exaggerated

fear from ionizing radiation. In order to quantify this fear, we executedseveral surveys using different exposure scenarios. Methods

These scenarios included internal and external exposure with lowdoses as well as one with a very high dose (direct exposure to the con-tent of a CASTOR container without screening). One survey has beenexecuted with over 100 highly educated adults form several Europeancountries; another with a representative sample of over 1000 peoplesfrom Switzerland. We asked to estimate the risk of death or of gettingcancer or to compare the risk from radiation with other risks or with nat-ural radiation. The answers were compared with risk estimations usingICRP risk models (linear extrapolation from high dose, LNT).Results and Conclusions

The surveys revealed an extremely undue risk perception. Most peo-ple overestimated the risk by a factor of up to a million or more, espe-cially in the case of low doses. The answers also showed an extremerange of perceptions, in some scenarios over 10 orders of magnitude. Asurprisingly large fraction of the people thought that even a very lowdose leads always to cancer, overestimating the risk billion-fold or more.Risk communication

In case of ionizing radiation, risks communication is very difficult, inparticular for low and very low doses. For the general public, units likeBq, Sv, Gy are meaningless. Comparisons with legal limits are not veryhelpful either. Absolute risk values are better suited, but also difficult to

interpret for non-experts. The best way is probably the comparison withother, well accepted or at least tolerated risks. Such risks may be: smok-ing one single cigarette, being killed by a meteorite, crossing a street,walking just one step and many others. One may also compare the riskof e.g. an exposure of 1 mSv to the beneficial effect of eating 1 kg offruits/vegetables or exercising for one hour. All such estimations arebased on rather well known risks or benefits of high “doses” and usingLNT (extrapolation to small doses).

Dirty bomb event in urban environment – radiation exposureof public and emergency personnel

F. Langeformer at GRS - Gesellschaft für Anlagen- und Reaktorsicherheit mbH,Cologne, Germany

Malicious acts with explosives are frequently carried out worldwide. Itis conceivable that radioactive material is added to the explosive therebycreating a “dirty bomb” or radiological dispersal device (RDD) with theintention to achieve dispersion and resulting contamination of larger sur-face areas, e.g. within urban environment. Authorities and specialisedinstitutions responsible for emergency response and radiological mitiga-tion are obliged to be prepared for such threats. This involves the analy-sis of such kinds of threat scenarios, the assessment and managementof resulting radiological situations in case of such an event, the imple-mentation of appropriate exercises and the training of operational per-sonnel.

Widespread fear in the population of radioactive material and associ-ated ionizing radiation may lead to distorted perception of the resultinghazard situation, thereby requiring enhanced competence of emergency

authorities and involved support institutions for crisis management. A major problem will arise from anxieties caused within the public and

also among emergency personnel to implement technically adequate andjustified protection and remediation measures which are in accordancewith the ALARA principle.

Based on examples with unfavourable but not extreme conditions of adirty bomb event associated radiological situations will be illustra ted. Var-ious contamination scenarios and exposure conditions regard ing emer-gency personnel and affected population will be covered and required

protection measures addressed. For gamma-emitting radionuclides andfor beta-emitting radionuclides wirh higher decay energies the radiationexposure of persons is dominated by external radiation. Due the orders ofmagnitude higher radio-toxicity of alpha-emitting radionuclides whenincorporated by inhalation compared with beta-/gamma emitters soliddata are needed regarding magnitude and time dependence of resus-pension from contaminated surfaces. Radiation exposure from inhalationof airborne particulates following resuspension of deposited radioactivedust and required respiratory protection will be covered in some detail.

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3. Radiation emergency medical preparedness and response

Software architecture of the BOOSTER comprehensiveapproach to crisis management in case of dirty bomb attacks

T.O. MüllerInstitut für Kern- und Energietechnik, Karlsruher Institut für Technologie,Eggenstein-Leopoldshafen, Germany

The BOOSTER project (BiO-dOSimetric Tools for triagE to Respon-ders) funded by the EU aims to effectively support and coordinate firstresponders in case of a terroristic attack with a Radiological DispersalDevice (dirty bomb) on a large crowd (e.g. a soccer game). The systemconsists of a wide variety of hardware devices and software methods fortriage, i.e. to fast and efficiently determine the contamination of manypeople in a relatively short time. This includes a gamma camera todetermine areas of contamination, sample based bio-dosimetric sensors,hand held real time radiological measurement devices, sensors withspectroscopic means to identify radionuclides, rugged PDAs or smart-phones for the on-site data collection etc. The information is managedby an adapted Commanded and Control (C2) software based on SIMA-COP which is developed by the University of Valencia. Linked to this C2component is a decision support system (DSS) that provides tools likechecklists, geographic information system (GIS), evaluation of bio-dosi-metric measurements, dispersion calculation, etc. The methodologicalsupport is based on the TMT Handbook (Triage, Monitoring and Treat-ment-Handbook of people exposed to ionizing radiation following amalevolent act). A laptop with the SIMACOP installation serves as thecentral component of the system. The sensor devices are connectedusing an ad-hoc WLAN. The DSS consists of a web server and a webclient. The DSS web server is installed on a virtual machine (emulatedPC). For security reasons SIMACOP and DSS share a virtual private net-work (VPN). As the DSS client is based on generated JavaScript (GWT)any standard web browser is suited to act as frontend. Hence the DSSclient is usable at the triage stations, the command center, on mobiledevices like rugged PDAs or smartphones. All processed data has to besynchronized with the SIMACOP dedicated database. To prevent directaccess web services were established between the two systems.

Presentation title: Integrating Evidence-basis into NationalPreparedness for Medical Response to a Nuclear Detonation

J. KoernerAssistant Secretary for Preparedness and Response, US Department ofHealth and Human Services, Washington, DC, USA

A nuclear detonation in a major city will create unique challengesrelating to the large number, presentation, and distribution of casualties.Healthcare capacity and capabilities for road, power, and communica-tion infrastructure will be severely impaired. Planning for such a cata-strophic incident requires an approach that joins applicable evidencefrom relevant disciplines to lessons learned from other natural and man-made catastrophic incidents. We developed a systematic approach tointegrate evidence-basis, response lessons, and preparedness principlesto better inform and coordinate medical response with broader emer-gency management requirements. This system prioritizes maximum

lives saved in an environment with severe scarcity of medical resources.Included are national planning strategies for preparedness andresearch, clinical and technical guidance, diagnostics development andsurge, and tools to build knowledge and facilitate planning for all users,disciplines, and levels. This presentation will provide background onhow this is conducted in the US, summarize existing resources, andintroduce new resources. Included will be a functional discussion of anintegrated suite of web-based resources: “Radiation Emergency MedicalManagement (REMM)”; the “State and Local Planners Playbook forMedical Response to a Nuclear Detonation”; and the just published,”Medical Planning and Response Manual for a Nuclear Detonation: APractical Guide”. Additionally this discussion will propose a novel “Inte-grated Clinical Diagnostics System” and a concept to enhance coordi-nated hematology surge as a potential means to build coordinated ana-lytical and data management capacity for lymphocyte depletion kineticsas a biodosimetry tool for triage. This content represents the personalviews of the author and does not express the opinion or policy of theU.S. Department of Health and Human Services or its components andno statement should be construed as an official position.

BARDA Development of Medical Countermeasures AgainstRadiological and Nuclear Threats

R. ManningBiomedical Advanced Research and Development Authority, Washing-ton, DC, USA

The BARDA (Biomedical Advanced Research and DevelopmentAuthority) mission is to ensure the availability of medical countermea-sures to address public health emergencies. One of the threat areas ofinterest is Radiological-Nuclear (Rad/Nuc). At BARDA we are employ-ing a comprehensive portfolio approach to development and acquisitionof products. BARDA occupies a unique niche in U.S. Government bio-medical research and development activities. We emphasize mid- tolate-stage medical product development. We aim to work with industryto move product candidates through the drug development pipeline.BARDA employs staff with experience in product development andmanufacturing. Short Term goals include the repurposing of pharma-ceuticals already in use for related indications. We also intend toexpand Advanced Research and Development (AR&D) support of drugswhich demonstrate efficacy. Emphasis will be on blood and burn prod-ucts. We would like to achieve active Rad/Nuc AR&D contracts for allfor Acute Radiation Syndrome (ARS) subsyndromes. An importantactivity will be to maintain support for the development of well character-ized animal models. We will also continue to partner with our U.S. inDOD (CBMS, DTRA) and NIH. Mid Term we will focus on collectingdata which support pre-Emergency Use Authorization by the FDA. Wewant to increase our partnerships with Big Pharma and with Interna-tional partners. Long term we will strive to meet requirements withmulti-purpose drugs (using a stock rotation plan) which have a threatagent clinical indication. We intend to leverage international resourceswherever possible. We will also need to address polypharmacy efficacy

issues and drug-drug interactions. In this talk the current BARDA portfo-lio of projects will be discussed in detail.

MULTIBIODOSE: multi-disciplinary biodosimetric tools tomanage high scale radiological casualties - results andconclusions

A. Wojcik1, H. Romm2, U. Oestreicher 2, H. Thierens3, A. Vral3, K.Rothkamm4, E. Ainsbury4, M. Bendertitter5, F. Barquinero5, P. Fat-tibene6, C. Lindholm7, L. Barrios8, S. Sommer9, K. Woda10, H.Scherthan11, C. Beinke11, B. Vojnovic12, F. Trompier13 A. Bajinskis14, A.Jaworska14

1SU, Sweden, 2BfS, Germany, 3UGent, Belgium, 4HPA, UK, 5IRSN,France, 6ISS, Italy, 7STUK, Finland, 8UAB, Spain, 9INCT, Poland,10HMGU, Germany, 11UULM, Germany, 12UOXF, UK, 13EURADOS,14NRPA, Norway

In the event of a large scale radiological emergency biological dosime-try is an essential tool that can provide timely assessment of radiationexposure to the general population and enable the identification of thoseexposed people, who should receive immediate medical treatment. Anumber of biodosimetric tools are potentially available, but they must beadapted and tested for a large-scale emergency scenario. These meth-ods differ in their specificity and sensitivity to radiation, the stability ofsignal and speed of performance. A large scale radiological emergencycan take different forms. Based on the emergency scenario different bio-dosimetric tools should be applied so that the dosimetric informationcan be made available with optimal speed and precision.

The aim of this multi-disciplinary collaborative project was to analyse avariety of biodosimetric tools and adapt them to different mass casualtyscenarios. The following biodosimetric tools were established, improvedand/or validated: the dicentric assay, the micronucleus assay, thegamma-H2AX assay, the skin speckle assay and the blood serum pro-tein expression assay. In addition EPR/OSL dosimetry in components ofpocket electronic devises is investigated. The assays were chosenbecause they complement each other with respect to sensitivity, speci-ficity to radiation and the exposure scenario as well as speed of perform-ance.

The project involved the key European players with extensive experi-ence in biological dosimetry and finished in April 2013. Training wascarried out and automation and commercialisation pursued. An opera-tional guide was developed and disseminated among emergency pre-paredness and radiation protection organisations. The final deliverable ofthis project was establishment of a biodosimetric network that is fullyfunctional and ready to respond in case of a mass casualty situation.The main achievements and conclusion from the project will be pre-sented.

The project was funded by the FP7 Security program. URL:http://www.multibiodose.eu.

RENEB – Realizing the European Network of BiologicalDosimetry

U. Kulka1, E.A. Ainsbury2, M. Atkinson3, J.F. Barquinero4, C.Bassinet4,L. Barrios5, C. Beinke6, A. Cucu8, F. Darroudi9, P. Fattibene10,O. Gil11, E. Gregoire4, V. Hadjidekova12, S. Haghdoost13, R. Herranz14,A. Jaworska15, C. Lindholm16, R. Mkacher17, S. Mörtl3, A. Montoro18, J.Moquet2, M. Moreno14, A. Ogbazghi17, U. Oestreicher1, F. Palitti19, G.Pantelias20, I. Popescu8, M.J Prieto14, H. Romm1, K. Rothkamm2, L.Sabatier17, S. Sommer21, G. Terzoudi20, A. Testa22, H. Thierens23, F.Trompier4, I. Turai7, V. Vandersickel23, P. Vaz11, P. Voisin4, A. Vral23, F.Ugletveit15, A. Wieser3, C. Woda3, A. Wojcik13

1Bundesamt fuer Strahlenschutz (Germany), 2Health Protection Agency(United Kingdom), 3Helmholtz Centre Munich (Germany), 4Institut deRadioprotection et de Sûreté Nucléaire (France), 5Universitat Autonomade Barcelona (Spain), 6Bundeswehr Institut für Radiobiologie / Univer-sität Ulm (Germany), 7National Research Institute for Radiobiology &

Radiohygiene (Hungary), 8Institutul National de Sanatate Publica(Romania), 9Leiden University Medical Center (The Netherlands), 10Isti-tuto Superiore di Sanità (Italy),11IST/ITN Instituto Superior Técnico, Uni-versidade Técnica de Lisboa (Portugal), 12National Center for Radiobiol-ogy and Radiation Protection (Bulgaria), 13Stockholm University(Sweden), 14Servicio Madrileño de Salud - Hospital General UniversitarioGregorio Marañón, 15Norwegian Radiation Protection Authority (Nor-way), 16Radiation and Nuclear Safety Authority (Finland), 17Commis-sariat à l´Énergie Atomique (France),18Fundacion para la Investigationdel Hospital Universitario la Fe de la Comunidad Valenciana (Spain),19University of Tuscia (Italy), 20National Centre for Scientific ResearchDemokritos (Greece), 21Institut Chemii i Techniki Jadrowej (Poland),22Agenzia Nazionale per le Nuove Tecnologie, L´Energia e lo SviluppoEconomico Sostenibile (Italy), 23Universiteit Gent (Belgium)

The RENEB project (www.reneb.eu) aims at establishing the Euro-pean network of biological and retrospective dosimetry. Started in Janu-ary 2012 and supported by the European Commission for 4 years (grantagreement no. 295513) a total of 23 organisations from 16 EU countriesjointly realize the project.

Following large-scale radiological accidents or malevolent actions themedical and radiological triage of patients according to the degree oftheir injuries and the level of their radiation exposure will be required inthe shortest possible time. Besides individuals, who were actuallyexposed to doses of ionizing radiation that may cause acute healtheffects, there will be the huge number of ‘worried well’, i.e. persons whoare extremely distressed but have not actually received radiation doseslikely to cause any acute health effect. A lesson learned from previousincidents is the high importance to identify those “worried well” in orderto prevent the healthcare infrastructure being overwhelmed and to avoidsocio-economic harm. In both contexts, biological dosimetry is an essen-tial tool to estimate an actual absorbed dose without being influenced bytemporal or individual variations in blood counts or confounding factorssuch as chemical agents or psychogenic reactions. Since in large-scaleradiological emergency scenarios the number of people that may needto be screened could easily exceed the capacity of a single or even anumber of laboratories, biodosimetry networking has been recognized asa sensible and important emergency response strategy. Thus, a wellorganised and harmonised cooperative action between the RENEB labo-ratories will offer a realistic chance for a rapid and trustworthy doseassessment urgently needed in an emergency situation. As a conse-quence, implementing such a network in national and international radi-ation emergency preparedness and response systems will significantlyimprove the capabilities in case of a large-scale radiological emergency.

Advances in a Framework to Compare and IntegrateBiodosimetry Methods for Planning Triage

A. Barry FloodDept. of Radiology, Community & Family Med., and The DartmouthInstitute of Health Policy and Clinical Practice, The Geisel School ofMedicine at Dartmouth, NH, USA

In a major event involving individuals who may have been exposed toionizing radiation, the capacity to rapidly and accurately determinewhether or not exposure warrants further medical evaluation or immedi-ate treatment for acute radiation syndrome is crucially important foreffective medical management and conservation of available manpower.Current guidelines are inadequate to address the response to thousandsof people in a compromised medical infrastructure and need to be re-evaluated and advanced methods need to be considered using a com-parative framework and integrative plan. Biodosimetry methods need tobe evaluated for their capacity to effectively meet the needs for triagedecisions in disasters including: the validity and accuracy of the doseestimate, the eligibility of the impacted population to be measured (e.g.,feasibility and timeliness of obtaining a sample or measurement, specialconsiderations in combined injury, and mitigating circumstances unre-

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lated to the disaster), and the capacity to deliver timely patient-specificof information.

Our framework for comparative effectiveness for triaging a large popu-lation assuming a major radiation disaster includes: obtaining updatedand new information for assessing the effectiveness, efficiency and fea-sibility of the framework; modeling the data in a variety of scenarios; andbuilding an integrated approach to triage and updated data on advancesin the biodosimetric methods.

We reexamined 3 dosimetry methods commonly used in plans formedical triage (time-to-emesis, lymphocyte depletion rate, dicentricchromosome analysis) and 3 advanced methods under development(two types of cytogenetic changes: block micronucleus and γ-H2AX andin vivo electron paramagnetic resonance tooth dosimetry. We concludethat the comparative framework, modified to take into account inade-quate and compromised healthcare and transportation infrastructures isuseful to evaluate alternatives.

Rapid clinical triage of radiation injured patients usingstandard blood cell counts within 3 days after whole bodyradiation exposure

M. Port, T. Knie, H. Dörr, B. Pieper, A. Ganser, D. Graessle, V.Meineke and M. AbendHematology, Hemostasis, Oncology and Stem Cell Transplantation, Han-nover Medical School, Hannover, GermanyAim

To establish easy and rapid clinical triage of radiation injured patientswe examined the relevance of blood cell counts on day one to three afteraccidental or planned whole body irradiation. With this for physician’s

easy to use procedure we want to examine whether prediction of theclinical outcome is possible.Material and Methods

Blood cell counts (BCC) from radiation accident victims and for valda-tion from irradiated patients were correlated with their clinical outcome(severity scores H1-4). Unirradiated controls (H0) were taken from anout-patient facility for model construction and for independent valida-tion. Binary categories comprising (1) H0 versus H1-4, (2) H0-1 versusH2-4 and (3) H0-2 versus H3-4 severity scores were discriminatedemploying logistic regression analysis. The test sample contained 454BCC from 267 individuals. The validation step was performed on a sec-ond independent group again chosen from radiation accident victims,from hospital patients and from outpatients.Results

It is possible to separate H0 from H1-4 score using BCC of the firstthree days after irradiation due to a developing lymphopenia and granu-locytosis. With increasing irradiation and haematological severity scorethe difference became more prominent. Validation of the models allowedan almost complete discrimination of HO versus radiation exposed indi-viduals (negative predictive value >94%) for all three days while the trueidentification of radiation exposed individuals increased from day 1 (pos-itive predictive value 78 -89%) to day 3. The models allowed no predic-tion for 10.9% of the test samples. Conclusion

In the first 3 days after radiation exposure clinical outcome of radia-tion injured patients can be rapidly and reliably predicted by the use ofperipheral blood cell counts. Detailed results and limitations will be -presented.

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4. National, international and global radiation accident management

Improvement of Radiation Emergency Medical ResponseSystem of Korea

M. Cho, M.D. Seung-suk Lee, M.D. Hyo-rak Lee, M.D. Jin-kyung Lee,M.S Wi-ho Ha and colleaguesRadiation Emergency Medical Team, National Radiation EmergencyMedical Center in Korea Institute of Radiological & Medical Sciences,Seoul, Republic of KoreaBackground

There are 23 nuclear reactors in operation at 4 sites and 9 are underconstruction in Korea. To respond to radiation disaster, Korea govern-ment have designated National Radiation Emergency Medical Center(NREMC), 9 primary and 12 secondary institutes as a national responsesystem. The Fukushima combined disaster have aroused interest incapacity of national radiation emergency medical preparedness systemof Korea.Purpose

This research aims to analyze the capacity of each radiation emer-gency medical institute and to plan the long-term strategy for \'establish-ment of regional preparedness system about each nuclear power plantsite\'Methods

NREMC developed survey, evaluation index of infrastructure level,prediction program for medical demand according to radiation disasterscenario, short- and long-term plan. Seven-point scale is applied in thesurvey. The evaluation index of infrastructure level is graded on a sixpoint scale. The field of evaluation consists of on-site response, emer-gency room, psychological support, radiation burn, bone marrow trans-plantation, internal contamination and acute radiation syndrome.Results

If 1000 patients appear in the situation of combined disaster, accord-ing to simulation analysis the medical demand exceed the capacity of

national radiation emergency medical response system. If 250 patientsappear in case of radioactivity leakage accident, it is expected to havesome difficulty within capacity of regional response system, but it is pos-sible to respond within national level.

Conclusion : For the last ten years NREMC have established nationalradiation emergency medical response system. The current level can beevaluated by comprehensive indicators and it is possible to plan the fur-ther development.

Radiation exposure case management after incorporation ofradioactive materials

G. Goulko, G. and V. MeinekeBundeswehr Institute of Radiobiology affiliated to the University of Ulm,Munich, Germany

The wide use of radioactive material in medicine and industry makesan accidental incorporation, as well as misuse for terroristic purposespossible. The preparedness for such a realistic scenario includes thedevelopment of an Information and Communication System to assist arapid estimation of exposure doses, effectiveness of countermeasuresand prognosis of health effects. The estimates of internal doses arebased on measurements of the activity in the body or in excreta. Theinterpretation of measured values requires dosimetric and biokineticmodels describing the metabolism of incorporated radionuclides. Forthis purpose a dedicated computerized support tool has been devel-oped. It allows to assess the amount and distribution of radionuclides inthe body and to calculate resulting radiation doses over specific timeperiods. Because the ionizing radiation energy deposited in organs fromradionuclides incorporated in the body cannot be measured directly,internal doses are estimated principally from in vivo or in vitro bioassay.To make a reliable estimate of the dose a number of parameters mustbe known: incorporated radionuclides (including chemical-physical

characteristics), route of intake, elapsed time after intake, criticalorgans, radioactivity distribution pattern, biokinetic parameters. The aimof this study is the design and implementation of user friendly Med-A-Defence-Information-Communication-System to support decorporationmeasures and risk assessment after incorporation of radioactive materi-als. The following input information for the calculations is needed: age,gender, incorporated radionuclides, intake scenario, bioassay data(probe type, activity), biokinetic parameters (pathway depending), meas-urement time. This information as well as calculated doses will be docu-mented. It includes following modules: Personal Data and Case Descrip-tion, Dosimetry (exposure type, bioassay data, whole body and organdoses at measurement time and prognosis), Decision-making (recom-mendation of countermeasures based on estimated actual and futureexposure doses, assessment of effectiveness of countermeasures) andRisk Estimation. The output information is: exposure dose at the time ofmeasurement, prognosis of future dose without countermeasures andeffectiveness of proposed countermeasures in terms of averted dose.

Improving of public involvement in European nuclearpreparedness and safety

A. Fucic1, P. Dorfman2

1Warwick Business School, University of Warwick, UK.2Institute for Medical Research and Occupational HealthAbstract

Nuclear plants, Uranium storages and military sources are currentlysites of potential nuclear accidents in Europe (EU). Major nuclear acci-dents at Chernobyl and Fukushima point to the importance of ongoingand active public involvement in nuclear accident preparedness andimplementation of nuclear accident management protocols. RecentEuropean “stress tests” reported in COM (2012)571 evaluated risk andsafety assessments for 132 nuclear reactors located on 58 sites. Stresstests identified major concerns on the level of facilities (lack of seismicinstrumentation) and not coherent methodologies in assessment of thesafety implications. Further, aside from normal access to publishedreports, there was no direct public involvement in the stress testprocess. This results in the perpetuation of the same approach in whichEuropean radiation protection organizations and agencies predominantlycommunicate horizontally, with vertical two-way communication withgeneral public being largely missing. Currently, EU level nuclear emer-gency education information provision is very limited. Post-Fukushima,effective management of any future nuclear accidents will need clearinformation provision to the general public about radiation risk andemergency planning response, and this information should be availablein an ongoing basis. General public awareness of emergency planningand radiation protection preparedness will contribute to panic-reduction,thereby enabling improved emergency response. In the event of anemergency, there is a need to establish a strong and on-going risk communication media network in order to prepare and disseminateinformation. Additionally, local and regional community emergency

response facilitators should be able to act in tandem with statutoryorganizations. Structured and coordinated communication about nuclearrisk and emergency planning management with the general publicshould be a very high priority for the EU. It is unfortunate to reflect thatcurrently national agencies in EU are not oriented towards general populations on this topic.

Improvements in Medical Care and Treatment of IrradiatedIndividuals in Switzerland

D. StorchRadiological Protection, Federal Office of Public Health, Bern, Schweiz

Japan found itself after the devastating earthquake of 11 March 2011confronted to a very complex crisis with tens of thousands of deaths andinjuries, devastating damage to infrastructure and a number of big technological accidents, as in the nuclear power plants in Fukushima.

In May 4th 2011, the Federal Council installed a interdepartmentalworking group called IDA NOMEX, which should examine, based on theexperience of Japan, if and how the emergency response in Switzerlandcould be improved.Improvements in Radiation protection

a better coordination among the federal agencies within the organiza-tion of sampling and measurement, and the evaluation of the radiologi-cal situation; harmonization of exposure limits and reference values tothe international standards; review of the treatment of irradiated peopleand the concept of \"Kontaktstelle\" for affected persons.Care and treatment of heavily irradiated individuals

Today it is not clear who is responsible for the coordination of care ofstrongly irradiated persons. Several federal agencies are involved, but noone has the lead. It needs to be checked, which measures should betaken to ensure the care of radiation victims in accordance with interna-tional standards (WHO REMPAN).Kontaktstelle

The “Kontaktstelle” is a reception center which helps to increase incapacity of the highly stressed health care in an event with increasedradioactivity. The hospitals and doctors\' offices should, if possible,relieved. The “Kontaktstelle” acts as assessment of radiological condition, triage and psychological care to persons of the population inan accident with increased radioactivity.Decontamination-Hospitals

In Switzerland currently 16 existing hospitals are referred as deconta-mination hospitals with the following goals:

n Prevention of contamination of the hospital (protecting people[patients, staff, visitors, etc.] and infrastructures in the hospital).

n Providing patients from any further damage from prolonged exposureof pollutants and radioactivity. Best possible medical care should beprovided (triage, life-saving measures).These kind of hospitals require a wide range and large quantities of

various antidotes for different events.

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5. External exposure assessment

External dose assessment in a radiological mass casualtyscenario using OSL on mobile phones

C. Woda1, C. Bassinet2, P. Fattibene3, E. Bortolin3

1Helmholtz Zentrum Muenchen, Germany2Institut de Radioprotection et de Sûreté Nucléaire, France 3Istituto Superiore di Sanità, Italy Background

Within the framework of the EU-FP7 project MULTIBIODOSE, a com-plementary approach is being implemented to assess the external doseof potential radiation victims by using either EPR of display glass and/or

OSL of electronic components on the circuit board of mobile phones.Here we describe and present results of the OSL assay. Methods

Electronic components, comprising ceramic chip resistors, inductorsand capacitors were extracted from seventy-five different mobile phonesand analyzed by OSL in three laboratories. Main properties such asoccurrence, dose response, detection limit and fading were studied. Results

Resistors were present in all, inductors in most and capacitors inabout 50% of the mobile phones investigated. All components showed a

linear dose response up to 10 Gy, with mininum detectable doses beingon average 30, 8 and 200 mGy for resistors, inductors and capacitorsrespectively. A second common feature was signal fading with storagetime, only resistors, however, showed a comparable homogeneous fadingrate and were thus chosen as the material of choice for further investiga-tions. Two measurement sequences were developed, optimized for eitherspeed of response (dose measurement in under five minutes) or accuracy.The protocols were validated in blind tests among the three laboratories.

ConclusionsAlumina-rich resistors on the circuit board of mobile phones have

high potential for rapid individual dose assessment in a radiologicalmass casualty scenario.ACKNOWLEDGEMENTS

The research leading to these results has received funding from theEuropean Union\'s Seventh Framework Programme (FP7/2007-2013)under grant agreement n° 241536.

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6. Decontamination measures and monitoring

Radiation monitoring of larger numbers of people viaRadiation Monitoring Units

N J Thompson, M J Youngman, J Moody, N P McColl, D R Cox, AWilton, J Astbury, S Webb and S L ProsserHealth Protection Agency, Didcot, UK

A variety of incidents may require people to be monitored for radioac-tive contamination. Experience in the UK after Chernobyl (1986), theLondon polonium incident (2006) and Fukushima (2011) showed thatthere may be significant demand for early, sustained and scalable deliv-ery of appropriate screening for radioactive contamination with associ-ated provision of information. At a local level in the UK, many plans tomonitor people following an incident are in the primary stages of devel-opment. Producing a people monitoring plan involves blending scientificrequirements, crowd management, patient confidential data acquisitionand logistical needs of different organisations and large numbers of peo-ple. A need to provide guidance on how to consistently initiate and oper-ate a facility for radiation monitoring was identified by the HPA. Guid-ance for radiation monitoring of large numbers of people at one venue,known as a Radiation Monitoring Unit (RMU), has been developed. Thisguidance has recently been implemented in two local level emergencyplans and trialled at exercises involving members of the public. Lessonshave been learned from the production of these plans and the associ-ated exercises. The HPA guidance along with lessons learned and nextsteps in developing people monitoring in the UK will be presented.

Combined drug and surgery treatment of Plutoniumcontaminated wounds: indications obtained using a rodentmodel

N. Griffith

Laboratoire de RadioToxiciologie, CEA/DSV/iRCM, CEA, Arpajon, FranceBackground

There is an important requirement following accidental actinide con-tamination of wounds to limit the dissemination and retention of suchalpha-emitting radionuclides. One approach is resection of the woundsite but it has been hypothesized that this may lead to contaminantrelease and possible increased organ retention. This study in the rataddresses this question.Methods

Anesthetized rats were contaminated with plutonium nitrate followingwounding by deep incision of hind leg muscle. Resection of the contam-inated site was performed 7 days after with or without DTPA treatment(30 µmol/kg i.v.). Pu urinary excretion was then measured for a further 7days and animals were euthanized at 14 days after contamination. Tis-sue samples were evaluated for Pu levels and histological changes. Results

At 7 days after contamination around 50 % of the initial activityremained at the wound site. An average of 16% of this activity was thenremoved by surgery. Surgery alone resulted in increased urinary excre-tion suggesting release from the wound site but no subsequentincreases in organ retention (bone, liver) were observed at 14 days.Indeed organ Pu levels were slightly reduced. However combination sur-gery and DTPA increased Pu excretion and reduced tissue levelsmarkedly. Conclusion

This is the first report in an experimental rodent model of resection ofPu-contaminated wound. It provides evidence of activity release as aresult of surgery but this does not lead to increased organ retention. Thecombination of surgery and DTPA administration is highly recommended.

7. Biological dosimetry and EPR

Development Challenges for Radiation Biodosimetry: U.S.Department of Defense/AFRRI Subject Matter ExpertsPerspectives

W. F. Blakely, N. I. OssetrovaBiological Dosimetry Research Program, Armed Forces RadiobiologyResearch Institute (AFRRI), Uniformed Services University of the HealthSciences (USUHS), Bethesda, MD 20889-5603 USA

The current diagnostic approach for assessment of individuals sus-pected of radiation exposure involves use of multiple parameter radiationexposure and injury assessment. The U.S. Department of Defense(DoD) operational doctrine supports the needs for capabilities to obtainboth an exposure “dose of record” as well as bioindications of radiationinjury that would contribute to early- as well as late-phase medical man-agement treatment decisions. In this area, significant gaps exist to pro-vide robust capabilities particularly in the case of an accident or terroristattack with potential exposure of large population to radiation. No singledevice or capability is sufficient to fill this gap. A systems approach solu-tion is envisioned where selected capabilities are assembled to respond

to a specific radiation exposure scenario in an integrated concept ofoperations. The planned use of these biodosimetry devices or assaysneeds to be defined in their scope of operations for triage of radiationexposure and early medical-treatment decisions. Components of thissystem must meet appropriate regulatory standards for reliability andaccuracy of physical measurement of internal radionuclide contamina-tion and radiation exposure as well as biodosimetry-based measurementof radiation effects to support medical treatment decisions. Currently,there are no radiation biodosimetry devices approved to use by the Foodand Drug Administration. Devices to measure absorbed dose using radi-ation detectors and biophysical/biological samples are useful for the pur-pose of “dose-of-record” assessment, however various exposure condi-tions (i.e., total- vs. partial-body exposure, radiation quality, dose rate orfractionation, etc.) and potential confounders (i.e., combined injury, age,gender, intrinsic radiosensitivity, etc.) also need to be understood formedical management decisions. Many radiation dose biomarkers ofradiation dose absorbed can also be used for assessment of radiationbioeffects. In this case, biomarkers that provide early prognostic risk

indications of acute radiation sickness (ARS) would complement thecurrently accepted ARS severity scoring system, Medical Treatment Pro-tocols for Radiation Accident Victims – METREPOL. [The viewsexpressed do not necessarily represent AFRRI, USUHS, or DoD.]

Protein Biomarkers for Enhancement of Radiation Dose andInjury Assessment in Animal Models

N.I. Ossetrova, D.J. Sandgren, D.P. Condliffe, K. Krasnopolsky, P.H.Ney, A. Rahman, and W.F. Blakely.Armed Forces Radiobiology Research Institute (AFRRI), UniformedServices University of the Health Sciences (USUHS), Bethesda, MD,USA

Development and validation of early-response radiation injury bio-markers are critical for effective triage of irradiated individuals. Earlymedical management of suspected radiation exposed individualsrequires effective triage tools for first-responders and medical providersto initiate early cytokine therapy in individuals exposed to life-threateningradiation doses and at risk for bone marrow acute radiation syndrome(ARS). We previously established animal (Mus musculus, Macacamulatta) radiation models to evaluate radiation biomarkers for futureapplication to humans and reported that protein expression profilemeasured in samples collected 1-3 days after total-body irradiation (TBI)can predict the dose in photon-exposed animals.

Herein, we present further evaluation results for protein and hemato-logical profiles in CD2F1 male mice and male and female rhesusmacaques (n=30) total-body irradiated with 60Co γ-rays (0.55 Gy/min)over broad dose ranges from 1 to 14 Gy and from 1 to 8.5 Gy, respec-tively. Data include results of hematopoietic cytokines (Flt-3 Ligand,IL - 6, G-CSF, TPO, and EPO) and acute phase proteins (CRP and SAA)and their combinations with hematological profile for radiation dose andinjury assessment up to 5 - 7 days. Results show that the dynamicchanges in the levels of CRP, SAA, IL - 6, G - CSF, and Flt - 3 Ligandreflect the time course and severity of acute radiation sickness and mayfunction as prognostic indicators of ARS outcome. In rhesus macaques,combination of lymphocytes, ratio of neutrophils to lymphocytes, CRP,SAA, and Flt-3 Ligand gives better estimations for given TBI doses thanany one biomarker alone. In the mouse radiation model, results for radiation dose assessment in “blinded” tests will be presented.

These research findings will contribute to bridging a gap that exists inthe current capabilities to rapidly and effectively identify and assess radiation exposure early after a radiation event, especially after a mass-casualty radiological incident. Research supported by AFRRI under project RAB4AU and DTRA under grant CBM.RAD.03.10.AR.002. Theviews expressed do not necessarily represent the AFRRI, the USUHS or the DoD.

Development of a model for the coordinated use ofbiodosimetry methods after a nuclear incident

J.M. Sullivan1, J.F. Koerner1, C.N. Coleman, MD1,2

1Office of Preparedness and Emergency Operations, Office of the Assis-tant Secretary for Preparedness and Response, Department of Healthand Human Services, Washington, DC, USA2Radiation Research Program, Division of Cancer Treatment and Diag-nosis, National Cancer Institute, Bethesda, MD, USA

Given the size and complexity of a nuclear detonation in a US city,biodosimetry assessment will be an essential component of both med-ical triage and overall medical management of the incident. Distinguish-ing those who have received a dose of radiation that necessitates imme-diate medical intervention (>2 Gy) from those who can be managed withdelayed treatment or only require long-term follow-up, will allow for moreeffective use of available resources. This could potentially save thou-sands of lives and will enhance the efficacy of the overall response. Aspart of the national preparedness and planning for a nuclear or radiolog-ical incident, we examined the capabilities, limitations, and require-

ments of the biodosimetry methods currently available or under develop-ment for use in the initial and secondary triage of patients. Here, wepresent a model for how these methods could be used as part of themedical response to a nuclear incident. Due to the time and dose con-straints of the currently available biodosimetry methods we advocate foran integrated system of complementary tools that when used togetherwould give relevant information in a short time span. In order to havesufficient capacity for this multiparametric approach, we propose thedevelopment of clinical and laboratory networks to increase the numbersof technicians and scientists available to perform testing and interpreta-tion of the results.

