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No. of bitstreams: 2 Dissertação - Thimoteo Pereira Cruz - 2015.pdf: 1759692 bytes, checksum: 9a1c443cc287d952ad07bd3750bc78b2 (MD5) license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) Previous issue date: 2015-12-22 / This research talks about the formation in psychology based in life story of three graduates psychologists at the Universidade Federal de Goiás, Regional Catalão (UFG/RC). We propose thinking the formation based on the question: How did the formation of the psychologist at UFG/RC, in the perspective of egresses. The general objective is to analyze the formation of the psychologists at UFG/RC, from 2007 to 2014, in the perspective of psychology degree egresses. The specific objectives are: 1 - to understand how the process of formation lifelong and formation of psychologists is; 2 - analyze experiences lived lifelong of the egresses that reverberate in their professional formation; 3 - discuss what tell egresses from psychology degree on formation experiences and how mean the formation of psychologists. To execute this research, we use the method of life stories, which includes subjective processes as a source for knowledge production. We did semi-structured interviews and autobiographical writings to highlight the formation pathways and life experiences of three egresses of Psychology Degree of UFG/RC. In the data analysis, we used the thematic analysis proposed by Bertaux (2005). In general lines, it was possible to perceive that the formation of psychologists in UFG/RC afford a diversity of experience and knowledge that allows the student to cycle through different trajectories and to elaborate concepts and modes of thinking and acting, according as it experiencing the knowledge and learning devices at your disposal. / Esta investigación aborda la formación en psicología basada en la historia de vida de tres psicólogos egresados de la Universidade Federal de Goiás – Regional Catalão (UFG/RC). Proponemos pensar la formación basada en la siguiente cuestión: ¿Cómo sucede la formación del psicólogo de la UFG/RC, en la perspectiva de los egresados? El objetivo general es analizar la formación del psicólogo de la UFG/RC, de 2007 a 2014, en la perspectiva de los egresados de lo curso en Psicología. Los objetivos específicos son: 1 - entender cómo es el proceso de formación durante toda la vida y de la formación de psicólogos; 2 - analizar las experiencias vividas a lo largo de la vida de los graduados que reverberan en su formación profesional; 3 - discutir lo que cuentan los egresados del curso de la psicología de las experiencias de formación y cómo significan la formación de los psicólogos. Para hacer esta investigación, utilizamos el método de las historias de vida, que incluye procesos subjetivos como fuente para la producción de conocimiento. Hicimos entrevistas semi-estructuradas y escritos autobiográficos para destacar los itinerarios de formación y experiencias de vida tres egresados de lo curso de Psicología de la UFG/RC. En el análisis de los datos, se utilizó el análisis temático propuesto por Bertaux (2005). En términos generales, comprendemos que la formación de lo psicólogo en UFG/RC ofrece una diversidad de experiencias y conocimientos que permite a los estudiantes ir a través de diversos caminos y desarrollar conceptos y formas de pensar y de actuar, en la medida en que experimenta los dispositivos de conocimiento y de aprendizaje a su disposición. / Esta pesquisa trata da formação em psicologia a partir da história de vida de três psicólogos/as graduados/as na Universidade Federal de Goiás – Regional Catalão (UFG/RC). Propomos pensar a formação tendo como base a seguinte questão: Como se deu a formação do psicólogo na UFG/RC, na perspectiva dos/as egressos/as. O objetivo geral é analisar a formação do psicólogo na UFG/RC, no período de 2007 a 2014, na perspectiva dos/as egressos/as do curso de psicologia. Os objetivos específicos são: 1 -compreender como ocorre o processo de formação ao longo da vida e de formação do/a psicólogo/a; 2 - analisar experiências vivenciadas ao longo da vida dos/as egressos/as que reverberam na sua formação profissional; 3 - discutir o que contam os/as egressos/as do curso de psicologia sobre as experiências de formação e como significam a formação do psicólogo. Para realizar esta pesquisa, utilizamos o método das histórias de vida, que inclui os processos de subjetivação como fonte para produção de conhecimento. Realizamos entrevistas semiestruturadas e escritas autobiográficas para ressaltar os percursos de formação e as experiências de vida de três egressos/as do Curso de Psicologia da UFG/RC. Na análise dos dados, empregou-se a análise temática proposta por Bertaux (2005). Em linhas gerais, foi possível perceber que a formação do psicólogo na UFG/RC proporciona uma diversidade de experiências e de conhecimentos que permite ao aluno percorrer trajetórias diversas, bem como elaborar concepções e modos de pensar e agir, na medida em que experimenta os dispositivos de conhecimento e aprendizagens à sua disposição.

