Listeria monocytogenes est une bactérie pathogène responsable de la listériose chez l’homme. Cette bactérie est capable d’envahir et de se répliquer dans des cellules phagocytaires et non-phagocytaires de l’hôte. De fait, Listeria interagit et interfère avec de nombreux composants cellulaires au cours de sa réplication. Mon projet de thèse s’est focalisé sur les altérations du protéome des cellules infectées par Listeria. Je me suis en particulier intéressé à l’activation de protéases de l’hôte et aux dégradations de certains facteurs cellulaires en réponse à l’infection.Une analyse protéomique de cellules humaines traitées avec la Listeriolysine O (LLO), une toxine secrétée par Listeria formant des pores dans les membranes cellulaires, nous a permis d’identifier plusieurs dizaines de facteurs cellulaires dégradés en réponse à l’exposition à cette toxine.Nous avons pu confirmer la dégradation de ces protéines dans le contexte d’une infection par Listeria in vitro (dans un modèle de cellules en culture) et in vivo (dans un modèle animal). Nous anticipons que la dégradation de ces protéines, impliquées dans différentes fonctions cellulaires, joue un rôle au cours de l’infection par Listeria en modifiant la physiologie des cellules hôte.Nous avons en parallèle mis en évidence que Listeria interfère avec le fonctionnement des lysosomes. Nous avons montré que la LLO déstabilise les lysosomes des cellules hôtes et induit leur perméabilisation. Ceci permet la libération de certaines protéases lysosomales, telles que les cathepsines, dans le cytosol des cellules hôte. Ces cathepsines, une fois libérées, pourraient alors cliver différentes protéines cytosoliques et ainsi modifier la physiologie des cellules infectées.L’ensemble de mes travaux a permis de montrer que Listeria interfère avec le fonctionnement de nombreuses protéases de l’hôte et induit la dégradation de plusieurs facteurs cellulaires. Ces cibles dégradées constituent une nouvelle classe de protéines dont nous pouvons maintenant tester le rôle potentiel dans l’infection. Mes travaux ont d’autre part permis de montrer que d’autres toxines bactériennes, similaires à la LLO, induisent également des altérations du protéome des cellules hôtes. Ceci suggère que les mécanismes identifiés avec Listeria sont conservés entre différentes classes de pathogènes bactériens. / Listeria monocytogenes is a foodborne pathogen responsible for human listeriosis which invade and replicate in both phagocytic and non-phagocytic cells. Listeria intracellular life cycle involves interference with host cell components. My PhD project aims to characterize host cell proteome modifications in cells infected by Listeria. I focused my research on host proteases activation and the degradation of host cell proteins in response to infection. A proteomic analysis performed on cells treated with purified Listeriolysin O (LLO), a pore forming toxin secreted by Listeria, identified more than 90 proteins degraded in response to the toxin. We validated the degradation of these proteins using both in vitro and in vivo models of Listeria infection. We anticipate that the degradation of these proteins can significantly impact the infection process through the modification of host cell physiology. In parallel, we identified that Listeria impairs lysosomal functions. We demonstrate that extracellular Listeria, via LLO secretion, alter lysosomal integrity in epithelial cells. LLO induces lysosomal membrane permeabilization and the release of lysosomal proteases in the host cytosol. The release of such proteases, which remain transiently active, may alter cellular physiology by degrading different cytosolic factors. Altogether, my results highlight how Listeria reshapes the host proteome by altering the activation or localization of host proteases and by inducing protein degradations. My data also establish that other bacterial toxins close to LLO also induce proteome modifications, thus unveiling that these mechanisms are shared among different class of bacterial pathogens.

