The general public is at risk of ionising-radiation exposure. The development of high-throughput methods to triage exposures is warranted. Current biodosimetry techniques are low-throughput and encumbered by time and technical expertise. Although there has been an emergence of gene-profiling tools for the purpose of photon biodosimetry, similar capacities do not exist for alpha-particle radiation. Herein is the first genomic study useful for alpha-particle radiation biodosimetric triage. This work has identified robust alpha-particle induced gene-based biomarkers in isolated, ex-vivo irradiated leukocytes from multiple donors. It was found that alpha-particle and photon radiation elicited similar transcriptional responses, which could potentially be distinguished by aggregate-signature analysis. Although no distinct genes were sole indicators of exposure type, clustering algorithms and principal component analysis were able to demarcate radiation type with some success. By comparing the biological effects elicited by photon and alpha-particle radiation, significant contributions have been made to the field of radiation biodosimetry.

A specific feature of ionising radiation is the formation of clustered DNA damage, where two or more lesions form within one to two helical turns of the DNA induced by a single radiation track. The complexity of ionising radiation-induced DNA damage increases with increasing ionisation density and it has been shown that complex DNA damage has reduced efficiency of repairability. In mammalian cells, base excision repair (BER) is the predominant pathway for the repair of non-DSB clustered DNA lesions and is split into two sub-pathways known as short patch (SP) BER and long patch (LP) BER. SP-BER is the predominant pathway, especially in the repair of isolated DNA lesions. However, LP-BER is thought to play a greater role in the repair of radiation-induced clustered lesions. In this study, cell lines were generated stably expressing the fluorescently tagged BER proteins, XRCC1-YFP (marker for SP-BER) or FEN1-GFP (marker for LP-BER). The recruitment and loss of XRCC1-YFP and FEN1-GFP to sites of DNA damage induced by both ultrasoft X-ray (USX), a form of low linear energy transfer (LET) radiation, and near infrared (NIR) laser microbeam irradiation (‘mimic’ high LET radiation) was visualised in real-time and the decay kinetics of the fluorescently-tagged proteins determined. The half-life of fluorescence decay of FEN1-GFP following USX irradiation was longer than that of XRCC1-YFP, indicating that LP-BER is a slower process than SP-BER. Additionally, the fluorescence decay of XRCC1-YFP after NIR laser microbeam irradiation was fitted by bi-exponential decays with a fast component and a slow component, reflecting the involvement of XRCC1 in the repair of different types of DNA damage. In contrast to USX irradiation, where the XRCC1-YFP fluorescence decay reached background levels by 20 min, XRCC1-YFP still persisted at some of the NIR laser induced DNA damage sites even after 4 hours. This is consistent with the fact that the laser induces more complex damage that presents a major challenge to the repair proteins, persisting for much longer than the simple damage caused by low LET USX irradiation. Persistent, unrepaired DNA damage can potentially lead to mutations and replication-induced DSBs if it persists into S-phase. PARP1 inhibition reduced the recruitment of XRCC1 to DNA damage sites. However, a considerable amount of XRCC1 was still detected at the DNA damage sites, leading to the conclusion that there is a subset of DNA damage that requires XRCC1 but not PARP1 for repair. Understanding how clustered damage is repaired by the BER pathway can aid the design of future therapies which can be used in combination with radiotherapy to enhance the radiosensitisation effect. Knockdown of FEN1 was investigated and found to radiosensitise A549 (adenocarcinoma) cells, possibly as a result of an excess of unrepaired radiation-induced lesions requiring LP-BER for repair, although FEN1 knockdown alone induced cell death in non-cancerous BEAS-2B cells.