This content represents the personal views of the individual authorsand does not necessarily express the opinion or policy of the U.S.Department of Health and Human Services or its components. Nostatement should be construed as an official position of the U.S. Depart-ment of Health and Human Services or its components

Validation of semi - automatic scoring of dicentricchromosomes after simulation of 3 different irradiationscenarios

H. Romm1, E. Ainsbury2, S. Barnard2, J.F. Barquinero3, L. Barrios4,,C. Beinke5, M. Deperas6, E. Gregoire3, U.Kulka1, C. Lindholm7, J.Moquet2, R. Puig4, U. Oestreicher1, K. Rothkamm2, S. Sommer8, H.Thierens9 , A. Vral9, V. Vandersickel9, A. Wojcik6

1Bundesamt für Strahlenschutz (Germany), 2Health Protection Agency(United Kingdom), 3Institut de Radioprotection et de Sûreté Nucleaire(France), 4Universitat Autonoma de Barcelona (Spain), 5BundeswehrInstitute of Radiobiology (Germany), 6Stockholm University (Sweden),7Radiation and Nuclear Safety Authority (Finland), 8Institute of NuclearChemistry and Technology (Poland), 9University of Ghent (Belgium)

Mass casualty scenarios of radiation accidents require high through-put techniques of biological dosimetry for population triage in order toidentify individuals for whom clinical treatment is indicated. The dicen-tric assay is the gold standard technique, but it is very time consumingand needs well trained scorers. To increase the throughput of samplessemi-automation of dicentric scoring was investigated in the frameworkof the MULTIBIODOSE EU FP7 project and dose effect curves wereestablished in six biodosimetry laboratories.

To validate the dose effect curves, dose estimates were calculated fromblood samples, irradiated to simulate three different exposure scenarios:acute whole body, partial body and protracted exposure. Blood samplesfrom 33 healthy donors (> 10 donors / scenario) were irradiated in vitrowith gamma rays, simulating the three different types of exposure andincluding three different doses each. All the blood samples were irradiatedat the University of Ghent, Belgium and then shipped to the participatinglaboratories. The blood samples were set up by each lab using their ownstandard protocols and metaphase slides were prepared to validate thecalibration curves established by semiautomatic dicentric scoring. In orderto achieve this, 150 or 300 metaphases per slide were captured and thedoses were estimated using the newly formed dose effect curves.

After acute uniform exposure all laboratories were able to distinguishbetween 0 Gy, 0.5 Gy, 2.0 and 4.0 Gy (p < 0.001) and in most cases,the dose estimates were within a range of ± 0.5 Gy of the given dose.After protracted exposure, all laboratories were able to distinguishbetween 1.0 Gy, 2.0 Gy and 4.0 Gy (p < 0.001), and also in this case alarge number of the dose estimates were within ± 0.5 Gy of the irradia-tion dose. After simulated partial body exposure, all laboratories wereable to distinguish between 2.0 Gy, 4.0 Gy and 6.0 Gy (p < 0.001).Over-dispersion of the dicentric distribution enabled the detection of thepartial body samples; this effect was clearly dose dependent. In addition,improvement in this feature could be achieved with higher cell numbers.

The new method of semi-automation of the dicentric assay was suc-cessfully introduced in a network of six laboratories and can be used asa high throughput screening tool in a large scale radiation accident.

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The research leading to these results has received funding from theEuropean Union\'s Seventh Framework Programme (FP7/2007-2013)under grant agreement n° 241536.

Cytokinesis-Block MicroNucleus assay by manual andautomated scoring: calibration curves and dose prediction

S. De SanctisToxicology and Immunology Section, Army Medical and VeterinaryResearch Center, Rome, Italy

The cytokinesis-block micronucleus (CBMN) assay in peripheralblood lymphocytes is one of the best standardized and validated tech-niques for dose assessment after radiation exposure. This method hasbeen proposed as an alternative to the dicentric chromosome assay(DCA), the “gold standard” because requires less time and cytogeneticexpertise. Nevertheless for biodosimetry tool in large scale nuclear orradiological accidents CBMN assay needs further strategies, as theautomation of scoring, to speed up the analysis. As an essential prereq-uisite for radiation dose assessment is to establish a dose-effect curve,in this study blood samples of a healthy subject were irradiated withseven increasing doses of X-ray ranging from 0.25 to 4.0 Gy to generatemanual and automated calibration curves. Then the quality of the cali-bration curves was evaluated by determination of the dose predictionaccuracy. Biological doses of 10 blind samples of the same subject irra-diated at different X-ray doses were estimated by calculating frequencyof MN observed in binucleated cells by manual and automated scoringmode. The comparison between the two scoring approaches wasreported.

Dose Estimation For Dicentric Chromosome Assay AndCytokinesis – Block Micronucleus Assay: ComparisonBetween Manual And Automated Scoring In Triage Mode

A. De AmicisImmunology and Toxicology Section, Army Medical and VeterinaryResearch Center, Rome, Italy

In cases of an accidental overexposure to ionizing radiation, it isessential to estimate the dose. For this purpose biological dosimetry canbe performed to confirm, complement or even replace physical dosime-try when this could be not informative. The most validated biodosimetrytechniques for dose estimation are dicentric chromosome assay (DCA)as gold standard and cytokinesis – block micronucleus assay (CBMNassay). However both assays are time consuming and require skilledscorers. In case of large-scale accidents, the triage approach for visualscreening and the automation scoring are used to increase cytogeneticanalysis capabilities. Both these strategies require less time for estima-tion but are not always accurate. In this study we compared the accu-racy of triage mode dose assessment for DCA and CBMN in manual vs.automatic scoring. For dose estimation by DCA we analysed a numberof metaphases ranging from 20 to 50 for visual scoring and ranging from20 to 500 metaphases for automatic scoring. For dose estimations of thesame ten blind samples by CBMN assay we analysed binucleated cellsranging from 100 to 2,000 for manual scoring and various binucleatedcells for automatic scoring. The DCA and CBMN results were analysed consideringthe accuracy of dose estimation versus the clinical relevance for each scoringapproach. Furthermore, an intra-assay comparison was performed to elucidate thepower of dose discrimination between DCA and CBMN to identify the betterapproach in case of mass casualty accident.

Cell-free urine and serum miRNAs as sensitive radiationbiodosimeters

N. JacobRadiation Oncology, The Ohio State University Medical Center, Colum-bus, Ohio, USA

Radiation emergency management protocols include rapid doseassessment of the affected population for identification and prioritization

of individuals who require immediate medical attention. In addition toclinical observations, lymphocyte depletion kinetics and the dicentricchromosome assay are currently used. Dose estimation by lymphocytedepletion kinetics requires multiple days of readings and dicentric assayis highly technical, and expensive. The available biologics such asserum proteins and urine metabolites have limitations because of vary-ing normal tissue toxicity, depending on individual’s genetics, immunestatus and other confounding factors. We have investigated the time anddose dependent changes in small non-coding RNAs, microRNAs (miR-NAs), detectable in body fluids and exhibited robust changes followingexposure of mice to acute doses of gamma-radiation. An amplification-free, hybridization based quantitative assay utilizing the nCounter multi-plex platform developed by nanoString Technologies was used to com-pare the changes in over 600 miRNAs after radiation. Our studies haveidentified several evolutionarily conserved responsive miRNAs in serumand urine collected from two strains of mice, following exposure to radi-ation in a dose range relevant for triage. Development of a novel normal-ization strategy using multiple spike-in oligonucleotides allowed accuratemeasurement of changes in cell-free urine and serum miRNAs afterwhole body irradiation. Several markers sensitive to incremental changesin radiation dose, and stable for days after exposure were identified. Inserum, miR-200b and miR-762 showed a dose dependant increase,while miR-150 exhibited dose and time dependant decrease and mark-ers such as miR-23a did not change after radiation. Following exposureto acute doses of 1, 2, 4, 6 and 8 Gy, a 30 %, 38 %, 48 % 70 % and72 % statistically significant reduction in serum miR-150 was observedat 24 hrs, which was further reduced at later time points. Cell-free urinemiRNA markers with dose response include miR-804, miR-378 andmiR-1224, however exhibited different kinetics. Our studies have devel-oped a panel of novel biomarkers for rapid assessment of radiation doseusing non-invasive or minimally invasive methods, for triage purpose incase of a radiological incident.

DNA damage as indicator of external and internal radiationexposure

Scherthan H.11Bundeswehr Institute of Radiobiology affiliated with the Univ. of Ulm,Neuherbergstr. 10, D-80937 Munich, Germany

Ionizing radiation (IR) can induce cellular and tissue damage, ofwhich DNA double strand breaks (DSBs) are a severe threat to cellularsurvival and systemic outcome. DSB formation elicits a DNA damageresponse (DDR) that involves DSB recognition, damage signalling andrepair. One significant IR-induced DDR outcome is the phosphorylationof histone H2AX (a histone H2A variant) and the accumulation of the53BP1 protein around a DSB, creating microscopically visible gamma-H2AX (γH2AX) and 53BP1 foci. In the low dose range IR-inducedγH2AX foci correlate with the expected amount of DSBs, while the lossof foci over time indicates DNA repair. Particularly in non-cycling periph-eral blood lymphocytes and other tissues γH2AX and 53BP1 focus for-mation has proven a sensitive bio-indicator of radiation exposure. In lowdose rate in vivo scenarios, such as after incorporation of radionuclides,e.g. I-131 in the wake of thyroid tumour treatment, DNA damage focusenumeration shows a linear correlation with blood dose in early timepoints (hrs) as determined by automatic and manual fluorescencemicroscopy for focus formation. DNA damage focus assays can rapidlyindicate an external and internal radiation exposure, even at droppingdose rates 72h post exposure. However, a large inter-individual variationof the DDR renders individual dose reconstruction in such scenarios dif-ficult. In acute external partial body exposure events, like in the Göttin-gen minipig skin model, the focus assay can reveal high foci values inepidermal cells shortly after 50Gy γ exposure with the damage signalbeing near saturation. The focus assay regains resolution power withgrowing (repair) time post IR, leaving significantly elevated foci levels indifferentiated keratinocytes even weeks after exposure. In all, the DNA

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repair-associated focus assay is a versatile tool to study the DDR in dif-ferent exposure scenarios from nuclide incorporation up to externalexposures in high and low dose scenarios. In particular, DDR foci in ter-minally differentiated tissues like the epidermis may serve as an indica-tor of previous acute irradiation events even weeks after high dose expo-sure. Finally, new insights regarding the transfer γH2AX detection in abiochemical lateral flow chromatography format will be discussed.

Correlation of nuclar abnormalities in myeloid cells withestimated whole-body dose of radiation received during theY-12 criticality accident

R. Goans, C. Iddins, D. Christensen, W. Wiley, T.M. Fliedner,N. DainiakRadiation Emergency Assistance Center/Training Site, Oak Ridge, Ten-nessee, USA;

University of Ulm, Ulm, Germany; Yale New Haven Health-BridgeportHospital and Yale University School of Medicine, New Haven, CT, U.S.

Individual biological measures of cumulative whole-body radiationdose include the lymphocyte dicentric assay and the rate of decline andnadir of the circulating absolute lymphocyte count. Although radiationexposure has been associated with other changes in the hematopoieticsystem, including the appearance of internuclear bridges, binuclearity,multinuclearity, karyolysis and nuclear fragmentation (Fliedner et al.,Blood 1964; 23 :471), the relationship between such changes and radi-ation dose has not been assessed. To determine whether morphologicalalterations appearing during myeloid differentiation can be used as adosimetric measure, smears of bone marrow and peripheral blood wereanalyzed from 8 individuals exposed to ionizing radiation (estimatedwhole-body dose range of 0.2 - 4.6 Sv) and from three control individu-als. Five of the 8 individuals received doses exceeding 2 Sv, all of whomrecovered from the hematopoietic syndrome with supportive care thatwas appropriate for the time period. Approximately 4.3 % of myeloidcells from control individuals (correlating with a “zero” dose) containedbridges. Among smears from exposed individuals, we observedincreased inter-lobar stranding/bridging in nuclei of the myeloid ele-ments, many of which were bilobed and morphologically similar to thePelger-Huet (PH) variant of PMNs. The percentage of myeloid elementsfrom exposed individuals containing such bridges was determined incells with or without the PH variant. Raw data and data averaged in binsof 0 -1 Sv, 1 -2 Sv, 3 -4 Sv, and 4 -5 Sv, and plotted at the midpoint eachbin, were graphed as a function of dose. The frequency of cells fromeach group containing bridges (range of 4% to 8.7%) increased linearlywith estimated radiation dose. The relationship between per cent bridgesand dose in Sv is best characterized with a simple linear regressioncurve (N = 8; regression slope = 0.74 %/Sv; p = 0.01). Together, thesefindings raise the possibility that examination of smears from peripheralblood or bone marrow may provide individual dosimetric information.Prospective assessment of bridges in myeloid elements relative to doseestimates is indicated, and molecular mechanisms underlying radiation-associated augmentation of bridge formation requires definition.

Assessment of total- and partial-body irradiation in a baboonmodel: a kinetic multiparameter study of chromosomalaberrations, biological and clinical parameters.

Francis Hérodin, Nancy Grenier, Josiane Denis, Patrick Martigne,Philippe Arvers, Stéphane Baugé, Hervé Chaussard, Michel Drouet,Marco ValenteRadiobiology, Institut de Recherche Biomédicale des Armées, LaTronche, France

In this biodosimetry study baboons were irradiated to determine theefficacy of a kinetic multiparameter (clinical, physical and biological)approach in discriminating partial-body irradiation (PBI) from total-bodyirradiation (TBI) in terms of dose, heterogeneity of radiation exposureand severity of injury. Anesthetized Papio anubis were unilaterally

exposed to gamma-rays as follows: 5 Gy TBI and partial-irradiated equiv-alents (10 Gy 50 % PBI, 7.5 Gy 50 % / 2.5 Gy 50 % TBI, 6.25 Gy 80 %PBI, 5.55 Gy 90 % PBI and 15 Gy 30 % PBI); 2.5 Gy TBI and its equiv-alent 5 Gy 50 % PBI (two animals per exposure condition). Blood sam-ples were collected before exposure and from one hour until 200 daysafter irradiation. More than 50 parameters were analyzed including clini-cal status, blood cell count, biochemical parameters and the biodosime-try gold-standard dicentric assay. A partial least square discriminantanalysis showed that TBI could be distinguished from several equivalentPBI situations. Thus, four groups of baboons irradiated to a 5 Gy equiv-alent whole-body dose (i.e. 5 Gy TBI, 10 Gy 50 % PBI, 6.25 Gy 80 %PBI and 5.55 Gy 90% PBI) were clearly separated. Moreover, 5 Gy TBI,10 Gy 50 % PBI, 2.5 Gy TBI and 5 Gy 50 % PBI could also be distin-guished. Here, shielding as little as 10 % of the bone marrow preventedaplasia and therefore hematologic parameters contributed to discrimi-nate PBI from TBI. Moreover, parameters such as neutrophils to lym-phocytes ratio, c-reactive protein and citrullin levels showed some dis-criminating power as biodosimetric markers and neutrophil count,platelet count, hemoglobin and Flt-3 ligand levels revealed to be poten-tial markers of injury. When considering seven potentially confoundingirradiation situations, sequential assessment of clinical and biologicalstatus is required for reliable evaluation of radiation heterogeneity andinjury. Ongoing international collaborations will contribute to comprehen-sive interpretation.

Launching a Cytogenetic Biodosimetry Laboratory for theAssessment of Accidental Radiation Exposure in Saudi Arabia

G. AlsbeihKing Faisal Specialist Hospital & Research Centre, Riyadh, Saudi Arabia

The continuous expansions in the applications of nuclear technologiesin various aspects of life increase the probability of overexposure due toinvoluntary or premeditated nuclear accidents. National radiation-pro-tection preparedness response plan requires adequate estimate of dosereceived for efficient medical assistance of victims. Cytogenetic bio-dosimetry is a proven, ISO and IAEA standardized biotechnology tech-nique for assessing medically relevant radiation doses. We havelaunched a project to establish a reference biological dosimetry labora-tory to help the nation\'s ability to respond to sporadic and mass radia-tion casualty incidents by assessing the magnitude of radiation overex-posure. Accurate calculation of radiation doses received will result inevidence based treatment decisions and better management of valuableemergency resources. It will also contribute to the \"National RadiationProtection Program\" by playing a role in nuclear emergency plans. Thecytogenetic method is standardized and scalable. In addition to diagno-sis of overexposure, it provides triage capability for rapid stratification ofpatients who need more specialized medical care. It can also detectfalse positives and false negatives exposure particularly in cases of legalallegations.

A national standard dose-response calibration curve, pre-required toestimate doses received in case of accidental radiation overexposure isbeing established. Peripheral blood lymphocytes are collected fromhealthy Saudi volunteers and irradiated with different doses ranging from0 to 5 Gy of 320 KeV X-Rays. Stained cytogenetic slides are preparedfrom cultivated lymphocytes according to the IAEA protocol. The Metafersystem (MetaSystem, Germany) is used for automatic metaphase find-ing and assisted scoring of dicentric chromosomes. Results are fit to thelinear-quadratic dose-effect model. The obtained dose-response calibra-tion curve for the induction of dicentric chromosomal aberrations inSaudi Arabia is comparable to those described in other population. Cur-rently, more volunteers and experiments are being conducted; gammarays and neutrons will be used for irradiation. The laboratory will alsoseek accreditation from IAEA and WHO, and cooperation with interna-tional biodosimetry network. It is expected that the various activities ofthe biological dosimetry laboratory will add depth to information for

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decision-makers and public health officials who assess the magnitude ofpublic, medical, occupational and accidental radiation exposures inaddition to providing a platform for advanced education, research anddevelopment in Kingdom of Saudi Arabia and neighbouring countries.Supported by KACST under the Long-Term Comprehensive NationalPlan for Science, Technology and Innovation (Project No. 9 MED749-20;RAC#2110 005).

EPR Dosimetry for Triage of Large-Scale Radiation EventsH. Swartz

Geisel School of Medicine at Darmouth, Hanover, NH, USAThe potential exposure of large numbers of individuals to levels of

radiation that could lead to acute clinical effects is a major concernbecause of the possibility of radiation accidents, acts of terrorism, ornuclear warfare. In order to cope medically with such an event, it isessential to identify those individuals who probably have had clinicallysignificant exposures so that they can be entered into the medical sys-tem and to identify those who do not need medical intervention so thatthe response system is not overwhelmed. The capabilities of electronparamagnetic resonance (EPR) to measure radiation-induced paramag-netic species along with the persistence of such species in certain tis-sues (i.e. teeth, fingernail, toenails) has led to EPR becoming a promi-nent method for making these measurements in potentially affectedindividuals. Results in the last year demonstrate the high potential ofthese techniques and progress towards practical devices. With tooth bio-dosimetry, the device has now obtained excellent dose response curvesin human subjects undergoing total body irradiation (TBI) with cleardelineation of appropriate thresholds for triage. Plans for regulatoryapproval have advanced through discussions with the FDA. We alsostarted planning on a variant of the tooth dosimeter that will enablemeasurements be made suitably for subjects with combined injury, withthe patient in a recumbent position, no need to be able to cooperate,and improved dose resolution. The nail techniques have made signifi-cant new progress, which will allow improved assessment of heterogene-ity in the exposure. The technique using clipped nails has had a break-through, with elimination of much of the scatter in results with irradiatednail clippings; these results are being verified with clipped nails irradi-ated in vivo during TBI. The approach measuring nails in vivo withoutclipping had also had advances, with data being obtained on nails irradi-ated in situ on isolated fingers.

EPR retrospective dosimetry with fingernails: report on firstapplication cases

F. Trompier1, F. Queinnec1, E. Bey2, T. De Revel3, I. Clairand1, J.F.Bottollier-Depois1

1Institut de Radioprotection et de Sûreté Nucléaire, IRSN, BP17, 92262Fontenay-aux-Roses, France2Hôpital d\'Instruction des Armées Percy, Service d\'Hématologie, BP410, 92141 Clamart Cedex, France3Hôpital d\'Instruction des Armées Percy, Service de Chirurgie Plastique,BP 410, 92141 Clamart Cedex, France

For localized irradiation to hands, in case of sources accidentally han-dled, most of the time it is very difficult to estimate the dose distribution.Doses may reach several tens of grays and the dose distribution is usu-ally very heterogeneous. Until recently, dose could be estimated only byanalysis of bone biopsies using Electron Paramagnetic Resonance (EPR)spectroscopy. This technique was previously used on surgical wastes orafter amputation of a finger. Moreover, the dose information was avail-able in a few locations on the hand only, due to the limited number ofbiopsy fragments usually collected.

The idea to measure free radicals (FRs) induced by radiation in nailsto estimate a dose is not new, but up to now, no application cases werereported. As a matter of fact, the EPR analysis of nails is difficult due tothe presence of intrinsic signals and parasitic signals from mechanicalstress (when nails are collected) which overlaps the radio-induced com-ponents. In addition, the entire identified radio-induced FRs are unsta-ble and very sensitive to humidity. In these conditions, it was difficult toforesee any application for dosimetry with nails.

A study conducted at IRSN, based on the use of high frequency EPR,allows the identification and the characterisation of a stable radio-induced FRs in nails and the definition of a protocol for dose assess-ment. This protocol has been applied for fingernails samples from vic-tims of 3 different radiological accidents that occurred between 2008and 2012. The doses found in nails are in very good agreement with thedose in bone and clinical signs. For the most recent accident (Peru,January 2012), a first dose estimation on nails was available before theappearance of clinical signs.

Until recently, the dose evaluation to hands which was not possible, atleast in the early management of the victims, is now possible. This newtechnique can provide valuable information to the medical staff to opti-mise the therapeutic strategy.

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8. Radiation health effects and medical countermeasures

Medical countermeasures for radiation injury, where we areand where`s the way ahead

Viktor MeinekeBundeswehr Institute of Radiobiology affiliated to the University of Ulm,GermanyBackground

Medical RN Countermeasures include all activities related to MedicalRadiation Accident Management. The term MCM in RN scenariosshould therefore not be limited to or be understood as one isolatedaspect in radiation accidents, such as only a new drug with a certainDRF, or a single biodosimetry approach. MCM development means amost complex task. From the military perspective MCMs have always tobe related to Military Mission Support. In civilian mass casualty scenar-ios, such as the detonation of a 10 Kt Improvised Nuclear Device (IND),the rationale for the development of MCMs shows similarities but alsostriking differences. There will be the need to diagnose radiationexposed patients in both military and civilian scenarios. Therefore thedevelopment of specific tools needed should be done in a jointapproach. But essential differences need to be taken into account, such

as patient ages, the need to treat special populations and specific mili-tary requirements, just to mention a few.

Resources for the provision of therapeutic approaches and handling ofa larger number of irradiated patients still remain critical. This fact refers tothe principal limitation of controlled studies for a validation of drugs forhuman use. There is a strong need for alternative pathways of develop-ment and approval of drugs or biological products to treat radiationinjuries. These aspects will be discussed in this presentation. But pitfalls inmedical management to a large extent also relate to organizational issues,such as hospital treatment capacities, or training of medical personnel andemergency personnel in basics of radiation biology and radiation medicine.Conclusion

The best way ahead to achieve an optimal preparedness for radiationmass casualties will be a collaboration on an international level. Onlysharing of knowledge, training and treatment capacities and otherresources for radiation victims as well as in MCM research will ensurestate of the art treatment of radiation victims. Moreover it will also becrucial to revisit clinical models, such as radiation oncology for their suitability to act as validation tools.

HemaMaxTM Development as a Frontline, Single DoseRadiation Medical Countermeasure

Timothy K. Gallaher1, Zoya Gluzman-Poltorak1, Chris Lawrence1,Mamata Gokhale1, Jamie Tom1, Sarita Mendonca1, Simmy Thomas1,Joseph Miller2, Dolph Ellefson1, Lena Basile1

1Neumedicines Inc., Pasadena, CA, 2Keck School of Medicine, Univer-sity of Southern California, Los Angeles, CA

HemaMax™ (recombinant human interleukin-12; rHuIL-12) is beingdeveloped as a post-exposure (+24 hr) medical countermeasure (MCM)against the hematopoietic syndrome of acute radiation syndrome(HSARS). One single dose of HemaMax subcutaneously administered24 hours after lethal total body irradiation (TBI) exposure increases sur-vival in non-human primates (rhesus monkeys; NHP) in the absence ofany additional supportive care. These properties support conceptualoperations (ConOps) in which HemaMax as MCM can be administeredto persons in a mass casualty event who have been exposed or poten-tially exposed to ionizing radiation. This use of HemaMax is expected todecrease the number of casualties, even if no supportive care were to beavailable for protracted periods of time

Efficacy of a single dose of HemaMax administered after 24 hourswithout supportive care was established in NHP after lethal whole bodyirradiation (WBI) (LD50/30) with single subcutaneous doses of 100ng/kg and 250 ng/kg in initial HSARS survival efficacy experiments(PLoS ONE 7(2): e30434). Neumedicines subsequently conducted alarger, blinded, survival study in NHP under good laboratory practice(GLP).

Neumedicines GLP blinded NHP study established that one singlesubcutaneous dose of HemaMax administered 24 hours after lethalradiation exposure of 7 Gy WBI (LD90/60) significantly increased sur-vival in NHP compared to control (n=18) (p < 0.05 Logrank). Statisti-cally significant efficacious doses ranged from 500 ng/kg down t o 50ng/kg. This latter dose corresponds to an approximate unit dose inhumans of 1.2 ug.

The same HemaMax GMP drug product used in NHP studies wasadministered to healthy volunteers in two Phase I safety trials in healthyvolunteers. In the First-in-Human (FIH) single ascending dose study in32 subjects, HemaMax was found to be safe and well tolerated at 2, 5,10 and 12 ug doses. A follow-up phase 1b safety study is ongoing in 60subjects to further confirm the safety and tolerability.

Overview of Use of G-CSF in the Treatment of Acute RadiationInjury

G. ReevesApplied Research Associates (ARA), Inc., Arlington, USA

Depression of hematopoietic elements due to significant levels ofwhole-body or partial-body irradiation due to radiation-induced sup-pression of mitosis in the stem and progenitor cells can result in life-threatening injury. Successful administration of intensive care ofpatients experiencing acute radiation sickness (ARS; also called acuteradiation syndrome) is dependent upon the ability to stimulate therecovery of surviving hematopoietic stem cells (HSC), assuming thenon-hematopoietic injuries are also survivable with treatment. To datethere have been a number of studies involving radiation accidentswhere patients were treated with cytokines. Although the data overallseem to indicate that the period of neutropenia is shortened and sur-vival prolonged, so far there is no statistically significant proof thatcytokine administration actually decreases mortality in radiation-injuredhumans. Some studies have shown no improved survival when used ina mouse model; however, studies in canines and primates have shownimproved survival. Based on review of the human experience with G-CSF and GM-CSF, as well as some animal studies, current consensusopinions support the prompt administration of these materials topatients suffering significant bone marrow depression from exposure toionizing radiation.

Planning for response to a major radiation event must also involveknowledge of where stocks of CSFs are and how they are to be distrib-uted to medical treatment facilities.

CSF therapy is considered a valuable adjunct to treatment with antibi-otics and strict hygiene controls in certain irradiated patients. However,CSFs are not without side effects, and this fact needs to be taken intoaccount when prescribing them. Further work on modeling the effects ofG-CSF may help better define what range of radiation exposures wouldbenefit most from cytokine treatment. It appears that these drugs doshorten the periods of neutropenia in irradiated patients, and must beconsidered part of the therapeutic armamentarium in the treatment ofARS in a mass casualty situation.

The efficacy of G-CSF to mitigate survival following LD50/60total body irradiation (TBI) is dependent upon the timing of itsinitiation.

AM Farese1, CP Smith1, CR Brown1, A Bennett1, B Katz2, K Prado1,TJ MacVittie1. 1Radiation Oncology, University of Maryland, Baltimore, MD, 2Division ofBiostatistics, Indiana University, Indianapolis, IN

The identification of a mitigator and its effective administration proto-col(s) is required for any medical countermeasure (MCM) that may beemployed to treat individuals exposed to high dose radiation. The effi-cacy of filgrastim (Neupogen®), a potential MCM, to improve survivalwhen initiated either at 24 hours (hr) or 48hr following TBI in a nonhu-man primate (NHP) model of the hematopoietic syndrome (HS) of theacute radiation syndrome (ARS) was investigated. Two studies wereperformed in rhesus macaques. NHP were exposed to TBI, bilateralexposure, total midline tissue dose of 7.5 Gy, (target LD50/60) deliveredat 0.80 Gy / min, using linear accelerator-derived 6 MV photons. AllNHP were administered medical management. Following TBI on day (d)0, filgrastim (10 µg / kg / d) or the control (5 % dextrose in water) wasadministered subcutaneously (SC), daily through effect (absolute neu-trophil count = 1,000 / µL for 3 consecutive days). Study one (n = 46)was powered to demonstrate a 30 % improvement in survival followingthe administration of filgrastim or control when initiated at 23±3hr post-TBI. Survival analysis was conducted on the Intention-to-Treat (ITT)population using a one-tailed null hypothesis at a 5% significance level.The second study (n=80) was powered to demonstrate a 25% improve-ment in survival following the administration of filgrastim or controlbeginning at 48 ± 4hr post-TBI. Survival analysis was conducted on theITT population using a two-tailed null hypothesis at a 5 % significancelevel. Survival was significantly increased by 38.3 % over controls whenfilgrastim was administered daily, beginning at 24hr following lethal TBI(P = 0.004). However, when filgrastim was initiated 48hr after TBI, sur-vival was not improved (2.5% increase, P=0.8230).

These data demonstrate that efficacy of a MCM to mitigate lethality inHS-ARS can be dependent on how quickly the mitigator can be admin-istered after exposure.

This work was supported by the NIAID contractsHHSN266200500043C and HHSN272202000046C

The influence of G-CSF administration on the absoluteneutrophil: lymphocyte count (ANC: ALC) ratio in nonhumanprimates (NHP) following exposure to total- (T) or partial-body irradiation (PBI).

TJ MacVittie, A Bennett, AM Farese, W Blakely*, N Ossetrova*, CSmith, A Gibbs. University of Maryland, School of Medicine, Department of RadiationOncology, Baltimore MD; *Armed Forces Radiobiology Research Inst.,Uniformed Services University of the Health Sciences, Bethesda MD

Efficient and timely triage subsequent to a large-scale nuclear eventwill likely be based on symptoms using METREPOL analysis and vali-dated biomarkers. A possible confounding variable may be the early use

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of medical countermeasures (MCM) during the triage process. Granulo-cyte colony-stimulating factor (G-CSF), a candidate MCM may beadministered within 24 or 48 hrs post exposure. Therefore, we investi-gated if the administration of G-CSF affects use of the hematologic radi-ation biomarker ANC: ALC for effective triage over the early time coursepost TBI or PBI.

NHPs (male, rhesus macaques) were exposed to midline tissue dosesof TBI, (dose range for 2 cohorts, 7.1-8.9 Gy; 9.0-14.0 Gy) or PBI (doserange, 10-12.5Gy) utilizing LINAC-derived photons (0.80Gy/min). Dose-response relationships were established to define the hematopoietic (H)and gastrointestinal (GI) syndromes. All NHPs were administered medical management under IACUC-approved protocols. G-CSF (neu-pogen, 10 ug/kg, once daily through effect, SC) was administered toNHP exposed to either TBI (7.5 Gy) initiated at 24 hrs or 48 hrs post-TBI, or PBI (10 Gy) initiated at 24 hrs post-exposure. Complete bloodcounts (CBC) were taken at pre-dose and daily through 21 d, then inter-mittently throughout the study. The pre-irradiation value for ANC: ALCwas 0.77 ± 0.03 SEM.

TBI cohort: The ANC: ALC increased from 0.77 to an average ratio of20 and 40 at d1, and 12 and 22 at d2 for the H-ARS and GI-ARS doserange respectively; for an approximate 2-fold differential between the H-ARS (n=102) and GI-ARS (n=69).

PBI cohort: The ANC: ALC increased from 0.77 to 30 and 15 at d1and d2, respectively in NHP (n=80) exposed to the GI-ARS dose range.

G-CSF administration: G-CSF injected at 24 or 48hrs post-TBI shiftedthe ANC: ALC peak values to the right, in time, at 24hrs post- injection,i.e., d2 or d3 respectively. This effect skewed the time response patternof ANC: ALC. A similar effect was noted in the PBI model. These datawill permit a more definitive analysis of the value of ANC: ALC alone andwith multiple biomarkers in the realistic, nuclear triage scenario.

G-CSF effects on ANC: ALC values may confound subsequent radia-tion injury and dose assessment. These data will permit a more defini-tive analysis of the value of ANC: ALC alone and with multiple biomark-ers in a realistic, nuclear triage scenario.

This work was supported by the NIAID contractHHSN272202000046C

A Basic Fibroblast Growth Factor Analog for Protection andMitigation Against Acute Radiation Syndromes

P. OkunieffRadiation Oncology, University of Florida, Gainesville, Florida, USA

FGF-2 was the first angiogenic factor discovered and remains amongthe most powerful. It is distinguished from most angiogenic factors bythe absence of associated inflammation. It is an absolute requirementfor most or all tissue engineering work and for prevention of prematurematuration of stem cells. Among growth factors it is consistently found tobe a useful radiation protector and mitigator. FGF-2 however isextremely difficult to synthesize and extremely sticky, leading to difficultyobtaining quantities and administering the agent even in small rodents.

We have developed a peptide alternative to FGF-2. FGF-P was similarto FGF-2 in all tests performed. Mechanistically, FGF-P appears to havesimilar or better receptor binding to the FGFR1 receptor, and it inducesERK phosphorylation and proliferation of many cell lines to a degreesimilar to that of FGF-2. When given systemically, it demonstrates miti-gation effects that are similar to or better than those of native FGF-2. Forunknown reasons human FGF-2 is of little benefit to the C57BL/6 strain,but FGF-P works in all murine strains; this result is consistent with itsexcellent receptor binding. Based on these studies, FGF-2 and FGF-Pare mechanistically identical with the exception that FGF-P is morespecies-independent than FGF-2. In addition to many of the benefitsdescribed above, similarities and advantages of FGF-P over FGF-2include the following: n Bleeding time improved when given after irradiation.n Platelet recovery improved when given after irradiation.

n Specific binding to cell-surface receptor was shown in competitionassays and Amnis cell sorting with a biotin-labeled agent.

n FGF-P increases known beneficial cytokines (i.e., G-CSF) and lowersdeleterious radiation-induced cytokines (i.e., KC, MCP-1, TNF, andIL-6), which can be used as biomarkers for pharmacodynamics.

n Preliminarily, the therapeutic window of FGF-P is ˜200 times theeffective dose, and we have detected no immunogenicity. FGF-P has promise as a protector and as a mitigator of radiation g -

astrointestinal, cutaneous and bone marrow syndromes.