Submitted by Carla Santos (biblioteca.cp2.carla@bahiana.edu.br) on 2018-10-19T17:54:47Z No. of bitstreams: 1 Disserta????o vers??o FINAL Marina Ferraz (3).pdf: 1502315 bytes, checksum: e98dc9fa59178eb9cbf8525fb6e678e7 (MD5) / Approved for entry into archive by JOELMA MAIA (ebmsp-bibliotecacp2@bahiana.edu.br) on 2018-10-19T19:08:14Z (GMT) No. of bitstreams: 1 Disserta????o vers??o FINAL Marina Ferraz (3).pdf: 1502315 bytes, checksum: e98dc9fa59178eb9cbf8525fb6e678e7 (MD5) / Made available in DSpace on 2018-10-19T19:08:14Z (GMT). No. of bitstreams: 1 Disserta????o vers??o FINAL Marina Ferraz (3).pdf: 1502315 bytes, checksum: e98dc9fa59178eb9cbf8525fb6e678e7 (MD5) Previous issue date: 2017-10-30 / este estudo objetiva avaliar a fun????o da m??o em pacientes com Mucopolissacaridose (MPS). M??todos: estudo anal??tico cross-seccional em dois grupos com idades de 8 e 21 anos: pacientes com MPS e indiv??duos sem MPS. Dados cl??nicos e demogr??ficos foram coletados de 15 indiv??duos com MPS e de 44 participantes sem MPS. A avalia????o da fun????o da m??o foi baseada na vers??o brasileira do MHQ. Para avaliar essa fun????o, foram apresentadas a pontua????o global, assim como as pontua????es de cada dom??nio, considerando os grupos. Resultados: a m??dia de idade do grupo MPS foi de 11,73 (4,39) e a idade do grupo comparativo foi 12,80 (3,96). Todos os indiv??duos avaliados foram capazes de completar o question??rio MHQ. O grupo sem MPS teve suas principais caracter??sticas sociodemogr??ficas semelhantes ao grupo sem MPS. O grupo com MPS demonstrou comprometimento importante da fun????o da m??o. O dom??nio Fun????o da m??o direita de 66,00 ?? 23,05 (versus 93,18 ??11,46); Fun????o da m??o esquerda 62,33 ?? 20,86 (versus 83,91 ?? 17,58); AVD conjunto 45,60 ?? 29,88 (versus 95,78 ?? 8,49); Escola 56,00 ?? 32,68 (versus 95,68 ?? 11,08); Dor 20,67 ?? 30,87 (versus 98,77 ?? 3,58); Apar??ncia m??o direita 77,80 ?? 11,16 (versus 71,59 ?? 21,37); Apar??ncia m??o esquerda 77,80 ?? 11,16 (versus 71,51 ?? 20,92); Satisfa????o m??o direita 74,47 ?? 27,12 (versus 94,21 ?? 14,94); Satisfa????o m??o esquerda 74,67 ?? 27,24 (versus 98,77 ?? 3,59). Conclus??o: indiv??duos afetados pela MPS apresentam pontua????es mais baixas na avalia????o da fun????o da m??o, atividades da vida di??ria, atividades escolares e dor. Apenas nos dom??nios de Est??tica e Apar??ncia os grupos possu??ram resultados similares. Trata-se de uma avalia????o de m??o com um question??rio subjetivo de aplicabilidade ampla, validado para portadores de MPS e com grupo compara????o usando popula????o an??loga.