Le virus de l’hépatite C (VHC) dépend de facteurs cellulaires pour accomplir son cycle viral et persister dans l’hôte. L’une des stratégies de notre laboratoire consiste à étudier de manière approfondie le réseau d’interactions virus-hôte, afin d’identifier de nouvelles cibles thérapeutiques cellulaires et de développer des antiviraux plus efficaces pour vaincre la résistance virale. Durant ma thèse j’ai étudié deux facteurs cellulaires importants pour le VHC. Le premier est la protéine ribosomale RACK1. Nous avons montré que cette protéine est spécifiquement requise pour la traduction IRES-dépendante du VHC, et non pour la traduction coiffe-dépendante. Le deuxième facteur est une protéine de surface des gouttelettes lipidiques appelée TIP47. Nous avons montré que cette protéine est importante à la fois pour l’assemblage et pour l’export des particules virales. L’ensemble de ces travaux montre que de nouvelles cibles thérapeutiques pourraient être envisagées pour lutter contre le VHC. / The hepatitis C virus (HCV) relies on cellular factors to complete its life cycle and persist in its host. One of the strategies employed by our laboratory is the in-depth study of the network of virus-host interactions to identify new therapeutic cellular targets and develop more effective antivirals to overcome viral resistance.During my PhD, I studied two cellular factors involved in the HCV life cycle. The first factor is the ribosomal protein RACK1. We have shown that this protein is specifically required for the HCV IRES-mediated translation but not for the cap-mediated translation. The second factor is the lipid droplets binding protein TIP47. We have shown that this protein is important for both assembly and export of viral particles. This work shows that new therapeutic targets could be considered in the fight against HCV.

O objetivo do presente estudo é analisar as percepções relacionadas ao impacto gerado pelo mestrado profissional no desempenho profissional e na carreira dos egressos. Para tanto, foi realizado um estudo no Programa de Mestrado Profissional em Gestão e Desenvolvimento Regional (MGDR) da Universidade de Taubaté (UNITAU), compreendendo o período desde a sua criação no ano 2000 até a última turma formada, que concluiu o mestrado em 2011, sendo a população composta por 253 alunos e por 14 docentes, dos quais 5 são coordenadores no programa. A pesquisa é do tipo descritiva com abordagem quantitativa e qualitativa e com delineamento de pesquisa de campo. Foi realizada uma análise documental dividida em duas partes. A primeira parte destinada à apresentação do MGDR e a segunda parte, composta por dados sócio-demográficos dos egressos de todas as turmas do MGDR. Foram distribuídos questionários a toda população docente e discente, dos quais foram obtidas respostas de 8 docentes e 50 discentes, compondo-se, portanto, a amostra por acessibilidade, de acordo com a disponibilidade dos participantes de responder o questionário. Foram realizadas entrevistas semi-estruturadas com 3 coordenadores, e com uma amostra de 4 discentes. Para análise dos dados das entrevistas foi utilizado o método da análise de conteúdo. Para análise dos dados obtidos via questionários foram utilizadas técnicas quantitativas como tabulação e análise percentual. Resultados permitem constatar que as percepções de impactos puderam ser classificadas entre os níveis Bom e Muito Bom, demonstrando, de maneira geral, que os egressos percebem bons impactos em suas carreiras e desempenho após a conclusão do mestrado. Os docentes mostraram-se, de maneira geral, mais otimistas que os discentes em relação aos impactos do mestrado. As questões apresentaram percepção de importância equivalente ou superior ao grau `Importante , confirmando a relevância dos critérios adotados. / The aim of this study is to analyze the perceptions related to the impact generated by the professional master on the performance and career of its graduates. Therefore, a study was conducted in the Professional Master Program in Management and Regional Development (MMRD) from the University of Taubate (UNITAU), covering the period from its inception in 2000 until the last graduating class that completed the Masters in 2011 and the population was composed of 253 students and 14 teachers, of which 5 are program coordinators. The research is descriptive with a quantitative and qualitative approach and has a field research design. An documental analysis was made, which was divided into two parts. The first part was devoted to the presentation of MMRD and the second part, consisting of socio-demographic data of graduates from all classes of MMRD. Surveys were distributed to all teachers and student population, of which responses were received from 8 teachers and 50 students, composing, therefore, the sample by accessibility, according to the availability of the participants to answer the survey. Semi-structured interviews were conducted with 3 coordinators, and with a sample of 4 students. To analyze the data of the interviews the content analysis method were used. To analyze the data obtained by surveys quantitative techniques as tabulation and percentage analysis were used. Results assisted to determine that the perceptions of impacts could be classified between Good and Very Good levels, showing that, in general, the graduates perceive good impacts on their careers and performance after finishing the course. Teachers showed up, in general, more optimistic than the students about the impacts of the masters. The issues presented perceived importance degree equivalent or superior to `Important, confirming the relevance of the criteria.

