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Analyse du vieillissement d'un adhésif silicone en environnement spatial : influence sur le comportement électrique / Analysis of a silicone adhesive aging in the space environment : influence on the electrical behaviorRoggero, Aurélien 24 November 2015 (has links)
Cette thèse s'inscrit dans la thématique technologique des décharges électrostatiques sur les panneaux solaires des satellites de télécommunication en orbite géostationnaire. Son objectif est de déterminer les évolutions des propriétés électriques d'un adhésif silicone commercial en environnement spatial et de les corréler aux évolutions de sa structure chimique. Les principaux constituants du matériau ont été identifiés et des échantillons dépourvus de particules de renfort - assimilés à la matrice polymère isolée - ont été élaborés. Afin d'évaluer l'influence des particules, ils ont été systématiquement comparés aux échantillons nominaux dans l'ensemble de ces travaux. La structure physico-chimique du matériau à l'état initial a été caractérisée en étudiant ses relaxations enthalpiques, mécaniques et en pratiquant des analyses chimiques. Son comportement électrique (relaxations dipolaires et conductivité) a été sondé grâce à une approche expérimentale inédite croisant la technique de relaxation de potentiel électrostatique de surface, la spectroscopie diélectrique dynamique et l'analyse des courants thermo-stimulés. Le vieillissement du matériau en environnement spatial a été simulé expérimentalement par l'exposition des échantillons à des flux élevés d'électrons de haute énergie. Les analyses chimiques, notamment en RMN du solide, ont montré la prédominance d'un processus de réticulation du matériau sous irradiation, et des mécanismes de dégradation à l'échelle microscopique ont été proposés. Le comportement électrique du matériau est fortement impacté par l'évolution de sa structure chimique : sa résistivité augmente considérablement avec la dose ionisante. Il est suggéré que la résistivité de ce matériau soit directement liée à son degré de réticulation, influant sur la mobilité des porteurs dans le cadre du hopping et de la percolation électrique. Cette augmentation est beaucoup plus marquée en présence de particules, ce qui a été attribué à la formation de nœuds de réticulation matrice-particules qui constituent des pièges plus profonds pour les porteurs de charges. Ces travaux apportent une meilleure compréhension des phénomènes de vieillissement des élastomères silicones en environnement spatial. Ils permettront d'anticiper des évolutions structurales qui pourraient mettre en péril leur fonction d'adhésion, ainsi que des évolutions de résistivité électrique intrinsèque, facteur décisif dans le déclenchement de décharges électrostatiques. / This PhD thesis falls within the technical field of electrostatic discharges occurring on the solar arrays of communications satellites in the geostationary orbit. Its main objective consists in assessing the evolutions of a space-used commercial silicone adhesive's electrical properties, and to correlate them with the evolutions of its chemical structure. The main components of this material have been identified, and neat samples (deprived of fillers) were elaborated so as to study the isolated polymer matrix. In order to assess the influence of filler incorporation, neat samples were systematically compared with the commercial ones in this study. The material's physicochemical structure in the initial state was characterized by studying its enthalpy relaxations, mechanical response, and by performing chemical analysis. Its electrical behavior (dipole relaxations and conductivity) was investigated thanks to an original experimental approach combining surface potential decay measurements, broadband dielectric spectroscopy, and thermally stimulated depolarization currents. Aging in the space environment was experimentally simulated by exposing the samples to high fluxes of high energy electrons. Chemical analysis (solid state NMR in particular) revealed the predominant crosslinking tendency of this material under ionizing radiations, and allowed to suggest degradation mechanisms at the microscopic scale. These structural evolutions also strongly impact its electrical behavior: a great increase in electrical resistivity has been observed with increasing ionizing dose. It is believed that electrical resistivity directly depends on the degree of crosslinking, which affects charge carrier mobility, in the theoretical frame of hopping and percolation models. The increase in resistivity is considerably more pronounced in the filled material, which could be associated with crosslinking occurring at the matrix-particles interface. Such crosslinks are thought to represent deeper traps for charge carriers. This work brought better understanding of aging phenomena in silicone elastomers exposed to the ionizing space environment. This knowledge will help predicting structural evolutions that may compromise vital properties such as adhesion, and the evolutions of intrinsic conductivity, a critical factor involved in the triggering of electrostatic discharges.
