Thesis advisor: Stephen J. Pope / The aim of this dissertation is to expand a contemporary multidimensional discourse on the nature of eating disorders to encompass also a moral dimension. Eating disorders are complex phenomena which include biomedical, psychological, and sociocultural components. This dissertation brings the psychosocial literature on eating disorders and body dissatisfaction into dialogue with contemporary studies in Thomistic moral theology, and argues that such a multidisciplinary dialogue can illuminate new insights both for the study of eating disorders and for recent efforts to recover Thomistic moral theology in a contemporary context. Beginning empirically, the dissertation examines recent evidence showing that exposure to &ldquo;thin-ideal images&rdquo; in the mass media is positively correlated with an increase in body dissatisfaction and eating disorder symptomatology. Socioculturally, the explanation for this phenomenon is called &ldquo;thin-ideal internalization,&rdquo; and basically measures the extent to which individuals &ldquo;buy into&rdquo; the validity of images using ultra-thin female models as a paradigm of beauty. Women who have a high level of internalization desire to conform to a thin-ideal, and behave accordingly, even when they are rationally aware of the unrealistic and unhealthy nature of such an ideal. Turning to Thomas Aquinas' moral theology, the dissertation argues that thin-ideal internalization is a form of connatural knowledge, an affective form of knowing (<italic>per modum inclinationis</italic> or <italic>ex instinctu</italic>) which is at the very basis of Aquinas' moral theology, both in explaining the operation of habits and in explaining the role of grace in the moral life through charity and the gifts of the Holy Spirit. This dissertation argues that Aquinas' theory of connatural knowledge provides a relevant and constructive contribution to the study of eating disorders, especially on the relationship between body dissatisfaction and eating disorder symptamatology. Additionally, the incorporation of the psychosocial literature on eating disorders into Thomistic moral theology provide a valuable contribution to Thomistic moral theology in the effort to understand the role of the affections in moral deliberation, the development of habits, and the importance of Christian practices in the moral life. / Thesis (PhD) — Boston College, 2011. / Submitted to: Boston College. Graduate School of Arts and Sciences. / Discipline: Theology.

Early research on motivation draws connections to what are now understood as aspects of sensemaking, however research does not explicitly draw a connection to the theory of sensemaking itself. Therefore, the sensemaking perspective is applied to the motivational theory of self determination in order to gain a deeper understanding of how and why motivation develops through sensemaking. This is accomplished through a qualitative approach using semi-structured interviews for empirical data collection, followed by thematic analysis. The research concludes with a representation of the sequential process that demonstrates the interconnectedness of sensemaking and motivation, demonstrating the connection between environmental change, meaning creation and internalization, need satisfaction, motivation and action. Consequently, this research provides insight to the co-existence of sensemaking and motivation, facilitating opportunities to reach a more complete understanding of how individual motivation actually exists and develops.

Les nanoparticules (NP) sont de plus en plus utilisées en médecine (imagerie, vectorisation de médicaments). Au sein de l’EA 3452, des NP polymériques chargées en donneurs d’oxyde nitrique •NO (S-nitrosoglutathion GSNO) sont développées afin d’améliorer la stabilité et la biodisponibilité de •NO. Le travail de thèse comprend 2 parties : premièrement : évaluer les effets des NP polymériques d’Eudragit® RL vides avec deux lignées cellulaires humaine (THP-1) et de rat (NR8383). La modification de l’expression de certains gènes impliqués dans des voies de souffrance cellulaire a été étudiée par RT-qPCR après 4h d’exposition aux NP. L’expression d’ATG16L, BCL2 et TNFA est augmentée dans NR8383, ce qui est en faveur d’une induction des processus autophagiques et inflammatoires. La diminution de l’expression de NCF1, NFKB et IL1B dans THP-1 peut expliquer l'augmentation de la viabilité et de la croissance cellulaires. Deuxièmement : évaluer les effets des mêmes NP chargées ou non en GSNO sur les THP-1 par une analyse des transcriptomes ce qui a montré une variation significative de l’expression des gènes associés aux clusters « prolifération », « structure cellulaire » pour les NP vides et « mitochondrie », « processus métabolique » pour les NP chargées en GSNO. Ces résultats sont cohérents avec ceux des tests