L’irradiation (IR) est utilisée dans le traitement de plusieurs cancers et désordres hématologiques, en particulier dans les protocoles de conditionnement précédents les transplantations de moelle osseuse. L’emploi de doses réduites d’IR semble favoriser le succès de la prise de greffe. Cette observation soulève un point de plus en plus discuté dans la littérature, soit l’importance de l’intégrité du microenvironnement pour la transplantation et le bon fonctionnement de l’hématopoïèse. L’IR induit la sénescence des cellules stromales de la moelle osseuse in vitro. Ce mécanisme de défense cellulaire entraînant un arrêt de prolifération permanent est également observé in vivo dans différents systèmes, mais n’a pas encore été étudié dans le contexte de la niche hématopoïétique. Les travaux présentés dans cette thèse ont pour objectif de déterminer si l’IR induit la sénescence des cellules stromales de la moelle osseuse et si une telle induction altère les fonctions hématopoïétiques. Nos résultats ont permis de démontrer pour la première fois qu’une IR corporelle totale induit effectivement la sénescence des cellules stromales de la moelle osseuse. En outre, cette altération du microenvironnement affecte la lymphopoïèse B de façon Ink4a/Arf-dépendante (1er article). De plus, les modifications systémiques qui résultent de l’IR compromettent l’homéostasie osseuse en augmentant la résorption de l’os, sans toutefois diminuer la formation de celui-ci (2e article). Ces données nous permettent de mieux comprendre les effets de la sénescence des cellules stromales de la moelle osseuse sur les fonctions hématopoïétiques. Par ailleurs, elles suggèrent que l’emploi de drogues et/ou de procédés n’induisant pas la sénescence des cellules stromales de l’os offrirait un meilleur pronostic à long terme pour les patients. / Ionizing radiation (IR) is used in the treatment of several cancers and hematological disorders, especially in conditioning regimens for bone marrow transplantation. Reduced doses of IR seem to favor the success of engraftment. This observation supports the growing evidences suggesting the importance of the microenvironment integrity for the success of bone marrow transplantation and hematopoiesis maintenance. IR induces senescence of bone marrow stromal cells in vitro. This defense mechanism which leads to a permanent cell growth arrest is also observed in different organs in vivo but has not yet been studied in the hematopoietic niche. The objectives of this doctoral thesis are to determine whether IR induces senescence of bone marrow stromal cells and whether such induction alters hematopoietic functions. Our results have demonstrated for the first time that total body IR actually induces the senescence of bone marrow stromal cells. Furthermore, this alteration of the microenvironment affects B lymphopoiesis in an Ink4a/Arf-dependent manner (paper #1). In addition, the systemic changes associated with IR compromise bone homeostasis by increasing bone resorption without reducing bone formation (paper #2). All together, these data enhance our knowledge related to the effects of IR-induced senescent bone marrow stromal cells on hematopoietic function. Moreover, our results suggest that using drugs and/or procedures inducing no senescent bone marrow stromal cells would provide a better long-term prognosis for patients.

Thesis (M.S.)--Uniformed Services University of the Health Sciences, 2005. / Typescript (photocopy).