Sensitivity and specificity of medical tests mended by IAEApublications for early diagnosis of acute radiation syndromein a radiation accident

V. KrasnyukFederal Medical Biophysical Center, Moscow, Russian FederationIntroduction

In 1998 -2005 IAEA released series of publications on the proceduresfor an early medical response during a nuclear or radiological emer-gency[1, 2]. Particularly there were some instructions for doctors on diag-nosis of radiation injuries in the publications. Some countries which hadno clinical experience in treatment of accidentally exposed victimsaccepted the publications like a last handbook for local doctors. How-ever, there is no evidence on sensitivity (Se) and specificity (Sp) of diag-nostic approaches that were recommended by IAEA. Aim

Using the evidence-based medicine approaches we would like toevaluate the sensitivity and specificity of methods for early diagnosis ofacute radiation syndrome (ARS) on the Russian data base of acute radi-ation syndrome. Methods

Early clinical disorders and blood lymphocyte counts were analyzed intwo groups of patients that had been hospitalized at Hospital 6 inMoscow after the Chernobyl accident (145 persons)< as well as afterflash exposures in some accidents before it (100 persons). Amongthem 168 were sick with acute radiation syndrome. Others 77 wereexposed to ionizing radiation at doses of 0.25 -0.8 Gy. Totally clinical andlaboratory tests on 245 accidentally exposed persons were analyzed.The sensitivity and specificity for early diagnostic approaches for acuteradiation syndrome recommended by IAEA[1, 2] have been calcul a ted[3].Findings

Among investigated clinical and laboratory tests the best sensitivityand specificity for diagnosis of acute radiation syndrome have belongedto two tests: vomiting development and blood lymphocyte count. Bothsymptoms are recommended by IAEA as the most important for theearly diagnosis. Vomiting development test at patients with acute radia-tion syndrome after the Chernobyl exposure has demonstratedSe = 95 % and Sp = 90 % for all degrees together. The same test afterother radiation accidents has shown Se = 97 % and Sp = 82 %. Vomitingdevelopment for 30 min time after flash exposure has given an opportu-nity to diagnoses very severe cases of ARS with Se=63 % andSp=100%. Vomiting development for 1h time from the beginning of theChernobyl exposure for a diagnosis of very severe ARS showedSe=90% and Sp=89%. Lymphocyte count for the first 24h after expo-sure for diagnosis of ARS has demonstrated not so high results(Se = 73 % and Sp = 75 %), and for 24 - 48 h time after exposure:Se = 84 % and Sp = 89 %. Lymphocyte count for diagnosis very severeARS for the first 24 h time after exposure has shown Se = 38 % andSp = 90 %, and for 24 - 48 h time after exposure: Se = 43 % andSp=86%. Conclusion

The calculations of Se and Sp of clinical and laboratory tests recom-mended by IAEA for early diagnosis of ARS have shown that the testsshould be divided into two kinds. The first kind of tests with high Seshould be used as screening tests for separation people who may be

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sick with ARS. Another kind of tests with high Sp is suitable for the con-formation of the diagnosis. A new approach to use lymphocyte count forARS diagnosis is suggested for increasing both Se and Sp.References1. Generic procedures for medical response during a nuclear or radio-

logical emergency. IAEA, Vienna, 2005; 287 P.2. Diagnosis and treatment of radiation injuries. Safety Reports Series

No.2 IAEA, Vienna, 1998; 49 P.3. D.G. Altman, J.M. Bland Statistic Notes: Diagnostic tests 1: sensitivity

and specificity. BMJ 1994; 308: 1552.

Acute Radiation Syndrome management: towards newstrategies to mitigate hematopoietic and non hematopoietictoxicity

M. DrouetRadiobiology - Cell therapy unit, Institut de Recherche Biomédical desArmées, Bretigny sur Orge, France

Following exposure to high doses of ionizing radiation (accident ornuclear attack), Acute Radiation Syndrome represents the clinicalresponse of key radiation-sensitive tissues among which thehematopoietic syndrome represents the first therapeutic challenge(Drouet and Hérodin Int J Radiat Biol 2010). Medical management ofhematopoietic syndrome would consist in allogeneic hematopoieticstem cell grafting, if bone marrow has been severely depleted, withunsatisfactorily solved concerns regarding morbidity and mortality.Regarding extrahematological toxicity, no efficient standardized treat-ment has been proposed yet. Thus identifying new drugs and devel-oping new strategies remain priorities, especially in order to signifi-cantly raise up the hematopoietic stem and progenitors cellstransplantation threshold and to prevent/cure the radiation-induced(RI) multiple organ failure syndrome.

Synthetic small interfering ribonucleic acids (siRNAs) may representnew tools to achieve this goal. In order to in vivo mitigate RI-apoptosiswe set up a mouse model in which B6D2F1 mice were lethally irradiated(9 Gy gamma, LD 90 % 30 days) and then injected at 2 hours followingirradiation with siRNAs directed towards p53 gene (0.5 nmol/mouse,Dharmacon). Preliminary results confirm that the efficacy of such astrategy will depend on optimized cell penetration and rigorous gene tar-get(s) selection.

As a second approach our group has proposed the transient genetherapy strategy based on short term secretion of morphogene(s) tostimulate residual stem cells and favour microenvironment repair. Thuswe investigated the hematopoietic response in rhesus monkeys (n=4) toa single intra-osseous injection of xenogenic multipotent mesenchymalstem cells (ASCs) transduced with a Sonic-hedgehog (Shh) pIRES2plasmid and grafted 2 days after a 8-Gy gamma irradiation. Thrombocy-topenia and neutropenia duration were reduced in Shh-ASC injectedanimals when compared with mock-ASC injected controls (n=4) andrecovery accelerated.

Finally adult somatic cell reprogramming (induced pluripotent stemcells) represents today a new frontier in regenerative medicine whichmay offer new opportunities to restore irradiated tissues.

Mesenchymal stromal cells (MSC) therapy for theregeneration of radio-incuced proctitis

M. Benderitter1, A. Chapel1, C. Linard1, N. Mathieu1, J.M. Simon2, J. Voswinkel3, J.J. Lataillade 4, N.C. Gorin3

1Institute of Radioprotection and Nuclear Safety (IRSN), DRPH /SRBE/LRTE, Fontenay-aux-Roses, France.2Department of radiation Oncology, La Pitié-Salpétrière Hospital APHP,Paris, France3Department of Hematology, Saint Antoine Hospital APHP, Paris, France4Military Blood Transfusion Centre (CTSA), Research Department andCell Therapy Unit, Clamart, France.

The radiation oncology accident at the Public General Hospital inÉpinal was the highest in France. The accident affected 425 patientstreated for prostate cancer and the clinical consequences weresevere with sequelae classified from grade 2 to 5 on the CTCAE 3.0scale. The management of severe proctitis in patient is challenging.The fibro-necrosis, fistulae, chronic inflammation and haemorrhageinduced by pelvic over-irradiation have an impact on morbidity. Ourgroup has demonstrated in different preclinical animal models(rodents and mini-pig) that systemic and repetitive MSC injectionseffectively reduced inflammation and fibrosis and is a promise thera-peutic approach. We have shown that MSC migrate to damaged tis-sues and restore gut functions after irradiation. MSC treatment favorsthe re-establishment of cellular homeostasis by both increasingendogenous proliferation processes and inhibiting apoptosis of radia-tion-induced small intestinal epithelial cells. Our experiments showedan involvement of Wnt pathway activation in MSC effects on epithelialcell proliferation. MSC might regulate the epithelial stem/progenitorcells directly or indirectly, that is stem cell niches that provide andmaintain an optimal microenvironment for stem cell function. We alsocarefully study side effects of stem cell injection for further applica-tion in patients. The clinical status of four first patients suffering fromsevere pelvic side effects was improved following MSC injection in acompationnal situation. MSC therapy may represent a safe therapeu-tic measure for patients developing very severe rectitis.

An Amino Acid Mixture Mitigates Irradiation InducedGastrointestinal Toxicity

S. VidyasagarRadiation Oncology, University of Florida, Florida, USA

Radiation targets rapidly dividing cells of the cancer and cells withrapid mitotic activity in close proximity to the tumors such as the intes-tinal epithelial cells. Electrolytes and nutrients absorption occurs in vil-lus epithelial cells and its damage results in reduced electrolyte andnutrient absorption, a major reason for gastrointestinal (GI) toxicity.Studies were therefore undertaken to study 1) Alterations in glucoseand amino acid absorption across the ileal tissues after irradiation and2) Formulate a nutrient mixture based on their absorptive capacity.Methods: NIH Swiss mice were irradiated (0, 1, 3, 5 or 7 Gy) using 137-cesium source at 1 Gy/minute. Ussing chambers were used for measuring trans-epithelial short circuit current (Isc) and isotope flux forelectrolyte movement. Paracellular permeability was assessed using conductance measurements and dilution potential and comparedto plasma endotoxin and IL-1 levels. All 20 amino acids were classi-fied based on improved electrolyte absorption, mucosal barrier mechanism and survival in animals. Survival studies were done to calculate LD50/7. Amino acids were mixed to form a drink and given as a gastric-gavage (200 µl/day). Results: There is a significantdecrease in glucose absorption after 0, 1, 3, 5, 7 Gy (140.7±32.2,71.8±7.6, 43.3±6.7, 16.0±2.5, 3.2±0.8 A.cm-2.h-1). Some amino acids showed decreased absorption while some showed increased absorption. Glutamine, showed decreased absorption after irradiation (41.4±23.3, 23.9±1.1, 10.9±4.2, 8.5±1.7,8.1±1.3 A.cm-2.h-1). Unlike glutamine, lysine absorption was significantly increased following irradiation (2.1±0.4, 15.1±2.0,15.3±4.5, 20.2±4.1, 52.2±16.6 A.cm-2.h-1). Amino acids lysine,aspartic acid, glycine, isoleucine, threonine, tyronsine, valine, trypto-phan and serine decreased plasma endotoxin (120 ± 8 vs 62.1 ± 6.7EU/ml), IL-1 ± 6 vs 46.1 ± 4.3 pg/ml) levels andshowed increased anion selectivity with dilution potential experiments in7 Gy irradiated animals compared to non-irradiated mice. Conclusions:Irradiation altered electrolyte and nutrient transport. Amino acidsimproving electrolyte absorption and /or improving the mucosal barriermechanism formed an optimal mixture to effectively overcome GI toxicityfollowing radiotherapy.

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Evaluation of Citrulline as a Biomarker for Radiation-InducedGut Damage

J. JonesSchool of Pharmacy - Pharmaceutical Sciences, University of Maryland,Baltimore, MD, USA

The potential risk associated with deliberate and/or accidental radiologicalcatastrophes underscores the urgent need for medical preparedness andeffective medical countermeasures (MCM). To date there are no approvedMCM to treat severely irradiated personnel. The National Institute of Allergyand Infectious Diseases (NIAID)-sponsored Consortium, Medical Counter-measures Against Radiological Threats (MCART), is charged with develop-ing MCM to treat the key sequelae of the acute radiation syndrome (ARS)and the delayed effects of acute radiation syndrome (DEARE).

Identification of biomarkers that accurately model radiation-induceddamage is critically important. An ideal biomarker has the potential toprovide early indication of a radiation-induced syndrome prior to theonset of organ-specific damage. Furthermore, the ideal biomarker dis-plays a tethered relationship not only between radiological dose but alsodose over time and dose over time over intervention. Recently, Citrullinehas been identified as a possible biomarker for radiation-induced gutdamage (Lutgens 2007).

Acute radiation syndrome in the gastrointestinal system (GI-ARS) isvery early onset (due to the rapid rate of cell turnover in the tissue) but isinduced by higher doses than those causing the haematological syn-drome (H-ARS). In the mouse model GI-ARS is manifest at doses typi-cally above 12Gy, but with diarrhea evident as early as day 4/5 post irra-diation. At day 4 small intestinal crypt loss is greatest, with measurablerecovery evident by day 6. We therefore quantified the circulatingplasma citrulline levels using LC-MS/MS in 10 -12 week C57Bl/6 mice 4and 6 days following total body irradiation (300 kV X-ray) at 8-15Gy.

Our preliminary data indicate a dependent relationship between cit-rulline concentrations and radiation dose. Further, there was a correla-tion with the level of small intestinal epithelium present (number x sizeof the regenerating crypts). On day 4 there was a good dose response at8 - 11Gy, but at the higher GI-ARS doses the dose response was lessobvious. By day 6, however, the correlation extended to 14Gy.

On-going efforts will determine the relationship between plasma cit-rulline concentration and radiological dose over time and intervention viaMCM for the GI-ARS mouse and non-human primate models.

Downwinder\'s Syndrome; Implications for the Public andHealthcare, then and now.

S. Baack, R. KeeneDepartment of Veterans Affairs-Healthcare, Robinson, TX, USA

The threat of nuclear incidents has precipitously increased as worldaffairs grow turbulent, and natural disasters seem to have become morefrequent and virulent. Such events put the public at risk for exposure tonuclear accidents, as the world witnessed during the tsunami aftermathat Fukushima nuclear power plant. It is imperative to have an informedpublic and a healthcare workforce knowledgeable in the assessment,triage and care for victims of direct and indirect radiologic exposure.Downwinders Syndrome refers to individuals who lived and/or workeddownwind of nuclear testing facilities from January 1951 -October 1958.These individuals were exposed to fallout from the atmospheric detona-tion of nuclear devices at the Nevada test site in the United States.Those who have not yet died from associated diseases are presentlyplagued by illnesses that have been passed on to their children andgrandchildren. Healthcare workers must be knowledgeable and able torecognize the signs and symptoms associated with this syndrome inorder to expedite treatment.

They must also be prepared to respond to present day threats such asdetonation of a nuclear weapon, the meltdown of a nuclear reactor,explosion of a large radiologic dispersal device also known as a “dirtybomb”, or even a radiation exposure device strategically placed in a

densely populated area (Wolbarst et al 2010). Nursing comprises thelargest segment of the healthcare workforce. But, are they adequatelyprepared to respond to such looming threats, were one to occur today?The literature suggests that nurses are not prepared to manage the com-plexity associated with the variation that such events would incur. If thenursing workforce were prepared, would they respond? While research islimited, Veenema et al (2008) suggest that most would not respond iftheir personal of safety were in question. In a survey of 620 nurses(Baack & Alfred, 2012), 80 % were not familiar with decontaminationprocedures or felt they had the ability to identify exacerbation of expo-sure to chemical or biologic agents. It is incumbent upon Public Health,Military entities and private healthcare to ensure that their workforce iscapable and willing to respond in an effective manner to such an event.

Database SEARCH: Radiation induced skin reactions andgastrointestinal signs and symptom as prognostic factors ofthe acute radiation syndrome.

H. Dörr, T. Baier, M. Hoebbel, V. MeinekeBundeswehr Institute of Radiobiology affiliated to the University of Ulm,Munich, Germany

The databank system SEARCH (System for Evaluation and Archivingof Radiation Accidents based on Case Histories) contains 824 clinicalcases from 81 radiation accidents in 19 countries from 1945 to 2001.This exceptional collection of clinical data from accidentally radiationexposed persons are analysed regarding the clinical course and insearch of parameters allowing an early prognostic estimation of theacute radiation syndrome.

Radiation induced skin reactions and gastrointestinal signs and symp-tom were analysed in terms of their meaning as prognostic factors of theacute radiation syndrome.

The main focus of this investigation was the relation of the occurrenceof different signs and symptoms and their combination for the prognosis.Since most of the gastrointestinal signs and symptoms occur early andare easy to detect they can be used as early predictors of prognosis inmass casualty scenarios.

Radiation induced skin reaction appeared to be an independent prog-nostic parameter beneath the well-known categories regarding thehematopoietic system. Within the group of patients with potentiallyreversible hematopoietic damage the percentage of affected skin surfaceis crucial for the outcome of the patient.

Stem cell technologies in treatment of radiation burnsKotenko K.V., Bushmanov A.Y., Eremin I.I., Moroz B.B., Galstyan I.A.,

Nadezina N.M. State research Centre Burnasyan Federal Medical Biophysical Center ofthe FMBA of Russua, Moscow.

The wide use of diagnostic and therapeutic radiation exposure inmedicine leads to the formation of a new group of patients with radiationexposure aftereffects. The group injured with a medical irradiation ischaracterized by progression of II-IV severity local radiation injuries (LRI)which require operative treatment methods administration. However thesevere somatic status of the patient may interfere these interventions.

Isolated conservative therapy (both general and local) in the similarseverity LRI is usually ineffective. Therefore cellular technologies may bethe therapy of choice for this group of patients.

In numerous experimental studies in LRI animal models with 140Gy90Sr/90Y source of beta radiation use the strength of mesenchymalstem cells (LRI) in the early periods after irradiation has been proven.Subcutaneous cells injection around the affected area later 8 days afterexposure (moist dermatitis formative period) led to radiation ulcers heal-ing within the 2 months after irradiation, whereas ulcers in the controlgroup did not heal after 4 months.

Within the framework of scientific and research study and after ani-mals testing with the protocol research approval by ethics committee

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and after informed consent signing by the patient were performed lim-ited clinical trials.

Five patients with chronic radiation ulcers arising as a complication ofX-ray and radiotherapy, were transplanted autologous mesenchymalbone marrow cells, grown ex vivo.

Conservative treatment regimen was ineffective whereas surgical treat-ment is impossible for medical conditions. Surface of radiation ulcers ofthe all patients was cover with mesenchymal stem cells with fibrin glueand stem cells intake around the lesion was administer.

There was complete healing of the deep radiation ulcers, intractableby conservative therapy, as a result.

The use of MSCs could be useful in the treatment of severe local radi-ation injuries, burns of different etiology and non-healing wounds.

Gene therapy: a new approach to mitigate CutaneousRadiation Syndrome

Diane Riccobono1, Diane Agay1, Fabien Forcheron1, HarryScherthan2, Viktor Meineke2, Michel Drouet11IRBA-antenne La Tronche-CRSSA, La Tronche, France2Institut für Radiobiologie der Bundeswehr in Verbindung mit der Uni-versität Ulm, Munich, Germany

Cutaneous radiation syndrome (CRS) caused by accidental/therapeu-tic irradiations is characterized by iterative inflammatory waves, incom-plete wound healing and poor revascularisation. Adipose derived stemcells (ASCs) have been reported to favour wound healing and paracrineprocesses are thought to be crucial in this context. Ex vivo manipulationof ASCs in order to transiently secrete ad hoc factors may increase theirpotential. Based on its capacity to induce proliferation of a variety of celltypes and its pro-angiogenic activity we chose to evaluate the SonicHedgehog (Shh) morphogen as a first candidate.

Minipig ASCs were isolated as previously described (Agay et al Exphematol 2010) then electroporated (Amaxa®) using a PIRES2 plasmidcoding for Shh.

In a first step the biological activity of Shh-ASCs was evaluated in vitro.Cells were culture for 7 days in essential medium with 10 % fetal calfserum and culture media (CM) was harvested at the end of the culture.Minipig fibroblasts were irradiated (25Gy gamma) then immediatelyincubated in CM and analyzed at different time (30 min, 6 hours(H),H24 and H48 post irradiation) for apoptosis (annexin V staining), senes-cence (ß-galactosidase staining) and gene expression using RT-qPCR.Globally, Shh-CM improved fibroblasts survival from H6 to H24 (p<0.05)and senescence rate was significantly lower at H48. In addition weobserved a sustained production of pro-angiogenic factors such asVEGFa and ANGPT1 at H24. Work is going on to characterize the Shh-ASCs activity upon DNA damages and fibroblasts secretome.

Secondly, we evaluated the biological activity of Shh-ASCs in vivo.Growth factor reduced BD Matrigel® loaded with Shh-ASCs or ASCswere subcutaneously injected in NOD/SCID mice. One week postimplantation, mice were euthanized and BD Matrigel® were analysedusing Drabkin method (spectrophotometry at 540nm) for haemoglobincontent. We observed a two fold increase in the Shh-ASC group whencompared with ASCs counterpart. Finally we injected autologous Shh-ASCs in locally irradiated minipigs as previously described. Injection waswell tolerated and preliminary results suggest that cell dose may repre-sent a crucial parameter.

Further studies will establish whether Shh transient gene therapy mayrepresent a valuable strategy to mitigate CRS.

Development and refinement of murine model (s) of radiationpneumonitis/ fibrosis to link with non-human primate andhuman pulmonary responses to radiation

Isabel L. Jackson1, Puting Xu1, Andrew Zodda1, Pheris Karanja1, Chiwei Hung1, Julian Down2, Cynthia Calley3, Barry P. Katz3, ZeljkoVujaskovic1

1Division of Translational Radiation Sciences, Department of RadiationOncology, University of Maryland School of Medicine, Baltimore, MDUSA; 2Massachusetts Institute of Technology, Cambridge, MA USA:3Department of Biostatistics, Indiana University School of Medicine,Indianapolis, IN USA

Approval of radiation countermeasures under the FDA Animal Rulecriteria requires pivotal efficacy screening against radiation toxicity in twospecies that react similarly to humans with respect to radiation dose-response, temporal onset, and histopathology. For the past four years,we have systematically characterized the dose-response relationship formorbidity/survival due to radiation pneumonitis/fibrosis in three murinestrains, CBA/J, C57L/J, and C57BL/6J.

In these studies, we refined our dose response curves using a doserange that covers sublethal to supralethal pulmonary tissue responsesfor three murine strains based on our previous data. In these studies,CBA/J, C57L/J, and C57BL/6J mice (n=20/group) were irradiated with auniform single dose ranging between 9-17 Gy WTLI (320 kVp, 1.0 mmCu HVL, dose rate 69 cGy min-1) and followed for up to 180 d post-radi-ation for survival. During this time, temporal changes in respiratory func-tion and body weight were assessed and lung weight recorded at thetime of necropsy. Linear mixed model analysis was performed on respi-ratory function parameters. Logistic regression/ Cox proportional hazardswere performed to determine the association between respiratory func-tion changes and survival. Pneumonitis and fibrosis were assessed inlung tissue harvested at the time of major morbidity or at the pre-deter-mined endpoint of 180 d. Further, we compiled data from irradiatedcontrol animals (non-MCM treated) over the past four years (radiationdosimetric parameters consistent across all studies evaluated) to furtherredefine our dose response curves and compare pulmonary responsesamong rodent strains to non-human primate (NHP) and human pul-monary responses (published data) irradiated to the whole thorax with asimilar range of radiation doses.

The overall goal of these studies was to establish a murine model that“links” in temporal onset, dose response, and pathology to NHPs andhumans to satisfy the FDA Animal Rule. Acknowledgements

This project has been funded in whole or in part with Federal fundsfrom the National Institute of Allergy and Infectious Diseases, NationalInstitutes of Health, Department of Health and Human Services, underContract No. HHSN266200500043C, Aeolus Pharmaceuticals, Inc(Mission Viejo, CA) and, Federal funds from the Biomedical AdvancedResearch and Development Authority, Dept. of Health and Human Serv-ices, under Contract No. HHS0100201100007C.

Development of CLT-008, a universal myeloid progenitor celltherapeutic for the treatment of neutropenia after exposure toradiation

H. KarsunkyCellerant Therapeutics, San Carlos, CA, USA

Cellerant has developed a novel universal cellular therapeutic for thetreatment of the hematopoietic subsyndrome of the Acute RadiationSyndrome (ARS). CLT-008 human Myeloid Progenitor Cells (MPC)derived ex vivo from human adult hematopoietic stem cells has the abil-ity to generate functional neutrophils in vitro and in vivo. We previouslyreported that in pre-clinical in vivo studies in mice using an animal ana-log product myeloid progenitor cells (MPC) prevent fungal and bacterialinfection, provide protection from lethal radiation and enable engraft-ment of low doses of stem cells. We have demonstrated that mouseMPC can be administered five to seven days after exposure to lethaldoses of radiation (9 Gy and above) with significant survival benefit. Thefunction of MPCs is not MHC restricted and no matching was necessary.Its human counterpart named CLT-008 is being developed as an off-the-self product for the treatment of ARS under the U.S. Food and DrugAdministration’s Animal Rule. Efficacy for approval will be demonstrated

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in two animal models and the safety will be demonstrated in humanclinical trials. We have developed a scalable culture system for thehuman product using animal product-free medium and a definedcytokine to generate myeloid progenitors that are significantly depletedof long term reconstituting HSCs and lymphoid cells. Further, we havedemonstrated that cryopreserved MPC maintain their colony formationpotential and produce mature neutrophils when exposed to G-CSF invitro. These neutrophils are indistinguishable by phenotype from periph-eral blood neutrophils, respond to physiologic stimuli, release reactiveoxygen species and possess phagocytotic activity. Using a mousexenograft model we also have shown the potential of MPCs to generatehuman neutrophils in vivo. This human product CLT-008 is currently intwo Phase 1/2 clinical trials: (a) patients undergoing cord blood trans-plantation after chemotherapy and radiation for the treatment of hema-tological malignancies and (b) patients receiving high-dose chemother-apy for the treatment of Acute Myeloid Leukemia. We have dosed overthirty patients to date. CLT-008 offers a novel approach to mitigate con-ditions of severe neutropenia and potentially provide life saving therapyto victims that have been exposed to radiation.

Inhibition of activation of signal transduction and increasesin cytokine and sepsis limits ionizing radiation combinedinjury

J. KiangRadiation Combined Injury, Armed Forces Radiobiology Research Insti-tute (AFRRI), Bethesda, MD, USA

Radiation exposure combined with traumatic tissue injury (radiationcombined injury, as abbreviated RCI) could be the central health effectsissue in many exposure scenarios. RCI can produce injuries at themolecular, cellular, tissue, and organ levels that are unique and morepronounced than either insult alone. The mechanisms of these synergis-tic interactions remain largely unclear.

In dog, pig, rat, guinea pig, and mice, radiation exposure combinedwith burns, wounds, or bacterial infection results in greater mortalitythan radiation exposure alone. In our laboratory, we found thatB6D2F1/J female mice receiving 60Co-γ photon radiation combinedwith 15 % total body surface area wounding reduced the LD50/30 to8.95 Gy (CL: 8.74, 9.11) from the 9.65 Gy (CL: 9.51, 9.82) determinedin mice receiving radiation alone. We found this non-lethal woundingenhanced radiation responses including γ-H2AX increases and survivindecreases in bone marrow cells, circulatory blood cell losses, increasesin serum cytokine concentrations, and activation of nuclear factor-kappaB, nuclear factor–IL6 and iNOS pathway in ileum and skin. RCIdecreased cadherin - 6, increased MMPs and TLRs in ileum and skin,and produced an earlier onset of bacterial infection in liver, spleen, andheart blood. In ileum, cell death occurred via apoptosis (as determinedby TUNEL assay) and via autophagy (as determined by immunoflures-cence staining against LC3-II).

Treatment protocols for RCI are being investigated. Preliminary datafrom our laboratory have shown that administration of mesenchymalstromal cells, G-CSF, Alxn4100TPO, ciprofloxacin, or ghrelin significantlyimproved survival after RCI. To manage the complex physiologicalresponses after RCI, we are assessing the efficacy of coordinated use ofseveral potential treatment protocols. (Supported by NIH/NIAID YI-AI-5045-04 and R21/33 AI080553;The views expressed do not necessarilyrepresent NIH, AFRRI, USUHS, or US DoD.)

Preclinical development of a bridging therapy for radiationcasualties

Vijay K. SinghArmed Forces Radiobiology Research Institute, Bethesda, MD, USA,Department of Radiation Biology, F. Edward Hébert School of Medicine, Uniformed Services University of the Health Sciences, Bethesda, MD, USA,

Victims of a terrorist attack presenting with the hematopoietic syn-drome resulting from exposure to excessive levels of ionizing radiationwill succumb to sepsis if not adequately treated. The probability of sur-vival is increased substantially if the victim’s immune system is allowedto recover before sepsis sets in. We report here preclinical developmentof a new bridging therapy which will allow the victim’s immune system torecover from damage caused by ionizing radiation.

CD2F1 mice were irradiated with lethal, whole-body doses of cobalt-60 gamma-radiation and then transfused intravenously (retro-orbitalsinus) with whole blood, peripheral blood mononuclear cells (PBMC) orplasma from tocopherol succinate (TS) and AMD3100-injected miceafter irradiation. Survival was monitored for 30 days after transfusion ofwhole blood, PBMC or plasma. Intestinal and splenic tissues were har-vested after irradiation and cells of those tissues were analyzed for mark-ers of apoptosis and mitosis. Bacterial translocation from gut to heart,spleen, and liver in TS-mobilized PBMC-treated and irradiated mice wasevaluated by bacterial culture.

The infusion of PBMC from TS- and AMD3100-injected mice signifi-cantly enhanced survival after irradiation, inhibited apoptosis, increasedcell proliferation in the analyzed tissues of recipient mice, and inhibitedbacterial translocation to various organs compared to mice receivingcells from vehicle-mobilized cells. TS and AMD3100 mobilized progeni-tors into peripheral circulation and the infusion of mobilized progenitor-containing blood or PBMC acted as a bridging therapy for immune-sys-tem recovery in mice exposed to high, potentially fatal doses of ionizingradiation. We suggest this novel bridging therapeutic approach thatinvolves the infusion of TS-mobilized hematopoietic progenitors followingacute radiation injury might be applicable to humans as well.

Use of Clara Cell Specific Protein as a Biomarker of LateRadiation-Induced Lung Effects

J. WilliamsRadiation Oncology, University of Rochester Medical Center,Rochester,NY, USAPurpose/Objective

Death as a result of radiation pneumonitis and fibrosis is frequentlyseen in lethally-irradiated personnel and forms part of the characteristicspectrum of events that make up the radiation-induced multi-organ dys-function syndrome (RI-MODS). We determined to identify possiblemarkers of lung injury that may be detected with sufficient time to miti-gate these lethal effects.Material/methods

Using an integrated preclinical testing system in both an adult andneonate mouse model, we have investigated the delayed radiationresponse in lung following near-lethal irradiation, with or without an addi-tional challenge (influenza) in order to identify patterns of exacerbatedtissue remodelling.Results

Following previous data that suggested that RI-MODS was linked toimmune deficiency, we have subsequently shown an associationbetween increased radiation-induced pulmonary toxicity and declin-ing levels of Clara Cell Specific Protein (CCSP) mRNA. Clara cells arelocated principally in the epithelium of the proximal or central por-tions of the pulmonary acinus, where their function is orientedtowards protection of the respiratory tract; of note, the influenza virushas been shown to preferentially target epithelial cells, whereas theeffects of radiation on this population are relatively unknown. We willprovide evidence of a differential radiation-induced effect on thephysiology of the Clara cells and expression levels of CCSP in twostrains of mice that demonstrate alternative pulmonary responses toradiation injury.Conclusion

Subsets of CCSP-expressing cells have been identified in both lungand bone marrow compartments and have been described as a progen-

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itor/stem cell pool involved in airway regeneration and alveolar home-ostasis. Therefore, radiation-induced damage within this compartmentmay have profound downstream long-term effects on lung developmentand response. Importantly, identification of Clara cells as a culprit in the

progression of radiation injury leading to pulmonary fibrosis may providea target for mitigation; furthermore, expression levels of CCSP may pro-vide an early biomarker of disruption in this critical cell population andan early indicator of potential late effects.

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9. Effects of low dose ionizing radiation

Low dose radiobiologyM. Atkinson

Institute of Radiation Biology, Helmholtz Zentrum München, Neuher-berg, Germany

Exposure to low doses of ionizing radiation is an unavoidable facet ofmodern society. Moderate to low dose exposures, ranging between tensto hundreds of mGy may accrue from medical procedures, high altitudetravel, the environment and the workplace. Extrapolations of the risksobserved at high doses suggest that an increased risk of cancer andnon-cancer diseases at low doses cannot be excluded.

The accepted paradigm of the dose response relationship is of a lin-ear non-threshold DNA damage response (DDR) prone to infrequenterrors that allow clonal expansion of cells bearing. This mechanism can-not explain non-cancer effects and may be complemented or replacedat low doses by other, non-linear, processes that are collectively termednon-targeted effects.

We are re-examining the biological responses in irradiated cells andtissues involved in non-cancer effects using a combination of proteomic,metabolic and transcriptomic technologies. From these studies we haveestablished that the cellular response to radiation include:n Post-transcriptional regulation of survival and death promoting pro-

teins by the non-coding RNA transcriptome.n Adjustment of the mitochondrial respiration pathway to increase reac-

tive oxygen production.n Activation of cellular processes involved in inflammation and motility.n Triggering of genomic instability in radiation-susceptible individuals.

The integration of these additional pathways into the radiationresponse at low doses will allow the construction of more biologically rel-evant models of the radiation responses relevant to disease. This in turnwill improve the fitting of epidemiological and molecular epidemiologicaldata sets that are used in predicting low dose risk.

The research leading to these results has received funding from theEuratom Seventh Framework Programme under grant agreement295823 (PROCARDIO) and from the Bundesministerium für Bildungund Forschung (BMBF 02NUK007A).

Low dose ionizing radiation effects - effects of thoron inhalationC.K.K. Nair, Aditya Menon, R.Indu and Tiju Chacko

Pushpagiri Institute of Medical Sciences and Research Centre, Tiruvalla689101, Kerala, IndiaIntroduction

A major fraction of background radiation is contributed by inhalationof radon, thoron and their progenies. Although, effects due to exposureto radon and its decay products have been studied extensively, fewerstudies are reported on exposure to thoron and progeny concentrations.Unlike in the case of radon progeny nuclides, 212Pb, a daughternuclide from thoron, is having a long half life (10.6 h) hence, the knowl-edge of 212Pb deposition in the inhalation path way and its migration toother organs are important informations in computing the whole bodydose. 212Pb though a beta emitter, decays to an alpha emitter 212Povia 212Bi (T½=60 m), another beta emitter in the series. 212Po has thehighest alpha energy (8.76 MeV) in the decay series of 232Th and isexpected to give maximum dose to cells where the atoms get deposited.The work is aimed to explore the effect of thoron inhalation in various tis-sues of swiss albino mice.

MethodologyM ice were exposed to thoron inside a fabricated Thoron exposure

chamber containing Thorium oxide (2 kg, obtained from Indian RareEarths) and having an average radiation dose of 65 mSv/hr. Swissalbino mice were kept in the chamber for various time intervals. Ani-mals were removed from the chamber at various time intervals and sac-rificed to study the effect of exposure on different tissues such as extentof cellular DNA damage, tissue antioxidants, serum parameters such aslevels of Glucose, Cholesterol, SGOT, SGPT, ALP, Total protein, Albumin,Globulin, Urea, Creatinine, and LDH, blood count, histopathology onselected tissues, etc. Genotoxic effect of thoron exposure on peripheralblood leukocytes and bone marrow cells were analysed by employingthe techniques of alkaline single cell gel electrophoresis, micronucleusassay etc. Results

Studies on hematological parameters suggested that the alteration inhematological parameter induced by thorn exposure was transient andthere was recovery after 9 days. The DNA comets of the bone marrowcells of the control and thoron exposed mice were similar while those ofthe blood cells of the exposed animals revealed slight increase in DNAdamage. The DNA damage in lung cells could be detected as early as 4hrs exposure in the chamber which is equevalent to averl low dose of~260 mSv. The DNA comets from the cells of the lung tissue were typical of cells undergoing apoptosis by the presence of fan shapedcomets. This would suggest that the inhalation of thoron induces severeDNA damage in the lung cells of the exposed mice which would causeapoptotic elimination of these damaged cells. In studies on micronucleiinduction, it was found that after 70 days of thoron exposure, micronu-cleated reticulocyes did not change from the normal value indicative ofinability of thoron inhalation to produce genomic instability at the pres-ent dose. Normal levels of micronucleated reticulocytes establish thatthoron doesn’t produce genomic instability in bonemarrow. Histology oflungs showed areas of abnormalities. Conclusions

Presence of fan shaped comet confirms the induction of apoptosis inlungs. The damage to leukocytes may be due to the location of heart tothe proximity of lungs. Lack of micronucleated reticulocytes confirmsthat, thoron does not migrate to bonemarrow in large quantities to elicitany effect. Lungs is the primary organ affected due to exposure ofthoron. The observed DNA damage in the lung cells of the animals keptin Thoron chamber could be due to the damage in the DNA of the lungcells induced by high energy radiation emanating from radioactive decayof thoron and its progeny, particularly the alpha radiation of 8.8MeVfrom 212Po. Further studies are needed to confirm this.

Human Lymphocytes Responses to Low and High Doses of131I

A. Cebulska-WasilewskaInstitute of Nuclear Physics, Polish Academy of Sciences Krakow,Poland

In human body iodine is specially concentrating in a thyroid, thus,131I is used for diagnostic or therapy. An emerge of the Iodine-131 inambient air might be a sign of nuclear accident or threat. The aim of ourstudy was to explore how low and high doses of 131I influence on bio-markers levels associated to efficiency of DNA repair process and to

susceptibility to other exposure and health risk. Study groups consistedof 30 healthy, 41 individuals exposed to low 131I dose (Aav=3MBq),and 37 persons exposed therapeutic (Aav=497MBq). Cytogenetic bio-markers (CA, SCE, MN, FISH) were studied in vitro in cells before andafter challenging X-rays dose with methods for retrospective dosimetryand human monitoring. The standardized DNA repair competenceassay was used to study efficacy of repair process. Although, mean val-ues of all biomarkers after diagnostic treatment were slightly lower thanin control group, though, a strong variation between individualsresponses were different at low from that observed after high doses of131I. Moreover, five weeks after the therapeutic dose of 131I, lower effi-ciency of DNA repair and notably elevated cytogenetic damage inmetaphase and interphase[1] were detected suggesting increased healthrisk. Strong variability in levels of molecular and cellular were observedin responses to I-131 in subgroups, stratified according to reports on

cancers in immediate family (CiF - factor). When responses to low orhigh doses of I-131 of CiF+ subgroup are compared to group with nocancer reports, a higher level of sister chromatid exchanges, allied tohomologous recombination are observed. However, in this subgroup,23 % decrease of chromosome aberrations was observed and 35 %decrease in frequency of micronuclei in binucleated cells. Diverse, atlow versus high dose, dependence on CiF factor are observed for notrepaired DNA damage and number of cells with chromosomal damage.Moreover, dose response curves for percent of aberrant cells express thesaturation level, that is changing in the range from 0.5 - 4Gy dose fromhigh to low LET radiation. Concluding, our results, show that alteration ofDNA repair process due to genetic or radiation character and cellularvulnerability to exposure, might affects level of detected cytogeneticdamage and reliability of estimated dose and risk.