cellulaires. L’analyse du transcriptome obtenu avec le GSNO libre a permis de proposer un mécanisme d’activation de l’immunité non spécifique contre Leishmania au sein du macrophage. Ainsi, l’étude génomique devient un outil indispensable pour mieux comprendre les interactions entre les NP et les cellules et a ainsi toute sa place en toxicologie / Although nanoparticles (NP) are more and more used in medicine (diagnosis, imaging, drug targeting), few studies on their possible undesirable effects have been conducted. The aim of this work was to evaluate the effects of polymeric NP Eudragit® RL loaded or not with S-nitrosoglutathione (GSNO), a nitric oxide donor, using two different cell lines: monocytes/macrophages of rat (NR8383) and human (THP-1). First, our results show that exposure to blank NP Eudragit® induced an up-regulation of ATG16L, BCL2 and TNFA and a down-regulation of PDCD4 in rat cells, what are in favor of a repression of apoptosis and an induction of autophagy. On the other hand, NCF1, NFKB, and IL1B were down-regulated in human cells, which may contribute to explain the increase of cellular viability. Second, we evaluated the effects of these NP loaded or not with GSNO on THP-1 cells. The results show that NP Eudragit® were internalized by cells using likely the clathrine and caveoline endocytic pathways. Transcriptome analyses by microarray of THP-1, exposed to either GSNO-loaded or empty NP, have shown a significant dose and time-dependent variation of the expression of genes belonging to the clusters “proliferation”, “cell structure” for the empty NP and “mitochondria” and “metabolic processes” for the loaded ones. These results are consistent with those of cellular assays. Transcriptome analyses of free GSNO allowed to propose a mechanism of non-specific immunity activation against Leishmania in the macrophage. Thus, (i) the cytotoxicity of NP is cellular model dependent and (ii) mechanistic toxicology should be the corner stone for the evaluation of NP harmfulness

O presente trabalho teve por objetivo avaliar alterações devido à exposição a Nanotubos de Carbono de Parede Simples (NTCPS) em duas linhagens celulares epiteliais (BBnt e HK2) e em uma linhagem celular monocítica (THP-1), enfocando os mecanismos de internalização e os efeitos sobre o ciclo celular. Foi avaliada a ação dos receptores scavenger na internalização dos NTCPS nas células HK2 e THP-1 e a interferência de duas concentrações de NTCPS sobre os elementos do citoesqueleto e no ciclo celular, nas células HK2 e BBnt. As concentrações utilizadas foram equivalentes as permitidas pelo The National Institute for Occupational Safety and Health: 2,4 e 24 mg/cm2. A análise de expressão de mRNA por RT-PCR para receptores scavenger, mostrou que a internalização do NTCPS pode ocorre por endocitose. Sendo que os receptores SCARA5 e SRA são os responsáveis pela internalização nas células THP-1, enquanto MARCO e SRA realizam o processo de internalização nas células HK2. Observou-se que em ambas as concentrações, as células BBnt apresentaram amplificação centrossômica, com a ocorrência de 25,38% e 28,46% de mitoses alteradas para cada concentração, respectivamente. Não houve interferência significativa na progressão do ciclo celular em ambas as linhagens. O estudo da interação dos NTCPS com vesículas lipídicas não apresentou evidencias de alterações ou danos na membrana das vesículas, porém as vesículas apresentaram-se associadas umas às outras após o tratamento com 24 mg/cm2. / This study aimed to assess changes due to exposure to of Single-wall Carbon Nanotubes (SWCNT) in two epithelial cell lines (BBnt and HK2) and a monocytic cell line (THP-1), focusing on the mechanisms of internalization and effects on the cell cycle. The action of scavenger receptors in the internalization of SWNTC in HK2 and THP-1 cells and the interference of two concentrations of SWNTC about elements of the cytoskeleton and the cell cycle, in BBnt and HK2 cells was evaluated. The concentrations used were equivalent to those allowed by The National Institute for Occupational Safety and Health: 2,4 to 24 mg/cm2. Analysis of mRNA expression by RT-PCR for scavenger receptors showed that the SWNTC internalization can occurs by endocytosis. Being that SCARA5 and SRA receptors are responsible for internalization in THP-1 cells, while MARCO and SRA perform the process of internalization in HK2 cells. It was observed that at both concentrations, the cells showed centrosome amplification in BBnt cells, with the occurrence of 25.38% and 28.46% of mitosis changed for each concentration, respectively. There was no significant interference with cell cycle progression in both strains. The study of the interaction of lipid vesicles with SWNTC showed no evidence of change or damage the membrane of the vesicles, but the vesicles were associated with each other after treatment with 24 mg/cm2.