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Développement de nouveaux complexes organométalliques de métaux de transition polyvalents pour la scintillation et la chimie médicinale / Development of new versatile organometallic complexes for scintillation and medicinal chemistryElie, Margaux 06 October 2017 (has links)
Deux nouvelles familles de complexes de cuivre(I) cationiques, de formules [Cu(NHC)(N^N)][X] et [Cu(P^P)(N^N)][PF6], ont été synthétisées avec des ligands 2,2’ bis pyridyl pontés, chélates à six chaînons, facilement modulables. Ces complexes présentent des émissions à l’état solide centrées entre 455 et 520 nm (bleu à vert), avec de larges décalages de Stokes et des rendements quantiques pouvant atteindre 0,86. De plus, l’émission via un phénomène de fluorescence retardée activée thermiquement (TADF) a été prouvée pour les complexes [Cu(NHC)(N^N)][X]. Les premiers scintillateurs plastiques dopés avec des complexes de cuivre(I) détectant les radiations nucléaires de type gammas ont été obtenus avec des complexes de formule générale [Cu(P^P)(N^N)][PF6]. Les complexes de formule [Cu(NHC)(N^N)][X] ont permis l’obtention des premières Cellules Electrochimiques Luminescentes (LECs) émettant dans le bleu et incorporant des complexes de cuivre(I). Enfin, les complexes de formule [Cu(NHC)(N^N)][X] à ligand 2,2’ dipyridylamine présentent une activité cytotoxique envers différentes lignées de cellules cancéreuses et apportent la possibilité d’une action ciblée sur les cellules tumorales via l’ajout d’un vecteur. La polyvalence de ces complexes de cuivre(I) repose sur les ligands 2,2’-bis-pyridyl pontés, chélates à six chaînons, dont la synthèse est facile d’accès et les propriétés électroniques et structurales sont modulables. / New cationic copper(I) complexes of general formula [Cu(NHC)(N^N)][X] and [Cu(P^P)(N^N)][PF6] were developed with 6-membered-ring 2,2’-bis-pyridyl derivatives as ligand. These complexes exhibited blue (420 nm) to green (520 nm) emissions in solid state, with large Stokes shifts and photoluminescence quantum yields up to 0.86. Furthermore, the emission of the [Cu(NHC)(N^N)][X] complexes via a thermally activated delayed fluorescence (TADF) was demonstrated. The first plastic scintillators incorporating copper(I) complexes and detecting gamma radiations were obtained with [Cu(P^P)(N^N)][PF6] complexes. Application of the [Cu(NHC)(N^N)][X] complexes to the LEC technology led to the first copper(I)-based blue emitting device. In the last chapter, we also demonstrated that copper(I) complexes [Cu(NHC)(N^N)][X] bearing a 2,2’-dipyridylamine as N^N ligand exhibited high cytotoxycity against different cancer cells lines. These complexes paved the way for the design of a new type of copper(I) anti-cancer agents with the opportunity to increase the selectivity against cancer cells via a vectorization of the N^N ligand. The versatility of these copper(I) complexes demonstrated in this work relied on the easy to handle and highly modular 2,2’-bis-pyridyl ligands.
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Étude et modélisation numérique de l’effet des radiations spatiales sur l’évolution des propriétés physiques et électriques des matériaux embarqués / Study and numerical modelling of the space radiations effects on the evolution of the physical and electrical properties for embedded materialsPacaud, Rémi 13 December 2018 (has links)
Ma thèse consiste à établir un modèle numérique 1D qui permettra d'approfondir nos connaissances dans la compréhension des mécanismes physiques régissant le transport de charges dans les matériaux diélectriques comme le Kapton ou le Téflon soumis à des irradiations hauts-flux/hautes-énergies. Ce modèle est implémenté sous l'environnement Eclipse. Ensuite, les résultats numériques seront comparés aux résultats expérimentaux pour contrôler le bon fonctionnement du code une dimension (1D). A plus ou moins long terme, cette thèse permettra de déboucher sur une bonne compréhension du transport de charges dans les polymères embarqués en environnement spatial, ce qui permettra de comprendre l'origine des décharges électriques qui se produisent sur les panneaux solaires des satellites utilisés en orbite géostationnaire. / I have to establish a 1D numerical model that enables to better understand the physical mechanisms that steer charge transport in dielectric materials such as Kapton or Teflon under high fluxes and high energy electron beams. This model is implemented in Java under the Eclipse environment. Then, numerical results will be compared to experimental results in order to verify whether the 1D model is functional or not. In the near future, this phd will allow to better understand charge transport in satellite embedded polymers. We will then be able to understand the origin of electric discharges that occur on satellite solar panels used in geostationary orbit.