[1] Cebulska et al,J.of Radiat.Industry, 5(4) 331.

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Malignant neoplasms incidence among residents of Ozersk,Russia

I. MartinenkoSouthern Urals Biophysics Institute, Ozersk, Russian Federation

The safety of living in the vicinity of nuclear plants draws public andscientific attention, although large scale accidents (which for examplerequire evacuation of local residents) are rare. Information on possiblehealth effects among long-term residents of the areas close to objects ofnuclear industry should be available for trouble-free operation period ofa nuclear complex, this is even more important for periods of develop-ment due to possible technology shortcomings, in particular of wastemanagement system.

The city of Ozersk is located in the vicinity of the Mayak PA – nuclearcomplex commissioned in June 1948. During the period of early opera-tion (1948-1965) the technology was lacking gas purification system,which caused increase of background radiation in the city due to atmos-pheric releases, although the city is windward to the Mayak PA.Releases of noble gases and I-131 were the main contributors to the cityterrain.

Population of Ozersk (founded at the same time with Mayak PA)increased over time from about 50 000 people in 1950-ies to 85 000currently, in the same time number of children decreased during sameperiod by a factor of 2 and number of individuals older than 40increased by factor of 5. Ozersk cancer registry includes all city resi-dents diagnosed with malignant neoplasms in 1948-2008, and servesas the source of information about malignant neoplasms incidence.

There are 10939 cancer cases were diagnosed in the specifiedperiod. Solid cancers contributed 94 %, hemoblastoses-5.8 %. Thestudy presents structure of all malignant neoplasms diagnosed, identify-ing prevailing sites. Malignant neoplasms diagnosed in the workers ofthe Mayak PA main plants are given separately, including comparison ofcancer incidence structure to the remainder of Ozersk population. Thestudy also presents the malignant neoplasms incidence as number ofcases per 100 000 of population. The current study does not includeanalysis of radiation contribution to cancer risk, since estimation ofdoses resulting from population technogenic exposure is incomplete.

Biological indication and dosimetry of chronic exposureTamara Azizova1, Viktor Meineke2

1Southern Urals Biophysics Institute (SUBI), Ozyorsk, Russia2Bundeswehr Institute of Radiobiology (BIR), Munich, Germany

The objective of this research is identification of biological markers ofchronic external and/or internal exposure and development of the bio-logical dosimetry system for estimation of external or internal doses from

chronic radiation. Major study tasks are as follows: selection of Mayakworkers occupationally exposed to chronic external and/or internal radi-ation, and control individuals for the study; clinical and biophysical stud-ies; analysis of structural chromosomal damages; analysis of proteinexpression (activators and inhibitors of apoptosis, cytokines,immunoglobulin, heat shock proteins etc.); whole genome microarrayanalysis of gene expression (apoptosis genes, genes of reparation /prolif-eration, signaling genes, metabolic genes etc.); identification of biologi-cal markers of chronic exposure (bioindication); analysis of relation ofbiomarkers identified to external and/or internal dose.

The study was performed in 100 Mayak workers divided into 3groups:n Mayak workers with external gamma-exposure at total external-

gamma-dose from 0.5 Gy to 2.0 Gy (10 individuals);n Mayak workers with external gamma-exposure, total external gamma-

dose was more than 2 Gy (8 individuals);n Mayak workers with combined exposure: total external gamma-dose

was more than 1.0 Gy; 239Pu body burden was more than 0.8 kBq(82 individuals).Control group consisted of 50 individuals matched to individuals from

the main groups by gender and age never exposed occupationally neverinvolved in any cleanup operations following radiation accidents, neverlived at contaminated areas. To perform the study following methodswere applied: mFISH, telomere analysis, immune-enzyme analysis, flowcytofluorometry (analysis of different subsets of peripheral blood lym-phocytes), and molecular and genetic analysis (analysis of gene expres-sion based on miRNA microarray followed by gene expression test usingreal-time qPCR). Preliminary results showed statistically significant rela-tion of chromosomal aberrations (translocations), some components ofprotein expression (concentration of T-helpers and T-NK-lymphocytes,IgM and IgA) and gene expression (40 expressed miRNA, approximately350 dysregulated genes) to external dose from gamma-rays and/or inter-nal dose from alpha-particle radiation. Results of the study are to be pre-sented in details in the report.

The study was funded by Bundesministerium der Verteidigung, San-itätsamt der Bundeswehr under the contract M / SAB X / 9A001.

Gene expression analysis in Mayak workers withoccupational prolonged exposure

Abend M.1, Mueller K.1, Doerr H.1, Kreppel H.2, Rusinova GG3,Glazkova IV.3, Vyazovskaya NS.3., Schmidl D.4, Unger K.4, Meineke V.1,Azizova TV.31Bundeswehr Institute for Rabiobiology affiliated to the Univesity of Ulm,Munich, Germany, 2Bundeswehr Medical Office, Department IX 1,

10. Radiation epidemiology

CBRN Med Defence, Dachauer Str. 128, D-80637 Munich 3ClinicalDepartment Southern Urals Biophysics Institute Ozyorskoe shosse 19Ozyorsk, 456780, Russia, 4 Research Unit of Radiation Cytogenetics,Integrative Biology Group, Helmholtz Center, Munich, Germany

We evaluated gene expression in the peripheral blood in relation tooccupational exposure in Mayak workers. Workers were exposed toeither combined internal alpha-radiation from incorporated 239Pu(239Pu burden >0.7 kBq) and external -rays (> 1.01 Gy, n=82) or toonly external -rays (> 0.5 Gy, n=18) to the red bone marrow (n total =100). Gene expression of the exposed groups was examined in relationto 50 unexposed individuals with all of them living in Ozyorsk and alivein 2011. Peripheral blood aliquots using PaxGene tubes were taken upto 5 decades after exposure. RNA was isolated (mirVana Kit, Life Tech-nologies), converted into cDNA and stored at -20°C. A two stage studydesign was performed focusing on examinations on the transcriptional(mRNA) and posttranscriptional level (microRNA). In the first stage weidentified 40 samples for screening purposes and selection of candidategenes. Three exposure groups and one group comprising unexposedindividuals were identified and 10 RNA samples per group were used.For examinations on the transcriptional level we hybridized 40 RNAsamples on 40 whole genome microarrays (Agilent, 4x44K). For exami-nations on the posttranscriptional level we measured about 760 differentmicroRNA species simultaneously using qRT-PCR (LDA type A/B, LifeTechnologies). Candidate genes were assessed by (1) introducing a 2-fold difference in gene expression over the reference group and (2)showing a significant p-value using the Kruskall-Wallis or Man-Whitneytests. From about 40,000 transcripts hybridized on the wholegenome microarray we selected 376 candidate genes (80 up regu-lated and 296 down regulated relative to the reference group).Expression of almost all of the genes (70 - 98 %) appeared significantlyassociated with internal alpha-radiation from incorporated 239Pu and toa lesser extend associated with the external -rays (2 -30%). Associationsin the same direction were found for 46 microRNAs. A selection of these

candidate genes are currently processed for the second phase of ourtwo stage study design where we validate our results using qRT-PCR onthe remaining RNA samples. Results will be presented at the meeting.

Chronic radiation syndrome (CRS) in residents of the Techariverside villages

A. AkleyevUrals Research Center for Radiation Medicine, Chelyabinsk, RussianFederation

In 1950s 940 cases of CRS were diagnosed in residents of the Techariverside villages exposed to radioactive contamination due to thereleases of liquid radioactive wastes of Mayak PA. Patients during theperiod of pathology formation most frequently demonstrated hematologicand neurologic changes, different functional visceral disorders, ostealgicsyndrome and asthenic manifestations. Thereby, in clinical picture ofCRS specific symptoms were not observed. And indeed, as the results ofthe long-term follow up of the patients with CRS demonstrate in mostcases overdiagnosis occurred mainly due to the absence of individualdosimetry data and information about the initial health status of thepatients. Most often clinical picture of the CRS was imitated by hemato-logic, infectious and some other diseases. In some cases on the basis ofthe results of the long-term follow up the diagnosis CRS was verified.The obligatory procedure of the verification process was differentiateddiagnosis of the diseases that have similar clinical symptoms and theexistence of CRS course dependence on exposure dose rate and dose tocritical organs (bone marrow and nervous system). The analysis of theclinical symptoms dynamics and laboratory changes gave the possibilityto conclude that CRS at the initial stage is a “dysregulatory” pathologydue to the disorders of the regulatory systems of the human body (ner-vous, endocrine, immune and hematopoietic systems). The presentationwill dwell upon the following issues: CRS epidemiology, exposure dose tocritical organs assessment, pathogenesis, clinical manifestations during theperiod of pathology formation and recovery, and also late effects of CRS.

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The military use of magnesium/ thorium-alloys and radiationprotection of workers

U. Schenk1, A. Schirmer2, U. Warnecke2

1German Ministry of Defense, IUD II 4, Bonn, Germany, 2Northern Mili-tary District Administration - Radiation Measuring Laboratory, Munster,Germany

Due to their low density and large creep resistance up to high temper-atures Magnesium/Thorium-alloys have frequently been used for theconstruction of aircraft parts, especially aircraft engines. The averageconcentration of Thorium is about 1.7 %, the specific Th-232-activityranges between 45 Bq/g and 70 Bq/g. When the alloy is cast, Th-232and Th-228 are separated from their decay daughters, which dominatethe Gamma-dose rate at the surface of the material. The Gamma-doserate then builds up and may reach about 20 µSv/h at the surface of alarge component. Although Thorium is nowadays replaced in the pro-duction, many components are still in military use. Here the issues ofthe radiation protection of workplaces in aircraft-engine maintenanceand maintenance training are addressed. Dose estimates for the externalexposure are given and incorporation pathways are discussed.

Radiation protection in a mixed contaminant context, riskassessment methodologies

Ivica PRLIC1, Marija SURIC MIHIC1, Mladen HAJDINJAK21Unit for Radiation Dosimetry and Radiobiolgy, Institute for MedicalResearch and Occupational Health, 10000 Zagreb, Republic of Croatia2Haj-Kom d.o.o., 10000 Zagreb, Republic of Croatia

Increasing industrialisation and population density have led to situa-tions where humans and the environment are exposed to a multitude ofpotential stressors. Often little is known about the mid- and long-termhealth and ecological consequences of these especially when they occurin synergy, as mixtures. The consideration of chronic low-level mixedexposures presents considerable challenges for methodology and datainterpretation. To fully understand the effects of multiple stressors onlife-history responses such as growth, reproduction and survival requireschallenging experimentation and a multidisciplinary approach. X- rays isa specific stressor present in our lives for more than a century. It´s use,except for medical purposes, spreads very fast in civil use, airport secu-rity, state office security etc. The special agencies dealing with homelandsecurity are being established after the furious terroristic attacks all overthe world. The people employed in such agency use sophisticated imag-ing equipment. They need to be well educated, although their overalleducation is sufficient to be secondary grade. Radiation protection edu-cation is needed. These new working places are loaded with totally newpsychological and technological stressors which are certainly a newresearch challenge in occupational health. Population as a whole will beexposed to these new stressors. The need to develop and use improvedassessment tools and novel models, to reduce uncertainty in current riskassessment and screening methodologies, for example by improving thescientific basis for setting safety factors exists. This will facilitate humanand ecosystem health monitoring by providing the link with informationconcerning the health condition of the population and environment whileusing the new technology in an unselective manner.

11. Radiation protection

Radiation risks of potential cancer following the CTdiagnostic radiation exposure

V. V. Kashcheev, V. K. Ivanov, S. Yu. Chekin, A. N. Menyaylo, E. A.Pryakhin

National radiation-epidemiological registry, Medical RadiologicalResearch Center of the Ministry of Health of the Russian Federation(MRRC), Obninsk, Russian Federation

The current study quantify the magnitude of the difference betweenradiation risks evaluated with the use of organ or effective doses particu-larly when planning pediatric and adult CT examinations. The radiation-induced cancer risks from medical CT examinations have been evalu-ated as a function of the sex and age. Lifetime attributable risk values ofCT scanning have been estimated with the use of ICRP (Publication103) risk models and Russian national medical statistics data. For pop-ulations under the age of 50 the risk evaluated using organ doses usu-ally differs from the risk using effective doses by less than 30%. In olderpopulations the difference can be up to a factor of 3. Calculated valuesof lifetime attributable cancer risk and attributable risk fraction for par-ticular organs are presented as well.

Pharmacological strategies for the development ofradiomitigators

Gábor Tigyia, Gyöngyi N. Kissa,f, Renukadevi Patilb, James Fellsa,Shuyu Ea, Sue-Chin Leea, Yuko Fujiwaraa, Karin Emmons-Thompsonb,Charles R. Yatesb, Duane D. Millerb, Louisa Balazsc, Wenlin Dengd, JurStrobosd, Koen Van RompayeDepartments of Physiologya, Pharmaceutical Sciencesb, Pathologyc,University of Tennessee Health Science Center, 894 Union Avenue,Memphis TN, 38163 USA, RxBio Inc., 3N Dunlap St, Suite C303, Mem-phis TN, 38163, California National Primate Research Centere, Univer-sity of California, Davis, CA 95616, Department of Biochemistry andMedical Chemistryc, University of Pécs, Szigeti ut 12, Pécs, 7624 Hun-gary

A decade ago, based on earlier evidence establishing the prosurvivaland anti-apopotic actions of lysophosphatidic acid (LPA), our groupembarked on studies aimed at the evaluation of LPA and its analogs forthe attenuation of radiation injury. This research gave us two newinsights: First, we elucidated fundamental cell signaling mechanismsthat are activated by radiomitigators and second, we developed a classof compounds that selectively activate a subtype of LPA receptors.Octadecenyl thiophosphate (OTP) is a metabolically stabilized mimic ofLPA and a full agonist of the LPA2 receptor. The plasma half-life of OTPin mice and nonhuman primates is ~ 10 -14 h. LPA and OTP, adminis-tered to mice irradiated with lethal levels of γ-irradiation (6 – 12 Gy) toelicit the hematopoietic or the gastrointestinal acute radiation syn-dromes, reduced the number of apoptotic cells in the crypt-villus unitsand decreased mortality. The crypt cells of wild type mice and LPA1 KOmice were protected by LPA or OTP administration from GI-ARS. Bycontrast, neither LPA nor OTP showed any protection in LPA2 KO miceagainst radiation injury. OTP also protected crypt cells and decreasedmortality in a nonhuman primate model of GI-ARS. In vitro, the radio-protective effect of OTP was present only in cells that expressed theLPA2 receptor subtype. LPA2 forms a macromolecular signaling com-plex via its C-terminal type 1 motif recognized by the PDZ-motif bindingprotein NHERF2. LPA2, but not LPA1 or LPA3, can interact with anumber of zinc finger proteins though a C-terminus-mediated interac-tion. Ligand-activated binding of LPA2 to Siva-1 is of particular impor-tance in this regard because it is a target of p53, an early response geneactivated by DNA-damage that promotes apoptosis. Siva-1 binds up theantiapoptotic Bcxl-XL protein making the mitochondrial outer membranemore prone to apoptosis. Siva-1 recognizes a C311-x-x-C motif in the Cterminus of LPA2. Activation of LPA2 promotes the complex formationbetween this receptor subtype and Siva-1. Once this complex is formed,it is withdrawn for the endocytotic receptor recycling and instead it is

polyubiquitinated and degraded. The overall effect of LPA2 activation isthe depletion of the cell for Siva-1, which in turn leads to the attenuationof apoptotic signaling. We have extended our research to uncover thenext generation of LPA2 agonists. LPA2. Using GRI977143, a non-lipidspecific agonist, as a scaffold to derive a series of selective agonists,yielded highly effective new compounds reducing death in miceexposed to 15.6 Gy partial body γ-irradiation with 5 % bone marrowshielding and also from HEM-ARS elicited by 8.5 Gy total body irradia-tion. In mice the therapeutic window extends to +72 h postirradiation.Supported by NIAID 80405, 087550, and RxBio Inc.

Oxidative lipodomics of mitochondrial responses to radiationand design of new radioprotectors/radiomitigators

V. KaganEnvironmental/Occupational Health, University of Pittsburgh, Pittsburgh,USA

Radiation injury is mostly due to oxidative reactions triggered byimmediate radiolysis of water leading to DNA damage and host oxidativestress responses in different tissues. Lipid molecules are among themost vulnerable targets of these oxidative reactions. We have pioneeredthe field of oxidative lipidomics and established that different cell deathmechanisms engage specific lipid oxidation pathways leading to thegeneration of time- and tissue-selective “oxidation-specific” epitopes.We performed detailed analysis of lipid oxidation products accumulatingin vivo in several tissues (eg, small intestine, lung) of mice exposed to alethal dose of irradiation (9.25 Gy) as well as in the brain of rats aftercompact cortical impact. Lipid peroxidation did not follow the profile pre-dicted by stochastic involvement of lipids in the oxidation process butdisplayed a highly selective pattern. Two anionic phospholipids - mito-chondrial-specific cardiolipin (CL) and extramitochondrial phos-phatidylserine (PS) - were the major substrates of peroxidation reactionswhereas more abundant and highly polyunsaturated phosphatidyl-choline and phosphatidylethanolamine molecular species remainednon-oxidized. We established that this pattern of phospholipid oxidationis associated with the execution of apoptotic program and subsequentclearance of apoptotic cells by professional phagocytes. Notably,cytochrome c (cyt c) turned to be the major catalyst of oxidation reac-tions whereby the formation of high affinity complexes of the hemopro-tein with CL and PS was the major cause of selectivity of the oxidationprocess towards these anionic phospholipids. We designed and testedseveral novel small molecule inhibitors that specifically affect the peroxi-dase activity of cyt c complexes with CL and PS or deprive cells ofH2O2, a required source of oxidizing equivalents for the peroxidationreactions. These inhibitors were effective in preventing peroxidation ofCL and PS and acted as protectors against radiation-induced cell death.Thus, selective inhibitors of specific peroxidation reactions, catalyzed byredox-enzymes, represent new targets for mechanism-based “antioxi-dant” interventions.

Radioprotective Effectiveness of BASIC PENTACYSTEINEanalogs on Radiation-induced Skin Damage

Fuad Fares1, Steen Jensen2

1Department of Human Biology, Faculty of Natural Sciences, Universityof Haifa, Mount Carmel, Haifa 31095, Israel. 2Deparment of Cancer Surgery, Hammersmith Hospital, London, UK.

Radiation-induced skin damage is a well-known complication of radi-ation therapy. Cutaneous radiation injury (CRI) can occur with radiationdoses as low as 2 Gray (Gy) or 200 rads and the severity of CRI symp-toms will increase with increasing doses. Most cases of CRI haveoccurred when people inadvertently came in contact with unsecuredradiation sources from food irradiators, radiotherapy equipment, or welldepth gauges. In addition, cases of CRI have occurred in people whowere overexposed to x-radiation from fluoroscopy units. The aim of thepresent study is testing the efficacy of two BASIC PENTACYSTEINE

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analogs, BP-C2 or BP-Cx-1, in protecting mice from skin injures induce byhigh dose of radiation therapy (RT). Mice will be anesthetized with anintraperitoneal injection of ketamine (100mg/kg) and xylazine (8 mg/kg).After ten minutes, the animals, as many as 2 at a time, will be positioned inmetal jig that allowed radiation exposure of the right posterior leg. The doserate from a 6 MV linear accelerator will be 4 Gy/24h for 8 times (total of 32Gy) and 8 Gy/24 h for 4 times (total 32 Gy), using 75 cm source to the sur-face distance. A 2.0 cm tissue equivalent bolus will be used to bring themaximal dose to the skin surface. Dose will be prescribed to the Dmax andmice received the fractionated schedule (24 hours apart) as indicated foreach experiment. Doses will be confirmed using micro-TLD dosimetry.Experiments will be approved by and in accordance with the guidelines ofthe Institutional Animal Care and Use Committee. In order to clarify themechanism by which BPC-analogs protect radiation-induced skin damage,normal fibroblast skin cells will be irradiated with 2. 4, 8, 16, 24 and 32Gy and DNA damage, cell cycle analysis and apoptosis will be detected.

BP-C2 is an oral radio-protective agent combining molybdenum and aglobular polydentant ligand known for its detoxifying, free-radical scaveng-ing and anti-oxidant activity, while molybdenum is a constituent of a num-ber of important detoxifying enzymes. Animal trials have demonstrated thatthe administration of BP-C2 results in highly significant survival advantagein animals receiving either lethal or sub-lethal doses of gamma irradiation.No adverse side effects using BP-C2 have been observed in vitro (LDHrelease) or in vivo.

BP-C2 is complemented in the Meabco radiation concept with BP-Cx-1 that also possesses protection properties along with no adverseside efffects. However BP-C2 has a slightly higher radio-protection effec-tiveness.

The current dosing window has an LD50/30 for BP-C2 at 6.56 Gy andfor BP-Cx-1 at 6.28 Gy.

The results of the studies will be ready in due time for presentation at the Conference.

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12. Radiation biology/radiation physics

Emerging issues in radiobiology and cancer research – theimpact of non-targeted effects

Carmel MothersillMcMaster University, Hamilton, Ontario, Canada; Email:[email protected]

Since the acceptance that non-targeted effects (NTE) can be meas-ured in unirradiated cells or distant progeny of irradiated cells, the dis-cussion has developed about the relevance of these effects for radiobiol-ogy and radiation protection since they increase the complexity of theradiation response and allow for outcomes which are not as predictableas they were under the “old rules”. Specific examples will be presentedand analysed which challenge accepted paradigms. 1. Data show thatbystander mechanisms are either on or off in cells and that the “on”threshold appears to be at a very low dose (mGy range). 2. Data suggestthat adaptive responses may be induced not only in neighbouring cellsbut in organisms which receive bystander signals. 3. Data show thatchronic exposures to alpha or gamma irradiation lead to complexresponses in organisms which can be adaptive and protective. 4. Evi-dence suggests that mixed contaminant exposures which include radia-tion can have sub-additive or synergistic effects. A key consequence offindings in NTE biology is that at any given level of organization, fromgene to ecosystem – communication of stress signals and heritability ofstress adaptations provide the bridges linking one hierarchical level tothe next and enable the rapid propagation of change triggered by stressat one level, resulting in change at a higher (or lower?) level. Evolutioncould thus be regulated through communicated signals between cells,individuals, and populations which control and optimize responses coordi-nating the emergence of exquisitely tuned systems which can adapt rap-idly to micro or macro environmental change. A current view of cancer isthat it is a “systems level” disease which can not be understood or treatedby looking at individual genes or pathways in the traditional way. Rather, asystem approach is required with looks at the environment at both cellularand organismal levels to understand what has been perturbed. We sug-gest that bystander mechanisms operate to coordinate responses inorganisms and that in cancer a fundamental dysregulation and disfunctionof these mechanisms underlies the onset of carcinogenesis.

Use of archival material to study miRNAsGayle Woloschak, Sumita Raha and Tatjana Paunesku

Northwestern University, USIrradiated animals tissue archive hosted at Northwestern University is

a repository of samples and data collected in the course of radiation ani-mal mega studies between 1950’s and 1990’s in several different

national laboratories and institutes in the USA. These archival samples(including dogs, mice and rats) can be used for investigation of proteinexpression, DNA content and expression of micro RNAs (miRs). In amost recent set of studies on miRs a custom array with 40 miRs wasused to interrogate murine spleen samples from controls and 24 specificradiation exposure regimens (from acute to daily exposures, and totaldoses between 0.9 and 24.6 Gy gamma rays or 0.009 to 0.7 Gy neu-trons). Expression patterns of 40 miRs were recorded for each sampleand the results were analyzed computationally leading to a grouping ofeach sample-array combination into 20 possible clusters. Investigationof cluster distribution shows that the majority of sample-array data frommice with any type of splenic disease results in four clusters (cluster IDs7, 9, 11 and 18), while the majority of the data from healthy mice, irre-spective of radiation most frequently belongs to a different set of fivecluster (cluster IDs 1, 3, 4, 15 and 16). However, many of the mice thatshow no signs of splenic disease, but were exposed to high doses ofgamma rays (24.6 Gy gamma rays over 300 fractions or 3.99 or 5.45 Gyas a single exposure) or neutrons (0.18 or 0.37 Gy neutrons as a singlefraction) show the miR expression pattern that puts them into sameclusters where animals suffering from spleen disease belong. We areinvestigating at this time if these miR expression patterns signify onset ofgenomic instability (as a prerequisite for the development of spleen diseases).

Radiation-induced alterations in the proteome and miRNAomeof the endothelial cells

Sriharshan A, Barjaktarovic Z, Kraemer A, Boldt K, Azimzadeh O, Sar-ioglu H, Hieber L, Tammio, H., Hakanen A, Leszczynski D, ZitzelsbergerH, Ueffing M, Moertl S, Atkinson MJ, Tapio S.Department of Radiation Sciences, Helmholtz Gesellschaft München,Neuherberg, Germany

High doses of ionising radiation damage the endothelial cells leadingto vascular dysfunction. Epidemiological data suggest that even moder-ate doses (> 500 mGy) may increase the risk of cardiovascular disease.At lower doses, endothelial cell stress and vascular damage may stilloccur, but the relevance of these effects for long-term tissue damage isunknown. The aim of this study was to investigate immediate radiation-induced changes in the protein and microRNA (miRNA) levels ofendothelial cells.

Proteome and miRNA alterations were analysed in the endothelial cellline EA.hy 926 at 4 h and 24 h after exposure to a radiation dose of 2.5Gy (Cs137-γ). The proteomic studies were carried out using SILAC and2D-DIGE strategies whereas miRNA alterations were analysed using

TaqMan-based low density array technology. After irradiation a total of59 and136 proteins as well as 22 and 17 miRNAs were found to be sig-nificantly differentially regulated at 4 h and 24 h, respectively. Directinteractions (negative-regulations) were predicted between severalderegulated proteins and miRNAs using the Ingenuity software.

Further, investigation of protein alterations in primary human coronaryartery endothelial cells after a single 200 mGy radiation dose (Co60)using 2D-DIGE technology revealed 28 significantly deregulated pro-teins. For six miRNAs known to be radiation responsive or involved incardiovascular diseases single assays were performed. The expressionof miR-21 and miR-146b showed significant radiation-induced deregu-lation. Using miRNA target prediction, three proteins found differentiallyexpressed in this study were identified as putative candidates for miR-21regulation. A negative correlation was observed between miR-21 levelsand the predicted target proteins, desmoglein 1, phosphoglucomutaseand target of Myb protein.

We conclude that the high-dose alterations might result in endothelialcell damage further leading to endothelial dysfunction. This study showsfor the first time that also a low-dose exposure has a significant impacton miRNA expression that is directly related to protein expression alter-ations. The data presented here may facilitate the discovery of biomark-ers of radiation-induced cardiovascular damage.

Local high-dose cardiac irradiation induces disturbed energymetabolism associated with impaired PPAR alpha activity inC57BL/6 mice

O. AzimzadehRadiation Proteomics; Institute of Radiation Biology, Helmholtz ZentrumMünchen, Neuherberg, Germany

Radiation exposure is associated with a markedly increased risk ofcardiac morbidity and mortality with a latency period of several decades.Although different studies have confirmed the damaging effect of ioniz-ing radiation on the myocardium and cardiac endothelial structure andfunction, the molecular mechanism is not yet fully elucidated. Peroxi-some proliferator-activated receptor alpha (PPAR alpha), a transcrip-tional regulator of lipid metabolism in the heart tissue, has recentlyreceived great attention in the development of cardiovascular disease.The goal of this study was to investigate the possible role of PPAR alphain the cardiac radiation response.

C57BL/6 mice were locally irradiated on the heart at the age of 8weeks using X-ray doses of 8 and 16 Gray. The mice were sacrificed 16weeks after irradiation and the cardiac proteome changes were quanti-fied using Isotope Coded Protein Label (ICPL) method followed by ESI

LC-MS/MS and Proteome Discoverer software analysis. The proteomicsdata were further validated by serum lipid profiling, immunoblotting,transcriptomics and bioinformatics.

The proteomics analysis indicated significant alterations in the cardiaclipid metabolism and oxidative phosphorylation. Ionizing radiationmarkedly changed the phosphorylation and ubiquitination status ofPPAR alpha. This was reflected as decreased expression of its targetgenes.

This study suggests persistent alteration of cardiac metabolism due toimpaired PPAR alpha activity in the irradiated hearts.

Age-related increases in inflammatory cytokines areexacerbated in aged murine survivors of acute total bodyirradiation exposed in early adulthood

Chua HL.1, Wolfe HR, Plett PA, Sampson CH, McVittie TJ, OrschellCM.1Indiana University School of Medicine, Indianapolis, Indiana, USA

We have previously shown that exposure to lethal, high dose rate, totalbody irradiation (TBI) results in severe, life-long damage to hematopoieticstem cell (HSC) function and a 15-20-fold increase in aggressive lym-phoma in a murine model of the hematopoietic syndrome of the acuteradiation syndrome (H-ARS). Given that acute radiation exposure resultsin a surge of inflammatory cytokines, and that chronic inflammation isimplicated in genomic instability and cancer initiation, we examined thepresence and magnitude of several inflammatory cytokines throughoutlife in murine survivors of H-ARS. To this end, C57Bl/6 mice wereexposed to the LD50/30 dose (8Gy) of 137Cs radiation (60-70cGy/min)at 3mo of age (TBI mice). Surviving TBI mice were assayed at varioustime points up to 22mo of age, along with age-matched non-TBI controls,for serum cytokines using a multiplex assay (BioPlex). In general, an age-associated increase in several inflammatory cytokines was observed inboth TBI and non-TBI mice, including IL1a, IL1b, IL2, IL6, IL12, GM-CSF, MCP1, MIP1a, IFNg and TNFa. The anti-inflammatory cytokine IL10also showed an age-associated increase. Of these inflammatory mole-cules, TBI mice exhibited 1.3- to 2.1-fold higher levels of IL1a, IL12, andof the pro-tumorigenic cytokines IL1b and MCP1 compared with non-TBImice. Anti-inflammatory cytokines IL3 and IL5 remained unchangedthroughout life in both groups. Taken together, these results demonstratethe marked presence of several pro-inflammatory cytokines in aged sur-vivors of acute lethal radiation exposure, suggesting a possible linkbetween TBI-induced chronic inflammation exacerbated by old age withthe observed loss of HSC potential, increased incidence of malignancy,and other detrimental delayed effects of acute radiation exposure.

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13. Non-ionizing radiation

Fibrillation risk of Taser weaponsN. Leitgeb

Institute of Health Care Engineering with European Notified Body ofMedical Devices, Graz University of Technology

The debate on potential health hazards associated with deliveringelectric discharges to incapacitate subjects, in particular whether elec-tric discharge weapons are lethal, less lethal or non-lethal is still contro-versial. The cardiac fibrillation risks of Taser weapons, type X26, X3 andXREP have been investigated by experimentally determining the deliv-ered high-tension pulses in dependence on load impedance. Cardiacexposure to electric charge densities has been assessed by numericalsimulation at MRI-based detailed anatomical models (NORMAN andVisible Man) with a spatial resolution of 2 x2x2 mm voxels. Since exper-imental data on fibrillation thresholds are sparse and mainly available forpower frequency sinusoidal electric currents it was necessary to deter-

mine the specific fibrillation threshold of the delivered short Taserpulses. This was possible by mathematical simulation at numericalcellular models of endocardial, myocardial and epicardial cells. Thegenerated data on volume-dependent cardiac exposure and their rela-tionship with the location of pulse delivery together with fibrillationthresholds of Taser pulses allowed assessing fibrillation risk of singleworst-case hits in dart mode and in contact mode application. Theoverall risk of Taser application is depending on the probability of crit-ical hits. Therefore, spatial hit distributions have been determined byassessing hits generated in trainings of police officers under realisticscenarios. The results allowed estimating the probability of hitting tar-get areas critical for cardiac fibrillation and the potential of furtherreducing risks by adequate weapon use. The analysis of the resultsshowed that fibrillation risk of Taser application is different for contactmode and dart mode and that it is low but existent.

Poster PresentationsConRad 2013, 13.–16.5.2013, Conference, Munich

Measurement of radionuclides at Karlsruhe Institute ofTechnology‘s in vivo monitoring laboratory after theFukushima incidents

Breustedt B.1,2, Mohr U.1, Marzocchi O.1, Biegard N.2, Cordes G.2

Karlsruhe Institute of Technology – KIT 1Institute for nuclear waste disposal – INE , 2Safety management – KSM

The in-vivo monitoring laboratory of KIT is an approved lab for individ-ual monitoring for incorporation according to German regulation andoperates two whole body and two partial body counters. The lab isaccredited to DIN EN ISO 17025:2005 and per-forms approx. 2000measurements per year.

After the incidents at Fukushima NPP radionuclides were released tothe environ-ment. Persons which have been in Japan at these times andlater had a chance to incorporate these radionuclides via different path-ways. At KIT 41 persons which re-turned from Japan have been meas-ured up to now (12/2012). Besides the naturally occurring K-40 whichwas identified in all measurements as excepted, only in two measure-ments the fission products Cs-134, Cs-137, I-131, I-132 and Te-132were identified. Efficiency factors for quantifying non-routine nuclideswere interpolated from existing calibrations and later on verified byMonte-Carlo simulations. Doses were calculated using dose conversionfactors provided by the federal office for ra-diation protection in Ger-many (BfS). These factors assume for the first phase after the release aninhalation at the first day of the releases (March 12th). For later timesthe ingestion pathway will be dominant and needs to be taken intoaccount. Defining the intake scenario will be the difficulty then.

KIT’s in-vivo measurement lab quickly assembled handouts generallyunderstand-able also for the interested public, based on existing texts,

tables and figures. Infor-mation and values for comparison wererequested especially concerning the dose levels. A comparison with nat-ural levels of radiation proved to be very helpful here. Unlike expectedduring the first days, fortunately the number of these measurementsstayed low so that the routine operation could be continued unaffectedly.For the lab a good and quick response to the events in Japan was possi-ble by the teamwork within the laboratory and the good collaborationwith other German laboratories and BfS.

Operation Tomodachi (\"Friend\") Radiation Exposure RegistryK. Haines

US Air Force, Armed Forces Health Surveillance Center, Silver Spring;MD, USA

This presentation will cover important aspects of the U.S. Departmentof Defense’s force health protection mission following the Japanese earth-quake and tsunami which occurred on March 11, 2011. The earthquakeand tsunami resulted in severe damage to the Fukushima Nuclear PowerStation, which resulted in the subsequent release of low level radiationexposure to the nearly 70,000 members of the DoD-affiliated population.The presentation will provide information on the following: DoD’s health-related mission in the region; Internal and external radiation monitoringaccomplished; the determination of the radiation monitoring period to beused for dose assessments; how the annual dose estimates were calcu-lated; the maximum total effective doses calculated; and a familiarizationwith the construction of an exposure registry and the development of anassociated Registry website for communications with the affected popula-tion, as well as the general public, and where location-based doses aremade available along with their medical interpretation.