O presente trabalho teve por objetivo avaliar alterações devido à exposição a Nanotubos de Carbono de Parede Simples (NTCPS) em duas linhagens celulares epiteliais (BBnt e HK2) e em uma linhagem celular monocítica (THP-1), enfocando os mecanismos de internalização e os efeitos sobre o ciclo celular. Foi avaliada a ação dos receptores scavenger na internalização dos NTCPS nas células HK2 e THP-1 e a interferência de duas concentrações de NTCPS sobre os elementos do citoesqueleto e no ciclo celular, nas células HK2 e BBnt. As concentrações utilizadas foram equivalentes as permitidas pelo The National Institute for Occupational Safety and Health: 2,4 e 24 mg/cm2. A análise de expressão de mRNA por RT-PCR para receptores scavenger, mostrou que a internalização do NTCPS pode ocorre por endocitose. Sendo que os receptores SCARA5 e SRA são os responsáveis pela internalização nas células THP-1, enquanto MARCO e SRA realizam o processo de internalização nas células HK2. Observou-se que em ambas as concentrações, as células BBnt apresentaram amplificação centrossômica, com a ocorrência de 25,38% e 28,46% de mitoses alteradas para cada concentração, respectivamente. Não houve interferência significativa na progressão do ciclo celular em ambas as linhagens. O estudo da interação dos NTCPS com vesículas lipídicas não apresentou evidencias de alterações ou danos na membrana das vesículas, porém as vesículas apresentaram-se associadas umas às outras após o tratamento com 24 mg/cm2. / This study aimed to assess changes due to exposure to of Single-wall Carbon Nanotubes (SWCNT) in two epithelial cell lines (BBnt and HK2) and a monocytic cell line (THP-1), focusing on the mechanisms of internalization and effects on the cell cycle. The action of scavenger receptors in the internalization of SWNTC in HK2 and THP-1 cells and the interference of two concentrations of SWNTC about elements of the cytoskeleton and the cell cycle, in BBnt and HK2 cells was evaluated. The concentrations used were equivalent to those allowed by The National Institute for Occupational Safety and Health: 2,4 to 24 mg/cm2. Analysis of mRNA expression by RT-PCR for scavenger receptors showed that the SWNTC internalization can occurs by endocytosis. Being that SCARA5 and SRA receptors are responsible for internalization in THP-1 cells, while MARCO and SRA perform the process of internalization in HK2 cells. It was observed that at both concentrations, the cells showed centrosome amplification in BBnt cells, with the occurrence of 25.38% and 28.46% of mitosis changed for each concentration, respectively. There was no significant interference with cell cycle progression in both strains. The study of the interaction of lipid vesicles with SWNTC showed no evidence of change or damage the membrane of the vesicles, but the vesicles were associated with each other after treatment with 24 mg/cm2.