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Etude et développement d’une chaine de traitement analogique du signal dédiée à la détection de particules en environnement sévère / Development of an analog processing chain dedicated to particles detection in harsh environmentBen Krit, Sabrine 17 December 2015 (has links)
Le travail présenté dans ce mémoire est basé sur l’étude et le développement d’une chaine analogique de traitement du signal destinée à la détection de particules issues d’un réacteur nucléaire. Le détecteur de radiations relatif à cette chaine, étant placé au cœur du réacteur (hautes températures et radiations), deux scénarii d’intégration de la chaine analogique en environnement sévère ont été étudiés. Tandis que le premier scénario consiste à placer le préamplificateur et l’amplificateur de mise en forme loin du détecteur, le deuxième est basé sur l’intégration de ces blocs analogiques en environnement contraint. Bien que le premier scénario ait démontré de bonnes performances en termes de linéarité et sensibilité, l’amélioration de la résolution digitale fournie par le système nous a mené à investiguer le placement de la chaine tout près du détecteur. Plusieurs études, basées sur l’évaluation de la fiabilité de la structure vis-à-vis des contraintes de l’environnement sévère ont été donc élaborées. De plus une solution innovante permettant une meilleure caractérisation de la particule incidente a été présentée et détaillée. / The work presented in this thesis is based on the study and development of an analog processing chain dedicated to the detection of particles delivred from a nuclear reactor. The radiation detector related to this chain, being placed at the heart of the reactor (high temperatures and radiation flux), two scenraii related to the integration of the analog chain in harsh environment were studied. While the first scenario consists on placing the preamplifier and the shaping amplifier away from the detector, the second is based on the integration of these analog blocks in harsh environment. Although the first scenario has proven good performances in terms of linearity and sensitivity, the improvement of the digital resolution provided by the system leaded us to investigate the placement of the chain close to the detector. Several studies based on the evaluation of the structure reliability under harsh environment constraints were thus elaborated. In addition, an innovative solution providing better characterization of the incident particle was presented and detailed.
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Física das radiações: uma proposta para o Ensino Médio / Radiations physics: a proposal for the High SchoolSousa, Wellington Batista de 14 August 2009 (has links)
A sociedade vem questionando os conteúdos presentes no currículo de ciências, quanto ao \"por que\" e \"para que\" ensiná-los. Isso ocorre também dentro do ambiente escolar. Atualmente, o ensino de ciências tem acontecido de maneira bastante desconexa da realidade de nossos alunos, afinal, este ensino não tem apresentado inovações ou trazido temas atualizados para as discussões que ocorrem no ambiente da sala de aula. Apresentamos neste trabalho uma proposta de atualização curricular do ensino de ciências através da inserção de tópicos de Física Moderna e Contemporânea, com base na proposta curricular inserida nos PCNEM e PCNEM+ para a disciplina de Física. Nossa proposta consiste em uma seqüência didática de um curso de Física das Radiações voltado ao Ensino Médio, onde se utilizam as noções de \"marcadores\" e \"marcadores-estruturantes\", desenvolvidas no contexto da teoria da Transposição Didática, para delinear as atividades que acreditamos serem essenciais para que esta nova seqüência de ensino seja exeqüível. / The society has questioned what should be taught in science learning, including both \"why\" and \"what for\" some subjects should be taught. This kind of discussion occurs also inside the educational environment. Presently the science teaching is not connected to the reality of the student, because teaching has not related to scientific innovations, no updated themes are discussed in the classroom. In this work the curricular updating is inserted through the inclusion of Modern and Contemporary Physics topics, as suggested in the PCNEM and PCNEM+. This proposal consists of a didactic sequence of a course on Radiations Physics appropriated to the high school teaching. The concept of \"markers\" and \"structurating-markers\" is discussed within the Didactic Transposition Theory. \"Markers\" are established through the proposed activities, which are supposed to be the essential ones to the learning sequence.