L'insuffisance cardiaque (IC) touche 1-2% des adultes dans les pays développés. On distingue classiquement 2 formes d’IC : l’IC chronique, dont la mise en place est lente, et l’IC aigue dont l’apparition est rapide ou due à l'aggravation des symptômes et/ou des signes de l’IC. Quelles soient chroniques ou aigues, certaines formes d'IC n'ont pas de traitement efficace. L'insuffisance cardiaque à fraction d'éjection préservée (ICFEp) devient la forme prédominante d'IC chronique mais la compréhension de sa physiopathologie reste encore incomplète. Un modèle de rat transgénique sur-exprimant le récepteur β3- adrénergique humain au niveau endothélial a été développé dans l'équipe. Comme les patients, ce modèle développe une ICFEp avec l’âge qui se manifeste notamment par une dysfonction diastolique et une intolérance au stress. Dans le cadre de ma thèse et sur la base d'une étude transcriptomique, j'ai évalué les altérations du cycle du calcium, du métabolisme ainsi que l'apparition de fibrose et d'inflammation. Le choc septique est considéré comme une IC aigue particulièrement violente. Le deuxième axe de ma thèse a été de développer des modèles pertinents de choc septique et d’évaluer le potentiel thérapeutique d’une modification post-traductionnelle : la O-GlcNAcylation, déjà connue pour ses effets bénéfiques dans des formes d’IC aigues. Ainsi, une augmentation des niveaux de l'O-GlcNAc, grâce à un agent pharmacologique, le NButGT, améliore la fonction cardiovasculaire et la survie des animaux en choc septique. / Heart failure (HF) affects 1-2% of adults in developed countries. There are two distinct forms of HF: chronic HF, with a slow development, and acute HF that appears rapidly or is due to an aggravation of the symptoms and/or signs of HF. For some forms of HF, acute or chronic, no effective treatment exists. Heart failure with preserved ejection fraction (HFpEF) is becoming the predominant form of chronic HF but the understanding of its pathophysiology remains incomplete. A transgenic rat model over-expressing the human endothelial β3-adrenergic receptor has been developed in the team. As in patients, this model develops HFpEF with age, and is characterized by diastolic dysfunction and stress intolerance. During my thesis and based on transcriptome data, I evaluated alterations of the calcium cycle, metabolism as well as the appearance of fibrosis and inflammation. Septic shock is considered to be a form of severe acute HF. The second axis of my thesis was to develop relevant models of septic shock and to evaluate the therapeutic potential of a post-translational modification: O-GlcNAcylation, already known for its beneficial effects in acute HF forms. Indeed, an increase in levels of O-GlcNAc, through a pharmacological agent, NButGT, improved cardiovascular function and survival of animals in septic shock.

Thesis (doctoral)--Technische Hochschule Karlsruhe, 1904. / Includes bibliographical references.

Thesis (M.A.) B.Y.U. Religion Dept. / Electronic thesis. Also available in print ed.

Thesis (M.A.) B.Y.U. Religion Dept.

Les cellules humaines sont soumises à des stress induisant des cassures double-brin de l’ADN principalement réparées par la voie NHEJ, où la kinase DNA-PKcs joue un rôle central. L’activité de DNA-PKcs, régulée par de nombreuses phosphorylations, est cruciale pour le maintien de l’intégrité génomique. Plus récemment, il a été montré que cette protéine était également modifiée par l’O-GlcNAcylation dans la lignée COS7. Sachant l’équilibre existant entre phosphorylation et O-GlcNAcylation, nous avons étudié le rôle de cette nouvelle MPT dans la régulation de l’activité de DNA- PKcs. Nous avons montré que DNA-PKcs est O-GlcNAcylée dans les cellules HeLa. Puis nous avons montré que la modulation de l’O-GlcNAcylation de DNA-PKcs impacte son autophosphorylation en Ser2056, suggérant l’existence d’une balance O-GlcNAcylation /phosphorylation, ainsi que la capacité des cellules à réparer les DSBs par la voie NHEJ. De plus, nos résultats nous laissent envisager que cette modification puisse jouer un rôle dans la stabilité de la protéine. DNA-PKcs est une cible potentielle dans les stratégies de lutte contre le cancer. Nous avons étudié l’impact d’un composé sur DNA-PKcs. Cette