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1. Fukushima

2. Radiation risk perception and communication

Parallel societies in radiation protection?Dieter F. Regulla Research Unit Medical Radiation Physics and Diagnostics, Helmholtz

Zentrum München, Neuherberg, GermanyIonizing radiation and its impact to man is judged by the vast majority

of the people first and foremost from the consequences of the atomicbombs of Hiroshima-Nagasaki as well as from the nuclear incidents ofChernobyl and Fukushima. At the same time, however, ionizing radiationis accepted part of everyday lives. In diagnostic radiology for instance, X-ray based imaging, such as computer tomography, PET/CT, angiographyand interventional radiography, represents an essential element of up-to-date medical diagnostics and considered prerequisite aiming to preservehealth and prolonging life. But the revolutionary imaging techniquesbecame, over the last few decades, largest man-made radiation sourcewith some hundred million examinations annually, most of them inhealth-care level 1 countries. One single CT exam, for instance, turns-out to administer an effective dose of hundred, thousand or more con-ventional film/screen exams executed in series. Apart from the hundredmillion high-level exposed patients annually, the radiation workers

among them experience incomprehensible inconsistency as comparedwith the situation found at their daily occupational level. While at work-place radiation workers profit from legally regulated radiation protectionunder guidance of expert protection officers, medical supervision andwith the exposure limited and documented in registries for decades,none of these protective mechanisms is offered to them once they arepatients in diagnostics – which situation creates a kind of parallel soci-eties. Is patient exposure unlimited in medicine - and is it high time torecognize this and change?

The many million patients undergoing high-level diagnostic examina-tions could however, at least in future, serve for an urgently needed eval-uation of a human-centered radiation risk at so-called “low-doses”, i.e.around and below 100 mSv separately for men, women and children –and that particularly for X-rays; due to the performance limits of epi-demiology and radiation biology the radiation risk for carcinogenesis ofman has so far not yet been clarified. Realistic radiation risk, especiallyfor X-rays, could be basis of strategic changes in applied radiation pro-tection, to the benefit of human health and in balancing technical effortsand economic expenses. Details will be presented and discussed.

Conception of radiation hazard of possible accidents atworking nuclear power plants in the regions of the RussianFederation

G. Avetisov, S. Goncharov, G. KiporMedical Care Management in Radiation Accidents, Organisation: All-Russian Centre for Disaster Medicine, Moscow, Russian Federation

Conception of radiation hazard of possible accidents at workingnuclear power plants (NPP) in the regions of the Russian Federation,developed considering experience of Chernobyl NPP accident response,is proposed. The conception is based upon the postulate that in theresult of NPP large-scale radiation accidents radiation hazard for thepopulation is possible at a distance up to 1000 km from the accidentsite. At present international experts estimate the probability of a radia-tion accident comparable with the Chernobyl accident as 70%. Accord-ing with the given conception on the RF territory the zone of high radia-tion hazard is marked out (RF territories where NPPs are situated), thezone of heightened (increased) hazard (RF regions on the territoriesneighboring with the zone of high radiation hazard) and the zone ofpotentially radiation hazard (RF regions situated farther than 100 km butin the limits of 1000 km from NPP). For each zone the tasks of RF dis-aster medicine service are considered concerning medical provision ofpopulation in case of a large-scale NPP accident. Complex of protectivemeasures is differentiated depending on the distance from NPP. For RFregions of high radiation hazard zone during planning medical provisionin case of radiation accident it is necessary to provide all protectivemeasures for the population of the whole region. For RF region ofincreased radiation hazard the same measures should be given for pop-ulation living in 100 km zone from NPP. For the population of the restpart of the territory it is sufficient to provide protection against iodine-131 and other radionuclides intake into human body. For RF regions ofpotentially radiation hazard zone it is sufficient to provide protectionagainst iodine-131 and other radionuclides intake into human body.

Real Experience Training in Radiation Protection ScenariosP. Hickmott

Argon Electronics (UK) Ltd, Luton, UKA clear, current and demonstrable understanding of the operation of

radiological dosimetry, survey, and spectrometry instruments is an obvi-ous pre-requisite to their use, but there are inherent difficulties in train-ing and verification of competency with such devices. The necessity toutilise radioactive sources and materials to gain experience of theresponse of real detectors can conflict with the requirement to minimiseoperator exposure. Increasing regulatory and economic restriction in thecontrol of radioactive sources can be addressed in practice by theimproved use of simulation training systems.

The use of simulators is not new in the RP field. This paper shalldescribe currently available systems and provide examples of best prac-tice use includingn Types of radiation source/radioactive material simulation? ultrasound,? magnetic,? fluorescent.

n Types of simulator? generic model replacement, ? replication models,? substitution probes.New developments in simulator technology shall be described including

nThe development of personal dosimeter simulators and the integrationof their response with survey meter simulator readings;

n Integration of simulators with teledosimetry systems;

n Remote monitoring of simulator readings and control of simulatedsources;

n The introduction of integrated spectrometer simulators;n Wide area classroom (table top) and field exercises using simulation

instruments.

Armed Forces Radiobiology Research Institute: MilitaryMedical OperationsB. LivingstonMilitary Medical Operations, Armed Forces Radiobiology Research Insti-tute, Bethesdam MD, USAThe Armed Forces Radiobiology Research Institute (AFRRI) inBethesda, Maryland, which is under the Uniformed Services Universityof the Health Sciences, is the only radiobiology research facility in theUnited States Department of Defense. With program areas in combinedinjury, internal contamination/heavy metals, biodosimetry, and agentdefeat, the AFRRI performs world-renowned research to increase theprobability of survival in the event that a patient in contaminated orexposed with ionizing radiation.The Military Medical Operations (MMO) ia an AFRRI directorate with themission of educating the Department of Defense personnel on theeffects of ionizing radiation. Additionally, MMO has a deployable capabil-ity and subject matter experts that can advise on radiation during a radi-ation incident/accident.The education mission is accomplished by teaching the Medical Effectsof Ionizing Radiation course and associated medically relevant radiationcourses to audiences worldwide. MMO also creates educational prod-ucts that have been distributed throughout DoD, including a new onlinecourse that teaches radiological effects and response in a short 6-hoursession.The Medical Radiobiology Advisory Team (MRAT) is comprised of healthphysicists and radiation-trained physicians who are the DoD’s leadingexperts on effects, protection, and treatment from a radiological incidentor accident. They provide vital response assets to the protection oftroops and families in Japan during the 2011 Fukushima nuclear inci-dent. The MRAT team participates in training and exercises to be readyand able to respond 24/7 to nuclear or radiological incidences.

Risk Management Implementation in Military MedicalRadiation Accident ManagementHotz M. and Meineke V.Bundeswehr Institute of Radiobiology affiliated to the University of Ulm,MunichRisk assessment and risk management are essential components in themanagement of medical radiation accidents. Applied risk assessmenteffectively increases preparedness. Furthermore, the implementation ofrisk management strengthens sustainability of emergency services andimproves the logistics as well as communication and informationprocesses in all contingencies. In principle, risk assessment in medicalcontingency planning is a scenario-dependent valuation of at least twosettings, the optimized best case and the worst case. The differencebetween both cases is the operational overall risk, which is to be kept aslow as possible. This overall risk is influenced by multiple factors, whichmust be identified and evaluated as closely as possible. For instance,NBC risks must be considered relative to the other hazards of combat.For all medical operations a fundamental MASCAL planning shouldexist, which schedules the general responsibilities. At the operationallevel the detailed planning can be based on three main-phases: „Estab-lishing the context“ to define the external and internal parameters to betaken into account when managing risk, “risk assessment“ and “risk

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ConRad 2013, 13.–16.5.2013, Conference, Munich Poster Presentations

3. Radiation emergency medical preparedness and response

treatment”. Furthermore, risk assessment is the overall process of riskidentification, risk analysis and risk evaluation. Besides all these main-phases communication and consultation among all involved partiesshould take place during all stages as well as monitoring and reviewshould be a planned part of the risk management process and shouldinclude regular checking or surveillance.All these components are to be taken into account in risk managementimplementation, described in the internationally recognized ISO31000:2009 (Risk management - Principles and guidelines). This stan-dard contains the essential steps in the implementation and subsequentmastery of the risk process. Numerous tools and methods are availablefor risk evaluation and identification. These have continuously beenimproved and refined in recent years. Some risks are quantifiable, oth-ers, however can only be described in terms of quality. For all establish-ments/facilities conducting risk management, information on the overallrisk in relation to risk bearing ability is vital. In conclusion, ISO31000:2009 is an international standard which offers an excellent basisin military medical planning and could play a key role in multi-nationaloperations to work with a homogeneously concept even in the case ofmedical radiation accident management.

Sustained Controversy of the Internal Contamination withUranium Isotopes in the Radioactive Warfare in RecentConflicts in Iraq, Afghanistan, and Gaza

Durakovic, Asaf (MD, PhD, DSc, FACP) , Klimaschewski, Frank (MSc,PhD cand.)UMRC, NY, USA, UMRC, NY, USA, www.UMRC.netPurpose

Since 1997, the Uranium Medical Research Centre (UMRC) hasconducted the quantitative analysis of four uranium isotopes 234U,235U, 236U and 238U and their ratios in military personnel returningfrom (1.1) the Persian Gulf regions after Gulf War I, Desert Storm;(1.2) Iraq after Gulf War II, Operation Iraqi Freedom (OIF); (2)Afghanistan after Operation Enduring Freedom and in local civilianpopulations including civilians from (3) the Gaza Strip after the use ofuranium containing weapons during Operation Cast Lead (12/2008 –01/2009). The research hypothesis is that the uranium dust set freeby uranium weapons has lead to its inhalation and internal contami-nation of both the military and civilian populations in these areas ofconflict manifesting clinically as a variety of symptoms, such as inca-pacitating fatigue, musculoskeletal and joint pains, headaches, neu-ropsychiatric disorders, affect changes, confusion, impaired vision,memory loss, lymphadenopathies, respiratory system impairment,impotence, and urinary tract morphological and functional alterations.The chemical toxicity of uranium has been recognized for more thantwo centuries. Animal experiments and human studies are conclusive about meta-bolic adverse affects and nephrotoxicity of uranium compounds. Radiation toxicity of uranium isotopes has been recognized for over 100 years.

MethodologySubjects were chosen based on the correlation of the onset of symp-

toms with the time of exposure to uranium aerosols. Twenty-four hoururine samples were collected under controlled conditions. The environ-mental pollution of soil and water with uranium isotopes was alsoassessed. All isotopic measurements except for samples from the GazaStrip were conducted in specialized radiochemistry laboratories withmass spectrometry, including pre-concentration of urine by co-precipita-tion, oxidation of organic matter, uranium purification by ion-exchangechromatography, and ICP-MS double focusing Thermo Finnigan Nep-tune multi-collector. Soils (fraction < 150 micrometer) were separatedand leached in hot agua regia over 48 hours. Water samples were evap-orated, organic matter oxidized, and the residue dissolved in concen-trated nitric acid at 120°C over 48 hours. Uranium was separated andpurified in all samples by ion-exchange chromatography followed by thesame isotopic ICP-MS analysis. Genomic assessments for 5 returningsoldiers were conducted using spectral karyotype (SKY) imaging.Results(1.1) Persian Gulf War I:

The internal contamination with depleted uranium (DU) isotopes wasdetected in the urine samples of 27 British, Canadian, and United StatesGulf War veterans, as late as nine years after inhalational exposure toradioactive dust. The presence of DU was confirmed in 14 of 27 sam-ples, with the 238U:235U ratio > 207.15 which is significantly differentfrom natural uranium as well as from the DU shrapnel analysis, with22.22 % average value of DU fraction. DU isotopes were also identifiedin a Canadian veteran’s autopsy samples of lung, liver, kidney, and bone. (1.2) Gulf War II:

Seven symptomatic civilians were chosen from northern Iraq’s Bagh-dad area exposed to aerial bombings and tank battles. Further 12 symp-tomatic Iraqi civilians from southern Iraq were selected from similar sitesof combat in the cities of Nasiriyah and Al Basra. 236U was present inthe urine of all DU positive subjects with a mean concentration of totaluranium of 30.68 ± 19.67 ng/l in DU positive and 22.08 ± 13.35 ng/l inDU negative subjects. The uranium concentration in the soil fine-fractionfrom these areas varied from about 1 to 2,600 mg/kg which clearly cor-related with the DU 238U:235U ratio. All analyzed fine-fractions of soilsfrom those areas had a DU signature. 236U was present in all soil fine-fraction, from 0.1 ppt (parts per trillion) to 12.4 ppb (parts per billion).The urine from nine symptomatic US soldiers, the members of MilitaryPolice unit 442, deployed during the Iraq military operations in March2003, was later analyzed. The mean concentration of total uranium was3.2 ± 0.6 ng/l. Five of the nine soldiers had a 238U:235U ratio of natu-ral uranium. Three subjects of this group had detectable levels of 236U.Four soldiers were clearly identified as positive for depleted uraniumexcretion. Urinary 236U concentrations of these four individuals variedfrom 1.4 to 12.2 fg/l. Increased levels of genomic variation were found inall five participants with SKY testing at a rate suggestive of ten times anormal range. Furthermore, natural uranium and DU was found in watersamples from Iraq. The presence of elevated levels of uranium in soilsand 236U, indicative of traces of spent nuclear reactor fuel, could bemeasured in some of the water samples. (2) Afghanistan:

The concentration of uranium isotopes in the urine of civilians from 22male subjects from three districts of Kabul and Nangarhar-Jalalabadregion of Eastern Afghanistan in 2002 was found to be up to 100 timeshigher than the worldwide average range of various geographical areas.Results from the 6 civilians of the Bibi Mahro region of easternAfghanistan were even up to 200 times higher. There was a smalldetectable presence of 236U in seven samples from Kabul. Uraniumlevels in the soil samples from bombsites showed values two to threetimes higher than worldwide concentration levels of 2 to 3 mg/kg andsignificantly higher concentrations in water than the World HealthOrganization maximum permissible levels. The results demonstrate that

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the civilian population of Kabul and Jalalabad had a significant elevationof total uranium compared to world-wide urinary uranium concentrations(1-20 ng/l). Fourteen soil samples from Afghanistan had a mean con-centration of 4.95 mg/kg. The presence of elevated levels of uranium insoils and 236U could be measured in some of the water samples of theJalalabad and Kabul areas. Minimal trace amounts of DU were found inthe water samples. (3) The Gaza Strip:

The field team collected the urine samples of civilians in Gaza Stripexposed to the dust of the recent military conflict. Twelve subjects fromJabaliya, Beit Lahia, Rafah, and Gaza City, selected on the basis of theirhistory of exposure and the standard profile of atypical symptoms, had

their 24 hour samples of urine analyzed for the uranium isotopes at theLaboratories of the Harwell Science and Innovation Centre, England, UK,by alpha spectrometry (method HS/GWI/2055). However, neitherdepleted uranium nor man-made uranium isotopes were detectable inthe urine samples of symptomatic Gaza civilians.Conclusion

Our results confirm the presence of uranium isotopes in urine sam-ples of the civilian and military personnel after the recent radioactivewarfare in the Persian Gulf and Afghanistan. The concentrations and theratios of four uranium isotopes, including the presence of man-made236U, suggest their origin in the spent uranium fuel with somatic andgenetic implications.

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4. National, international and global radiation accident management

Emergency planning in Austria for the Treatment ofDeterministic Effects

A. ZieglerRettung sdienst der Stadt Wien, Austria

The focus of nuclear emergency planning in Austria has been so far onmitigating effects of widespread contamination (e.g. after NPP accidents)by bilateral and international agreements on information exchange and bya \"Radiation Early Warning System\" with 300 dose rate meter and 10 airsampler devices.

However there were no provisions on the medical management of anAcute Radiation Syndrome (ARS). In 2008 the author was contracted bythe Federal Ministry of Agriculture to create a \"Medical Radiation Emer-gency Plan\".

After an extensive literature search it was decided to base planning onthe British Institute of Radiology publication on the \"METREPOLapproach\", taking account also of publications by IAEA, REAC/TS, theRadiation Protection Commission of Germany, the Swiss-German Radia-tion Protection Association, UNSCEAR and ICRP.

The management of radiation accident victims requires a multidisci-plinary effort of many specialties. Given the rarity of the event, it appearsnot as prudent to create any specialised institutes or organisations forthis purpose.

The chosen approach foresees to combine the already availableresources (medical and non-medical, pre-hospital and intra-hospital) tointerdisciplinary networks, which include nuclear medicine and radiotherapy departments, haematology and bone marrow transplantationcentres, ICUs, surgery and dermatology as well as biological dosimetrylaboratories. For further consultation the IAEA/WHO REMPAN networkwould be used.

Using these resources, the planning foresees a multi-step approach.\"Basic\" hospitals need to prepare to properly refer patients while protect-ing themselves, \"regional\" and \"central\" hospitals (with radiation expert-ise by radiotherapy and nuclear medicine) are required to plan for provid-ing diagnostic and treatment as necessary.

As emergency response and health care issues are regulated on provin-cial level, the discussed federal-level \"Medical Radiation EmergencyPlan\" needs to be transferred into the emergency planning of theprovinces and into the emergency preparedness of the hospitals.

The described multi-step-approach appears appropriate for Austriancircumstances. Its practicability is still to be demonstrated by exercisesand experience.

One year medical follow-up of persons accidentally exposedto 60Co in Bulgaria.

J. Djounova-VelikovaRadiation medicine & emergency, National Centre of Radiobiology &Radiation Protection, Sofia, Bulgaria

A severe radiation accident occurred on 14 June 2011, in an indus-trial irradiation facilities for medical equipment sterilization with veryhigh-activity gamma sources. Five people were exposed for 5 to 10 min-utes to 60Co source (137 TBq). This accident was the first in Bulgaria,in which the whole body irradiation doses exceeded 1Gy and suggesteddevelopment of ARS. We put into practice the plans for medical careprovision in radiation accidents and the developed procedures for injuryseverity assessment, the decision-making algorithm regarding subse-quent treatment and therapy of persons affected. The activities per-formed for initial assessment of the severity of injury of irradiatedpatients was published in RPD journal 151(4): 640-644 2012.

Based on predictive assessments of the severity of radiation damage, it was decided that the expected development of ARS in vic-tims required hospitalization at a specialized haematology clinic. PercyHospital in Paris was chosen for this purpose, where treatment withgrowth factors to help haemopoiesis recovery was used.

The aim of this report is to present the results of one year follow-up of the victims.

After acute exposure to radiation, the recovery to normal content ofperipheral blood cells was observed in all victims. Nevertheless, therewere observed cases of thrombocytopenia, granulocytopenia and leuco-cytopenia at various times after exposure.

During the period of observation morphological changes in red bloodcells such as anisomicrocitosis, macrocytes, mégalocytes, and polychromatic erythrocytes were demonstrated. The one year observa-tion period all victims showed slight hypocellularity of bone marrow.

Critical parameters that influence efficient cooperation insidethe biological dosimetry network (RENEB) in an emergencysituation

Sylwester Sommer1, Octavia Montero Gil2, Alicja Jaworska3, UlrikeKulka4, Ursula Oestreicher4, Finn Ugletveit3, Pedro Vas2, Horst Romm4

1Institute of Nuclear Chemistry and Technology, Warsaw, Poland;2Instituto Superior Técnico, Universidade Técnica de Lisboa, Bobadela,Portugal;3Norwegian Radiation Protection Authority, Østerås, Norway;4Federal Office for Radiation Protection, Neuherberg, Germany;

RENEB – Realising the European Network of Biodosimetry is a Coordination Action project funded within the 7th EU frameworkEUROATOM Fission Programme (no. 295513). RENEB’s aim is to estab-lish a sustainable network of biological dosimetry in Europe. In total 23organisations, representing experienced laboratories from 16 EuropeanUnion countries participate in the project.

Aim of working package 4 of RENEB is to set up the hierarchical, com-municational and logistical infrastructure to establish an operational biodosimetry network in Europe. This include linking the network to national first responder and disaster management units, provide a -

long-term funding strategy for the network by connecting RENEB capabilities to the European research area and by establishing links to public health organisations. Finally will WP4 attempt to establisha legal framework for the network.

In order to find out which critical parameters are most important tomake the biodosimetry network operational, surveys on critical parameters that influence an efficient cooperation in an emergency situation were designed and posted among the RENEB partners. A total of 5 critical aspects were identified: to know about the contactdata of the partners (1), the emergency organisations and emergencyplans in each country (2), the logistic and organisational aspects of blood (or other material) sample transport in Europe (3), how the net-work should operate in an emergency situation and which researchshould be done in “silent” periods (4), the financial aspects of this kind of mutual assistance on international level (5).

The results are:(1) Contact persons were appointed and the relevant basic

data will be available on the RENEB internet platform.

(2) Radiological emergency plans exist at the national level in each of the 16 countries, and biological dosimetry is part of the emer-gency plan in most countries. Most, but not all RENEB partners areofficially appointed as organisations that perform biological dosimetryin the national emergency plans. The list of national organisationsresponsible for the radiological emergency plans and for the first med-ical response has been collated and actualized.

(3) The transport of a biological samples will be performed in general by commercial delivery companies. Many RENEB partnershave experience in biological sample shipment and have an agree-ment with commercial delivery companies; the most popular one isDHL. Experience concerning a timely delivery of samples by DHL has shownrecently, that most samples will arrive during 24 or at least 48 hours.But nevertheless, exceptions during exercises have shown that onecannot trust on this.

(4) and (5) The decisions on the organisational structure and finan-cial aspects of RENEB as a biological dosimetry network will be made during the following 3 years.

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5. External exposure assessment

Calibration and uncertainty evaluations of protection levelionizing chambers in photon reference radiations

I.I. Suliman 1*, A. Beineen2

*Corresponding Author1Medical Physics Section, College of Medicine & Health Sciences;Postal code 123; P.O. Box 35 Al-khod, Oman 2Radiation Safety Institute, Sudan Atomic Energy Commission, PostalCode 11111; Jamma Str.; P.O. Box 3001; Khartoum, Sudan

Calibration of radiation measuring equipment is required to achiev-ing dose standardization and traceability of measurand to the relevantSI unit. The aim of this article is to present some selective measure-ments performed to calibrate two protection ionization level chambersat the Secondary Standard Dosimetry Laboratory (SSDL) in Sudan.Nuclear Technology (NE) 600 cc former and Radcal 1800 cc ionchambers were calibrated against Ls-01 100 cc reference standardion chamber. Experimental measurements were carried out using137Cs and narrow spectrum X-ray radiation described in the interna-tional standard ISO 4037 [1]. Calibrations were performed using theInternational Atomic Energy Agency (IAEA) dosimetry protocol TRS277 base on air kerma standards [2]. Calibration coefficients andassociated uncertainties were evaluated. The percentage uncertaintyin calibration coefficients for NE 600 cc former ionization chamberevaluated at 95 % confidence level ranged 2.1 % at N40 to 7.1 % atN 60. The percentage uncertainty in calibration coefficients for Rad-cal 1800 ionization chamber evaluated at 95 % confidence levelranged 2.0 % at N40 to 8.5 % at N 60. The instrument responseswere surprisingly high 40 kV (33 keV) which could be due to scatterradiation. This can be reduced using addition beam limitingdiaphragm. Variability in air kerma measurements can be reducedusing monitor chamber as recommended [2]. References

[1]ISO 4037-1:1996. X and gamma reference radiation for calibrat-ing dosimeters and dose rate meters and for determining theirresponse as a function of photon energy d Part 1: Radiation charac-teristics and production methods. ISO, 1999.

[2]IAEA . Absorbed dose determination in photon and electronbeams. An International Code of Practice. IAEA TRS. No.277 Sec. Ed.IAEA, Vienna, 1997.

CONTEMPORARY RADIATION PROTECTION TRENDS IN IR ANDIC - NEW ELECTRONIC DOSIMETRY DEVELOPMENTS - Do weneed a new type of digital personal dosemeters to be used inmedicine and homeland security?

Ivica PRLIC1*, Marija SURIC MIHIC1, Tomislav MEŠTROVIC1, MladenHAJDINJAK2, Zdravko CEROVAC3

*presenting author: [email protected] for Radiation Dosimetry and Radiobiolgy, Institute for MedicalResearch and Occupational Health, 10000 Zagreb, Republic of Croatia2Haj-Kom d.o.o, Zagreb, Republic of Croatia3ALARA ltd. Zagreb, Rapublic of Croatia

Legal personal dosimetry is based on the use of passive dosemeterssuch as film, thermoluminiscent (TLD) or optically simulated (OSL)dosemeters. Modern technology, extensive use of radiation imaging inmedicine and industry imposes use of upgraded dosimetric deviceswhich could provide additional information on radiation exposure suchas information od exposure dose rates, data on the moment of the expo-sure, duration of the exposure, etc. Majority of available active electronicpersonal dosemeters (AEPD), due to a type of detector used, are notsuitable for measurements in pseudo-pulsed or pulsed radiation fieldsused in interventional radiology (IR) and cardiology (IC). An AEPD typeALARA OD, based on GM tube, was used to measure the levels andstructure of the occupational exposure of workers that are predominantlyexposed to scattered X-ray radiation of the continuous and pulsed radia-tion fields. The 3D H*´(10) isodose patterns were constructed. Secondset of measurements was performed at border crossings during the reg-ular customs x-ray and LINAC cargo screening procedures.

Energy response of radiation protection area survey meters tophoton reference radiations

I.I. Suliman 1,*, M.M. El-Hassan2

1Medical Physics Section, College of Medicine & Health Sciences; Postal code 123; P.O. Box 35 Al-khod, Oman 2Atomic Energy Council, Sudan Atomic Energy Commission, Postal Code11111; Jamma Str.; P.O. Box 3001; Khartoum, Sudan

*Author for correspondence Dosimetric measurements were made to study the energy response to

a set of 8 radiation protection area survey meters to low energy photon

radiation. Experimental measurements were carried out at the Second-ary Standard Dosimetry Laboratory in Sudan using photon referenceradiation described in the International Organization for Standardization(ISO) standard ISO 4037 [1]. Irradiations were carried out using Cea-sium-137 and X-ray beam qualities of: 48, 80, and 100 keV. Referencestandard ionization chambers were used to measure the air kerma rateat predefined calibration point. Appropriate, conversion factors wereused to determine ambient dose equivalent H*(10) as standard dosi-metric quantity of interest [2]. Parameters tested include the variation ofinstrument response with radiation energy and variation of instrumentresponse with dose rates. The responses of the survey meters understudy were within the limits of acceptable performance. Energy responseof survey meters: Tracerco, RDS and Radiagem ranged from 0.4 to1.04, (0.77-1.3), (0.85-1.4), respectively. Dosimeter responses werewithin the acceptable limits. The results confirmed that radiation pro-tection survey meter calibrated using Ceasium-137 (662 keV) could beuse for monitoring photon energies encountered in radiation protectionpractice with acceptable accuracy. References

[1]ISO 4037-1:1996. X and gamma reference radiation for calibratingdosimeters and dose rate meters and for determining their response asa function of photon energy d Part 1: Radiation characteristics and pro-duction methods. ISO, 1999.

[2]International Atomic Energy Agency. Calibration of radiation protec-tion monitoring instrument .IAEA Safety Reports Series.No.16.IAEAVienna, 2000.

DNA damage focus analysis in lymphocytes after acute partialbody gamma irradiation of minipigs

Lamkowski, A.1, Agay D.2, Drouet M.2, Meineke V.1 and Scherthan H.11Bundeswehr Institute of Radiobiology affiliated to the University of Ulm,Munich, Germany

2IRBA-antenne La Tronche - CRSSA, 24 Avenue des Maquis du Grési-vaudan, 38702 La Tronche, France

Past radiation accidents often involved acute partial body exposure with high doses, leading to victims with radiation sicknessand cutaneous radiation syndrome (CRS). Recently, CRSSA estab-lished a minipig model to study treatment options for CRS. Ionizingradiation (IR) can induce physiological responses at different levels,which eventually may lead to cell death. Cells that are not lethally irra-diated seek to repair IR-induced damage, which is particularly important for the DNA molecule, since repair restores genomicintegrity and ensures cellular survival. Since there are only a fewstudies that consider DNA repair in large animal in vivo models after acute IR exposure, we investigated the DNA repair response inminipig lymphocytes after acute partial body irradiation with 60Co γray dose of 50 Gy affecting a ~125 cm2 skin area in Göttingen minipigs (Agay et al. 2010). Previously we showed that keratinocytes of Göttingen minipig skin display a persistent DNA damage response (DDR) that, in some cells, can last for months after acute 50 Gy irradiation (Ahmed et al. 2012). Here, we investigatedwhether the DDR of peripheral blood lymphocytes (PBL) can be used as an indicator for acute partial body photon irradiation in this minipig model. Blood samples were taken 4, 24 and 168 hours post IR and subjected to γ-H2AX and 53BP1 immunostaining. The number of IR-induced DNA lesions was analysed by manualcounting. Automated DNA damage focus analysis was attempted with the Metafer image analysis system. In all, we find that PBL of PB irradiated minipigs display foci numbers that significantlydeceed the high foci numbers present in keratinocyte cell nuclei ofthe irradiated skin regions. Thus, PBI is a challenge when dosereconstruction shall be attempted by γH2AX DNA repair focus num-bers in lymphocytes. Hence, the γH2AX assay may not be suited toreveal a reliable dose indication after acute high dose partial body irradiation.

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6. Decontamination measures and monitoring

User interface features of information-analytical system forradiation safety of personnel on the example of NorthwestCenter for Radioactive Waste Management \"SevRAO\"

K. ChizovBurnasyan Federal Medical Biophysical Center of Federal Medical Bio-logical Agency, Moscow, Russian Federation

Algorithms for radiation protection services and regulators was devel-oped. They provide guidelines for choosing actions in planning the elim-ination of accidents. Method is based on work with a grid of radiationfield (this grid is with constant spacing along axes in the plane andevery point has the value of the dose rate), which is obtained by interpo-lation of the measured values. Based on these algorithms, a softwarewith a graphical representation of the original data and the results of cal-culations was developed. It allows us to quantitatively evaluate the qual-ity of information received from the monitoring team, search for the bestroutes on site and find points that make a maximum contribution to thecollective dose.

Has shown that an important condition for the enterprise softwareimplementation is that when facility use it, the operator\'s efficiencyincreases, while working time stay constant or reduces. This statementapplies for input data into the database, mathematical analysis andpresent the results in a report drawn up in accordance with the rules ofthe company. On the example of SevRAO shown that the installation ofinformation-analytical system (IAS) helped reduce the time for enteringand processing data on the radiation measurements. IAS has an oppor-tunity to automate the creation of reports.

Regulator should control about 250 parameters of the radiation situa-tion. Basically, this is a list of control points and the reference values ofdose rate at these points for the technical area, buildings and structureslocated at the industrial site. For each point are two control values - thedose rate at rest and during work. It is more important for regulator tocompare the radiation situation with control levels, but not only to seeradiation situation itself. So we have developed software that graphicallypresents information as areas on the map. Regulator with a glance at thescreen can see where levels have been exceeded.

Study on paramedics Attitudes and Compliance withprehospital decontamination Standard Precautions

D. ChoiEmergency Medicine, Disaster Medicine, Gyeongju-si, Republic of KoreaPurpose

The purpose of this study was to identify paramedics attitudes to, andcompliance with, standard precautions in the prevention of prehospitalinfections by enhancing their practice of standard precautions.

Methods: The participants were 80 paramedics who worked in Firestation in Gyeongju-si. The questionnaire was consisted of 18 questionsabout hand washing, personal protective equipment, sharps, linen and patient care equipment. Collected data were processed using SPSS 15.0 WIN.Results

Mean scores for attitudes to standard precautions and for com pliancewith standard precautions were 4.43 (±0.83) and 4.22 (±0.44)

respectively. This difference was statistically significant (t=3.368,p=.001). The paramedics compliance with standard precautions differed significantly according to the general characteristics of age (F=8.705,p<.001), total clinical experience (F=9.426, p<.001), current department experience (F=6.555, p<.001), and education experience

(t=0.616, p<.043). There was a positive correlation between attitudes to,and compliance with, standard precautions (r=.156, p=.025).Conclusion

The results of this study indicate that educational programs and policyon prehospital decontamination control and standard precautions forparamedics are needed.

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7. Biological dosimetry and EPR

Towards a common standard in radiation: impact ofautomated image analysis on duration, accuracy, andreproducibility of bio-dosimetry studies

C. SchunckMetaSystems GmbH, Altlussheim, Germany

Bio-dosimetry is a well established procedure to assess radiation dam-age in humans involved in mass radiation accidents. Several assays (e.g.the analysis of dicentric chromosomes or the micronucleus assay) areavailable to quantify DNA damage induced by irradiation, aiming to esti-mate the body or partial body dose of the victims in order to apply suit-able treatments.

Earlier we have described a system that is based on the microscopicslide scanning platform Metafer, allowing to automate analysis of thechromosomal aberration and dicentrics test, the micronucleus assay,and scoring of -H2AX foci. It was shown in several studies that the useof automated image analysis with Metafer is a remarkable time saver forthe above mentioned bio-dosimetry assays. Authors also state that, dueto higher numbers of cells being analyzed, data obtained with the auto-mated system are similar or even more reliable than manually obtainedresults. Recent investigations, however, aim towards a standardization ofanalyses between different institutes of different countries. A couple ofinternational projects are currently testing the possibilities of sharinganalysis work, with the objective to establish robust, decentralized logis-tics for emergency situations. Here we summarize the current proceed-ings of bio-dosimetry research with Metafer, with a focus on internationalcollaborations.

Radiation Bio-Dosimetry Automation with the Help ofAutomated Slide Scanning

C. SchunckMetaSystems GmbH, Altlussheim, Germany

Bio-dosimetry is a well established procedure to assess radiation dam-age in humans involved in mass radiation accidents. Several bio-dosimetry assays are available to quantify DNA damage induced by irra-diation, all aiming to estimate the body or partial body dose of thevictims in order to apply suitable treatments.

Several feasibility studies and unfortunately also experiences from realaccidents have shown that there are a couple of key requirements toexecute bio-dosimetry in a large-scale accident scenario:(a) Speed - It can be expected, that large numbers of cases have to be

accessed in short time, aiming to provide fast help for the victims.(b) Ease of Use - In the probably highly confusing situation after a large

radiation accident any delay caused by avoidable methodical short-comings may be precarious.

(c) Reproducibility - Data obtained for bio-dosimetry should clearly allowfor drawing the respective countermeasures, e.g. provide a dose esti-mation that should be as precise as possible.

(d) Comparability - It is most likely that in large accident scenarios morethan one institute will be involved in bio-dosimetry work. Therefore itshould be possible to merge data obtained from different labs with-out violating data integrity.

Unfortunately conventional bio-dosimetry assays involve long andtedious analysis steps, which are usually done manually and are, thus,

error prone and subject to biases. A feasible alternative is theautomation of image analysis, providing advantages such as definedanalysis environments, 24/7 operation mode, and transparent docu-mentation of all findings.

Based on the microscopic slide scanning platform Metafer we havetherefore developed tools to automate analysis of the chromosomalaberration and dicentrics test (DIC), the micronucleus assay (MN),the Comet assay , and the analysis of g-H2AX foci (Schunck et al.,2004).

Several studies have shown that the use of imaging automation withMetafer for bio-dosimetry has remarkable advantages over manualscoring. Though time saving is obviously one of the major benefits,authors also reported an increase in accuracy due to larger samplesizes, a better documentation of results, and higher reproducibility byavoiding scoring biases. In the frame of increasing cooperation andnetworking within the international radiation bio-dosimetry commu-nity, however, the possibility to exchange analysis parameters andresults is seen to be the most important plus factor.

Radiological accident in Peru: contribution of dosimetry inthe early medical management phase

F. Trompier1, C. Huet1, G. Gruel1, S. Roch-Lefèvre1, E. Gregoire1, T.De Revel2, E. Bey3, JJ Lataillade4, J.F. Barquinero1, M Benderitter1, I.Clairand1, F. Queinnec1, P. Voisin1 and J.F. Bottollier-Depois1

1 Institut de Radioprotection et de Sûreté Nucléaire, IRSN, BP17,92262 Fontenay-aux-Roses, France 2 Hôpital d\'Instruction des Armées Percy, Service d\'Hématologie, BP410, 92141 Clamart Cedex, France3 Hôpital d\'Instruction des Armées Percy, Service de Chirurgie Plas-tique, BP 410, 92141 Clamart Cedex, France4 Unité de Thérapie Cellulaire, Centre de Transfusion Sanguine desArmées Jean Julliard, BP 410, 92141 Clamart Cedex, France

In January 2012, a radiological accident involving a gammagraphydevice containing an iridium-192 source has led to the overexposureof three persons in the district of Chilca, Peru. In addition to thewhole body exposure, hands were severely exposed to radiation.