Radiations can be classified as either ionizing or non-ionizing according to whether it ionizes or does not ionize the medium through which they propagate. X-rays photons and gamma rays are the typical examples of ionizing radiations whereas radiowave, heat or visible light are examples of non ionizing radiations. UV photons have some features of both ionizing and non-ionizing radiation. Both ionizing and non-ionizing radiation can be harmful to living organisms and to the natural environment. Hence the detection and measurement of radiation is very important for the well being of living organisms as well as the natural environment. Not only for safety reasons, have radiation detectors found their applications in various fields including medical physics, nuclear and particle physics, astronomy and homeland security. Industrial sectors that use radiation detection include medical imaging, security and baggage scanning, the nuclear power industry and defense. Gas electron multiplier (GEM) is one of the most successful representatives of gaseous detectors used for UV photon and X-ray photon detection. Recently there is a growing demand for large area photon detectors with sensitivity reaching to the level of single photon. They are used in spectroscopy and imaging in astronomy high energy physics experiments etc. Thick GEM (THGEM) is a mechanical expansion of standard GEM. It has all the necessary requirements needed for large area detector and offers a multiplication factor that permits efficient detection of light. Hence, the development and performance study of THGEM based radiation detector is chosen as the topic of study in the present thesis. The initial part of the thesis contains simulation studies carried out for the understanding the working of the detector and the effect of various design parameters of THGEM for the above said applications. Different steps for the fabrication of THGEM and the technical challenges faced during the process are discussed. In the view of application of the fabricated THGEM for UV photon detection, cesium iodide photocathode is prepared using thin film technology and characterized. The performance of the photocathode under various operating conditions is studied in terms of its photoemission property. The effect of vacuum treatment on the photoemission property of the photocathode exposed to moist air is studied in detail. A major portion of this thesis focuses on maximizing the detection efficiency of the UV photon detector realized using the fabricated THGEM coupled with the cesium iodide photocathode. Simulations are used at different stages to interpret the experimental observations. The electron spectrum obtained from the detector under study was analyzed. The dependence of secondary effect like photon feedback on the operating parameters is also discussed. The last portion of the thesis deals with the application of THGEM as an X-ray detector. The performance is evaluated in terms of the gain and energy resolution achieved. The thesis is organized as follows: Chapter 1 is divided into two sections. Section A gives a general introduction to different types of radiation detectors found in the present day and their working principles. This is followed by discussion about gas ionization based detector and its working principle in detail. A brief literature survey of the different types of micropattern gas detectors is also given in this section. In Section B of this chapter GEM and THGEM are introduced with discussion about their working principle and areas of application. Chapter 2 deals with the simulation study of THGEM undertaken to have a clear understanding of the detector’s working. Section A of this chapter gives an overview of the simulation tools used for the present thesis in particular ANSYS and GARFIELD. Section B presents the results of the simulation study highlighting the effects of different geometrical and operating parameters on the electric field distribution in and around the THGEM aperture. The relevance of the study to the detectors performance is discussed vividly for all the cases. In Chapter 3, the details of the different steps involved in THGEM fabrication are given. Design aspects involved, fabrication of the THGEM using standard PCB technology coupled with photolithography technique are discussed in this chapter. This is followed by an elaborate description of the test setup used for all the performance study. Preface In the view of application of THGEM as a UV photon detector, cesium iodide photocathode was prepared and characterized. Chapter 4 discusses about the CsI photocathode preparation and its characterization for the above said application. Photoemission property of the photocathode was analyzed under various operating parameters. The effect of vacuum treatment on the photocathode performance is a new aspect of this thesis. Its correlation with the microstructure of the film is reported for the first time. Chapter 5 deals with the application of THGEM as a UV photon detector. The study mainly focuses on the improvement of the detection efficiency of the detector. The effect of drift parameters on the electron transfer efficiency and hence on the detection efficiency of the detector is a major contribution of this thesis. There are no literature available which discusses this aspect of a UV photon detector. The experimental study has been supported with simulation results. In addition to the study on detection efficiency, electron spectrum has also been acquired from the UV photon detector. The spectrum has been analyzed under various operating conditions. Discussions about secondary effects like photon feedback prevailing in the detector output are also present in this chapter. Chapter 6 presents the results of THGEM as an X-ray detector. The performance of the detector has been evaluated in terms of the effective gain and energy resolution achieved under different operating conditions. The gain instability with time and its uniformity across the THGEM area are also studied. The effect of drift field on the energy resolution and its correlation with ETE is a new aspect of this work. Chapter 7 summarizes the salient features of the work presented in this thesis. Also the scope of future work based on this thesis is discussed at the end of the chapter.