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Avaliação do dano radioinduzido, capacidade de reparo e morte cecular em células humanas tumorais (T-47D e MCF-7) e não tumorais (MCF-10) de mama / Evaluation of the radioinduced damage, repair capacity and cell death on human tumorigenic (T-47D and MCF-7) and nontumorigenic (MCF-10) cell lines of breastValgôde, Flávia Gomes Silva 25 July 2008 (has links)
Câncer de mama é considerado uma das malignidades mais comuns que acometem as mulheres, representando cerca de uma em cada três de todas as neoplasias femininas. Aproximadamente, 90% dos casos de câncer de mama são esporádicos, atribuíveis aos eventos somáticos e cerca de 10% estão associados com a história familial e destes somente 4-5% são decorrentes de fatores hereditários. Em clínica, a radiação ionizante é a principal ferramenta utilizada no controle do crescimento tumoral, além da intervenção cirúrgica e quimioterapia. Há, no entanto, poucas infomnações no que diz respeito a resposta celular frente à ação da radiação ionizante em células-alvo, isto é, em linhagens celulares originárias de câncer de mama. O presente estudo foi proposto para analisar a radiossensibilidade de células humanas tumorals (T-47D e MCF-7) e não tumorals (MCF-10), originárias de mama, submetidas a várias doses (0,5 a 30 Gy) de radiação y de 60Co (0,72 - 1,50 Gy/min). Para tanto, foram utilizados como parâmetros de radiossensibilidade, dano radioinduzido ao DNA, capacidade de reparo e morte celular, por meio das técnicas do micronúcleo, eletroforese de microgel (teste do cometa) e viabilidade celular. Os dados obtidos mostraram que as linhagens tumorais (T-47D e MCF-7) foram mais radiossensíveis que a linhagem não tumoral (MCF-10) para todos os testes utilizados. A linhagem T-47D foi a que apresentou uma maior quantidade de dano radioinduzido, um ciclo celular mais acelerado e uma maior taxa de morte celular. As três linhagens celulares apresentaram uma capacidade de reparo relativamente eficiente, tendo em vista que uma hora após a irradiação, todas elas exibiram uma redução considerável de dano radioinduzido quando comparadas logo após as exposições. Os testes empregados mostraram ser seguros, sensíveis e reprodutíveis e permitiram quantificar e avaliar danos induzidos ao DNA, capacidade de reparo e morte celular, nas três linhagens originárias de mama humana. / Breast cancer is one of the most common malignancies that account women, representing about one in three of all female neoplasm. Approximately, 90% of cases are considered sporadic, attributed to somatic events and about 10% have a family history and this only 4 - 5 % is decurrent of hereditary factors. In the clinic, ionizing radiation is a major tool utilized in the control of tumour growth, besides surgery and chemotherapy. There is, however, little information concerning cellular response to the action of ionizing radiation in the target cells, i.e., cell lines originating from breast cancer. The present study proposed to analyze the radiosensitivity of the human tumorigenic (T-47D and MCF-7) and nontumorigenic (MCF-10) cell lines, originating from breast and submitted to various doses (0.5 to 30 Gy) of 60Co rays (0.72 - 1.50 Gy/min). For this purpose, DNA radioinduced damage, repair capacity and cell death were utilized as parameters of radiosensitivity by micronucleus, single cell gel electrophoresis (Comet assay) and cell viability techniques. The data obtained showed that tumorigenic cell lines were more radiosensitive than nontumorigenic breast cells in all assays here utilized. The T-47D cell line was presenting the highest amount of radioinduced damage, a more accelerated proliferation rate and a higher rate of cell death. The three cell lines presented a relatively efficient repair capacity, since one hour after the irradiation all of them showed a considerable reduction of radioinduced damage. The techniques employed showed to be secure, sensitive and reproducible, allowing to quantify and evaluate DNA damage, repair capacity and cell death in the three human breast cell lines.