Different approaches were used to estimate the dose and its distribution in the organism of the victims. In addition to cytogeneticdicentric analysis and Monte Carlo calculations based on testimoniesand characteristics of the iridium-192 source, EPR analysis of sam-ples of tooth enamel and nails were performed with a high frequency EPR (Q-band) spectrometer. This new dosimetric approachusing EPR technique allows measuring with high sensitivity samplesof small volume (a few mm3). Thus, mini-biopsies of tooth enamelwith mass ranging between 2-5 mg were analyzed. Fingernails were analyzed also with Q-band EPR with a new protocol establishedrecently at IRSN.

An overview of the data obtained with different retrospectivedosimetry techniques will be given and the complementarities of thedifferent dosimetry techniques used in this expertise will be presented. The contribution and the relevance of the data obtained during the early medical management phase will also be discussed.

Cytogenetic biodosimetry for accidental radiation exposure: avirtual laboratory concept

Maznyk N.1, Fehringer F.2, Johannes C.3, Sipko T.1, Pshenichna N.1,Müller W-U.31Institute for Medical Radiology AMSU, Kharkov, Ukraine (IMR)2Institute for Radiation Protection of the BG ETEM and BG RCI (IfS),Cologne, Germany3University Duisburg-Essen, Essen, Germany

For radiation accident management cytogenetic biodosimetry is anessential tool. Radiation accidents are rare events and it is often not pos-sible to keep running biodosimetry laboratories in radiation protectioninstitutions.

We applied the idea of a virtual biodosimetry laboratory that becomespossible with advanced technologies. This idea allowed combining theefforts of experts both in cytogenetics and radiation protection fieldsfrom different institutions and countries.

The must have for a virtual laboratory is an image capturing system.The captured images are sent as files to scorers at various locations.This system required some different skills than microscopy analysis. Inorder to use image analysis data we validated them with microscopyscoring.

We completed the pilot study with comparison of cytogenetic datafrom 5 German NPP professionals and 5 Ukrainian uranium miners. Agood agreement was shown between microscope and image analysisresults for both groups where the chromosome aberration level washigher in uranium miners.

We conducted in vitro experiments for inter-laboratory comparison ofcytogenetic data for clinically significant whole body doses. The addi-tional goal was to compare the data from scorers with various cytoge-netic (non-biodosimetry) background. It was shown that image analysistook three to five times less time than microscopy for the equal numberof cells. The influence of software parameters on the data outcome aswell as inter-scorers variability will be discussed for the low and highdoses range biodosimetry.

Further work will comprise data comparison for partial body exposure.We can conclude that virtual laboratory or small network gave enough

flexibility to radiation protection institutions in building up biodosimetryservice including accidental occupational overexposure. The peculiari-ties of QA/QC procedures, image vs. microscope analysis data compari-son for the triage and expert mode will be discussed.

Laboratory Calibration Of Dicentric Chromosome Assay:Establishment Of Manual And Automated Standard Curves

A. De AmicisImmunology and Toxicology Section, Organisation: Army Medical andVeterinary Research Center, Rome, Italy

The risk of accidental human exposure is linked to the use of ionizingradiation sources in medical, research and industrial areas. Further-more, the possibility of terrorist attack using radiological or nucleardevices must be considered. Dose estimation is the first important stepfor medical treatment of subjects exposed to ionizing radiation. For thispurpose, clinical signs /symptoms and biological dosimetry are the twomain approaches to assess radiation exposure. Biodosimetry is amethod to measure the ionizing radiation dose absorbed by an individ-ual using biological markers. This type of approach is useful when anindividual is accidentally exposed and physical dosimetry is not availableor uncertain. The most validated assay for biodosimetry and radiationinjury assessment is the gold standard Dicentric Chromosome Assay(DCA). Prerequisite for dose assessment is the establishment of a dose-effect calibration curve. Based on the principle of the comparability of invitro and in vivo irradiation effects, it is possible to generate calibrationcurves (dose-response) by in vitro exposure of peripheral blood lympho-cytes at sequential increasing doses. For this purpose, blood samplescollected from a healthy donor were no irradiated and irradiated with

increasing doses of X-rays (0.25-4Gy). Three scorers analysed wellspread metaphases with 46 centromeres at each dose to construct theown personal manual calibration curve. Each generated calibrationcurve was compared with each other to test the inter-scorers goodnessof fit. Then the three different curves were matched to generate a labo-ratory calibration curve for dose estimation purpose. Furthermore, thesame slides were used to generate the automated calibration curveusing the Metafer4 scanning system. All these calibration curves werevalidated evaluating the dose prediction accuracy through the analysis ofblind samples irradiated with single different doses of X-rays.

Radiation Dose Dependent Changes on the AbsoluteNeutrophil: Lymphocyte Count (ANC:ALC) in C57BL/6 micefollowing Total Body Irradiation.

PA Plett, HL Chua, CH Sampson, TJ MacVittie*, CM Orschell.Indiana University School of Medicine, Indianapolis, IN, USA. *Univer-sity of Maryland, School of Medicine, Dept of Radiation Oncology, Balti-more MD, USA.

Effective biodosimetry early after accidental or deliberate radiationexposure will be necessary to design and implement effective medicalcountermeasures (MCM). The highly radiosensitive nature of thehematopoietic system, as well as the ease of sampling from this tissue,makes assessment of blood cell parameters attractive for biodosimetryand triage. The absolute neutrophil count (ANC) relative to the absolutelymphocyte count (ALC) has been postulated as a valuable tool for earlyassessment of radiation dose. Confounding factors to the use of theANC:ALC, such as early administration of MCM such as G-CSF (Neu-pogen), as well as species differences in the radiation response, illus-trate the need to examine such parameters in multiple species for accu-rate modeling of the human situation. To this end, we have developed amouse model of the Hematopoietic Syndrome of the Acute RadiationSyndrome utilizing C57BL/6 mice exposed to 137Cs radiation (60-70cGy/min). Groups of mice were exposed to sublethal radiation (1Gy or4Gy), or to the LD50/30 (8Gy), and were administered Neupogen(125ug/kg, 8Gy only) beginning 24hr after irradiation until day 16. Sub-groups of mice were tail bled for daily CBC every 5th day. The ANC:ALCratio on day 1 post-exposure increased from 0.25 to 0.55, 0.85, and1.31 for 1, 4, and 8Gy, respectively. A similar pattern was noted on d2,with ANC:ALC ratio increasing to 0.5, 1.22, 1.63 for 1, 4, and 8Gy,respectively (n=3-9mice/dose). Similar to the non-human primate(NHP), the peak ANC:ALC in Neupogen-treated mice shifted from 24 to48hrs post-exposure, relative to irradiated controls. Other differencesbetween mice and NHP, specifically in the magnitude and timing ofpeak ANC:ALC, illustrate the need for further assessment of the value ofthe ANC:ALC as a reliable biodosimeter, with or without other radiationsensitive biomarkers such as cytokines. [Funded by NIAID, contract #HHSN272201000046C]

Biodosimetry of large populations after nuclear accident byFISH in suspension and telemedicine

A.Fucic1, J.N. Lucas2

1 Institute for Medical Research and Occupational Health, Zagreb, Croatia2 ChromoTrax Inc., Lorton, VA, USA.

Nuclear accidents at Chernobyl and Fukushima showed that despitethe differences in technological conditions, social and economical envi-ronment the number of causalities was higher than expected despiteprepared protocols and anticipated health risks. In both cases addition-allly to exposure to ionizing radiation, populations were exposed to differ-ent chemical pollutants and personal stress which significantly modu-lated final biological response. Ionizing radiation causes complexdamages on genome and non-genome levels including disturbances ofimmunological and cardiovascular systems which are gender and agerelated. Biodosimetry after nuclear accidents has to be fast and compat-

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ibile with other medical checks. Fluorescence in situ hybridization ofmetaphase chromosomes in suspension, S-FISH (He et al., 2001) enablesrapid counting of the painted human chromosomes and region-specificprobes. This method offers a new tool fast, early detection and screening ofboth balanced or unbalanced chromosomal exchanges for large popula-tions. This is because S-FISH can be combined with flow cytometry, allow-ing for fast precise detection based on flow fluorescent signals. Usingtelemetry and telemedicine results of biodosimetry S-FISH analysis can bemerged with other parameters (haematology) and interpreted by expertsfrom one central point for different geographical regions to operative groupsin the field. S-FISH requires small blood samples, which allows sampling ofall age groups including newborns. This approach can estimate genomedamage caused both by internal and external radiation and clastogenicchemical agents contrary to limited information gained from physical meas-urements of received radiation dose. As in Chernobyl and Fukushima, thelargest group of people were those who were exposed to doses that do notcause radiation disease but long term and transgeneration effects sug-gested approach is a novel tool in management of nuclear accidents.

Biological dosimetry based on dicentric analysis of differentnumber scored cells

V. Hadjidekova1, R. Hristova1, A. Staynova1, S Deleva1, E.A.Ainsbury2

1 National Centre of Radiobiology and Radiation Protection, Sofia, Bul-garia2 Health Protection Agency, Chilton, UK.

Biological assessment of the absorbed dose ionizing radiation wasdone for five victims of a radiation accident which occurred in Bulgariain 2011. The accident happened with an industrial gamma-irradiationfacility, during a planned technical service for changing the configurationof Co-60 sources, because of human mistake, when instead of the imi-tator one of the tubes containing the sources with ionizing radiation (totalactivity 137 TBq ) was taken out by hand. As a result, 5 subjects weresubjected to overexposure for about 10 minutes, before it was collectedback in to the protected container.

Dose estimation was carry out on cytogenetic data collected after dif-ferent numbers of metaphases were analysed for dicentrics for eachindividual during chromosomal analysis of peripheral blood lympho-cytes. The first dose assessment was made after cytogenetic analysis of50 metaphases in cultured lymphocytes, the second – after counting100 metaphases and the final dose assessment was done after comple-tion the slide scoring, i.e. from 210 - to 376 metaphases scored for eachof the investigated subjects.

The final dose estimates for the five patients was between 5.60 – 1.25Gy acute, whole body radiation exposure. There are no significant differ-ences in the dose estimates obtained after as few as 50 metaphases ormore than 200 metaphases were scored for dicentrics in peripheral lym-phocytes for all of the five patients investigated.

The results of cytogenetic analysis and biodosimetry performed in thisreal case of accidental exposure show that fifty metaphases is a relevantnumber cells to be scored for detection of human radiation exposureabove 1 Gy.

The practical experience of this study will be introduced to the EU-project RENEB (Realizing the European Network of Biodosimetry). Thusthe results can contribute to one of the priority objectives of RENEB toguarantee the highest efficiency in the processing and scoring of biolog-ical samples for fast and reliable dose estimation urgently needed inemergency management.

γ-H2AX in ratio of peripheral blood lymphocytes vsgranulocytes (RL/G) detected using flow cytometric as a newreliable biodosimetry for radiation exposure

Y. ChenRadiation Toxicology and Oncology, Beijing Institute of Radiation Medi-cine, Beijing, China

PurposeTo assess the use of RL/G of γ-H2AX (ratio of γ-H2AX in lymphocytes

to that in granulocytes ) in blood as a rapid method for population triageand doses estimation in events of large-scale radiation emergencies.

Materials and methods: Blood samples from healthy volunteersexposed to 0-10Gy of 60Co irradiation and cultured for 0-24h wereanalysed using flow cytometry assay to measure γ-H2AX levels in lym-phocytes and granulocytes. The basal levels of RL/G of γ-H2AX in healthhuman blood, the response of RL/G of γ-H2AX to ionizing radiation andits relationship to doses, time intervals after exposure, and individual dif-ferences were analyzed in our study.Results

The γ-H2AX levels in lymphocytes increased in a dose-dependentmanner after radiation, and the γ-H2AX levels in granulocytes were notaffected. A linear dose-effect relationship with low interexperimental andinterindividual variations was observed. A formula was constructed byregression for high throughput assessment of radiation doses of victims6 to 24 hours after radiation accidents.Conclusions

RL/G of γ-H2AX may be used as a biomarker for population triageand dose estimation in case of large-scale radiation emergencies if bloodsamples can be collected within 6- 24 hours.

Assessment of total- and partial-body irradiation in a baboonmodel: reliability of the dicentric assay.

Marco Valente, Nancy Grenier, Josiane Denis, Patrick Martigne, PhilippeArvers, Stéphane Baugé, Hervé Chaussard, Francis HérodinInstitut de Recherche Biomédicale des Armées (IRBA), La Tronche, France

The dicentric chromosome assay is the “gold standard” in radiationbiological dosimetry but is limited when distinguishing partial-body irra-diations (PBI) from their total-body irradiation (TBI) equivalent. Untilnow, studies focussing on inhomogeneous exposures have mainly usedin vitro exposures and/or small animal models. In this study, we irradi-ated an animal model with corpulence similar to humans in order toassess the reliability the dicentric chromosome assay would have in dis-tinguishing PBI from equivalent TBI.

Anesthetized Papio anubis were unilaterally exposed to gamma-rays asfollows: 5 Gy TBI and partially-irradiated equivalents (10 Gy 50 % PBI,7.5 Gy / 2.5 Gy TBI, 6.25 Gy 80 % PBI and 5.55 Gy 90 % PBI); 2.5 GyTBI and its equivalent 5 Gy 50 % PBI. Two animals were used per condi-tion. Blood samples were collected before exposure and at several timesafter irradiation: 1h, 6h, 1 day, 28 days, and 200 days. Dicentric fre-quency was determined for at least 150 lymphocytes per sample. Thisstudy was approved by the Army Medical Service Ethics Committee.

Our scoring results showed that, 24 hours post-exposure, 5 Gy TBI wasdistinguishable from the equivalent PBI conditions tested in terms of dicen-tric distribution. However, 2.5 Gy TBI was only distinguishable from theequivalent 5 Gy 50% PBI for one of two animals. 28 days after irradiation,the statistical results obtained were too unreliable to distinguish heterogenicexposures. Furthermore, compared to 5 Gy TBI, the equivalent PBI doseshave a decreased dicentric frequency most likely due to increased celldeath, which is another potential confounding factor. The cytogenetic dataanalyzed until now points to the need of alternative (or additional) bio-dosimeters / biomarkers for a more reliable TBI-PBI discrimination.

Cytogenetic Biodosimetry for Fukushima Travelers after theNuclear Power Plant Accident: No Evidence of EnhancedYield of Dicentrics

H.R. LeeDepartment of Emergency Medical Preparedness, Korea Institute ofRadiological and Medical Sciences (KIRAMS), Seoul, Republic of KoreaAim

Individuals who traveled to contaminated areas after the Fukushimanuclear accident have concerns about the health effects. However,

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medical follow up for any adverse health effects will be difficult withoutpersonal dose measurements. Cytogenetic biodosimetry is a reasonablemethod of assessing absorbed doses retrospectively. Methods

We analyzed dicentric chromosomes for 265 Fukushima travelers,mostly journalists and rescue workers, who had been dispatched tonortheastern Japan during nuclear emergency. As a control group, 37healthy volunteers who had not visited Japan since the accident wereenrolled. Yields of dicentrics and absorbed doses calculated from adose-response calibration curve for travelers and the control group werecompared. Results

The cut-off level for dicentric chromosomes in the control group was3.5 per 1000 cells. Of the 265 travelers, 31 had elevated number ofdicentrics (High-Dics group) while 234 were below the cut-off (Normal-Dics group). All but one of the individuals in the High-Dics group alsoreported a significantly higher number of medical exposures to radiationwithin the past three years compared with the Normal-Dics or controlgroups. The 225 travelers with no history of medical exposure showedno difference of dicentrics yield compared to control group. Conclusion

Our data indicated the Fukushima travel alone did not enhance theyield of dicentrics.

Assessment of the Whole Body Dose in Patients Treated with[131I]-NaI by Analysis of A Dicentric Chromosome Assay:Pilot Study and Further Planning.

M. Cordes1, C. Beinke2, H. Dörr2, H. Scherthan2, M. Abend2, T. Kuwert3, V. Meineke2

1 Radiologisch-Nuklearmedizinisches Zentrum, Nuremberg, Germany2 Bundeswehr Institute of Radiobiology affiliated to the University ofUlm, Munich, Germany3 Clinic of Nuclear Medicine, Friedrich-Alexander-University Erlangen-Nuremberg, Erlangen, Germany

The measurement of the whole body dose in patients treated with [131I]-NaI by physical means has some limitations. We therefore conducted apilot study in which dicentric chromosomes were assessed in 2 patientstreated with this ß- emitter.

The objectives of this study were a) to test the feasibility of this approachand b) to monitor the logistics between the sites of application and analysiswith a setoff of 150 km.

Two patients at the age of 79 and 68 years were included in the pilotstudy. Both patients had been diagnosed with a unifocal thyroid autonomy.For the radioactive iodine treatment 1020 MBq and 712 MBq [131I]-NaIwere administered, resp.. Using a [137Cs] calibration curve (dose rate 0.4Gy/min) whole body doses of 0.42 Gy and 0.023 Gy were calculated, resp..The monitoring of the logistics did not reveal any limitations in this setting.

Future planning includes the establishment of a calibration curve for thedicentric chromosome assay after in vitro exposure of whole blood to[131I]. A dosimetric comparison between the dicentric chromosome andthe gammaH2AX assays will be conducted which has already beenapproved by the Ethical Committee of the University of Erlangen, Germany.

Real-time PCR analysis of expression of DNA damageresponsive genes as a biomarker for biological dosimetry

Kamil Brzóska1, Maria Wojewódzka1, Tomasz Stepkowski1, MarcinKruszewski1,2

1 Institute of Nuclear Chemistry and Technology, Centre for Radiobiologyand Biological Dosimetry, Dorodna 16, 03-195 Warsaw, Poland2 Institute of Agricultural Medicine, Independent Laboratory of MolecularBiology, Jaczewskiego 2, 20-090 Lublin, Poland

In a large-scale radiologic accidents, fast identification of radiation-exposed individuals is crucial for triage and optimal medical manage-

ment. Current biological dosimetry methods are inadequate for the task,mainly because of their low throughput resulting from time consumingprocedures and requirement of highly trained and experienced person-nel. Our objective is to develop bioassays for biological dosimetry basedon molecular biomarkers such as gene expression signatures. To thisend we analyzed the expression of DNA damage responsive genes in anex vivo irradiated whole blood from three healthy donors. Analyzedgenes were as follows: GADD45A, CDKN1A, MDM2, BBC3, SESN2,BAX, DDB2, ATF3, PLK3, GDF15 and BCl2. Blood was X-irradiated witha doses of either 0 Gy; 0,6 Gy; or 2 Gy and gene expression was meas-ured by real-time PCR at 6, 12, 24, and 48 hours after irradiation. Thepreliminary data suggest that the analysis of expression profiles of theselected genes in whole blood may be very useful for fast identificationof irradiated samples and therefore is a promising molecular biomarkerfor radiation biodosimetry.

This work was supported by grant number SP/J/6/143 339/11

PCC methods in biological dosimetry: PCC fragments, PCCrings, unusually long PCC fragments, the Rapid InterphaseChromosome Assay (RICA)

Sylwester Sommer, Iwona Wewiór, Iwona Buraczewska, Teresa Bart-lomiejczyk, Irena Szumiel, Marcin KruszewskiInstitute of Nuclear Chemistry and Technology, Warsaw, Poland

Chemically induced Premature Chromosome Condensation (PCC) isuseful in biological dosimetry in the range of high doses (10 -30 Gy),where classical methods of biological chromosomes dosimetry, like scor-ing dicentrics chromosomes fail, because the damaged cells cannotenter the mitotic phase. Entering mitosis is necessary for dicentric scor-ing but not for PCC assays.

To prematurely condense chromatin we are using phosphataseinhibitors such as okadaic acid or calyculin A. Then, we can observeradiation induced morphological changes (aberrations) in G2 phasecells. It is possible to score the PCC fragments (excess in relation to thenormal number of chromosomes) or frequency of PCC rings, both dosedependent in the range of 0.5 -30 Gy.

Chromatin condensation by okadaic acid or calyculin A is a short timeprocess: 3 hours or 45 minutes, respectively. However, the cells have tobe in the cell cycle. Usually, biological dosimetry is performed on theperipheral blood lymphocytes, which in majority are in G0 phase. ForPCC-based dosimetry they have to be stimulated to division. Therefore,in the case of lymphocytes, we can obtain PCC spreads not sooner thanat 48 hours. Since time is crucial for biological dosimetry purposes,Prasana and Blakely (2000) introduced a new method: Rapid Inter-phase Chromosome Assay (RICA), which can be performed in unstimu-lated lymphocytes during 5 - 6 hours after exposure. The method is sim-ple: the additional chromosome territories stained with FISH wholechromosome probes are scored and their number is dose dependent.

In 2007 Wang et al. observed that while performing PCC in lympho-cytes after irradiation with high doses of heavy ions, unusually long PCCfragments can be found and the ratio of the longest chromosome to theshortest chromosome is dose dependent. In our lab, we observe suchextra long PCC fragments after high doses of X-rays and their frequencyis dose dependent, although sensitivity of that method is lower in the lowdose range as compared to scoring PCC fragments or PCC rings.

Dose Response of Plasma DNA as an Adjunct to WBC forBiodosimetry

P. OkunieffRadiation Oncology, University of Florida, Gainesville, Florida, USA

In the case of a radiological or nuclear event, rapid and precise triagethat can distinguish exposed individuals will be essential and a rapid,precise, simple, and cost-effective assay is urgently needed. To that end,we conducted fit-for-purpose studies of the use of ionizing radiation (IR)-induced alterations in plasma DNA as the basis of a retrospective

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biodosimetry assay system. In this study, 36 Rhesus monkeys (3-6 yearsold, 18 males/18 females, 6 primates/group) were exposed to TBI at dosesof 0, 1, 2, 3, 5.5, or 6.5 Gy using cobalt-60 γ rays (0.5 Gy/min). After TBI,blood was collected from each primate at days 1, 2, 3, 4, 5, 6, or 7 andanalyzed for plasma DNA levels with the QuantiDNA kit and for lymphocyteand total white blood cell (WBC) counts using a clinical blood analyzer.The results showed thatA. Plasma DNA concentrations increased on day 1 with good separation

between different doses and then decreased on days 2 and 3 whileremaining above the basal level.

B. On days 4-7, plasma DNA remained above basal levels in primatesexposed to >3 Gy.

C. Lymphocyte counts rapidly decreased to their lowest level on day 1and remained at that level from days 2-7 with good separationbetween exposure doses.

D. Total WBC counts generally exhibited a continuous decline, reachinga nadir at 6-7 days following irradiation to doses =3 Gy. On days 5-7,good dose-dependent separations in the WBC counts were obtainedat doses of 0, 1, 2, 3, and 5.5 Gy. Based on the data, we concluded that these 3 indices form a strong

combination of biomarkers that will distinguish between significantlyexposed (=3 Gy) primates within 1-7 days after TBI. From days 1-3,plasma DNA and lymphocytes were the dominant biodosimetric indices;from days 4-7, WBC and lymphocyte counts were dominant. With sensi-tivity analysis using a specificity of 95 %, the combined index achieved100% sensitivity at doses =3 Gy from 1-7 days, except for day 3. At thistimepoint, sensitivity was equal to 83 % for both 2 and 3 Gy and 100 %for higher doses. These results demonstrate the utility of the combinedbiomarker approach for retrospective dose estimation at doses of0-6.5 Gy.

This project was funded by BARDA (Biomedical Advanced Researchand Development Authority, Office of the Assistant Secretary for Pre-paredness and Response, Office of the Secretary, DHHS); Contract #HHSO100201000003C.

Retrospective dosimetry on the territories with highradioactive contamination

D. DubovMinistry of Health of Russian Federation, Federal State Iinstitution “Med-ical Radiological Research Center”, Obninsk, Russian Federation

The review of methods and results of retrospective estimates ofirradiation dose among population living on the territories with veryhigh radioactive contamination are presented. The methods used arethe thermoluminescence technique with the quartz inclusions inbricks (TL and OSL technology) and EPR dosimetry using humantooth enamel. The individualization of TL/OSL-dose was performedusing the data obtained by individual questioning regarding the indi-vidual behaviour, food consumption and detailed information aboutthe radioactive contamination. The results of comparison of dataobtained by different methods will be demonstrated in the report.

External dose assessment in a radiological mass casualtyscenario using EPR on mobile phones

P. Fattibene1, F. Trompier2, A. Wieser3, C. De Angelis1

1 Istituto Superiore di Sanità, Italy 2 Institut de Radioprotection et de Sûreté Nucléaire, France 3 Helmholtz Zentrum Muenchen, GermanyBackground

In the framework of the EU-FP7 MULTIBIODOSE project, a methodbased on EPR and OSL in display glass and circuitry board resistorsof mobile phones is being implemented as a tool to assess the exter-nal dose of potentially ionizing radiation exposed individuals. Here wedescribe the method based on EPR (the OSL method will be describedin another work of this conference).

MethodsThe glass from displays and touch screens of 75 mobile phones of

various brands and models were analyzed by EPR in three laboratories.The characteristics of the EPR spectrum before and after in-laboratoryirradiation and the stability of the EPR signal up to 10 days after irradia-tion were studied. Results

About 85 % of mobile phones presented a detectable radiationinduced EPR signal in glass. We distinguished five types of EPR signals,correspondingly to different main glass-forming constituents. One type ofglass, mainly present in touch screens, had lower background signaland longer signal stability than the other types. The EPR spectrum ofthis glass presented one stable and one unstable component. Theunstable component was canceled in about one day if the glass wasexposed to light. The response to radiation was linear between 0 Gy and10 Gy (the maximum investigated dose). In a blind dose test among thepartners, the dose was assessed within 20%. Conclusions

Touch screen glass showed properties of radiation sensitivity and timestability suitable for its potential use as a fortuitous dosimeter in radio-logical mass casualty scenarios.Acknowledgements

The research leading to these results has received funding from theEuropean Union\'s Seventh Framework Program (FP7/2007-2013)under grant agreement n° 241536.

Inter- and intra-individual variability of baseline andradiation-induced gamma-H2AX foci

J. Al-HafidhHealth Protection Agency, Chilton, Didcot, UK.

The gamma-H2AX assay has the potential to be a useful triage tool dur-ing a large scale radiation incident to help identify critically exposed indi-viduals and reassure the worried-well. However, it is important to knowthe extent of inter- and intra-individual variability and how this may affectdose estimations. Knowing baseline levels of gamma-H2AX foci within ahealthy, non-exposed control population is important to determine mini-mum detectable doses and quantify associated uncertainties. Similarly,intra- and inter-individual variations in the yields and kinetics of radiation-induced gamma-H2AX foci have to be considered as important potentialmodifiers for uncertainties associated with dose estimations. Here we haveanalysed 339 samples from 32 healthy donors of working age to deter-mine spontaneous and ex vivo X-ray-induced foci yields at different dosesand time points, with up to four independent repeat samples from thesame donor. Multivariate analysis of variance showed that foci yields werea function of dose (p < 0.001) and time point (p < 0.001), but not theindividual donor (p = 0.786), with similar levels of intra- and inter-individ-ual variability observed among these healthy donors. Smoking status, gen-der and age had no significant effect on spontaneous or radiation-inducedgamma-H2AX foci in this small cohort. We conclude that genetic or envi-ronmental factors do not appear to significantly modify baseline and radia-tion-induced gamma-H2AX foci yields in healthy adult donors. The identi-fied range of variation will be used to refine uncertainties associated withgamma-H2AX-based dose estimations.

The research leading to these results has received funding from theEuropean Union’s Seventh Framework Programme (FP7/2007-2013)under grant agreement n° 241536 and from the UK Home Office.

Cigarettes as fortuitous dosimeters in case of radiologicalemergency

E. Bortolina,b, C. Boniglia, R. Gargioloa, Nucetellia,b, P. Fattibene a,b

a Istituto Superiore di Sanità, Viale Regina Elena 299, Roma, Italyb Istituto Nazionale di Fisica Nucleare, Gruppo Collegato Sanità, VialeRegina Elena 299, Roma, Italy

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The use of silicates as dosimetric material is well known and rangesfrom the identification of irradiated food to the monitoring of environmentaldose. The quartz extracted from bricks and tiles, for example, has beenwidely used for dose reconstruction using thermoluminescence technique.Recently the possibility to collect silicates from the dust present on per-sonal objects for individual dosimetry in case of radiation accident or radi-ological attack, was successfully investigated. Silicate dust can be easilycollected by “washing” the objects using laboratory small equipments asan ultrasound bath and a density separation, as described in the CEN-2001-EN1788 standard for extraction of silicates from food. The extractionof silicates can be performed contemporary on different samples, whichallows to carry out several tens of analysis at the same time. As mineraldebris occur ubiquitously on everything has been exposed to air, silicatescan be collected from dust particulate extracted from any personal items,e.g. coins, keys, fingernails, tobacco of cigarettes, etc.,. Among them,tobacco offers the unquestioned advantage that is considered of no valuefor people who could donate it easily in case of a radiological emergency.

The aim of this work was to study the dosimetric characteristics of sili-cates extracted from tobacco of cigarettes in view of their possible use inemergency dosimetry. The results are encouraging, as just one cigaretteprovided amounts of silicates sufficient to reveal doses as low as 100 mGy.Silicates showed a background signal (BGS) that partially overlaps theradiation induced signal and might reduce the accuracy of the low dosesevaluation. BGS might due to the natural radiation or to radioactive ele-ments in tobacco. Further investigation on the background origin as wellas on its influence on the dose reconstruction, are in progress.Keywords

Retrospective dosimetry, TL, silicates, tobacco, cigarettes.References

Is dust a suitable material in retrospective personal dosimetry? RadiationMeasurements, 45 (2010) 753–755.

Use of In Vivo and Ex Vivo EPR Measurements in Finger/ToeNails as the Basis of a Radiation Biodosimetry Method

Steven G. Swarts2, Jason W. Sidabras3, Oleg Grinberg1, Andres E.Ruuge1, Dmitriy S. Tipikin1, Michael Mariani1, Jean P. Lariviere1, Xiaoming He1, Dean E. Wilcox1, Jiang Gui1, Shiv Varanasi3, Stephen D.P.Marsh2, Benjamin B. Williams1, Ann B. Flood1, Harold M. Swartz1

1Geisel School of Medicine at Dartmouth, Hanover, New Hampshire,03755; 2Department of Radiation Oncology, University of Florida,Gainesville, Florida, 32610; 3Medical College of Wisconsin, Milwaukee,Wisconsin, 53226

Methods that can rapidly and accurately determine individual exposureto radiation will be needed for screening (triage) of populations and guidemedical treatment in the emergency response to an accidental or masscasualty radiological/nuclear event. One such method uses X-band Elec-tron Paramagnetic Resonance (EPR) to measure a radiation-induced sig-nal finger and/or toe nails. Two methods are presented: ex vivo in nailclippings, and in vivo on the nail plate using specially designed resonators.The key to the success of the ex vivo approach is addressing the majorchallenge of signal overlap between the radiation-induced signals (RIS)and a mechanically-induced signals (MIS) formed when the nail is har-vested. This is made possible by utilizing proper modeling of the MIS andRIS spectral elements, obtaining a robust fitting of these spectral elementsto the experimental EPR spectra of the irradiated nail clipping, and defin-

ing the variability of the background signal. The use of a multi-componentEPR spectral analysis of irradiated (0-6 Gy) clipped nail spectra acquiredfrom a large dataset (90 samples from 15 donors) shows a high degree ofconsistency between the pure RIS and the estimated RIS computed fromspectral analysis. In addition to the success in RIS estimation, we havealso achieved linear dose response for all individuals in this study, wherethe radiation dose of interest ranges from 0-6 Gy.

In the in vivo EPR approach, two types of resonators are being devel-oped to provide large sampling volumes of the nail plate while limiting themicrowave field to avoid the underlying lossy soft tissues. The SurfaceArray Resonators (SRA) consist of 6-7 parallel elements which limit depthsensitivity primarily to the nail plate, whereas the Aperture Resonators(AR) makes use of a modified conventional resonator (e.g. TE102) with adielectric insert to enhance the sensitivity at one or more apertures (5 mmaperture that varying in width from 1.0-2.5 mm). Initial testing of the SRAand AR prototypes in tissue-equivalent nail models show that these res-onators are capable of achieving detection sensitivities within the rangerequired for measuring RIS to at least a 2 Gy dose. Proof-of-concept stud-ies have progressed to testing in irradiated cadaver fingers. This in vivoapproach will compliment the ex vivo nail EPR approach for providing ret-rospective dose estimation.

Protein candidates in ELISA-based dosimetryME Hotz, V Meineke

Bundeswehr Institute of Radiobiology affiliated to the University of Ulm,Munich, Germany

Ionizing radiation (IR) exposed individuals exhibits protein changes intheir serums. Among those proteins ataxia telangiectasia mutated(ATM) is one of the major DNA damage sensors. The most harmfulform of DNA damage is the double-strand break (DSB). The first eventsmarking the DSB is the phosphorylation of the histone H2A isoformH2AX, also referred to as gamma-H2AX, which is mediated by ATM.Furthermore, ATM is characterized by hypersensitivity to IR. IR-inducedDNA damage is known to rapidly upregulate ATM kinase activity/phos-phorylation events in the control of cell cycle progression and otherprocesses.

To examine the effect of IR on different cell types (primary blood lym-phocytes and different human cell lines) the production of the proteinmarker phospho-ATM (p-ATM) was measured in vitro after irradiation(240kV X-ray, 1 Gy/min). The intracellular and extra cellular concentra-tions of p-ATM were determined by sandwich-Enzyme-LinkedImmunosorbent Assay (ELISA) before and 8-48 hours post-irradiation.

The identification and characterisation of potential bioregulators canmonitor radiation syndrome at an early and late stage. Furthermore theknowledge of bioregulators can have applications for early triage andfollow-up medical assessments such as to support and optimise a pur-poseful and causal therapy of radiation exposed personnel.

First results demonstrate that p-ATM is present in variable and lowamounts in different cell types. Phospho-ATM levels can be rapidlyinduced by IR to saturable levels specific for different cell types.

It is expected that such protein-based biodosimeter applications willrequire only minutes to exhibit results, and with a resolution thatenables rapid triage to distinguish between the truly significantlyexposed and the expected very large numbers of worried wells. Even toassign exposed individuals to appropriate medical treatments.

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Stem cell therapy for the treatment of severe tissue damageafter radiation exposure

M. Benderittera, R. Tamarata, J.F. Bottolier-Depoisa, P. Gourmelona,E. Buglovab, J.J. Latailladec, E. Beyd

a Institut de Radioprotection et de Sûreté Nucléaire, Laboratoire deradiopathologie et de thérapies expérimentales, Fontenay-aux-Roses,France.b International Atomic Energy Agency, IEC, Vienna, Austria.c Centre de transfusion Sanguine des Armées, Clamart, France.d Service de Chirurgie Plastique, Hôpital d'Instruction des Armées

Percy, Clamart, France.Effect of radiation on healthy tissue. The French Institute of Radiopro-

tection and Nuclear Safety (www.irsn.fr) contributes to understand thebiological mechanism of initiation and progression of healthy tissuedamage, resulting both from radiological accidents or radiotherapy sideeffects. IRSN is strongly implicated in the field of regenerative medicineof healthy tissue after severe radiation damages.

Regenerative medicine based on stem cell injection as a medicalcountermeasure in case of severe radiation burns: a clinical transfer.Recently, in collaboration with the Percy hospital (Clamart, France) ourgroup have evidenced for the first time, the efficiency of MSC therapy inthe context of acute cutaneous and muscle damage following accidentalexposure. Treatment of severe radiation burns remains a difficult med-ical challenge. The most severe manifestations are highly invalidating.Although several therapeutic strategies (excision, skin grafting, skin or muscle flaps) have been used with some success, none haveproven entirely satisfying. The concept that stem cell injections could be used for reducing normal tissue injury has been discussed for anumber of years. Pre-clinical and clinical benefits of mesenchymal stemcell injection for ulcerated skin and muscle restoration after high doseradiation exposure have been successfully demonstrated. Up to tenpatients suffering from severe radiological cutaneous syndrome weresuccessfully treated in France based on autologous human grade mesenchymal stem cell injection combined to plastic surgery or skin graft.

The medical management of radio-induced aplasia: overviewof 8 patients (2006-2012).