The diploma thesis on "Radiation exposure of persons working at risk of ionising radiation at the Pilsen University Hospital" is divided into two parts. The first analyses personal effective doses of the employees of the Pilsen University Hospital in 20092013, and the second focuses on measurements of secondary radiation created after the activation of collimator systems of linear accelerators. The introduction of the theoretical part discusses the history of radiology, followed by a summary of information on the sources of ionising radiation, its characteristics, properties, methods of interaction with the environment and biological effects on organisms. The chapter on radiation protection focuses on basic legislative regulations relevant to the topic. In the Czech Republic, radiation protection of people working with sources of ionising radiation is mostly based on Act No. 18/1997 Coll., on peaceful uses of nuclear energy, and Regulation of the State Office for Nuclear Safety No. 307/2002 Coll., on radiation protection. The thesis also lists dose limits and basic information about personal dosimetry, and the theoretical section is concluded with information on the methods of protecting personnel and on provided healthcare. The thesis uses as its sources primary literature, internet links, current legislation and data from the database of the State Office for Nuclear Safety on records from personal dosimeters in the analysed period. The thesis had two objectives. The first was to compare levels of exposure at the Pilsen University Hospital. The analysis involved 579 persons in total; after arranging the data into tables and dividing employees into groups by the type of tasks they perform and their job, it was possible to compare median value indicators, confirming the hypothesis that persons carrying out intervention tasks are exposed to more ionising radiation. In order to compare the development of doses over time, a double-selection t-test was performed on data from 2009 and 2013, confirming the effectiveness of anti-radiation measures adopted at the University Hospital and a reduction in the exposure of employees in this period. The second objective was to confirm or deny the hypothesis that secondary radiation created by collimator systems, while measurable, is not sufficiently large to be detected by personal dosimeters. In order to achieve this objective, measurements were carried out at the Oncology and Radiology Clinic of the Pilsen University Hospital. The first measurements tried to determine the relationship between dose and time at various levels of energy, the second compared the relationship between dose and distance from the collimator plane and the third monitored the number of exposures until the collimator plates are saturated and dose is no longer increased. All results were arranged in tables and projected in charts. The results of the research confirmed the hypothesis that secondary radiation created during the use of high-energy equipment is just below the detection limits of personal dosimeters. For this reason, it is recommended to frequently rotate personnel working with the equipment.