molécule provoque une réduction de la quantité et de l’activité de DNA-PKcs, impliquant son ubiquitinylation et sa dégradation par le protéasome et menant à une sensibilisation des cellules à un traitement génotoxique. Dans ce contexte, nous avons développé une puce à anticorps pour évaluer le profil phosphoprotéique des voies de réparation de l’ADN et ainsi évaluer l’effet d’inhibiteurs de DNA-PKcs. L’ensemble de ces résultats contribuent à une meilleure compréhension de la régulation de DNA-PKcs. / Human cells are subjected to stresses inducing DNA double-strand breaks mainly repaired by the NHEJ pathway, where the kinase DNA-PKcs plays a central role. The activity of DNA-PKcs, regulated by numerous phosphorylations, is crucial for the maintenance of genomic integrity. More recently, it has been shown that this protein is also modified by O-GlcNAcylation in the COS7 cell line. Knowing the balance between phosphorylation and O-GlcNAcylation, we studied the role of this new PTM in the regulation of DNA-PKcs activity. We have shown that DNA-PKcs is O-GlcNAcylated in HeLa cells. We then showed that the modulation of DNA-PKcs O-GlcNAcylation affects its autophosphorylation on Ser2056, suggesting an O-GlcNAcylation/phosphorylation balance, as well as the ability of cells to repair DSBs by NHEJ pathway. Moreover, our results allow us to consider that this modification may play a role in protein stability. DNA-PKcs is a potential target in anticancer strategies. We studied the impact of a chemical compound on DNA-PKcs activity. This molecule causes a reduction in the amount and activity of DNA-PKcs, through its ubiquitinylation and its degradation by the proteasome and leading to sensitization of the cells to genotoxic treatment. In this context, we have developed an antibody microarray to evaluate the phosphoprotein level of DNA repair pathways and thus estimate the effect of DNA-PKcs inhibitors. All these results contribute to a better understanding of the regulation of DNA-PKcs.

Submitted by Sara Ribeiro (sara.ribeiro@ucb.br) on 2017-11-10T12:49:11Z No. of bitstreams: 1 ElianeFernandesLim??oDissertacao2017.pdf: 3384824 bytes, checksum: 48859c5e4218ac293f1014a67509fe14 (MD5) / Approved for entry into archive by Sara Ribeiro (sara.ribeiro@ucb.br) on 2017-11-10T12:49:36Z (GMT) No. of bitstreams: 1 ElianeFernandesLim??oDissertacao2017.pdf: 3384824 bytes, checksum: 48859c5e4218ac293f1014a67509fe14 (MD5) / Made available in DSpace on 2017-11-10T12:49:36Z (GMT). No. of bitstreams: 1 ElianeFernandesLim??oDissertacao2017.pdf: 3384824 bytes, checksum: 48859c5e4218ac293f1014a67509fe14 (MD5) Previous issue date: 2017-08-17 / The primary aim of this dissertation is to analyze the implementation and results of a differentiated and assisted assessment system implemented by Escola Amarela, located in Cidade Ocidental, Goi??s, seeking to contribute to and enable the development and learning of students who have been medically identified as having learning disabilities. A convenience sample consisting of 1 pedagogical coordinator and 5 teachers from private schools, 5 parents of students, and 3 specialists participated in the research. The study employed a qualitative approach and a participatory action research method. A case study technique with document analysis and semi -structured interviews was used for data collection. The results were analyzed through content analysis with the identification of thematic categories. The literature review addressed the concepts of learning disabilities and disorders, especially ADHD (Attention Deficit Hyperactivity Disorder), the teaching-learning relationship in Vygotsky's socio-historical theory, the medicalization of students with ADHD, and the assisted assessment of students with learning problems and/or medical conditions in the schooling process. The study suggests that the conceptions and practices of parents, teachers, and specialists regarding learning problems reinforce the process of medicalization in childhood. It is thus necessary to strengthen continuing education for teachers in order to enable a better understanding of the educational process, especially assisted assessment. With regard to parents, it is important to reinforce their participation and responsibility