Der cGMP-Signalweg ist als einer der Hauptregulatoren von diversen Funktionen in Eukaryoten bekannt; allerdings ist seine Funktionsweise in Protozoen wenig verstanden. Im Rahmen dieser Arbeit wurde eine Guanylatcyclase, gekoppelt mit N-terminalen P4-ATPase, in intrazellulären Parasiten Toxoplasma gondii gemeldet. Eine in silico-Analyse wies auf eine Aktivierung der Guanylatcyclase durch Heterodimerisierung ihrer Cyclasedomänen hin und ermöglichte wertvolle Einsichten in mögliche Funktionen ihrer ATPase-Domäne. Dieses Protein (477-kDa) bezeichnet als TgATPaseP-GC in dieser Studie, lokalisiert in der Plasmamembran am apikalen Pol des Parasiten. TgATPaseP-GC ist unempfänglich gegenüber genetischer Deletion und seine CRISPR/Cas9 unterstützte Spaltung beendet den lytischen Zyklus von T. gondii vorzeitig. Darüber hinaus reduzierte ein Cre/loxP-vermittelter Knockdown von TgATPaseP-GC die Synthese von cGMP im Tachyzoiten und inhibierte das Parasitenwachstum aufgrund von Beeinträchtigungen Motilitäts-abhängiger Prozesse des Austretens und Eindringens. Trotz seiner zeitlich beschränkten Funktion ist TgATPaseP-GC konstitutiv während des ganzen lytischen Zyklus exprimiert, welches eine post-translationale Regulierung des cGMP-Signalweges bedingt. Nicht zuletzt impliziert das Vorhandensein von TgATPaseP-GC-Orthologen in anderen Alveolata eine divergente Umfunktionierung der cGMP-Signalwege in Protozoen. Darüber hinaus wurde ein optogenetischer Ansatz verwendet, um den cGMP-Weg durch eine photo-aktivierte Rhodopsin-Guanylat-Cyclase (RhoGC) in T. gondii zu exprimiert. Dieses System erlaubte eine kontrollierte Erhöhung von cGMP durch Licht in einer schnellen und reversiblen Weise. Die Anregung von RhoGC stimulierte signifikant die Parasitenmotilität, deren Auswirkung auch mit erhöhten Eindringen und Austreten überwacht wurde; im Gegensatz zum genetischen Knockdown von TgATPaseP-GC. Das System ermöglicht die Vermittler des cGMP-Signalwegs durch Phosphoproteomics zu identifizieren. / cGMP signaling is known as one of the master regulators of diverse functions in eukaryotes; however, its architecture and functioning in protozoans remain poorly understood. In the scope of this thesis, an exclusive guanylate cyclase coupled with N-terminal P4-ATPase was reported in an obligate intracellular parasite Toxoplasma gondii. In silico analysis indicated an activation of the guanylate cyclase by heterodimerization of its two cyclase domains and offered valuable insights into possible functions of its ATPase domain. This bulky protein (477-kDa), termed in this study as TgATPaseP-GC to reflect its envisaged multifunctionality, localizes in the plasma membrane at the apical pole of the parasite. TgATPaseP-GC is refractory to genetic deletion, and its CRISPR/Cas9-assisted disruption aborts the lytic cycle of T. gondii. Besides, Cre/loxP-mediated knockdown of TgATPaseP-GC reduced the synthesis of cGMP in tachyzoites and inhibited the parasite growth due to impairments of motility-dependent egress and invasion events. Notably, despite its temporally restricted function, TgATPaseP-GC is expressed constitutively throughout the lytic cycle, entailing a post-translational regulation of cGMP signaling. Not least, the occurrence of TgATPaseP-GC orthologs in several other alveolates implies a divergent functional repurposing of cGMP signaling in protozoans. Furthermore, an optogenetic approach was utilized to induce cGMP pathway by a photo-activated rhodopsin-guanylate cyclase (RhoGC) in T. gondii. The system enabled a light-control of cGMP elevation on crucial steps of lytic cycle in a fast, spatial and reversible manner. Excitation of RhoGC significantly stimulated the parasite motility of which impact was also monitored with an increased host-cell invasion and egress; as opposed to the genetic knockdown of TgATPaseP-GC. Having an established optogenetic system in the parasite allows to identify downstream targets of cGMP signaling via phosphoproteomic analysis.