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Modelo computacional aplicado ao efeito Bystander radioinduzidoViviane Vitória Bento Braga 24 April 2013 (has links)
Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / Este trabalho apresenta um modelo computacional aplicado na quantificação do efeito bystander radioinduzido no qual células não irradiadas sofrem danos provocados por algum tipo ainda não totalmente esclarecido de sinalização que células diretamente atravessadas por radiação emitem no meio no qual elas se encontram. No modelo proposto, uma dada célula pode se encontrar em três estados: saudável (S), danificada não emissora de sinais (D) e danificada emissora de sinais (F). As transições podem ocorrer entre os estados S e D, S e F e D, F para S. Os sinais se propagam pelo meio através da equação de difusão discreta com decaimento em duas dimensões. Análises temporais estocásticas e determinísticas foram concordantes qualitativamente. O modelo foi aplicado a um cenário experimental no qual a fração de células sobreviventes decresce com a dose usando a hipótese que uma dada célula irradiada libera uma concentração de sinais dependente da dose para um dado intervalo. Os resultados simulacionais apresentaram boa concordância quantitativa com os resultados experimentais. / This work presents a computational model to quantify the radiation-induced bystander effect in which non-irradiated cells suffer damage caused by some kind of still unknown signals emitted by neighboring cells that were directly traversed by radiation. In the model, a given cell can be found in three states: healthy (S), damaged and not signal emitter (D) damaged and signal emitter (F). Transitions can occur between the states S and D, S and F and D, F to S. Signals propagate through the medium according to the discrete diffusion equation with decay in two dimensions. Temporal stochastic and deterministic analyzes were qualitatively consistent. The model was applied to an experimental scenario where the cells surviving fraction decreases with the dose using the hypothesis that a given irradiated cell releases a dose dependent signal concentration for a given interval of dose. The simulations show good quantitative agreement with the experimental results.
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Avaliação do dano radioinduzido, capacidade de reparo e morte cecular em células humanas tumorais (T-47D e MCF-7) e não tumorais (MCF-10) de mama / Evaluation of the radioinduced damage, repair capacity and cell death on human tumorigenic (T-47D and MCF-7) and nontumorigenic (MCF-10) cell lines of breastFlávia Gomes Silva Valgôde 25 July 2008 (has links)
Câncer de mama é considerado uma das malignidades mais comuns que acometem as mulheres, representando cerca de uma em cada três de todas as neoplasias femininas. Aproximadamente, 90% dos casos de câncer de mama são esporádicos, atribuíveis aos eventos somáticos e cerca de 10% estão associados com a história familial e destes somente 4-5% são decorrentes de fatores hereditários. Em clínica, a radiação ionizante é a principal ferramenta utilizada no controle do crescimento tumoral, além da intervenção cirúrgica e quimioterapia. Há, no entanto, poucas infomnações no que diz respeito a resposta celular frente à ação da radiação ionizante em células-alvo, isto é, em linhagens celulares originárias de câncer de mama. O presente estudo foi proposto para analisar a radiossensibilidade de células humanas tumorals (T-47D e MCF-7) e não tumorals (MCF-10), originárias de mama, submetidas a várias doses (0,5 a 30 Gy) de radiação y de 60Co (0,72 - 1,50 Gy/min). Para tanto, foram utilizados como parâmetros de radiossensibilidade, dano radioinduzido ao DNA, capacidade de reparo e morte celular, por meio das técnicas do micronúcleo, eletroforese de microgel (teste do cometa) e viabilidade celular. Os dados obtidos mostraram que as linhagens tumorais (T-47D e MCF-7) foram mais radiossensíveis que a linhagem não tumoral (MCF-10) para todos os testes utilizados. A linhagem T-47D foi a que apresentou uma maior quantidade de dano radioinduzido, um ciclo celular mais acelerado e uma maior taxa de morte celular. As três linhagens celulares apresentaram uma capacidade de reparo relativamente eficiente, tendo em vista que uma hora após a irradiação, todas elas exibiram uma redução considerável de dano radioinduzido quando comparadas logo após as exposições. Os testes empregados mostraram ser seguros, sensíveis e reprodutíveis e permitiram quantificar e avaliar danos induzidos ao DNA, capacidade de reparo e morte celular, nas três linhagens originárias de mama humana. / Breast cancer is one of the most common malignancies that account women, representing about one in three of all female neoplasm. Approximately, 90% of cases are considered sporadic, attributed to somatic events and about 10% have a family history and this only 4 - 5 % is decurrent of hereditary factors. In the clinic, ionizing radiation is a major tool utilized in the control of tumour growth, besides surgery and chemotherapy. There is, however, little information concerning cellular response to the action of ionizing radiation in the target cells, i.e., cell lines originating from breast cancer. The present study proposed to analyze the radiosensitivity of the human tumorigenic (T-47D and MCF-7) and nontumorigenic (MCF-10) cell lines, originating from breast and submitted to various doses (0.5 to 30 Gy) of 60Co rays (0.72 - 1.50 Gy/min). For this purpose, DNA radioinduced damage, repair capacity and cell death were utilized as parameters of radiosensitivity by micronucleus, single cell gel electrophoresis (Comet assay) and cell viability techniques. The data obtained showed that tumorigenic cell lines were more radiosensitive than nontumorigenic breast cells in all assays here utilized. The T-47D cell line was presenting the highest amount of radioinduced damage, a more accelerated proliferation rate and a higher rate of cell death. The three cell lines presented a relatively efficient repair capacity, since one hour after the irradiation all of them showed a considerable reduction of radioinduced damage. The techniques employed showed to be secure, sensitive and reproducible, allowing to quantify and evaluate DNA damage, repair capacity and cell death in the three human breast cell lines.