Submitted by Jailda Nascimento (jmnascimento@pucsp.br) on 2016-10-26T15:09:33Z No. of bitstreams: 1 Mônica Marques Zilli.pdf: 898413 bytes, checksum: b9ee4ead79b3cd989666db8bcff48caf (MD5) / Made available in DSpace on 2016-10-26T15:09:33Z (GMT). No. of bitstreams: 1 Mônica Marques Zilli.pdf: 898413 bytes, checksum: b9ee4ead79b3cd989666db8bcff48caf (MD5) Previous issue date: 2016-09-09 / The aesthetic experience has been fascinating people throughout history, being the target of several investigations in different fields of knowledge. In this work, the aesthetic encounter is conceived as an experience of transformation of the self, in so far as it constitutes an encounter with a transformational object, the effects of which are rooted in the experiences with the first caregiver objects. Based on W. Bion’s concept of réverie and on H. Kohut’s of transmuting internalization, the functional dimensions of the transformational object, from which the work of art operates, conveying an intrinsic sublimating potential, are investigated. This perspective legitimizes art as a complex and rich therapeutic device, hence its importance as a promoter of individual and collective health / Há séculos a experiência de fruição estética fascina o ser humano, sendo alvo das mais diversas investigações em diferentes campos do saber. Neste trabalho, busca-se pensar o encontro estético como experiência de transformação do self na medida em que se constitui um encontro com um objeto transformacional, cujos efeitos radicam-se nas experiências com os primeiros objetos cuidadores. Com aporte na teoria bioniana da réverie e kohutiana da internalização transmutadora, são elucidadas as dimensões funcionais do objeto transformacional a partir das quais a obra de arte, entendida como tal, operaria, veiculando um potencial sublimatório intrínseco. Esta perspectiva legitima a arte como complexo e rico dispositivo terapêutico, de modo que as transformações propiciadas pelo encontro estético na experiência do self dizem respeito à oferta de uma forma especializada de cuidado - a partir do que também se advoga a importância da arte e da cultura enquanto promotores de saúde individual e coletiva

Submitted by Cássia Santos (cassia.bcufg@gmail.com) on 2014-10-15T15:19:45Z No. of bitstreams: 2 Tese Marcia Santos Anjo Reis - Educacao.pdf: 3619898 bytes, checksum: 96bc827312052a74d40818e8e2f2822d (MD5) license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) / Approved for entry into archive by Jaqueline Silva (jtas29@gmail.com) on 2014-10-20T19:40:09Z (GMT) No. of bitstreams: 2 Tese Marcia Santos Anjo Reis - Educacao.pdf: 3619898 bytes, checksum: 96bc827312052a74d40818e8e2f2822d (MD5) license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) / Made available in DSpace on 2014-10-20T19:40:10Z (GMT). No. of bitstreams: 2 Tese Marcia Santos Anjo Reis - Educacao.pdf: 3619898 bytes, checksum: 96bc827312052a74d40818e8e2f2822d (MD5) license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) Previous issue date: 2014-02-25 / Fundação de Amparo à Pesquisa do Estado de Goiás - FAPEG / This work is linked to the research line "State, Policies and History of Education" of the Doctoral Program in Education at UFG and to the Nedesc/UFG. Discussing the subject “expansion and internalization of higher education”, we seek to understand the specificities and the uniqueness of the expansion process and internalization of higher education in Jataí, state of Goiás. It was adopted, from 1985 to 2010, a time marked by expansion and internalization politics of brazilian higher education, Goiás and Jataí. The centrality of the analysis was in the presidential managements of Cardoso (1995-2002) and Lula (2003-2010), highlighting the decrees, laws, programs and actions that contributed to the brazilian expansion, looking to establish a counterpoint for reflection and analysis of the expansion movement of Goiás and Jataí. For the development of the thesis, documentary and literature researches, statistical analysis and interviews were realized with 18 subjects. The theoretical framework was based on the following studies: Amaral (2009 ); Catani and Oliveira (2002); Cunha (2003 , 2004, 2006); Dourado (2001a, 2002, 2008, 2010, 2011); Frigotto (2009); Oliveira D. (2011a, 2011b); Oliveira J. (2000); Sguissardi (1993, 2004, 2006, 2008); Silva Jr. and Sguissardi (2001, 2005); Trow (2005), among others. The analysis of indicators centered on the data in relation to the institutions courses and vacancies of present graduation, that give expression to expansion and internalization categories. Reflections on the characteristics of the transformation of higher education were guided in the theoretical model developed by Trow (2005), which characterizes three phases: of elite, mass and universal access systems. The field of research focused in Jataí. Five IES were analyzed: CAJ/UFG; IFG - Campus Jataí; UEG - UnU Jataí; Cesut and Faja. FEJ was also investigated, important foundation in the creationand expansion process of higher education in Jataí. The process of internalization and expansion of higher education in Brazil, in Goiás and Jataí occurs in the confrontation between public and private, each one adopting its own dynamic. Like Brazil and Goiás adopted a policy of diversification and differentiation in