La sénescence est un important mécanisme cellulaire qui prévient la tumorigenèse et se caractérise par un arrêt permanent du cycle cellulaire orchestré principalement par les inhibiteurs des cycline-kinases dépendantes (i.e p16INK4a). La sénescence est une caractéristique importante du vieillissement, mais un déséquilibre dans son induction peut être délétère pour la régénération tissulaire et paradoxalement pour la progression tumorale. L'irradiation (IR) est couramment utilisée comme approche thérapeutique dans le cancer. Chez les enfants survivants du cancer, l’exposition à l’irradiation et à la chimiothérapie entrainent le développement d’importants effets secondaires, lesquels sont associés à une forme de vieillissement prématuré. La formation de cellules sénescentes, en inhibant la prolifération tissulaire et en sécrétant des cytokines proinflammatoires, pourrait être en être responsable. Notre groupe a précédemment démontré que le gène p16INK4a est augmenté de manière tardive (environ 8 semaines) suite à une exposition à l’irradiation. Il n'a pas encore été étudié si cette expression retardée survient en réponse aux dommages causés par l'irradiation sur l’homéostasie tissulaire ou à titre de mécanismes de suppression tumorale. Un objectif de cette thèse visait donc à déterminer s’il était possible de moduler/inhiber l’expression de p16INK4a dans le but d’accroitre la régénération tissulaire sans nécessairement accroitre les risques d’incidence du cancer. En effet, ceci pourrait être possible dans la mesure ou la sénescence induite par p16INK4a est également irréversible in vivo. Nos résultats ont démontré que l’inhibition de l’expression de p16INKa (suite à l’administration de tamoxifen chez les souris p16L/LCre), induit à la fois une augmentation de la régénération tissulaire mais malheureusement également une augmentation de l’incidence du cancer. Nous voulions également connaitre l’impact de l’accumulation de ces cellules sénescentes sur les tissus, plus spécifiquement sur la fonction des cellules immunitaires de la rate. Nous avons démontré que des altérations (dépendantes de p16INK4a) au sein du microenvironnement splénique pouvaient altérer les fonctions intrinsèques des macrophages, des cellules dendritiques et des lymphocytes T. En outre, l'élimination systémique des cellules p16INK4a positives (modèle de sourie p16-3MR) a conduit à une restauration partielle de la fonction de ces cellules immunitaires. La combinaison de ces données nous permet de mieux comprendre le rôle et la fonction du gène p16INK4a dans le processus de sénescence induite par l’irradiation. Nos résultats suggèrent qu’il est envisageable d’utiliser des agents pharmacologiques tels que des composés sénolytiques, capables d’induire l’apoptose chez les cellules sénescentes spécifiquement, afin de potentiellement diminuer les effets du vieillissement prématuré induit par la sénescence cellulaire chez les survivants du cancer. / Senescence is an important cellular mechanism that prevents tumorigenesis and is characterized by a permanent cell cycle arrest orchestrated by cyclin-dependent kinases inhibitors (i.e p16INK4a). Senescence is an important hallmark of aging and unbalanced levels of senescence is considered deleterious for tissue regeneration, and paradoxically for tumor progression. Irradiation (IR) is commonly used therapeutic approach in cancer treatment. Together with surgery and chemotherapy, it has helped to increase the life expectancy of patients and, in some cases, leads to complete remission. However, long-after therapy, children who survive cancer encounter alterations in the integrity of tissues/organs associated with premature aging. The accumulation of senescent cells may be responsible for this accelerated aging by limiting tissue proliferation and secreting pro-inflammatory cytokines. Our group has previously demonstrated that the p16INK4a gene is increased in a delayed manner (approximately 8 weeks) following exposure to IR. It has not yet been investigated whether this delayed expression occurs in response to IR-induce damage of tissue homeostasis or as tumor suppression mechanisms. One objective of this thesis was to determine whether it was possible to modulate / inhibit the expression of p16INK4a in order to increase tissue regeneration without necessarily increasing the risk of cancer incidence. Indeed, this may be possible since p16INK4a-induced senescence is also irreversible in vivo. Our results demonstrated that the inhibition of p16INK4a expression in conditional-p16INK4a null mice , induces both an increase in tissue regeneration but unfortunately also an increase in the incidence of cancer. We also wanted to know the impact of the accumulation of these senescent cells on the tissues, more specifically on the function of the immune cells in the spleen. We have demonstrated that alterations (p16INK4a-dependent) within the splenic microenvironment can alter the intrinsic functions of macrophages, dendritic cells and T cells. In addition, the systemic elimination of p16INK4a positive cells (mouse model p16-3MR) has led to a partial restoration of the function of these immune cells. The combination of these data allows us to better understand the role and function of the p16INK4a gene in the irradiation-induced senescence process. Our results suggest that it is conceivable to use pharmacological agents such as senolytic compounds, capable of inducing apoptosis in senescent cells specifically, in order to potentially reduce the effects of premature aging induced by cellular senescence in cancer survivors.