Lors de l'infection par le virus herpès simplex de type 1 (VHS-1), quatre types de capsides nucléaires sont créés : les procapsides et les capsides A, B, et C. Sur les quatre capsides, seules les capsides C contiennent de l'ADN viral et deviendront des particules infectieuses. Un niveau de régulation se produit lors de la sortie du noyau qui favorise la sortie d’es capsides C du noyau. Le mécanisme qui sous-tend ce phénomène est actuellement inconnu. Les recherches actuelles suggèrent que l'interaction entre la protéine virale pUL25 modifie la conformation de la couche hexamérique plane du complexe de sortie nucléaire (NEC) pour y introduire des pentamères et donc causer un arrondissement de la membrane et le bourgeonnement des capsides. Cependant, des questions subsistent quant à la manière dont les capsides A, B et C sont différenciées au sein du noyau pour assurer une sortie spécifique de la capside C puisque pUL25 se retrouve dans tous les types de capsides. Nous étudions ici comment les protéines de l'hôte peuvent agir dans la sortie nucléaire des capsides C. En se basant sur une étude précédente du laboratoire où la protéine hôte poly(C)-binding protein 1 (PCBP1) a été trouvée spécifiquement sur les capsides C par spectrométrie de masse, nous explorons le rôle de la PCBP1 dans l'infection par le VHS-1. À l'aide d’essaies de plaques, nous montrons que la PCBP1 est importante pour l'infection virale, car en son absence, les titres diminuent et lorsque la PCBP1 est sur-exprimée, les titres augmentent. Ce résultat ne semble pas être dû au fait que les PCBP1 affectent l'expression génique de sous-ensembles de gènes viraux immédiats précoces, précoces ou tardifs, ni qu'ils affectent la réplication du génome ou son encapsidation. La réduction des PCBP1 ne provoque pas d'accumulation de capsides ou de particules matures tel qu’évalué par la microscopie électronique, mais elle augmente le nombre de capsides B enveloppées dans l'espace périnucléaire (PNS). L'inhibition de PCBP1 diminue également le niveau de protéine pUL24, une protéine virale importante pour la sortie du virus du noyau. Nos résultats démontrent que la PCBP1 pourrait réguler l’activité de pUL24, de sorte que lorsque la PCBP1 est épuisée, pUL24 permet à plus de capsides B de se rendre dans l'espace périnucléaire. Cette recherche constitue un point de départ pour une analyse plus approfondie du mécanisme exact des PCBP1 dans les infections à HSV-1. En outre, elle pourrait fournir des indices importants pour élucider comment le pUL24 favorise la sortie du nucléaire. / During herpes simplex virus type 1 (HSV-1) infection, four types of nuclear capsids are made: procapsids and A-, B- and C-capsids. Of the four capsids, only C-capsids contain the viral DNA and will become infectious progeny. A level of regulation occurs during nuclear egress that ensures only C-capsids exit the nucleus. The mechanism that underlies this phenomenon is presently unknown. Current research suggests the viral protein pUL25 alters the conformation of the viral nuclear egress complex (NEC) that forms a flat hexameric coat on nuclear membranes by the introduction of pentamers and therefore the induction of membrane rounding and viral budding. However, questions remain for how A-, B-, and C-capsids are differentiated within the nucleus to ensure