M. Benderittera, J.F. Bottolier-Depoisa, P. Gourmelona, N.C. Gorinb,c,E. Buglovad, T. De Revele

a Institut de Radioprotection et de Sûreté Nucléaire, Laboratoire deradiopathologie et de thérapie expérimentale, BP 17, 92262 Fontenay-aux-Roses, France.

b European Cooperative Group for Blood and Marrow Transplantation(EBMT), Paris Office, Université Pierre et Marie Curie, Paris VI, Paris,France.

c Hôpital Saint-Antoine, service d'hématologie et de thérapie cellulaire,Paris, France.

d International Atomic Energy Agency, IEC, Vienna, Austria.e Service d’hématologie, Hôpital d'Instruction des Armées Percy, Cla-

mart, France.An European consensus concerning the medical management

of mass radiation exposure was obtained in 2005 under the aegis of the European group for Blood and Bone Marrow Transplantation, IRSNand University of Ulm. European consensus was that in case of ARS,HSC transplantation is not an emergency and that cytokine should be used as early as possible for 14-21 days. Physical dose reconstruc-tion combine to daily blood count follow-up is a crucial point for the evaluation of the existence of residual haematopoiesis. The possibility of HSC must be discussed only if severe aplasia

persists under cytokine. The 8 cases report (2006-2012) presented in this review were managed according to this European consensus and were a medical success in the treatment of radio-induced aplasia.

Mitigation of radiation lung injury in rats by a short course ofenalapril

1,2,3Meetha Medhora, 1Feng Gao, 1Jayashree Narayanan, 1Brian L.Fish, 2,3Elizabeth R. Jacobs, 1John E. Moulder1Radiation Oncology, 2Pulmonary Medicine, Medical College of Wiscon-sin, Milwaukee, WI 53226; 3Zablocki Veterans Affairs Medical Center,Milwaukee, WI 53295, USA.Introduction

We have demonstrated that angiotensin converting enzyme (ACE)inhibitors including enalapril mitigate radiation injuries to the lung and kid-ney when started after irradiation. Aim

Our goal is to optimize the dose and schedule of enalapril for mitigationof radiation pneumonitis and pulmonary fibrosis in a whole animal modelrelevant to a nuclear accident or radiological-terrorism event.Experimental procedures

Female rats (WAG/RijCmcr, 8-10 weeks) were treated with a single doseof 13 Gy X-irradiation to the thorax. Irradiated rats remained untreated orwere treated with enalapril (gift from Merck USA) at 30 mg/L (low dose) or60 mg/L (high dose) starting at 7 days, 35 days (just before pneumonitis),70 days (after pneumonitis), 105 days or 140 days (before fibrosis). Drugwas also stopped at 30, 60 and 90 days after radiation. Breathing ratesand newly synthesized lung collagen were measured. Survival was moni-tored in accordance with criteria of the Institutional Animal Care and UseCommittees. Summary of results

Enalapril mitigated pneumonitis and pulmonary fibrosis if started as lateas 35 days and continued. Enalapril (low and high dose) started at 7 daysand stopped at 90 but not 30 or 60 days improved survival through pneu-monitis. However starting at 7 days and discontinuing drug therapy at 90days reduced mitigation of radiation fibrosis at 210 days. Conclusions

We show that enalapril, even if started up to 35 days after radiationand continued, or started at 7 days and stopped at 90 days, improvessurvival from pneumonitis. At the high dose it also mitigates radiationfibrosis if started at 7 or after 35 days and continued. Thus a shortcourse of enalapril mitigates radiation lung injuries in a model relevant toa mass casualty event. Similarly, a short course of ACE inhibitors alsomitigated multiple-organ system failure after total body irradiation fol-lowed by a bone marrow transplant. Funding

NIH/NIAID USA, RC1 AI 81294, 81294-01S1 and U19-AI-67734.

Protection from radiation pneumonitis in rats by non-overlapping irradiation to 10% of the skin

Meetha Medhora1,3,4,5, Feng Gao1, Ashley Schock2, Brian Fish1,Jayashree Narayanan1, Elizabeth Jacobs3,4,5,6, John Moulder1, ZelmiraLazarova2

1Dept of Radiation Oncology, 2Dept of Dermatology, 3Pulmonary & Criti-cal Care, Dept of Medicine, Medical College of Wisconsin, 8701, Water-town Plank Road, Milwaukee, WI 53226, USA. 6Research Service, Deptof Veterans Affairs, Zablocki Veterans Affairs Medical Center, Milwaukee,WI.Introduction

After a radiological attack or accident, victims may suffer partial orwhole body exposures to radiation, with multiple organs being affected.

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8. Radiation health effects and medical countermeasures

The lung is a sensitive target, suffering acute interstitial pneumonitis at6-10 weeks. Injury to the skin is also a major cause of morbidity. Aim

To test the consequence of combined radiation injury to the lungsand skinMethods

Rats (female WAG/RijCmcr at 9-10 weeks of age) were randomizedinto 4 groups: (i) no irradiation (n=10) (ii) 13 Gy to the whole thorax(n=16) (iii) 30 Gy soft X-rays to ~10 % of the skin (n=12) (iv) 13 Gy tothe whole thorax followed by 30 Gy soft X-rays to ~10 % of the skingiven 3 hours after thorax irradiation (n=16). Irradiation to the skin waslimited to the full thickness of the skin only. All rats were regularlymonitored for morbidity and euthanized under criteria set by the Insti-tutional Animal Care and Use Committee (IACUC). Results

Irradiation to the lung (13 Gy, group ii) induced 90 % morbiditybetween 6-8 weeks post-irradiation. Combined irradiation (group iv) tothe lungs (13 Gy) and skin (30 Gy) induced 25 % morbidity between 7-10 weeks after irradiation. The combined lung and skin irradiation pro-tected rats from morbidity due to pneumonitis (p<0.0001, n=16/group). There was no morbidity in unirradiated rats (group i) orthose irradiated to the skin alone (group iii). These results are consis-tent with similar experiments combining 12 or 12.5 Gy irradiation to thethorax with 30 Gy to the skin, where neither thoracic nor skin injury wasexacerbated by combining them. Circulating cytokines were measuredin these rats by a Bio-Rad multiplex assay. Granulocyte colony stimu-lating factor (G-CSF) levels were lowest after 6 weeks in rats irradiated to the thorax alone.

Our data indicate that limited radiation-induced skin injury protectsand may delay radiation pneumonitis in a rat model of combined lungand skin injury. One mechanism for this may be by elevation of circu-lating levels of G-CSF.Funding

NIH/NIAID (USA) agreements U19-AI091036 & 67734 and RC1AI81294 & 01S1.

Contraindication of DTPA by uraniumC. Jones

Radiation Protection Group, AWE, Reading, UKDiethylene-triamine-penta-acetate (DTPA) is the drug of choice for

decorporation treatment (removal of radionuclides from the human sys-temic circulation) following intakes of plutonium, americium andcurium[i]. However, it is a common belief that DTPA should not beadministered for intakes of uranium because not only is it ineffective atpromoting the excretion of uranium, but it can actually enhance thenephro-toxic effects of uranium.

The question therefore arises of whether DTPA should be adminis-tered following a mixed intake of plutonium and uranium, for examplefrom a MOX fuel. The work being presented combines the OMINEX rec-ommended workplace exposure limit of 10 mg of uranium [ii] withresearch into the effects of combining DTPA and uranium, principallyfrom Muller et al. [iii], to suggest that DTPA can be administered with-out concern for nephro-toxic effects if there is less than 3mg of ura-nium in systemic circulation. If the level is greater than this, the prosand cons of treatment will have to be considered; for example, if thequantity of plutonium internalised is sufficient to cause tissue reactions,then it is suggested that DTPA should be given regardless of theamount of uranium present.

[i] NCRP Report No. 65, Management of Persons Accidentally Contam-inated with Radionuclides, NCRP, 1980

[ii] Stradling, N, et al. OMINEX report; industrial Uranium Compounds:Exposure Limits, Assessment of Intake and Toxicity after inhalation

[iii] MULLER, D., HOUPERT, P., HENGE NAPOLI, M., METIVIER, H.,and PAQUET, F., Synergie potentielle entre deux toxiques renaux: leDTPA et l’uranium,

Radioprotection 41(4): 413 – 420, 2006

HemaMax™ Simultaneously Enhances Near-Term Survivaland Long-Term Recovery from Lethal Total Body IrradiationDolph Ellefson1, Tim Gallaher1, Sarita Mendonca1, Simmy Thomas1,Zoya Gluzman-Poltorak1, Chris Lawrence1, Liron Shiri1, Lena Basile1

1Neumedicines Inc., Pasadena, CA, 2Keck School of Medicine, Univer-sity of Southern California, Los Angeles, CARadiation exposure often results in outgrowth of enteric bacteria translo-cated across the intestinal barrier. Radiation-induced immunosuppres-sion enhances mortality by allowing translocated bacteria to quicklybecome septic. In addition, radiation exacerbates mortality by denudingbone marrow progenitor populations essential to regeneration of periph-eral immunity. Mitigation of hematopoietic syndrome requires the sup-pression of bacterial outgrowth in the near-term until restoration of effec-tive peripheral immune surveillance. HemaMaxTM (rHuIL-12), currentlyin advanced development as a frontline drug for the treatment of thehematopoietic syndrome component of acute radiation sickness,enhances survival when administered 24hrs after exposure to lethal totalbody irradiation. HemaMax enhances long-term survival by generationof systemic multipotent recovery of neutrophils, lymphocytes, platelets,and other critical organ systems essential to survival and recovery fromlethal irradiation. In the near-term, HemaMax enhances survival by sup-pression of bacterial outgrowth by induction of innate and adaptiveimmune responses. Recent studies in non-human primates and micetreated 24 hrs after exposure to lethal total body irradiation display adiminished bacterial load relative to vehicle-treated controls. Cumulativeevidence from studies in humans, non-human primates, and micedemonstrate that HemaMax treatment, irradiation, and wounding upreg-ulate IL-12R2, the endogenous receptor for HemaMax, in neutrophils,NK cells, as well as regenerative cells in bone marrow and spleen.These data and others demonstrate that IL-12 is an essential require-ment for recovery following irradiation and injury. The HSARS project isfunded solely with federal funds from the Biomedical AdvancedResearch and Development Authority under ContractsHHSO100200800060C and HHS0100201100037C.

Differential Effects of Ionizing Radiation on Thyroid EpithelialCells Under the Influence of Radiation DosageM.J. ChaeRadiation Emergency Medical Response Team, Seoul, Republic of KoreaRadiation exposure is a well-known risk factor for thyroid cancer devel-opment. After Chernobyl and Fukushima accidents, the calcinogeniceffect of ionizing radiation (IR) on thyroid emerged as an important issuein the field of radiation protection. According to the reports from Cher-nobyl accidents, the exposed dosage of radiation is one of the mostimportant factors, as well as the age of subjects exposed. However, therehas been little known about the dosage effects of radiation exposure tothe thyroid cells. The purpose of this study was to investigate the bio-logic effects of different radiation dosages in thyroid cells by evaluatingcell proliferation and clonogenecity, or genetic changes. The normal thy-roid epithelial cell line (N-thy-ori-3-1) used in our study (ECACC, UK). N-thy-ori-3-1 cells were irradiated doses of 0.1, 1, 2, 4, and 10Gy withgamma cell irradiator (IBL 437C, CIS Bio International, Bangnols surCeze, France). Cell proliferation and survival were increased at low doseradiation (0.1 Gy). Those indicators were decreased at over 1 Gy. Cellmorphologic change was observed the dosage over 4 Gy. They arebizarre and multinucleate. We cannot detect any DNA fragmentationuntil 20 Gy, compared to positive control by IM-9 cell. DNA repair anddamage indicator with by γ-H2AX, foci of irradiated cells was peaked at1 h, and then dropped; there were positive correlation with foci number

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and dosage. These results suggest that thyroid epithelial cells havehighly radio-resistant characters, and lower dosages than 1 Gy of radia-tion did not induce significant biologic or genetic damages in thyroidepithelial cells within short term

Expression of plasma TGF beta in non-human primates (NHP)exposed to whole thoracic lung irradiation (WTLI)

Wanchang Cui, Michael C. Garofalo, Kim G. Hankey, Kaitlyn M. Kieta,Taymin O’Brien, Thomas J. MacVittieDepartment of Radiation Oncology, University of Maryland School ofMedicine, Baltimore, MD, USAIntroduction

There have been contradictory reports about the reliability of plasmaTGF beta as a predictor of radiation-induced lung injury. Here we stud-ied whether plasma TGF beta levels were affected by thoracic radiationand whether treatment with a candidate radiation mitigator (AEOL10150) would change the plasma TGF beta level in a NHP model. Wealso studied the expression of a TGF beta target PAI-1 in irradiatedlungs. Methods

NHP (Macaca mulatta) were exposed to a single fraction of WTLI atdoses of 9, 10, 11 or 11.5 Gy. In the 11.5 Gy group, half of the cohortreceived AEOL10150 (a superoxide dismutase mimetic) starting at 24hours after WTLI and continued once daily for 28 days, the controlcohort received no drug. Plasma was collected at multiple time pointspre and post radiation exposure and lung tissue was collected atnecropsy. Plasma TGF beta levels were measured using ELISA method.Pulmonary PAI-1 expression was assessed using Western Blottingmethod. Results

Both absolute and relative values (to each individual’s baseline value)of plasma TGF beta showed a time-dependent increase in the NHPexposed to 9, 10 and 11 Gy WTLI. Plasma TGF beta peaked at 6-8weeks and returned to baseline at 12 weeks following WTLI. PAI-1 hadan elevated expression in the irradiated lungs compared to that found inthe non-irradiated controls. Within the 11.5 Gy cohort, plasma TGF betalevels also peaked at 6-8 weeks post-exposure in the control group,while the AEOL 10150 group showed marginal increase. TGF beta levelswere lower at multiple time points in the AEOL 10150 group when com-pared with the control group.Conclusion

Our results support the hypothesis that plasma TGF beta levels arepredictive of radiation-induced lung injury in a NHP model of high-doseacute WTLI. In our model, the peak of plasma TGF-beta levels correlatewith the average time of onset of radiation-induced pneumonitis. Thelower level of plasma TGF beta in the AEOL10150-treated group sug-gests that plasma TGF beta levels reflect treatment benefits. Therefore,plasma TGF beta may prove to be a useful biomarker of radiation-induced lung injury.

This work was supported by BARDA contractHHSO0100201100007C, NIAID contract HHSN272202000046C andAeolus Pharmaceuticals Inc.

New radioprotective molecules: toward ThiolsP. Martigne

IRBA (french military biomedical research institute), LA TRONCHECEDEX, FranceAbstract

Since the sixties, many research studies on radioprotective moleculeshave been conducted, in particular by the Armed Forces RadiobiologyResearch Institute (AFRRI, USA) and by the Institute of BiomedicalResearch of the French Army (IRBA-CRSSA) (Weiss et Landauer, 2009).However, only one molecule obtained the FDA certification in the anti-cancer area, namely the amifostine or WR-2721 (Ethyol°), the precursor

of the active thiol, WR-1065. Unfortunately, the adverse effects of thisphosphorothioate – as a major hypotension potentially leading to a lossof consciousness, severe nausea and vomiting… – drastically limit its possible use, in radiotherapy as well as in chemotherapy. This is evenmore harmful if administered in military or civilian security missions during nuclear or pyrotechnic accidents, for example.

The naphthyl-methyl-imidazole (NMI) also showing a promising radioprotective activity (Laval and al., 1993, Prouillac and al., inSpotheim-Maurisot 2008), we thought about functionalizing the NMI by a phosphorothioate group. However, a complex and expensive syn-thesis of the N-substituted NMI target combined with first disappointingresults in terms of survival at LD100/30d on a mouse model led us to rethink both the binding and the length of the sulfur alkyl chain. The first results are promising since we got 100% survival at LD100/30d(global acute irradiation with a 60Co source at a dose of 8.5 Gy and adose rate of 0.6 Gy/min) after administration of NMI (30 mg/kg ip),which result is very close to that obtained with Ethyol° (400 mg/kg ip).

The partnership with NovAliX (Strasbourg-Illkirch, France), a companyoffering capabilities in chemistry and biophysics to support the pharma-ceutical industry’s needs, continues through a French research project(DGA/DGCIS). Among the challenges of this collaboration, the reductionof the NMI vasoconstrictor effect and the optimal structural modificationfor the introduction of the active thiol moiety are considered.

Biomarker Discovery Utilizing MALDI Imaging MassSpectrometry to Characterize Radiation-Induced TissueDamage

Alison J. Scott1, Jace W. Jones2, Maureen A. Kane2, Thomas J.MacVittie 3, Robert K. Ernst11University of Maryland School of Dentistry2University of Maryland School of Pharmacy3University of Maryland School of Medicine

Integral to the characterization of the radiation-induced tissue damageis the identification of unique biomarkers. Biomarker discovery is a chal-lenging and complex endeavor requiring both sophisticated experimentaldesign and accessible technology. The resources within the National Insti-tute of Allergy and Infectious Diseases (NIAID)-sponsored Consortium,Medical Countermeasures Against Radiological Threats (MCART) allow forleveraging robust animal models with novel molecular imaging techniques.

One such imaging technique, MALDI (matrix-assisted laser desorptionionization) imaging mass spectrometry (IMS), allows for the direct spatialvisualization of lipids, proteins, small molecules, and drugs/drug metabo-lites – or biomarkers - in an unbiased manner. MALDI-IMS acquires massspectra directly from an intact tissue slice in discrete locations across an x,y grid that are then rendered into a spatial distribution map composed ofion mass and intensity. The unique mass signals can be plotted to gener-ate a spatial map of biomarkers that reflect pathology and molecularevents. The crucial unanswered questions that can be addressed withMALDI-IMS include identification of biomarkers for radiation damage thatreflect the response to radiation dose over time and the efficacy of thera-peutic interventions. Techniques in MALDI-IMS enable integration of bio-marker identification among diverse animal models, including mouse andnon-human primate.

We will show preliminary data from a MALDI-IMS survey of mouse andnon-human primate for overall lipid composition. Data on phospholipidand flavanoid species within gut tissue demonstrates the ability of MALDI-IMS to provide information on spatial localization. Analysis of radiation-induced tissue injury samples from non-human primate lung tissue showsphospholipid signatures consistent with oxidized phospholipids that couldhave implications for observed tissue pathology. Additional MALDI-IMSanalysis for molecular discovery of radiation-induced tissue injury bio-markers are currently underway using murine and non-human primatetissues samples.

This work was supported by NIAID contract HHSN272202000046C

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Examination of gene expression, micro-RNA, cell death andproliferation after low dose ionizing radiation: an in vitro study.

M. Port, F. Monje, Annette Schmidt, V. Meineke, M. AbendIntroduction

Ionizing radiation is known to cause stochastic late effects like cancer.To evaluate long lasting epigenetic changes we exposed HL-60 cells tolow level ionizing radiation and examined transcriptional and posttran-scriptional changes over time after irradiation.Material and methods

HL-60 cell cultures were irradiated on five consecutive days with 0.01Gy, 0.05 Gy and 0.1 Gy resulting in a total exposure of 0.05 Gy, 0.25 Gyand 0.5 Gy. To examine biological effects on different time points up to 6weeks after irradiation we (1) quantified micronuclei, apoptotic andabnormal cells (MAA-assay), (2) screened the whole genome for geneexpression changes in cultures exposed with 0.5 Gy (microarray), (3)examined 380 mRNAs representing key regulators for cell death, prolif-eration and DNA repair (qRT-PCR, CLARCC-array) and (4) performedexpression analysis on 668 microRNA (qRT-PCR, low density array).Results

Significant changes in MAA-assay could not be detected. We did findcertain genes and microRNAs to be differentially expressed (>= 2-fold)using either the whole genome microarray (153 genes), the CLARCC-array (59 genes) and microRNA experiments (24 microRNAs), but nopersistent pattern versus time evolved.Conclusion

The negative results of our examinations could represent a featureinherent to cancer cells or caused by methodological variance of ourexperiments. Further examinations using primary cultures such asembryonal stem cells but no tumor cells are currently in progress.

Molecular and cellular effects of chronic low doserategamma-radiotion exposure in mice

A. OsipovBurnazyan Federal Medical and Biophysical Center of the Federal,Moscow, Russian Federation

While there are no doubts about negative biological effects of highdose ionizing radiation (IR), debates about whether low dose IR expo-sure is harmful or beneficial (hormetic) are still continuing among scien-tific community. There are many evidences that low dose-rate IR expo-sure induces a complex of molecular and cellular responses in animals.The main question is: whether those changes are consequences oforganism adaptation to increase in IR background, and whether lowdoses cause any significant genetic alteration? In the present report pro-vides an overview of the data obtained during the experimental studiesof chronic IR effects in mice. Changes in the levels of DNA single-strandbreaks (SSB), apoptosis, cytogenetic disturbances and reactive oxygenspecies (ROS) production rate were analyzed in blood system cells (lym-phocytes, leucocytes, spleen and bone marrow cells) of CBA male-micechronically exposed (up to 1 year) to gamma-radiation at a dose-rate of61-62.5 cGy/year. The results showed that the molecular and cellularresponses to chronic low dose-rate ionizing radiation exposure develop-ing with an increase in the exposure time (dose) of irradiation in a fewstages. Beginning from 120-th day of exposure (~ 20 cGy), statisticallysignificant increase in the SSB level, apoptotic cells frequency and ROSproduction rate was observed. Need to note that further prolongation ofexposure time to 1 year and, hence, increase of a total dose did not,lead to further increase in the yields of these indices. The effects wereaccompanied by elevated cell resistance to additional hydrogen peroxidetreatment in vitro indicated adaptive response development. No signifi-

cant increase in the cytogenetic disturbance frequencies was found untilexposure time at 300-365 days (50-62.5 cGy). Upon the whole, theseresults provide further support for the non-linear threshold model of lowdose-rate gamma-radiation genotoxic action.

Balanced Emergency Preparedness for Civil Protection -Implementing the Scorecard Concept in Public Managementto Develop Response Capabilities of Civilian Organisations forMass Casualties in Urban and Rural Scenarios in theEuropean Union in the Light of Disaster Response Failuresduring the Fukushima Nuclear Catastrophe in Japan 2011 – AConceptual Study

Frank Klimaschewski (MSc, PhD student) Helmut-Schmidt University Hamburg, Germany and Uranium MedicalResearch Centre (UMRC), USA.Purpose

The nuclear disaster in Fukushima, Japan 2011 clearly demonstratedto the member states of the European Union the vulnerability of a highlyindustrialized society. The ordinary command and control of publicadministration (linear management approach) and its emergency serv-ices were beyond capacity and unable to cope with mass casualties.The local population were not trained to organize themselves to deal withthe disaster and the implications of radiation exposure. Consequentlythe EU reviewed the technical safety of its nuclear reactors to develop animproved safety culture. No initiative, however, has been taken to estab-lish a significantly enhanced emergency preparedness culture thatactively trains and involves civilian organisations in preparation for masscasualties on a long term basis. The purpose of this work, therefore, isthe description of a new linear-reflective management system, i.e. theapplication of the balanced scorecard approach for emergency pre-paredness in public management, an adaptable strategy to meet civilianneeds in mass casualties and stimulate ongoing improvement of civilprotection.Strategy

The balanced scorecard is a pragmatic management tool that hasbeen successfully applied in public management after its successfulapplication in private businesses and corporations worldwide. Its top-down and bottom-up approach is unique. It enables a managementteam to setup and pursue a number of strategic objectives with quanti-tative indicators, thus reducing complexity and focussing on key strate-gic goals.

A city scorecard for emergency preparedness, for example, is beingdefined after initial risk assessments for the city. It sets an overall visionand mission for the local community’s disaster response from which alimited number of objectives and strategies are derived. A maximum of 5objectives are usually set for 5 standardized areas of organisationalactivity: (1) Citizens (safety/protection), (2) change processes (disasterresponse patterns, equipment, local resources), (3) finance (budgeting),(4) employees (learning and development of key staff), and (5) futuredevelopments (new objectives, standards and degrees of preparedness).Objectives for these areas can partly be standardized but must allowindividual flexibility. They, therefore, need to be assessed and scored todetermine deviations from ideal target values and performance.

Public management authorities in partnership with private enterprisetake the top-down initiative and have to support bottom-up develop-ments on various levels of society, such as schools, hospitals, privatebusinesses, households, etc. Schools in particular play a key role in thisstrategy being places of organized teaching and learning for the nextgeneration where a new preparedness culture and awareness can crys-

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9. Effects of low dose ionizing radiation

tallize. Each individual organisation’s preparedness scorecard formulatesa mission, vision, objectives, strategies, and ongoing cycles of planning,training, practice runs and evaluations which align with the city score-card. An online internet platform can provide state of the art supportwith standards, processes and training specifically maintained by thelocal authorities for the different types of organisations that can log onand register. This allows an overview of progress made and the identifi-cation of areas of weakness to be addressed. Conclusion

The long-term development of effective community preparedness formass casualties with a particular focus on radiation emergencies in theEU is warranted in view of its many risk potentials such as numerousactive nuclear reactors and especially because of the severe lack of bot-tom-up response strategies. The balanced emergency preparednessapproach described provides an innovative adaptable management toolwhich can help to structure, synchronize and synergize public and pri-vate resources. Existing organisations can be adapted to provide a civil-ian response to support emergency services that are challenged beyondcapacity during a large scale disaster.

Long term changes of hippocampal and cortical proteins afterbrain irradiation in young mice

S.-J. Kempf (p)1, O. Azimzadeh1, C. von Toerne2, S. Buratovic3, M.-J.Atkinson1, 4, P. Eriksson3, S. Moertl1, S. Pazzaglia5, M. Ueffing2, S.Tapio1

1 Institute of Radiation Biology, Helmholtz Zentrum München, GermanResearch Center for Environmental Health, Neuherberg, Germany 2 Department of Protein Science, Proteomics Core Facility, HelmholtzZentrum München, German Research Center for Environmental Health,Neuherberg, Germany3 Department of Environmental Toxicology, Uppsala University, Sweden4 Department of Radiation Oncology, Klinikum rechts der Isar, Technis-che Universität München, Munich, Germany 5 Laboratory of Radiation Biology and Biomedicine, Agenzia Nazionaleper le Nuove Tecnologie, l\'Energia e lo Sviluppo Economico Sostenibile(ENEA) Centro Ricerche (CR) Casaccia, Rome, Italy

Low-dose ionising radiation (LDIR) exposure from medical proce-dures such as X-ray computed tomography has increased >7-fold sincethe early 1980s and now comprises nearly 50 % of the average. Thereis a great concern about (i) the validity of the linear no-threshold modeland (ii) detrimental long-term effects of ionising radiation exposure.Children may be a particular target group with pronounced sensitivityfor these alterations as they have long-life expectancy under risk.Focusing on the brain, these children are even more susceptible to ion-ising radiation due to their immature brain. Further, there is a debate ifLDIR has a role in the aetiology of late-onset neurodegenerative dis-eases such as Alzheimer´s or Parkinson´s whereas learning and mem-ory deficits are well-known after radiotherapy of head cancer patients.Our aim is to enlighten these still marginally understood mechanisms ofionising radiation on the proteome level. We investigated the long-termprotein alterations in radiation-exposed mice hippocampi and cortices.NMRI mice were exposed to total body irradiation (60Co, gamma radia-tion) on postnatal day 10 with doses of 0 (sham-irradiated), 0.02, 0.1,0.5 and 1.0 Gy. 7 months post-irradiation, the hippocampus and cortexregions were isolated and the whole corresponding proteome wasquantified using Isotope Coded Protein Label approach; proteinchanges were analysed regarding to learning and memory. Hippocam-pal analysis showed an upregulation of cytoskeletal and cytoskeleton-associated proteins throughout all radiation conditions. Affected sig-nalling pathways analysed via several software tools indicatedalterations in RhoGDI, Rho family GTPase and Ephrin B signallinghypothesising deficits in correct assembly of the spine apparatus andaxonal growth cone. Further studies will include changes in the corre-sponding miRNAome to get more information about the protein dereg-

ulation; immunohistochemistry and behavioural tests will supplementthis study.

Status of Inborn Immunity during the Period of Late Somatic-Stochastic Radiation Exposure Effects

Akleyev, A.A.1, Dolugushin I.I.1, Grebenyukan A.N.21 Southern-Urals State Medical University of the RF Ministry of PublicHealth, Chelyabinsk, Russia 2 S.M. Kirov Military Medical Academy, St. Petersburg, Russia

The risk of leukemia and solid neoplasm development has beenidentified among the late effects of chronic exposure in man. Since thebasic function of inborn immunity is to provide anti-tumor protection tothe living organism it would be important to assess its current status forresidents the Techa riverside villages who have been exposed to radia-tion for multiple years due to releases of liquid radioactive waste fromthe Mayak PA. Taking into account the combined nature of the expo-sure: external (γ-radiation) and internal (mostly due to 90Sr) and a highradiosensitivity, the red bone marrow (RBM) was identified as the criti-cal organ. The maximum dose rate to RBM was registered in theseindividuals in 1951 (1.48 Gy/year). The dose rates were graduallydiminishing overtime, and since 1985 they have never exceeded thepermissible levels (1 mGy/year). In the early years of radiation exposurethe exposed individuals exhibited decreased levels of phagocytic activ-ity of blood neutrophils. As the dose rates decreased, the manifesta-tions of these changes subsided, but the functional reserve of theinborn immunity was decreasing too. The purpose of the study was toassess the status of inborn immunity for exposed persons 60 years afterthe onset of exposure. The study group was composed of 95 exposedindividuals. The average accumulated dose to RBM estimated for thisgroup was 0.93±0.06 Gy. The control group consisted of 66 unexposedpersons. The studies performed for both groups included analyses ofphagocytic and lysosomal activity, and intracellular oxygen-dependentmetabolism of neutrophils and monocytes, as well as CSF-GM andCSF-G concentrations in blood serum.

Health effects of the accident at the Chernobyl NPP femalecohort of the emergency accident workers

A. MenyayloNational Radiation and Epidemiological Registry, Medical RadiologicalResearch Center of the Russian Ministry of Health, Obninsk, RussianFederation

The National Radiation and Epidemiological Registry (NRER) wasformed by the Decree of the RF Government N 948 of 22.09.1993. TheNRER database has accumulated medical and dosimetric informationabout 190.9 thousand the Chernobyl emergency accident workers,among them 6.6 thousand females. The ratio of radiation risks formales and females is a topical problem (Russian \"Basic Sanitary Reg-ulations\", NRB-99/2009). For the first time the authors present analy-sis of radiation and epidemiological information about the female cohortof the Chernobyl emergency accident workers and provide cancer riskassessments using UNSCEAR reports, ICRP recommendations andmathematical methods of regression analysis.

Telomere length in Chernobyl accident clean-up workersJelena Reste1, 2, Gunda Zvigule3, Tija Zvagule1, 2, Natalja

Kurjane1, 2, Maija Eglite1, 2, Natalija Gabruseva2, Dace Berzina3,Juris Plonis3, Edvins Miklasevics31 Institute of Occupational Safety and Environmental Health, RigaStradins University, Latvia2 Centre of Occupational and Radiological Medicine, Paula StradinsClinical University Hospital, Latvia3 Institute of Oncology, Riga Stradins University, Latvia

The outcome of Chernobyl nuclear power plant (CNPP) accident in1986 became a huge number of persons exposed to ionizing radiation.

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During stay in Chernobyl for 1-3 months clean-up workers wereexposed both to external and internal radiation, moreover certainamount of long-living radionuclides accumulated in their body. Previousstudies of clean-up workers’ health condition showed high prevalenceof malignancies and age-dependent diseases. The aim of the study wasto evaluate the effect of low-dose radiation long-term exposure ontelomere length of peripheral blood leukocytes (PBL).Materials and methods

Relative telomere length (RTL) was measured in PBL by real timequantitative PCR. RTL was evaluated in 831 males (595 CNPP clean-up workers from Latvia and 236 age matched controls). Data on healthstatus, participation time and job tasks during stay in Chernobyl werereceived from the Latvian State Register for persons exposed to ionizingradiation in Chernobyl. Results

The age structure didn’t differ in both groups (mean age 55 years).High inter-individual variability of RTL was observed. RTL was slightlyhigher in CNPP clean-up workers than in controls (p<0.01). Signifi-cantly longer telomeres were found in 1986 year participants and“dirty” tasks performers (digging and deactivation) comparing with con-trol group (p<0.01). Slightly shorter than in previous subgroups(p>0.05) telomeres were in 1987-1991 year participants and anotherjob workers, but RTL in them was still longer than in control group(p<0.01). Conclusions

RTL in CNPP clean-up workers was significantly longer than in ageand gender matched control group (p<0.01). Probably this indicatesactivation of telomerase because of DNA damage by ionizing radiationand defects in telomerase regulation. Longer telomeres may be signifi-cant factor for carcinogenesis. Considering signs of premature aging inclean-up workers their long telomeres may be dysfunctional. It requiresfurther investigation.

Analyses of CT induced DNA damage to determine radiationsensitivity of different age groups, especially young children– a pilot study in the frame of the EU EPI – CT project

1Oestreicher U., 1Roessler U., 2Lang P., 2Neumaier K., 2Belka C.,3Lindholm C., 4Niemeyer M., 4Kiechle M., 5Hasbargen U., 5HübenerC., 6Kirlum H.-J., 1Kulka U., 7Baatout S., 8Kesmiene A. and 1Gomolka M. 1Bundesamt für Strahlenschutz, Germany, 2LMU, Klinik und Poliklinikfür Strahlentherapie und Radioonkologie, Germany 3Radiation andNuclear Safety Authority, Finland, 4TUM, Klinikum Rechts der Isar,Frauenklinik, Germany 5LMU, Klinik und Poliklinik für Frauenheilkundeund Geburtshilfe, Germany 6Praxis für Kinderchirurgie, Germany 7Bel-gian Nuclear Research Centre, SCK-CEN, Belgium 8IARC, InternationalAgency for Research on Cancer, France

The increasing use of paediatric computed tomography (CT) world-wide has raised the question of possible late effects from exposure toionising radiation. The European collaborative EPI-CT project aims atstudying the cancer risks and the underlying biological effects in aninternational cohort study. The project is coordinated by the Section ofEnvironment and Radiation at the International Agency for Research onCancer (IARC). Eighteen centres from Belgium, Denmark, Germany,Finland, France, Luxemburg, the Netherlands, Norway, Spain, Swedenand the United Kingdom are cooperating in this project to enrol approx-imately one million patients. The overall objective is to inform aboutdose reduction and optimisation in paediatric CT.

The aim of the biological part of the study is to compare different bio-markers for radiation exposure and to test their sensitivity in clarifying

the biological mechanisms behind low dose hypersensitivity observed inCT examined paediatric patients. The work is divided into a number ofdistinct and complementary tasks which will allow to study the effectsof CT exposure using a variety of approaches, including assessment ofDNA damage, mainly through monitoring chromosomal aberrations and y H2AX foci.

In this context, an in vitro feasibility study to investigate age-depend-ent radiosensitivity is conducted at the BfS in Germany in cooperationwith STUK (Finland). Blood samples from three different age groupsranging from newborns (umbilical cord blood), young children (2 – 5years) to adolescents (>18 years) are being collected and investigatedfor different radiation-induced DNA damage parameters. Blood sam-ples are being irradiated in a CT scanner. Chromosomal aberrations aswell as the induction and repair of DNA double strand breaks havebeen investigated using y H2AX foci analyses for the different agegroups (adults, umbilical cord blood and young children). Preliminaryresults give a clear dose-dependency according to the sensitivity of theassay used for all three groups. Age dependencies between the groupsare visible for both biological assays and have to be statistically con-firmed. Final results of this pilot study will be used for evaluating thefeasibility of conducting a larger study allowing sufficient statisticalpower for estimation of age and sex dependent radiosensitivity.

Funded by the EC (FP7 under Grant Agreement n◦ 269912)

The experience of biodosimetry and bioindication low dosesof radiation in different groups of irradiated persons.