higher education, Jataí followed this trend, but with the uniqueness marked by the predominance of public expansion. There was a significant expansion of public higher education in Jataí, from the 1990s, contrary to the national and the state of Goiás logic, however, with institutional conditions very different, including the public ones. The expansion and internalization of public IES of CAJ/UFG and IFG - Câmpus Jataí were guided on quality indicators, such as: public tender, for functional employment of 40 hours and DE, concerning with the teacher training (stricto sensu), adequate physical infrastructure. In contrast, the expansion of UEG - UnU Jataí revealed lack of attendanceto the quality basic parameters, with temporary teachers contract, hourly, where most are graduates or specialists. The private sector had more homogeneous expansion, although also marked by non-attendance to quality standards. The analysis on the characteristics of the transformations of higher education indicate that in 2010, despite the policies adopted in the FHC and Lula governments, Brazil remains as an elite system (13,9%). Goiás is entering the system mass (16,3%) and Jataí, with the net rate of 20,93%, is closer to the mass, that predicts the rate of 30 %. The research points to the complex process of expansion and internalization, analyzes the singularities of higher education in Jataí and at the same time shows the need for studies that uncover singularities on this education level in the articulation process to the national policies. / O trabalho encontra-se vinculado à linha de pesquisa “Estado, Políticas e História da Educação” do Programa de Doutorado em Educação da UFG e ao Nedesc/UFG. Ao abordar o tema “a expansão e a interiorização da educação superior”, busca-se compreender as especificidades e a singularidade desse processo no município de Jataí, do estado de Goiás. Adotou-se como recorte temporal o período de 1985 a 2010, momento marcado pelas políticas de expansão e interiorização da educação superior brasileira, de Goiás e de Jataí. A centralidade das análises foi nos governos de FHC (1995-2002) e de Lula (2003-2010), destacando-se as legislações, os programas e as ações que contribuíram para a expansão brasileira, procurando instituir um contraponto para a reflexão e análise do movimento expansionista de Goiás e Jataí. Para o desenvolvimento da tese, realizaram-se pesquisa bibliográfica e documental, análise estatística e entrevistas, com 18 sujeitos. O referencial teórico se pautou nos estudos de: Amaral (2009); Catani e Oliveira (2002); Cunha (2003, 2004, 2006); Dourado (2001a, 2002, 2008, 2010, 2011); Frigotto (2009); Oliveira D. (2011a, 2011b); Oliveira J. (2000); Sguissardi (1993, 2004, 2006, 2008); Silva Jr. e Sguissardi (2001, 2005); Trow (2005), dentre outros. A análise dos indicadores centralizou-se nos dados relativos a IES, matrículas, cursos e vagas da graduação presencial, que dão expressão às categorias expansão e interiorização. As reflexões a respeito das características das transformações da educação superior pautaram-se no modelo teórico desenvolvido por Trow (2005), que caracteriza três fases: os sistemas de elite, de massa e de acesso universal. O campo de investigação da pesquisa concentrou-se no município de Jataí. Foram analisadas cinco IES de Jataí: CAJ/UFG; IFG – Câmpus Jataí; UEG – UnU de Jataí; Cesut e Faja. Investigou-se também a FEJ, fundação importante no processo de criação e expansão da educação superior de Jataí. O processo de expansão e interiorização da educação superior, tanto no Brasil quanto em Goiás e em Jataí, se dá com embate entre o público e o privado, com dinâmica própria. Assim como o Brasil e Goiás adotaram a política de diversificação e diferenciação na educação superior, Jataí seguiu essa tendência, mas com a singularidade marcada pelo predomínio da expansão pública. Houve uma expansão da educação superior pública significativa no município de Jataí, a partir da década de 1990, na contramão da lógica nacional e de Goiás, no entanto, com condições institucionais muito díspares, inclusive entre as públicas. A expansão e a interiorização das IES públicas CAJ/UFG e IFG – Câmpus Jataí pautaram-se em indicadores de qualidade, como: concurso público, regime de trabalho de 40 horas e DE, ênfase na formação docente (stricto sensu), infraestrutura física adequada. Em contrapartida, a expansão da UEG – UnU de Jataí revelou o não atendimento aos parâmetros básicos de qualidade, por meio de contrato de docentes temporários, horistas, onde a maioria é graduada ou especialista. O setor privado teve pequena expansão, demarcada também pelo não atendimento aos padrões de qualidade. As análises a respeito das características das transformações da educação superior sinalizam que, em 2010, apesar das políticas adotadas nos governos de FHC e de Lula, o Brasil permanece como um sistema de elite (13,9%). Goiás está entrando no sistema de massa (16,3%), e Jataí, com a taxa líquida de 20,93%, está mais próximo da massificação, que prevê o índice de 30%. A pesquisa sinaliza para o complexo processo de expansão e interiorização, analisa as singularidades da educação superior em Jataí e, sinaliza para a necessidade de estudos que descortinem singularidades desse nível de ensino no processo de articulação às políticas nacionais.