C-capsid specific egress since pUL25 is found on all capsid types. Here we investigate how host proteins may play a role in nuclear egress of C-capsids. Based on the lab’s previous study where host protein poly(C)-binding protein 1 (PCBP1) was found specifically on C-capsids via mass spectrometry, we explore the role of PCBP1 in HSV-1 infection. Using plaque assays we show that PCBP-1 is important for viral infection, as in its absence titers decrease and when PCBP1 is over expressed titers increase. This result does not seem to be due to PCBP1 affecting gene expression of immediate early, early, or late viral gene subsets, nor does it seem to affect genome replication or encapsidation. PCBP1 knockdown does not cause an accumulation of capsids or mature particles as assessed by electron microscopy, but it does increase the number of enveloped B-capsids observed in the perinuclear space (PNS). Depletion of PCBP1 also decreases the level of pUL24, a viral protein implicated in viral nuclear egress. Our results suggest that PCBP1 could be regulating pUL24 for proper activity in nuclear egress, such that when PCBP1 is depleted, more B-capsids are able to bud through the PNS. This research constitutes a starting point for further analysis into the exact mechanism of PCBP1 in HSV-1 infections. In addition, it may provide important clues to elucidate how pUL24 supports nuclear egress.

[pt] Esse estudo propõe uma análise sobre acolhimento e desligamento institucional, entendendo-os como questões que dialogam entre si. Parte-se do princípio que é fundamental dar centralidade às vozes de quem vivenciou os efeitos práticos da medida de acolhimento institucional, o que poderá contribuir de modo significativo para a compreensão do tema. Ressalta-se que tal perspectiva é atravessada pelo direito à participação como fundamental também no contexto de institucionalização. Além disso, entende-se que essa condução seja potente para a construção de caminhos para o acolhimento institucional que respeitem os direitos destes sujeitos, bem como a formulação de um suporte mais adequado no processo de desligamento. Em termos metodológicos, elegeu-se a história oral como forma de escuta sobre temas sensíveis, valorizando as memórias e as perspectivas de jovens que tiveram o silenciamento ou a invisibilidade como realidades em suas trajetórias. Este estudo contou com as narrativas de duas jovens egressas da medida de acolhimento institucional, que aprofundam e enriquecem o conhecimento existente sobre o tema. / [en] This study proposes an analysis on acceptance and institutional dismissal, understanding them as issues that dialogue with each other. It is based on the principle that it is essential to give centrality to the voices of those who experienced the practical effects of the institutional reception measure, which could significantly contribute to the understanding of the theme. It is emphasized that this perspective is crossed by the right to participation as fundamental also in the context of institutionalization. In addition, it is understood that this conduction is potent for the construction of paths for institutional reception that respect the rights of these subjects, as well as the formulation of a more adequate support in the dismissal process. In methodological terms, oral history was chosen as a way of listening to sensitive topics, valuing the memories and perspectives of young people who had silencing or invisibility as realities in their trajectories. This study relied on the narratives of two young women who left the institutional care measure, which deepen and enrich the existing knowledge on the subject.