E. NeronovaLaboratory of genetical research and biodosimetry, The Nikiforov Russ-ian Center of Emergency and Radiation Medicine (NRCERM) EMER-COM of Russia, St.Petersburg, Russian Federation

Cytogenetical investigations related to radiation exposure were per-formed in people exposed mainly to low doses irradiation. Persons weredivided into several groups according to scenario of irradiation: group ofpersons who had a contact with radiation because of their profession(crew members of nuclear submarines, radiologists, participants ofnuclear tests, scientific workers), group of former residents of Semi-palatinsk nuclear testing region and the group of Chernobyl clean-upworkers, who worked at the station in 1986-90 years. The most of themtook part in recovery workers in 1986 and official doses of irradiation asa rule not exceeded 25 cGy. Analysis of stable aberrations (transloca-tions) by FISH-method was performed for former residents of Semi-palatinsk nuclear tests region. Increased level of translocations wasfound in some cases and the doses of irradiation were estimated.Analysis of unstable chromosome aberration was performed in period6-47 years after the irradiation and revealed increased level of differenttypes of chromosomal aberrations in all groups of investigated persons.It was shown that increased level of dicentrics could be found manyyears after low doses of irradiation. Frequency of dicentrics exceedingcontrol level was revealed in 33,3 % till 63,5 % persons from differentgroups even a long time after a contact with radiation factor. Moreoverstatistical analysis demonstrated correlation between frequency ofdicentrics, estimated 6-21 years after accident, and official dose of irra-diation in Chernobyl clean-up workers.

The experience of our laboratory demonstrate that cytogenetical tests – analysis dicentrics and stable aberration by FISH method couldbe an effective tool to receive relevant information concerning radiationexposure even a long time after low doses of radiation exposure. More-over the cytogenetical findings in different groups of irradiated personsin remote period after irradiation could demonstrate the phenomenonof radiation induced genomic instability.

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The cancer risk among workers of the nuclear center atSwierk, Poland

K. FornalskiNPP Project Development Department, PGE EJ 1 Sp. z o.o. (PGENuclear Energy), Warszawa, Polen

The complete dosimetric information concerning 4606 workers in nuclear center at Swierk (Poland), from 1956 to 2001, were collected. In this group 575 workers have their own medical statistics. The average cumulative doses per worker were 34 mSv (effective dose) and 17 mSv (equivalent dose to hands). There are no indicationsthat this level of exposure causes an increase of the cancer risk among irradiated workers.

Rate of still births among children whose fathers werepreconceptionally exposed at Mayak Production Association

N. KabirovaRadiation Epidemiological Laboratory, Southern Urals Biophysics Institute

Ozersk, Russian FederationThe interest in preconseptive radiation exposure effect in creases

with the increasing role of nuclear technologies within the community. The effect of father’s exposure on children has been investigated only in a small amount of studies. At the first Russian nuclear enterprise – Mayak Production Association male workers were exposed to radiation in a wide dose range. The objective of the present study is the analysis of natimortalityrate among children of Mayak PA male workers.

On the basis of the Ozersk Children’s Register database 75 956individuals born in 1949-2002 were selected, of them 602

children were stillborn. Two cohorts were formed from this amount. Thestudied cohort comprises 16 013 liveborn and stillborn children whose fathers had external gamma radiation dose at conception. Aver-age preconceptive dose, accumulated for the whole period of work was 29.2 cSv, average dose accumulated per year – 55.7 mSv. Control group comprises 26 617 liveborn and 165 stillborn children whose parents had no contact with ionizing radiation sources.

The mortinatality dynamics among the descendants of the ex posedfathers tends to excess that of the control group over almost all theperiod of observation, among both boys and girls.

The dynamics of mortinatality among offspring of exposed fathers-tends to excess that of the control group both among girls and boysalmost during the whole period of follow-up.

In the structure of causes of death of stillborn children, both in thestudied cohort and control, both among males and females, the leading-place was for diseases of newborns and inaccurately defined states, thesecond place - complications related to fetus condition and the third-place - congenital malformations.

The frequency of stillbirths in the cohort of workers is higher than in control group (7.8 and 6.2 per 1000 children, liveborn and still borncorrespondingly), that relates to significant increase of stillbirth rateamong males (8.8 and 6.3 per 1000).

Analyzing stillbirth rate depending on male preconceptive exposuredose both for the year prior conception and for the whole period ofemployment at Mayak PA, a definite tendency to the increase of stillbirths with the increase of dose is found.

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10. Radiation epidemiology

11. Radiation protection

The nuclear safety and radiation protection in PolandK. Fornalski

NPP Project Development Department, EJ 1 Sp. z o.o. (PGE NuclearEnergy), Warszawa, Polen

Poland has recently adopted PPEJ - the Polish Nuclear Power Pro-gram. Its implementation involves introduction of necessary amend-ments to the existing regulations regarding the nuclear, especially in thefield of radiation protection and nuclear safety. Considering that thenuclear research center in Świerk (near Warsaw) which operates smallresearch reactors has been running since 1958, the present situationrequires the Polish authorities improve the old, existing and train youngexperts. PGE EJ1 (PGE Nuclear), the company which will possibly bethe operator of the first Polish nuclear power plant in the future, is highlyinterested in developing nuclear safety and radiation protection stan-dards in Poland.

Dosimetry extremum tasksI.Kudrin1, I. Mazur1, K. Chizhov1, A. Simakov1, A. Tsovyanov1,

V. Kryuchkov1, I. Kemsky2, A. Krasnoschekov3, A. Kosnikov3, M. Sneve4

1 Burnasyan Federal Medical Biophysical Center of Federal Medical Bio-logical Agency, RF Ministry of Health and Social Development. 46,Zhivopisnaya St., Moscow, 123182, Russian Federation,+74991909405, [email protected] Regional management № 120 of the Federal Medical-BiologicalAgency, 184 682, Snezhnogorsk, Valentina Biryukova St., 5/1,+7(153)06-04-58, [email protected]

3 Northwest Center for Radioactive Waste Management \"SevRAO\" - abranch of the Federal State Unitary Enterprise \"Enterprise for Radioac-tive Waste Management\" RosRAO \"183017, Murmansk, Lobova st.,100, +7(152) 22-31-56,22-42-93,22-31-56, 22-42-93, [email protected] Norwegian Radiation Protection Authority, Postboks 55, 1332 Østerås,Norway,

(+47) 67 16 25 00, [email protected] report describes the application of graph theory in dosimetry. Sev-

eral of classical problems in graph theory in a reformulated form areapplicable to the field of radiation safety. We can represent a radiationsituation on some territory in a form of undirected graph. Vertices of thegraph form a regular grid, and edges connect neighboring vertices.Weight of each edge is equal to the dose, received by a person that haspassed along this edge with some, fixed for the entire graph speed. Thereport considers some problems of graph theory in their dosimetry interpretations.

The Shortest Path Problem answers the question how to go thru the radiation hazard area from point A to point B, and receive the lowest dose.

The traveling Salesman Problem determines order of passage thecontrol points to obtain the minimum dose. For example points ofplanned determination of radiation situation.

Euler path problem helps to plan transport network bypass, so as topass on every road once (where possible). This task is useful for deter-mining the radiation situation using automatic dosimeter with georefer-

ence. In this case, the vertices of the graph are the crossroads, and theedges are the roads.

Route Inspection problem helps to plan transport network bypass, ifthere is no Euler path. The problem of the shortest connection grid helps toconnect some fixed number of dosimeters to one network in optimal way.The problem of the critical path helps to choose the best option of a rangeof works, if the order of work is due by technological map of the enterprise.

Anschrift:

Experience of implementing of Information-Analytical Systemfor Radiation Protection staff on SevRAO

I. Mazur1, K. Chizhov1, A. Simakov1, A. Tsovyanov1, I.Kudrin1, V. Kryuchkov1,I. Kemsky2, A. Krasnoschekov3, A. Kosnikov3, M. Sneve4

1Burnasyan Federal Medical Biophysical Center of Federal Medical Bio-logical Agency, RF Ministry of Health and Social Development. 46,Zhivopisnaya St., Moscow, 123182, Russian Federation,+74991909405, [email protected] management № 120 of the Federal Medical-BiologicalAgency, 184 682, Snezhnogorsk, Valentina Biryukova St., 5/1,+7(153)06-04-58, [email protected] Center for Radioactive Waste Management \"SevRAO\" - abranch of the Federal State Unitary Enterprise \"Enterprise for Radioac-tive Waste Management\" RosRAO \" 183017, Murmansk, Lobova st.,100, +7(152) 22-31-56,22-42-93,22-31-56, 22-42-93, [email protected] Radiation Protection Authority, Postboks 55, 1332 Østerås,Norway,

(+47) 67 16 25 00, [email protected] to the Agreement between FMBC and NRPA (DRIVE and

DATAMAP projects) Information-Analytical System (IAS) for RadiationProtection (RP) of SevRAO staff in Andreeva Bay was developed. How-ever, the implementation of this system has encountered difficultiesrelated to the fact that RP services staff of SevRAO had to spend consid-erable extra time to input the information to the IAS database cause datainput in accordance with the regulations was different to IAS.

To motivate the staff for use the IAS the developers created an addi-tional interface. We called them the Mazur Interface. This Interfacegreatly simplifies the data input for radiation field in accordance with theregulations and save the information in the IAS RP database.

The presentation will detailed overview of features and functionality ofthe interface.

Measuring doses for deterministic effectsC. Jones

Radiation Protection Group, AWE, Reading, UKThe dose quantity “effective dose” is ubiquitously used in radiation pro-

tection. For routine monitoring it demonstrates compliance with dose limitsand when used for optimisation provides a conservative estimate of risk.

However, where interventions are required following an accident or inci-dent, it is important to have an accurate assessment of the deterministicdose. Effective dose includes weighting factors that reflect the detrimentfrom stochastic health effects. For deterministic effects, a different dosequantity with different weighting factors is required: the gray-equivalent.

The relative biological effectiveness (RBE) values required to calculatethe gray-equivalent dose have been published, for example Kutkov et al[i]. This makes calculating doses from external exposures relativelystraight forward, but calculating doses from internal exposures is still dif-ficult without the appropriate dose coefficients that provide Gy/Bq for dif-ferent nuclides, organs and radiation type. The work being presentedhas included calculating such dose coefficients for common uraniumand plutonium radionuclides. The poster then gives examples of theiruse, comparing the outcome in gray-equivalent to the equivalent dosefor the same intake.

Reference:[i] Kutkov, Buglova and McKenna, J. Radiol. Prot 31 (2011) 237-253

PENELOPE-2008 Monte Carlo Simulation of Gamma Exposurerelated to radioactive Contamination of Steel and radioactiveSources shielded by Metal Scrap

R. Merk, L. HornungBfS, Federal Office for Radiation Protection, Germany

The inadvertent radioactive contamination of steel scrap and steelproducts is a topic of growing concern. In 2009, a large amount ofCobalt-60 contaminated steel had to be seized in Europe. The origin ofthe steel was supposedly a steel mill in Far East. Also, recalls of con-sumer products represent an ongoing problem, for example the recentrecall of Kleenex boxes in the USA or bicycles in Japan. In Germany,these incidents triggered a discussion of the exemption value for Cobalt-60. The EU suggests in its new Basic Safety Standards to unify exemp-tion and clearance values. For Cobalt-60, this would amount to the intro-duction of a new activity concentration (AC) value of 0.1 Bq/g, originallyproposed by the IAEA in 2005. We plan to present PENELOPE-2008Monte Carlo simulations that support introducing the new AC value.PENELOPE-2008 is a state-of-the-art Monte Carlo computer code for thecoupled radiation transport of photons, electrons and positrons in matterand was developed at the University of Barcelona (Salvat et al., 2009).We also plan to present PENELOPE-2008 calculations related toradioactive sources shielded by metal scrap.Reference:

Salvat, F., Fernandez-Varea, J. M., Sempau, J. 2009. PENELOPE-2008: A code system for Monte Carlo simulation of electron and photontransport. NEA No. 6416, OECD Nuclear Energy Agency, Issy-les-Moulineaux, France.

Effects of ferulic acid on hematopoietic cell recovery inwhole-body gamma irradiated mice

Y. GaoDepartment of Pharmacology & Toxicology, Beijing, ChinaPurpose

The objective of this study was to investigate the mechanism for fer-ulic acid (FA)-induced radioprotection by evaluating the recovery ofbone marrow cells and peripheral blood hematology.Materials and methods

Balb/c mice were irradiated at a dose of 2.5 Gy using cobalt-60gamma resources. Following irradiation, FA was administered intragas-trically for seven consecutive days. Hematopoietic progenitor colony-forming cell assays were used to assess the reconstitution of bone mar-row after radiation-induced myelosuppression. Cytokine levels wereinvestigated using enzyme-linked immunosorbent assay and Westernblot analysis. Results

The results demonstrated that FA treatment enhanced hematopoieticprogenitor cell activity resulting in

accelerated blood cell recovery. FA administration increased levels ofgranulocyte-colony stimulating factor (G-CSF) and erythropoietin.Conclusion

These results suggest radioprotective efficacy by FA may be a result ofearly recovery of hematopoietic cells due to enhanced production of G-CSF and erythropoietin.

Identification of radioprotective compounds using a 3D-microtissue technology

N. AnastasovInstitute of Radiation Biology, Helmholtz Zentrum München, Germany

Two key issues in radiation therapy are the development of tumorresistance and the induction of toxic effects on normal tissue. In thissense new strategies are required to increase efficacy of radiation to

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improve the therapeutic impact and reduce toxicological side effects.The latter is of course also important for radiological protection ofexposed populations and individuals.

3D cell culture systems have been shown to provide a closer repre-sentation of tissue-level biology over classical 2D culture systems. Thishas led to the adoption of 3D systems for both drug discovery and toxi-cology. A highly reproducible hanging drop technology is able to gener-ate monotypic and heterotypic cell spheroids (microtissues) in a 96-wellformat. We have adopted this innovative 3D-microtissue plate technol-ogy for the screening of potential radioprotective small molecules. Wehave followed the response to radiation and drug-treatment of cells sta-bly transduced with a GFP-expressing lentiviral vector. High throughputquantification of 3D microtissue growth was assessed in real time usingthe Operetta microscopy platform and software ‘Harmony 3.0’(PerkinElmer, USA). In the next stage of the development we willdevelop multi-typic minitissues using up to four different fluorescent-dyeexpressing lentivirus transfected cell types.

Estimation of radiological consequences of RDD event on thebase of simple radiometric measurements

O. PavlovskiyRadiation Safety, Nuclear Safety Institute of the Russian Academy of Sci-ences, Moscow, Russian Federation

The appraisals of total individual effective dose as a result of external(from submersion in contaminated air and from contaminated surfaces)and internal (by inhalation of radionuclides from primary contaminatedair and from resuspended contamination) doses of radiation exposure ofpersons from population in the event of a radiological dispersal device(RDD) incident on the territory of megalopolis are presented. In theseestimations the total individual effective dose was the sum of the effectivedose equivalent from external radiation exposure and the committedeffective dose equivalent from internal radiation exposure. Calculationswere produced in the assumption that as a \"radioactive stuffing\" of RDDthe most widespread in the industry and medicine radioactive sources(Co-60, Sr-90, Cs-137, Ir-192, Pu-239 or Am-241) can be used. For thesubsequent purposes all calculated values of radiation doses for peoplewere normalized on a gross alpha-, beta- and gamma-contamination ofthe ground surface in kBq/m2 or gamma-exposure dose rate at height of1 m from a ground surface in mkSv/h. It has allowed to get a set of thesimple relationships between the recommended by ICRP values of refer-ence levels for emergency exposure situation (sheltering-in-place or evac-uation of the public in the first days after the RDD event) with real resultsof simultaneously made measurements of alpha-, beta- and gamma-fluxdensity or exposure dose rate of gamma-radiation, received by means ofmodern radiometric devices. These relationships have been calculated asfor a case when one of listed above 6 radionuclides has been involved inthe RDD, and in cases if more than one radionuclide have been used forthese purposes, and also when used radionuclide is at all unknown. It isshown that sensitivity of similar devices even in the mixed radiation fieldsallows to define reliably enough zones of radioactive pollution of the cityterritory in which on the early phase of protective action realization ofpopulation protection measures can be demanded.

Retrospective radiation dose audit from multi-detector-rowcomputed tomography (MDCT) scanner for adult bodycomputed tomography (CT) examinations using size-specificdose estimates (SSDE) method.

S. InkoomRadiation Protection Institute, Ghana Atomic Energy Commission, Accra,Ghana

A retrospective patient dose audit from a multi-detector computedtomography (MDCT) scanner was done. The conversion factors (CF) ofthe American Association of Physicists in Medicine Task Group 204 onsize-specific dose estimates (SSDE) in pediatric and adult body com-

puted tomography (CT) examinations were used in this study. These CFcan be applied to the displayed volume computerized tomography(CTDIvol) dose index to estimate patient dose. The factors take intoaccount patient size. The dose data was extracted from the dose reportof examinations of thorax, abdomen and pelvis for 50 standard adultpatients each from control console. The dosimetric parameters analysedwere CTDIvol and dose length product (DLP). The effective dose (E) wascalculated using the DLP and normalized values of effective dose perDLP based on International Commission on Radiological Protection(ICRP) publication 103 for the body regions imaged. Mean values ofCTDIvol were: thorax (18 mGy SSDE), abdomen (22 mGy SSDE) andpelvis (21 mGy SSDE). Similarly, the mean DLP values were: thorax (478mGy.cm), abdomen (620 mGy.cm) and pelvis (552 mGy.cm). The meanE values were: thorax (6.9 mSv), abdomen (9.5mSv) and pelvis (7.1mSv). The CTDIvol values from this study exceeded that of the EuropeanGuidelines for Multislice Computed Tomography and other recommen-dations. The DLP and E recorded variations with the reference levels.The inclusion of patient size in the estimate of CTDIvol by the SSDEmethod accounts for the high doses reported. This is due to thedependence of patient dose from a CT scan on both patient size andscanner radiation output. Regular patient dose audits during MDCTscanning and comparison with reference levels is recommended due tothe high dose burden of MDCT. This also offers a practical approachtowards optimization of patient protection.

Dose Reconstruction for F-104 Starfighter MaintenancePersonnel

T. KuipersForce Health Protection Expertise Centre, Ministry of Defence, Doorn,Netherlands

The F-104 Starfighter was in active duty and maintenance at theDutch Royal Airforce in the period 1962-1983. Maintenance personnelthat worked on the F-104 Starfighter J79 jet engine was occupationallyexposed to ionizing radiation, as the compressor casing alloy containsbetween 2 to 4 percent thorium and inherent daughter isotopes. Toquantify the external and internal effective dose received by the maintenance personnel, the following dose reconstruction methods have been used.

The quantification of the external effective dose is based on the combi-nation of two main parameters: 1) the yearly duration of exposure percategory maintenance personnel working in the vicinity of the compressorcasing and 2) a 2-dimensional effective dose profile of the compressorcasing. The information of the yearly duration of exposure was extractedfrom interviews with maintenance personnel and inspectors. The 2-dimensional dose profile was constructed by interpolating data fromrecently performed active and passive effective dose measurements.

The quantification of the internal effective dose is based on the com-bination of the yearly duration of exposure and four parameters for inter-nal exposure: 1) compressor casing wipe tests, 2) radon and thoronemanation estimates, 3) welding fumes, 4) machining dust particle dis-tribution estimates.

The reconstructed internal effective dose is significantly lower than theexternal effective dose per year mainly due to removal by suction ofwelding fumes. The yearly duration of exposure is the most influentialparameter in reconstructing the yearly effective dose but also the mostsensitive to recall bias. The reconstructed worst case total effective dosefor maintenance personnel is 17,5 mSv per year.

Mathematical modeling dynamic processes in chemicalbiological radiological nuclear research

A. AmiryanEngineering Academy of Armenia, Yerevan, Armenien

Mathematical modeling of dynamic processes has its own importancefor CBRN researches [1-8]. Mathematical modeling of dynamic

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processes very often leads to the system of differential equations. In thatcase it is very important to choose the right method of numerical inte-gration. Novel method of solving differential equations is presented.Comparative representation of implicit and explicit methods of solvingdifferential equation systems is discussed. Numerical calculations withthe usage of well known method of Euler, Runge-Kutta, Adams, Row4A,Row4S, ode15s, ode23s and new developed method are comparativelyrepresented for stiff and nonstiff systems. The calculation results of thenovel method are compared with the data of known methods. Programpackage for mathematical simulation complex process in CBRNresearches and real life examples are discussed [1-8]. Literature 1Artak Barseghyan, Anahit Amiryan Mathematical Modelling of ComplexReaction Systems for Computer-Aided Control and its Illustration onEnvironmental Problems // Economic Sustainability and EnvironmentalProtection in Mediterranean Countries through Clean ManufacturingMethods. NATO ATC – 984226. 3-7 October, Huelva, Spain, 2011, P 36-37.

2A.R. Barseghyan “Differential Equation Integration Numerical Methodsof Mathematical Simulation and their Comparative Representation onOzone Kinetics Problems” Proceedings of Engineering Academy ofArmenia.–Yerevan, 2007.–V4, №2.-P.276-280.

3A.R. Barseghyan “Problems of Solving Stiff Differential Equations forMathematical Modelling of Complex Processes” Proceedings of Engi-neering academy of Armenia.-2007.-V.2, №4.-P.647-651.

4A.R. Barseghyan Mathematical Modelling of Radiation Changes in theOzone Layer// Proceedings of the NATO Advanced Training Course onRadip Diagnosis in Population at Emergency and Risk, Krakow-Zakopane, Poland 19-24 October 2009, IOS Press 2010, P. 313-318.

5Barseghyan A.R. Mathematical Modelling of Complex Reaction Mecha-nisms for Computer-Aided Control// 13th ICQC -2009.- Helsinki, Finland, June 22–27.

6Artak R. Barseghyan “Mathematical Modelling of Complex ReactionMechanisms for Computer-Aided Control”13th ICQC International Con-gress of Quantum Chemistry ABSTRACTS AND PROGRAM June22–27, 2009, Helsinki, Finland 46 p.

7A. Barseghyan “Program Package for Kinetic Analysis of ReactionMechanisms and its Illustration on Ozone Kinetics” The 6th EuropeanConference on Ecological Modelling/ ECEM’07 conference proceed-ings.-Trieste, Italy.-2007, November 27-30, P.57-58.

8Barseghyan Artak, Martoyan Gagik Transportation and Storage of SpentNuclear Fuel: Security and Theory// Transport of Dangerous GoodsMethods and Tools for Reducing the Risks of Accidents and TerroristAttack. Proceedings of the NATO Advanced Research Workshop onRisk Prevention for Environment and Human Society against Danger-ous Goods Transport Accidents and Malicious Intents: Methods andTools Paris, France 5-9 July 2010.- Springer Science+Business MediaB.V. 2012.-P. 227-249.

Fast response mobile technology for water treatment andpurification from CBRN contaminationDr. Artak Barseghyana) b) c) , Dr. Gagik Martoyanb) c)

Yerevan, Armenia a)Engineering Academy of Armenia, b)“AREV” Scientific-Industrial CJSC,c)ECOATOM LLC [email protected]

March 11, 2011 earthquake and tsunami caused the worst nuclearaccident ever in Japan and the second-worst globally, after the 1986Chernobyl disaster. During the early phase of the emergency WHOurged people in the area to heed the advice of local authorities, as theyhave access to the latest measurements of radiation levels in water tocompare against the standards for adults and children. It is known thatrestrictions on tap water were applied in several villages [WHO 2011].

During similar and many other CBRN (chemical, biological, radiologicaland nuclear) disasters water resources often get contaminated andwater used from these sources must invariably be treated before use,otherwise this may lead catastrophic results. The localized drinking water treatment facilities may get wiped out ordisrupted, so there is great demand for Mobile technology for watertreatment and disinfection before distributing for human consumption oruse [1-8]. New unique technology -- specially designed separator-concentrators --employs coordinated use of an electrodialysis (ED) stack and a numberof electrolysis modules, each to output selected radionuclides (one or afew at a time) in the form of fine powder. The process is fully environmentally safe, has no emissions and pro-duces only selected elements. The new method’s extraction-separationefficiency averaged over radionuclides is 99.985%. The Processing Module presents a compact system, which meets majorsafety and efficiency requirements (The detailed information is pre-sented during the first meeting AREV SI CJSC / AREVA on 05.11.2010).After the processing the the level of radioactivity (U, Ra, Th, Po, Pb) willnot exceed 10 Bq/l. All desired features of the Mobile System can be assembled on a stan-dard 40-ft by 7-ft trailer driven by a heavy-duty truck, producing onlycleaned water and the entirely extracted radionuclides. In initial stage the system was designed to process liquid radioactivewaste at high, intermediate and low activity levels, at a rate of (10tone/hr for 5 g/l of salt concentration of radionuclides, 5 tone/hr for 10g/l of salt concentration of radionuclides, 1 tone/hr for 50 g/l of salt con-centration of radionuclides), but it can be used also for providing drink-ing water in crash areas, or providing safe water from sea, rivers, bore-holes and other sources [1-8]. The suggested mobile technology could be important tool for water pro-cessing from ionizing radiation and fast response during medical coun-termeasures providing safe water in mass casualty areas. Literature 1Artak Barseghyan Advanced Water Treatment System: Technologicaland Economic Evaluations // NATO Science for Peace and SecuritySeries C: Environmental Security Water Security in the MediterraneanRegion. An International Evaluation of Management, Control, and Gov-ernance Approaches.- Springer Science+Business Media B.V. 2011.-P.353- 362.

2Artak Barseghyan, Gagik Martoyan Transportation and Storage of SpentNuclear Fuel: Security and Theory// Transport of Dangerous Goods.Methods and Tools for Reducing the Risks of Accidents and TerroristAttack. Proceedings of the NATO ARW on Risk Prevention for Environ-ment and Human Society against Dangerous Goods Transport Acci-dents and Malicious Intents: Methods and Tools Paris, France 5-9 July2010.- Springer Science+Business Media B.V. 2012.-P. 227-249.

3Artak Barseghyan Stand-Alone Potable Water Making System Poweredby Solar Energy // 27th European Photovoltaic Solar Energy Conferenceand Exhibition (EU PVSEC ) 24 - 28 September 2012 Messe Frankfurt , Germany.-120 p.

Radiation protection relevant to Radiation emergency medicalpreparednessC.K.K. NairPushpagiri Institute of Medical Sciences and Research Centre, Tiruvalla,Kerala, IndiaOne of the major challenges faced with the widespread use of nucleartechnology is the fear of radiation exposure, weather planned orunplanned and protection of human beings from radiation hazards. Ion-izing radiation induces a wide range of molecular lesions in mammaliancells that can lead to diverse cellular responses such as cell inactivation,chromosomal rearrangements and mutations, eventually resulting incancer and hereditary diseases. Ionizing radiation generates reactive

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oxygen species in a biological system by radiolysis of water. The radio-protective compounds are of importance in medical, industrial, environ-mental, military and space science applications. The recent nuclearaccident in Fukushima, in Japan and the fear of nuclear terrorism havecreated a global awareness for emergency preparedness and need forfinding radioprotectors for human application and recent literaturereveals enormous interest in this field. An unfulfilled dream has been tohave a globally effective pharmacological agent that could be easilytaken orally, without any undue side effects, prior to a suspected orimpending nuclear/radiological event. Such an ideal radioprotectiveagent will greatly facilitate the emergency preparedmness. The ability ofantioxidants to reduce the cellular damage induced by ionizing radiationhas been studied in animal models for more than 50 years. On exposureto ionizing radiation a variety of symptoms are manifested, depending onthe dose of exposure. There may be immediate effects or delayed ones.Depending on the dose of radiation exposure this damage manifests ashaematopoietic syndrome, GI syndrome and CNS syndrome. The usesof plants in traditional medicine are widespread and still serve as leadsfor the development of novel pharmacological agents. As several of thesymptoms of radiation syndromes have some features in common withdiseases cured by medicinal plants, it was considered worthwhile toexamine the ability of plant products and their compounds to protectbiological systems from ionizing radiation. Many natural and syntheticchemicals have been investigated for their efficacy to protect radiationinduced damages in biological systems and found to protect cells, mem-branes and biomolecules such as DNA and proteins in vitro. Howevermany of these though show promising results in laboratory studies areof limited human application due to several factors such as toxicity, diffi-culties in administering required amounts, lack of preference for normal

tissues, limited availability etc. To avoid the problems of of toxicity andside effects of synthetic radioprotectors attention has been focused onphytochemicals, herbal medicines, dietary ingradents etc. Extracts ofseveral medicinal plants such as Acorus calamus, Glyzyrrhizia glabra, Ter-minalia chebla, Hemidesmus indicus, Rubia cordifolia, Centilla asiatica,Aconitum heterophyllum, Holarrhena atidysentrica etc and a variety ofcompounds- both natural and synthetic – with different molecular struc-tures, therapeutic activities such as curcumin, glyzyrrhizic acid, ferrulicacid, flavanoids, polyphenols, gallic acid, elagic acid; sesamol, andnutraceuticals such as vitamin A, E, C and their derivatives etc. havebeen found to possess good antioxidant, free radical scavenging andradioprotection property at molecular and cellular levels under in vitro andin vivo condtions. It was also found that oral administration of theAyurvedic rasayana preparations - Brahma Rasayana and Chyavanaprash– which are in use as tonics or supplements, for improving health, andare readily available over the counter in several of the Ayurvedic medicinaloutlets, prevented the deleterious effects of ionizing radiation exposure ina mammalian organism, mouse. Administration of these agents eitherprior to or following whole body gamma-radiation exposure of mice, resultsin enhancement of repair of radiation induced strand breaks in cellularDNA and reduction of radiation induced genomic instability monitored interms of micronuclei. The antioxidant status of various tissues of micewas also restored when these formulations were orally administered afterwhole body exposure to gamma-radiation. The talk will give an overview ofthe preclinical investigations on the prophylatic and therapeutic use ofsome of the phytoceuticals, nutraceuticals and herbals for protectingmammalian systems from the deleterious effects of lethal and sublethalionizing radiation exposures , the undrlying mechanisms and their possi-ble use in nuclear emergency preparedeness.

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12. Radiation biology/radiation physics

Telomeres and their influence on radiation induced genomicchanges

M. Eryilmaz, K. Greulich-BodeGenetics of Skin Carcinogenesis, German Cancer Research Center(DKFZ), Heidelberg, Germany

Ionizing radiation induces chromosomal aberrations. It has been pro-posed that in this process telomeres, play a central role. One known fac-tor hereby is the telomere length. We have additionally postulated atelomere-length independent mechanism of genomic instability, e.g.altered telomere organization/distribution. When the normal telomeredistribution in 3D interphase nuclei is disturbed, telomeric associationsand telomeric aggregates (TAs) are seen. Strikingly, TA induction isclosely associated with the establishment of new chromosomalrearrangements in two different system (c-Myc and UV).

To study the 3D telomeric organization in intherphase nuclei, 3D fluo-rescence in situ hybridization can be performed by probing for telomericDNA. To explore the chromosomal composition, for example after irradi-ation, metaphases are prepared and multicolor fluorescence in situhybridization (mFISH) performed.

Results of such investigations have previously been reported andshowed that upregulated c-Myc expression is able to induce TAs. Irradi-ating HaCaT-Myc cells with UV-C caused telomeric associations and TAsshowing highest TA numbers 48 to 72 hours after radiation. This was fol-lowed by the establishment of chromosomal aberrations as determined8 to 10 days after radiation.

To test if TA formation is also inducible by ionizing radiation, wetreated human fibroblasts with 2Gy. 72h post-irradiation we detectedonly a relatively low number of TAs, which increased up to 13d post-irra-diation. This effect is accompanied by chromosomal aberrations whenperforming mFISH after 13 days. Noteworthy, the timepoints for maximal

TA formation seem to differe between ionizing radiation and UV-irradia-tion, the final outcome, however, seems to be comparable.

We postulate from these results that TA-formation is a new indicator forradiation induced genomic instability. Thus, it is tempting to speculate thatTA formation can serve as a marker for exposure to ionizing radiation.

Serine protease inhibitors may exacerbate normal tissueresponses to radiation

Isabel L. Jackson1, Chirayu Goswami2, Laura Hale3, Barry P. Katz4,Puting Xu1, Zeljko Vujaskovic1

1Division of Translational Radiation Sciences, Department of RadiationOncology, University of Maryland School of Medicine, Baltimore, MDUSA; 2Department of Biostatistics, Indiana University School of Medi-cine, Indianapolis, IN USA; 3Department of Pathology, Duke UniversityMedical Center

Murine strains differ significantly with respect to pulmonary radiationresponse following whole thorax lung irradiation (WTLI). These differ-ences can be exploited to identify inherent differences among strainsthat may predispose them to increased risk for development of pneu-monitis and/or fibrosis following radiation exposure.

In these studies, C57L/J, CBA/J, C57BL/6J mice were irradiated with12.5 or 15 Gy WTLI (320 kVp, 1.0 mm Cu HVL, 69 cGy min-1 doserate). Microarray analysis was performed on lung tissue (n = 3/group) 24h after radiation. Sham-WTLI controls were included. Gene expressionwas validated by real-time RT-PCR and protein expression by westernblot (n = 4-5/group). Samples were not pooled for any of the analyses.Comparisons were made between radiation “sensitive” vs. “resistant”strains. Strains were designated as “sensitive” vs. “resistant” based onpre-determined dose response relationships between C57BL/6J (“resis-tant”) vs. C57L/J and CBA/J (“sensitive”).

Genes encoding for serine protease inhibitor, alpha-1 antitrypsin(A1AT), and a serine protease carrier, alpha-1 acid glycoprotein (AAG),were shown to be the top two differentially expressed genes among sen-sitive versus resistant strains (p < 1.4 x10-10 and p < 1.12 x 10-4,respectively). While differences in A1AT were not statistically significantat p<0.05 using real time RT-PCR, AAG showed a statistically significantincrease after radiation in CBA/J (*p<0.05) and C57L/J (**p<0.001). Ina separate study, rodents treated with serine protease inhibitor, camostatmesilate, showed increased normal tissue toxicity and decreased sur-vival following radiation to the abdomen/lung.

There is accumulating evidence to suggest A1AT and AAG may play arole in the development of radiation injury. A1AT and AAG have beenfound to correlate with fatigue and diarrhea in patients undergoing pelvicirradiation and to be associated with peak illness in rats and non-humanprimates following total body irradiation. Further investigation is war-ranted to determine whether serine protease inhibitors may not only playa role in radiation toxicity, but also may serve as potential biomarkers toidentify individual risk for development of radiation-related toxicity.Acknowledgements

This project has been funded in whole or in part with Federal fundsfrom the National Institute of Allergy and Infectious Diseases, NationalInstitutes of Health, Department of Health and Human Services, underContract No. HHSN266200500043C

The influence of fractionated X-irradiation on the nucleardamages of human hematopoietic stem/progenitor cells

Y. MariyaDepartment of Radiological Technology, Hirosaki University School ofHealth Sciences, Hirosaki, Japan

We have reported the influences of X-irradiation on the proliferationand differentiation of human CD34+ hematopoietic stem/progenitorcells. In this study, we estimated the influences of X-irradiation with avariety of fractionations on the cells. Highly purified CD34+ cells wereprepared from human placental/umbilical cord blood by usingautoMACS™ Pro Separator. X-irradiation was performed with the follow-ing fractionations: 0.5 Gy x 4 fr, 1 Gy x 2 fr, and 2 Gy x 1 fr. The inter-fractional interval was 2 h. Colony assay was performed by methylcellu-lose culture supplemented with appropriate cytokines. Alsocytokinesis-block micronucleus (MN) assay, gamma-H2AX expressionassay and comet assay were performed. Colony assay showed the signif-icantly decreased number of colonies in the irradiated cells, about 30-50% of the non-irradiated cells. However, there was no difference of thenumber between the 3 fractionation patterns. MN assay showed the sig-nificant increase of binucleated cells (BNCs) with MN in the cells irradi-ated with 2 Gy x 1 fr, although there was a tendency that the proportionof BNCs with MN negatively correlated with the number of fractionation.Gamma-H2AX expression was estimated 2 and 24 h after X-irradiation.After 2 h the number of gamma-H2AX foci was significantly increased inthe cells irradiated with 2 Gy x 1 fr, while it was slightly increased in theother fractionation patterns. After 24 h the number of foci was time-dependently decreased in the cells irradiated with 0.5 Gy x 4fr and 2 Gyx 1 fr compared with those after 2 h, the latter being significant. Neutraland alkaline comet assays demonstrated that both tail DNA and tailmoment significantly increased in the cells irradiated with 2 Gy x 1 fr,although not with the other fractionation patterns. These results suggestthat, although CD34+ cells were damaged by X-irradiation dependent onfractionation, cell death and loss of the clonogenicity are different storyin hematopoietic stem cells.

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