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Análise mecânica de réguas ósseas esterilizadas por radiação gama para uso em bancos de tecidos / Mechanical analysis of bone rulers sterilized by gama radiation for use in Tissue BanksLuis Otavio Carvalho Kosmiskas 06 February 2007 (has links)
Ao se trabalhar com materiais para a área de saúde, uma das principais considerações é evitar possíveis contaminações, dada a natureza da sua aplicação. Dentre os métodos desenvolvidos para esterilização, a radiação ionizante tem sido amplamente empregada por inúmeros setores da área de saúde pela sua eficácia em eliminar contaminantes biológicos de diversas origens. A problemática da utilização da radiação ionizante em materiais de origem humana, contudo, vai além da questão de sua eficácia como esterilizante, incluindo as possíveis alterações na estrutura do material de escolha. Desta forma, no presente trabalho, buscou-se avaliar qual a extensão das alterações biomecânicas causadas pela radiação em tecidos ósseos. Mais especificamente avaliamos as alterações no módulo de elasticidade, tensão de ruptura e porcentagem de deformação que decorrem do processo de irradiação. Como modelo de eleição foram escolhidas réguas ósseas retiradas da diáfase de fêmures bovinos, simulando as condições de ossos humanos armazenados em Bancos de Tecidos Biológicos. Estas foram congeladas a -70ºC e expostas a doses crescentes de radiação gama (0, 12.5, 25 e 50 kGy). Durante o trabalho foi desenvolvido um sistema de cortes para obtenção de amostras precisas e homogêneas. Os resultados demonstram que há uma diferença significativa entre as características observadas com o aumento da dose de radiação. / In the production process of health care products, contamination must be considered as one of the principal hazards to be avoided. Among the developed methods for sterilization, ionizing radiation has largely been used by many sectors in health care area as it is efficient in eliminating biological contaminants of several origins. The difficulty of deploying ionizing radiation in materials of human origin, though, includes which possible alterations it might cause in human tissue. In the present work, the extension of the biomechanical alteration generated by radiation in bone tissue was evaluated by biomechanical methods. More specifically, we evaluated alterations to the elastic modulus, rupture tension and percentage of deformation that are thought to be a consequence of the sterilization process. As a research model, bovine femur struts obtained from the diaphysis were used. The struts were frozen in a temperature of -70 ºC and irradiated with crescent doses of gamma radiation (0, 12.5, 25 e 50 kGy). During this work, a cutting system to obtain precision samples to use in such essays was developed. As results show that there is a significant different between the analyzed characteristics in the different doses of radiation.