Plus d'un siècle après la découverte des rayons X, les effets biologiques des radiations ionisantes restent encore méconnus. En particulier, une meilleure connaissance des phénomènes liés à la radiosensibilité individuelle permettrait une meilleure prédiction du risque radioinduit tant en ce qui concerne les réactions tissulaires que la formation de cancers.Dans le cadre des recherches menées par le Groupe de Radiobiologie de l'UMR 1052 Inserm (Centre de Recherche en Cancérologie de Lyon), l'accumulation de données radiobiologiques issues de patients radiosensibles a permis d'initier une théorie basée sur le transit cytonucléaire de la protéine ATM. Acteur majeur de la réponse aux radiations ionisantes ATM est muté dans l'Ataxie Telangiectasie, syndrome génétique rare associé à la plus forte radiosensibilité. Plus précisément, les chercheurs du Groupe ont proposé le modèle suivant : l'irradiation produit une monomérisation des formes cytoplasmiques de la protéine ATM. Les monomères d'ATM diffusent dans le noyau pour assurer la reconnaissance et la réparation des cassures double-brin de l'ADN (CDB), dommages-clés de la réponse aux radiations. Tout retard dans ce transit conduirait à une certaine radiosensibilité.Le but de cette thèse est d'identifier d'une part, les protéines (appelées X) qui freinerait ce transit en s'associant à ATM dans le cytoplasme ; d'autre part, les agents chimiques (métaux, pesticides) qui influeraient sur ce processus.Les protéines X identifiées dans le cadre de cette thèse sont notamment la huntingtine, la neurofibromine, la tubérine qui, lorsqu'elles sont mutées, causent respectivement la maladie de Huntington, la Neurofibromatose de type 1 et la Tubéreuse de Bourneville. Les métaux étudiés sont les chlorures d'aluminium, de cuivre, de zinc, de fer, de nickel, de palladium, de cadmium ainsi que le nitrate de plomb, le selenium et le chrome. Les pesticides sont l'atrazine, le glyphosate, la permetrine, le thiabendazole et le pentachlorophénol.Cette thèse introduit la notion de pro-, dys- ou anti-épiskévie, c'est-à-dire la capacité de certains agents, protéines ou drogues à accélérer, ralentir ou interdire le transit cytonucléaire de la protéine ATM / More than a century after the discovery of X rays, the effects of ionising radiation are still misunderstood. In particular, a better knowledge of individual radiosensitivity could lead to a better prediction of radio induced risk of cancer and acute reactions after radiotherapy. As part of the research conducted by the Radiobiology Group of UMR Inserm 1052 (Cancer Research Center of Lyon), the accumulation of radiobiological data from radiosensitive patients allowed to initiate a theory based on the ATM protein transit from cytoplasm to nucleus. ATM a the major actor in the response to ionising radiation and is mutated in Ataxia Telangiectasia, a rare genetic syndrome associated with the highest radiosensitivity. Specifically, the researchers of the Group proposed the following model: irradiation produces monomerization of cytoplasmic forms of ATM protein. ATM monomers diffuse into the nucleus to ensure the recognition and repair of DNA double-strand breaks (DSBs), the key damage response to radiation. Any delay in this transit would lead to radiosensitivity.The aim of this thesis is to identify in one hand, the proteins (called X proteins), which would slow the transit by interracting with ATM in the cytoplasm; on the other hand, chemical agents (metals, pesticides) that would affect this process.X proteins identified in this thesis include huntingtin, neurofibromin, tuberin which, when mutated, cause, respectively, Huntington's disease, Neurofibromatosis type 1 and Tuberous Sclerosis. Studied metals are aluminum, copper, zinc, iron, nickel, palladium and cadmium chlorides, lead nitrate, selenium and chromium. Pesticides are atrazine, glyphosate, permethrin, thiabendazole and pentachlorophenol.This thesis introduces the concept of pro-, dys or anti- episkévia, that is to say the ability of some agents, proteins or drugs to speed up, slow down or inhibit the the ATM nucleoshuttling

The public perception of ionising radiation focuses on its exploitation for energy production and medicine (diagnostics, therapy). In contrast, natural sources of ionising radiation are rarely considered, which contribute to more than half of the total radiation exposure. With regard to biological consequences, stochastic radiation effects (e. g. mortality of radiation-induced cancer, genetic effects), where the probability increases with dose, and deterministic radiation effects (pathological changes, e. g. changes of gonads, embryo and fetus) for which severity increases with dose, must be distinguished. Ionizing radiation is frequently linked to various risks (cancer, genetic damage, acute radiation syndrome). Usually, these risks are over-estimated. / Bei der öffentlichen Auseinandersetzung mit ionisierender Strahlung steht deren Nutzung im Rahmen der Energieerzeugung und der Medizin (Diagnostik, Strahlentherapie) im Vordergrund. Selten wird bedacht, dass ionisierende Strahlung auch natürlich vorkommt und damit mehr als die Hälfte der Gesamt-Strahlenbelastung bedingt. Die biologische Wirkung ionisierender Strahlung wird eingeteilt in stochastische Effekte, bei denen die Wahrscheinlichkeit eines Effektes mit der Dosis zunimmt (z. B. Tod durch strahleninduzierten Krebs, genetische Effekte), sowie deterministische Effekte (pathologische Veränderungen, z. B. an Fortpflanzungsorganen oder der Leibesfrucht), bei denen der Schweregrad dosisabhängig ist. Ionisierende Strahlung wird häufig mit verschiedenen Risiken (Krebs, Erbschäden, Strahlenkrankheit) in Verbindung gebracht, wobei diese in der Regel überschätzt werden.