Este trabalho investigou o processo de emergência de inglês em crianças de dezoito meses a três anos de idade em ambiente escolar de imersão através de uma pesquisa etnográfica. Em escola internacional, esse grupo de sete crianças (cinco brasileiros e dois sino-brasileiros), observado entre fevereiro e junho de 2008, deu os primeiros passos para a internalização da língua adicional através da interação dialógica entre seus professores e coetâneos, principalmente no momento da roda. Neste contexto, buscamos entender melhor os conceitos vygotskianos de atividades inter- e intramentais e a importância da convivência social para a internalização de conhecimento. Consideramos a relevância do gesto como primeira forma de expressão extraverbal da criança. Para analisarmos melhor a interação dialógica entre os integrantes da sala de aula, procuramos beneficiar-nos da área de estudos linguísticos, especificamente o conceito discursivo de tutela reflexiva e linguística proposto por Bitar (2002) e como a interação em pares, aplicada à realidade da criança pequena, pode proporcionar a apropriação e a internalização da linguagem. Pesquisamos sobre a importância da brincadeira livre para os desenvolvimentos linguístico e cognitivo da criança, através de estudos de Brougère (1993, 1995, 1999, 2000, 2004, 2005) e Kishimoto (1994, 1996, 1998, 2001, 2007). Esta pesquisa de abordagem interdisciplinar busca benefícios das áreas da Teoria Sociocultural, da Linguística Pragmática e da Educação. Após uma análise dos dados coletados, a roda mostrou-se um momento que proporcionou uma vivência positiva no contínuo plurilíngue de emergência em inglês pelas crianças, mas ainda era totalmente dirigido pelo professor. Ao final do trabalho, propomos um momento prévio de livre exploração pela criança que sirva para a criação de uma roda mais participativa, ou seja, uma roda social, a favor de proporcionar uma vivência mais completa pela criança pequena na língua adicional. / This study investigated the emergency process of English by eighteen-month-old to three-year-old children in an immersive school environment through ethnographic research. In an international school, this group of seven children (five Brazilian and two Chinese-Brazilian), observed from February to June 2008, gave the first steps towards the internalization of an additional language through a dialogic interaction among teachers and colleagues, meanly during circle time. In this context, we attempted a better understanding of the Vygotskian concepts of inter and intramental activities and the importance of social contact for knowledge internalization. We considered the relevance of gestures as the first childs extraverbal expression. To better analyze the dialogic interaction among the classroom participants, we benefited from the linguistic studies, specifically the discursive concept of reflexive and linguistic tutelage proposed by Bitar (2002) and how the interaction in pairs, applying it to the young child, may enhance language learning. We researched the importance of free play for the linguistic and cognitive development of the child, through Brougéres (1993; 1995; 1999; 2000; 2004; 2005) and Kishimotos (1994; 1996; 1998; 2001; 2007) studies. This research adopts an interdisciplinary approach since it gathers benefits from Sociocultural Theory, Pragmatic Linguistics and Education. After the collected data analysis, circle time seemed to be a moment that promoted a positive experience along the emergency continuum in English by the children but it was still a guided moment by the teacher. By the end of this paper, we suggest a previous moment of free exploration by the child, that would help create a more dynamic circle time concerning participation, in other words, a social circle time, in favor of providing a deeper childs experience in the additional language.