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Facteurs modulant la radiosensibilité : rôle des protéines PARP-1, PARP-2 et Cdk5 et implication de la chromatine / Radiosensitivity Modulating Factors : role of PARP-1, PARP-2 and Cdk5 proteins and chromatin implicationBoudra, Mohammed-Tayyib 14 December 2011 (has links)
Les modifications post-traductionnelles des protéines de réparation de l’ADN et des facteurs chromatiniens par poly(ADP-ribose)ylation et par phosphorylation sont essentielles pour le maintien de l’intégrité de l’ADN et de la chromatine, en particulier dans la réponse cellulaire aux dommages de l’ADN induits par radiation ionisantes (RI). Parmi les protéines impliquées dans ces deux processus nous trouvons, respectivement, la poly(ADP-ribose) polymérase-1 (PARP-1) et PARP-2, et la kinase dépendante des cyclines Cdk5 : PARP-1 et PARP-2 sont impliqué dans le mécanisme de réparationdes cassures simples brin (CSBs) de l’ADN (Single Strand Break Repair : SSBR ) et la déplétion de Cdk5 a été liée à l’augmentation de la sensibilité des cellules aux inhibiteurs de PARPs. Nous avons montré, en utilisant des cellules HeLa stablement déplété pour Cdk5 ou PARP-2, que ces deux protéines sont impliquées dans les deux sous-voies du SSBR, le short- (SPR) et le long-patch repair (LPR). L’absence de Cdk5 ou PARP-2 entraîne des modifications du fonctionnement du SSBR, notamment en termes de recrutement des protéines de réparation PARP-1 et XRCC1, impliquées dans le SPR, et PCNA, protéine clé du LPR, au site du photo-dommage. PARP-2 et Cdk5 agissent aussi sur la balance du niveau des poly(ADP-ribose) car en absence de Cdk5 une hyper-activation de PARP-1 a été montrée, et en absence de PARP-2 une diminution de l’activité de la protéine poly(ADP-ribose) glycohydrolase (PARG) a été aussi observée. Cependant, malgré ces changements les deux lignées cellulaires dépourvues de Cdk5 (Cdk5KD) ou de PARP-2 (PARP-2KD) réparent de façon normale les CSBs radio-induites, mais, intéressement et contrairement aux cellules PARP-2KD, les cellules Cdk5KDsont sensibles à l’effet létal des RI. De plus nous avons montré que Cdk5, PARP-2 et PARG sont toutes les trois impliquées dans la régulation du recrutement et de dissociation du facteur chromatinien ALC1 suggérant leur implication dans la régulation de la dynamique de la chromatine en réponse aux photo-dommages de l’ADN. Ces résultats avec l’observation de la diminution du recrutement de PARP-1 dans les cellules Cdk5KD et PARP-2KD, montrent l’apparition d’un réseau complexe de phosphorylation et de poly(ADP-ribose)ylation en réponse aux RI qui implique Cdk5 , PARP-1,PARP-2 and PARG et qui est fort probablement initié par l’activité kinase de Cdk5. / The post-translational modifications of DNA repair proteins and histone remodeling factors by poly(ADP-ribose)ylation and phosphorylation are essential for the maintenance of DNA integrity and chromatin structure, and in particular in response to DNA damaging produced by ionizing radiation (IR). Amongst the proteins implicated in these two processes are the poly(ADP-ribose) polymerase -1 (PARP-1) and PARP-2, and the cyclin-dependent kinase Cdk5: PARP-1 and 2 are involved in DNA single strand break (SSB) repair (SSBR) and Cdk5 depletion has been linked with increased cell sensitivity to PARP inhibition. We have shown by using HeLa cells stably depleted for either CdK5 or PARP-2, that the recruitment profile of PARP-1 and XRCC-1, two proteins involved in the short-patch (SP) SSBR sub-pathway, to DNA damage sites is sub-maximal and that of PCNA, a protein involved in the long-patch (LP) repair pathway, is increased in the absence of Cdk5 and decreased in the absence of PARP-2 suggesting that both Cdk5 and PARP-2 are involved in both SSBR sub-pathways.PARP-2 and Cdk5 also impact on the poly(ADP-ribose) levels in cells as in the absence of Cdk5 a hyper-activation of PARP-1 was found and in the absence of PARP-2 a reduction in poly(ADP-ribose) glycohydrolase (PARG) activity was seen. However, in spite of these changes no impact on the repair of SSBs induced by IR was seen in either the Cdk5 or PARP-2 depleted cells (Cdk5KD or PARP-2KD cells) but, interestingly, increased radiation sensitivity in terms of cell killing was noted in the Cdk5 depleted cells. We also found that Cdk5, PARP-2 and PARG were all implicated in the regulation ofthe recruitment and the dissociation of the chromatin-remodeling factor ALC1 from DNA damage sites suggesting a role for these three proteins in changes in chromatin structure after DNA photodamage.These results, taken together with the observation that PARP-1 recruitment is sub-optimal in both Cdk5KD and PARP-2KD cells, show that an intricate network of phosphorylation and poly(ADPribose) ylation occurs in response to IR that involves Cdk5 , PARP-1,PARP-2 and PARG and that is probably initiated by Cdk5’s kinase activity.
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