The aim of the study was to investigate the effect of X-rays used in micro X-ray computer tomography (µXCT) on the mechanical performance and microstructure of a variety of dental materials. Standardised bending beams (2 × 2 × 25 mm3) were forwarded to irradiation with an industrial tomograph. Using three-dimensional datasets, the porosity of the materials was quantified and flexural strength was investigated prior to and after irradiation. The thermal properties of irradiated and unirradiated materials were analysed and compared by means of differential scanning calorimetry (DSC). Single µXCT measurements led to a significant decrease in flexural strength of polycarbonate with acrylnitril-butadien-styrol (PC-ABS). No significant influence in flexural strength was identified for resin-based composites (RBCs), poly(methyl methacrylate) (PMMA), and zinc phosphate cement (HAR) after a single irradiation by measurement. However, DSC results suggest that changes in the microstructure of PMMA are possible with increasing radiation doses (multiple measurements, longer measurements, higher output power from the X-ray tube). In summary, it must be assumed that X-ray radiation during µXCT measurement at high doses can lead to changes in the structure and properties of certain polymers.

Ionisierende Strahlung verursacht in Abhängigkeit von den strahlenphysikalischen Eigenschaften der Radionuklide Zellschäden unterschiedlicher Komplexität. An humanen Lymphozyten wurde untersucht, ob die biologische Wirksamkeit von Alpha- und Betastrahlung sowie der Einfluss von Doxorubicin der Qualität des Strahlenschadens zugewiesen werden kann. Die DNA-Schäden und deren Reparatur wurden mit zellbiologischen Methoden quantifiziert.

Problems of dealing with radioactive materials, especially burnt out nuclear fuel, are a delicate and discussed topic in the Czech Republic and other advanced countries nowadays. In this work, the realization of radiation control of warehouse of burnt-out atomic fuel of nuclear power plant Temelín is analysed. Step by step there are stored packaging files filled with burnt-out fuel which is produced by the running of atomic reactors. In the introductory chapters individual packaging files are described. The shielding of stored burnt-out nuclear fuel and prevention of release of radionuclides beyond the protective casing of packaging files depend on their quality. Further the lo-cality of nuclear plant Temelín, where the warehouse of burnt-out nuclear fuel is placed including manipulations which are carried out with the packaging file, is described. The emphasis is placed on monitoring technological parameters during the storage of pack-aging files. The target of this work is to analyse the radiation control of warehouse of burnt-out nuclear fuel, precautions of radiation protection valid for this object and con-sider the possibility of optimalization of used measuring devices, suggest the adjustment of the way of carried out monitoring. For meeting the goals of this works it was neces-sary to carry out monitoring the current radiation situation in the storage boat which is partly filled with packaging spaces when according to the following calculations I ap-proached to the prediction of the level of radiation situation during the maximum filled storage boat. These calculations are at the same time the background for the proposal of adjustment of regime precautions in this object with the aim to optimise the professional radiation. One of the basic strategic targets of the operator of nuclear plant Temelín, company ČEZ a.s., is to ensure a safe operation of the biggest Czech nuclear plant, one of the basic conditions for meeting this target is to ensure the radiation control and pro-tection in the required quality. The results, which are the output of this work, will be given to the department Operation of radiation protection for other practical usage, pos-sible adjustment of a current operation documentation which solves the radiation control of the storage of burnt-out nuclear fuel.

Ionisierende Strahlung verursacht in Abhängigkeit von den strahlenphysikalischen Eigenschaften der Radionuklide Zellschäden unterschiedlicher Komplexität. An humanen Lymphozyten wurde untersucht, ob die biologische Wirksamkeit von Alpha- und Betastrahlung sowie der Einfluss von Doxorubicin der Qualität des Strahlenschadens zugewiesen werden kann. Die DNA-Schäden und deren Reparatur wurden mit zellbiologischen Methoden quantifiziert.