in the process of schooling their children and to promote a more effective communication between parents and schools. As for the specialists, a closer relationship with the schools would be a means of strengthening the professional ties among the pedagogical coordination, teachers, and specialists, so that they have a better understanding of the development and school achievement of the students and their evolution, allowing for a critical discussion about the medicalization of childhood and its effects on the subj ects' learning and developmental processes. / A presente disserta????o tem por objetivo principal analisar a implementa????o e os resultados de um sistema de avalia????o diferenciado e assistido implementado pela Escola Amarela, situada em Cidade Ocidental, Goi??s, visando a contribuir e a oportunizar o desenvolvimento e o aprendizado de alunos identificados com dificuldades de aprendizagem, com laudos m??dicos. Participaram da pesquisa 1 coordenador pedag??gico e 5 professores da rede privada de ensino, 5 pais de alunos e 3 especialistas, sendo que a escolha dos sujeitos se deu por amostra de conveni??ncia. A pesquisa se utilizou da abordagem qualitativa e o m??todo foi a pesquisa -a????o, de car??ter participativo, com a t??cnica do estudo de caso, tendo como procedimentos de coleta de dados a an??lise documental e a entrevista semiestruturada. Os resultados foram analisados por meio da an??lise de conte??do, com a elabora????o de categorias tem??ticas. A revis??o de literatura discutiu os conceitos de dificuldades de aprendizagem e de dist??rbio da aprendizagem, sobretudo o TDA/H (transtorno de d??ficit de aten????o e hiperatividade), a rela????o ensino e aprendizagem na teoria s??cio-hist??rica de Vygotsky, a medicaliza????o de alunos com TDA/H e a avalia????o assistida dos alunos com problemas de aprendizagem e/ou com laudos m??dicos no processo de escolariza????o. Concluiu-se, por meio da pesquisa, que as concep????es e pr??ticas de pais, professores e especialistas acerca dos problemas de aprendizado refor??am o processo de medicaliza????o na inf??ncia. ?? preciso, pois, fortalecer a forma????o continuada aos professores para possibilitar melhor compreens??o do processo educacional, especialmente da avalia????o assistida. Em rela????o aos pais, cabe refor??ar a sua participa????o e responsabilidade no processo de escolariza????o dos filhos e implementar uma comunica????o mais eficaz entre pais e escola. Em rela????o aos especialistas, uma maior aproxima????o com as escolas seria um meio de estreitar as rela????es profissionais entre coordena????o pedag??gica, professores e especialistas, de modo a que estes tenham uma melhor compreens??o sobre o desenvolvimento e o desempenho escolar dos alunos e sua evolu????o, oportunizando uma discuss??o cr??tica sobre a medicaliza????o na inf??ncia e os seus efeitos na subjetividade e nos processos de desenvolvimento e de aprendizagem dos sujeitos.

Submitted by Automa??o e Estat?stica (sst@bczm.ufrn.br) on 2015-11-25T20:56:13Z No. of bitstreams: 1 TarcilaMariaPinheiroFrota_DISSERT.pdf: 2847806 bytes, checksum: 145d4638a1cda51760a6a1f51ae48737 (MD5) / Approved for entry into archive by Elisangela Moura (lilaalves@gmail.com) on 2015-11-25T22:37:39Z (GMT) No. of bitstreams: 1 TarcilaMariaPinheiroFrota_DISSERT.pdf: 2847806 bytes, checksum: 145d4638a1cda51760a6a1f51ae48737 (MD5) / Made available in DSpace on 2015-11-25T22:37:39Z (GMT). No. of bitstreams: 1 TarcilaMariaPinheiroFrota_DISSERT.pdf: 2847806 bytes, checksum: 145d4638a1cda51760a6a1f51ae48737 (MD5) Previous issue date: 2009-07-24 / Durante o processo de produ??o de petr?leo ? comum o aparecimento de g?s e ?gua associados. A produ??o pode ser comprometida pela precipita??o de sais inorg? nicos tanto no reservat?rio como no po?o produtor, atrav?s da forma??o de incrusta??o , e ? apontada como prov?vel causa ao dano da forma??o. Nas condi??es de alta temperatura e alta press?o (HTHP), a precipita??o de sais insol?veis pode ser favorecida. Os tipos de incrusta??es mais comuns em campos de petr?leo s?o o carbonato de c?lcio e os sulfatos de c?lcio, estr?ncio e b?rio. Atualmente, novos tipos de incrusta??o v?m despertando aten? ?o especia l como o sulfeto de zinco e de chumbo . Esta s precipita? ?es