En France, le tritium est principalement rejeté sous forme d’eau tritiée (HTO) par les centres de production d’électricité et les centres de traitement des combustibles en fonctionnement normal. Les effets développementaux et reprotoxiques du tritium ont été étudiés. Cependant, peu d’études portent sur les effets moléculaires. Dans ce contexte, l’objectif de cette thèse est d’évaluer les effets de l’HTO sur les stades embryo-larvaire du poisson zèbre (Danio rerio). Deux débits de dose, 0,4 et 4 mGy/h, ont été testés. Un protocole pour la mesure de l’activité dans les organismes a été mis au point. Il a permis (i) de confirmer que l’internalisation de l’HTO est rapide, (ii) la discrimination des formes de tritium libre dans les tissues et lié à la matière organique, et (iii) le calcul de la dose permettant de relier les effets observés à une dose reçue. Les effets de l’HTO ont été évalués à différents niveaux d’organisation biologique. A l’échelle moléculaire, une analyse transcriptomique (mRNAseq) a montré la modulation de gènes impliqués dans la contraction musculaire et le développement de l’œil à 24hpf ainsi que dans le cycle circadien et la réponse au stress oxydant à 96hpf. Une modulation de gènes impliqués dans la réparation des dommages à l’ADN a été montrée. A des niveaux d’organisation supérieurs, quelques altérations des fibres musculaires ont été observées pour les deux débits de dose. Une diminution de la vitesse de nage a été montrée à 96 hpf après exposition à 0,4 mGy/h. Ces travaux ont permis de mieux appréhender l’internalisation du tritium, de caractériser la dose absorbée dans les organismes et de mieux comprendre les effets du tritium / In France, tritium is mainly released as tritiated water (HTO) by nuclear power plants and nuclear reprocessing plants. The developmental and reprotoxic effects of tritium have already been studied. However, few studies focus on molecular effects. In this context, the aim of this thesis is to evaluate the effects of HTO on the embryo-larvae stages of the zebrafish (Danio rerio). Two dose rates, 0.4 and 4 mGy/h, were tested. A protocol for the measurement of activity in organisms was developed. It has (i) confirmed that the internalization of HTO is rapid, (ii) allowed the discrimination of the tissue-free-water-tritum and organically bound tritium forms, and (iii) allowed the calculation of the dose permitting to link the observed effects to a received dose. The effects of HTO were evaluated at different biological levels. At the molecular level, a transcriptomic analysis (mRNAseq) showed the modulation of genes involved in muscle contraction and eye development at 24hpf, as well as in the circadian rythm and the response to oxidative stress at 96hpf. Modulation of genes involved in the DNA damage repair was shown. At higher levels of organization, some alterations of muscle fibers were observed for both dose rates. A decrease in swimming velocity was shown at 96 hpf after exposure to 0.4 mGy/h. This work allowed a better understanding of the tritium internalization, to characterize the absorbed dose in organisms and to better understand the effects of tritium

Obligate intracellular pathogens must invade host cells to survive and pose a global health risk. As such, internalization is a critical life stage and represents an excellent therapeutic target. Oxidoreductase exploitation is a thematic invasion strategy among obligate intracellular pathogens. Delineating the mechanisms and proteins mediating this exploitation could identify novel therapeutic targets for many important pathogens. Anaplasma phagocytophilum infects neutrophils by an incompletely defined mechanism, resulting in the emerging potentially fatal disease, human granulocytic anaplasmosis. The bacterial adhesin, Asp14, contributes to invasion by virtue of its C-terminus engaging an unknown receptor. Yeast two-hybrid analysis identified protein disulfide isomerase (PDI) as a putative Asp14 binding partner. Co-immunoprecipitation confirmed this interaction and identified the Asp14 C-terminus as critical to it. PDI reductase activity inhibition impaired bacterial infection of, but not binding to, host cells. A. phagocytophilum failed to productively infect myeloid-specific PDI conditional knock-out mice. This is the first demonstration of microbial PDI exploitation in vivo. Infection of PDI inhibited cells was rescued when bacterial, but not host surfaces were reduced with the reducing agent tris(2-carboxyethyl)phosphine (TCEP). Furthermore, TCEP restored bacterial infectivity after Asp14 inhibition using an antibody that reduces infection. Mutational analyses identified Asp14 residues critical for binding PDI. These data demonstrate that Asp14 binds and brings PDI to disulfide bonds within A. phagocytophilum surface protein(s) that it reduces, enabling infection. Targeting the Asp14 C-terminus could benefit approaches to prevent/treat granulocytic anaplasmosis. A similar approach would identify proteins from other obligate intracellular pathogens that could prove to be protective targets.