p ode m ocorrer nos poros da rocha - reservat?rio, na coluna de produ??o e nos equipamentos , provocando obstru??es e a conse qu ente perda da produ??o . Neste trabalho, estudou - se a influ ? ncia da profundidade, press?o e temperatura na composi??o das incrusta??es f ormadas, visando a otimiza??o de tratamentos de remo??o e seu comportamento ao longo da coluna. Foram selecionados 10 res?duos de uma coluna de produ??o (intervalo de 15,4 m a 4061,5 m), que apresentava problemas de corros?o/incrusta??o. Realiza ram - se ensa io s de dissolu??o em ?cidos org?nicos similares aos empregados em opera??es de remo??o desse tipo de incrusta??o. A composi??o majorit?ria foi definida e confirmada p elos testes de dissolu??o e por t?cnicas de Difratometria de Raios - X ( DRX ) , Espectrometria de Fluoresc?ncia de Raios - X ( EFRX ), Microscopia Eletr?nica de Varredura (MEV) . Foram observados res?duos com caracter?sticas distintas, em propor??es diferentes, mostrando uma tend?ncia de aumento e/ou diminui??o de massa com a profundidade. Os resultados obtidos indicam correla??es dos tipos de incrusta??es estudados com a profundidade e, consequentemente, com as condi??es termodin?micas de press?o e temperatura. / Water and gas is a common by - product of the oil production process. Production may be compromised by the precipitation of inorganic salts in both the reservoir and producing well, through scale formation. This precipitation is likely the cause of the formation damage. High temperatures and h igh pressures (HTHP) may favor the precipitation of insoluble salts. The most common types of scale in oil fields are calcium carbonate and calcium sulphate, strontium and barium sulphate. New types of scale formation have attracted special attention such as zinc sulphide and lead. This precipitation may occur in the pores of reservoir rocks, in the production string and in equipment, causing obstructions and consequent production losses. In this study, the influence of well depth on incrustation compositio n was investigated to design removal treatments and assess the behavior of these deposits along the string, through the analysis of pressure and temperature. Scale residues were recovered from the inside of the production string of an oil and gas well duri ng the string removal operation. A total of 10 samples from different depths (15.4 m to 4061.5 m) were obtained. Initially a dissolution test was conducted in weak acid, similar to that used in removal operations with this type of scale formation. Majority composition was defined and confirmed by dissolution tests using X - Ray Fluorescence Spectroscopy (XRF), X - Ray Diffraction (XRD) and Scanning Electron Microscope (SEM) techniques. Residues with distinct characteristics were observed in different proportion s, showing a tendency toward increased and/or decreased mass with depth. In the samples closest to the surface, typical sandstone residues were found, with calcium (45% Ca) as the metal of highest concentration. The obtained results indicate correlations o f the scale types studied with the depth and, consequently, with the thermodynamic conditions of pressure and temperature.

yes / Cardiovascular diseases (CVDs) are the leading cause of death globally. While the major focus of pharmacological and non-pharmacological interventions has been on targeting disease pathophysiology and limiting predisposing factors, our understanding of the cellular and molecular mechanisms underlying the pathogenesis of CVDs remains incomplete. One mechanism that has recently emerged is protein O-GlcNAcylation. This is a dynamic, site-specific reversible post-translational modification of serine and threonine residues on target proteins and is controlled by two enzymes: O-linked β-N-acetylglucosamine transferase (OGT) and O-linked β-N-acetylglucosaminidase (OGA). Protein O-GlcNAcylation alters the cellular functions of these target proteins which play vital roles in pathways that modulate vascular homeostasis and cardiac function. Through this review, we aim to give insights on the role of protein O-GlcNAcylation in cardiovascular diseases and identify potential therapeutic targets in this pathway for development of more effective medicines to improve patient outcomes.