The overall purpose of the present study was to examine the strategies used by older adults with regard to both foot placement and hand contact location as they enter (ingress) and exit (egress) a vehicle. A cross-sectional study was used to identify and, in turn, examine the participants’ ingress and egress strategies in relation to their physical mobility and other characteristics. All participants had a valid driver’s license and they completed the following: 1) demographic (self-report) questionnaires; 2) clinical measures of physical mobility (i.e. Timed Up and Go, Berg Balance Scale, One Legged Stance test, Rapid Pace Walk test). A physical mobility index was also developed using these clinical measures. During both ingress and egress, older drivers were more likely to use one-foot strategies, which are considered less stable. Participants who used one-foot ingress and egress strategies had better overall physical mobility. The armrest location was used by the majority of older drivers for both ingress and egress, and is used most often during one-foot strategies. Participants who used multiple hand contact locations during ingress/egress had lower physical mobility than participants who used a single hand contact location or made no contact with the vehicle. Finally, the results from this study can inform educational materials targeting this population and determining design changes to the automobile that can prevent injuries to older drivers. / Thesis / Master of Health Sciences (MSc)

Le virus herpès simplex de type 1 (HSV 1) affecte la majorité de la population mondiale. HSV 1 cause de multiples symptômes délétères dont les plus communs sont les lésions orofaciales usuellement appelées feux sauvages. Le virus peut aussi causer des effets plus sérieux comme la cécité ou des troubles neurologiques. Le virus réside de façon permanente dans le corps de son hôte. Malgré l’existence de nombreux traitements pour atténuer les symptômes causés par HSV 1, aucun médicament ne peut éliminer le virus. Dans le but d’améliorer les connaissances concernant le cycle viral de HSV 1, ce projet cible l’étude du transport du virus dans la cellule hôte. Ce projet aura permis la collecte d’informations concernant le modus operandi de HSV 1 pour sortir des compartiments cellulaires où il séjourne. Les différentes expérimentations ont permis de publier 3 articles dont un article qui a été choisi parmi les meilleurs papiers par les éditeurs de « Journal of Virology » ainsi qu’un 4e article qui a été soumis. Premièrement, un essai in vitro reproduisant la sortie de HSV 1 du noyau a été mis sur pied, via l’isolation de noyaux issus de cellules infectées. Nous avons démontré que tout comme dans les cellules entières, les capsides s’évadent des noyaux isolés dans l’essai in vitro en bourgeonnant avec la membrane nucléaire interne, puis en s’accumulant sous forme de capsides enveloppées entre les deux membranes nucléaires pour finalement être relâchées dans le cytoplasme exclusivement sous une forme non enveloppée. Ces observations appuient le modèle de transport de dé-enveloppement/ré-enveloppement. Deuxièmement, dans le but d’identifier des joueurs clefs viraux impliqués dans la sortie nucléaire du virus, les protéines virales associées aux capsides relâchées par le noyau ont été examinées. La morphologie multicouche du virus HSV 1 comprend un génome d’ADN, une capside, le tégument et une enveloppe. Le tégument est un ensemble de protéines virales qui sont ajoutées séquentiellement sur la particule virale. La séquence d’ajout des téguments de même que les sites intracellulaires où a lieu la tégumentation sont l’objet d’intenses recherches. L’essai in vitro a été utilisé pour étudier cette tégumentation. Les données recueillies suggèrent un processus séquentiel qui implique l’acquisition des protéines UL36, UL37, ICP0, ICP8, UL41, UL42, US3 et possiblement ICP4 sur les capsides relâchées par le noyau. Troisièmement, pour obtenir davantage d’informations concernant la sortie de HSV 1 des compartiments membranaires de la cellule hôte, la sortie de HSV 1 du réseau trans golgien (TGN) a aussi été étudiée. L’étude a révélé l’implication de la protéine kinase D cellulaire (PKD) dans le transport post-TGN de HSV 1. PKD est connue pour réguler le transport de petits cargos et son implication dans le transport de HSV 1 met en lumière l’utilisation d’une machinerie commune pour le transport des petits et gros cargos en aval du TGN. Le TGN n’est donc pas seulement une station de triage, mais est aussi un point de rencontre pour différentes voies de transport intracellulaire. Tous ces résultats contribuent à une meilleure compréhension du processus complexe de maturation du virus HSV 1, ce qui pourrait mener au développement de meilleurs traitements pour combattre le virus. Les données amassées concernant le virus HSV 1 pourraient aussi être appliquées à d’autres virus. En plus de leur pertinence dans le domaine de la virologie, les découvertes issues de ce projet apportent également de nouveaux détails au niveau du transport intracellulaire. / Herpes simplex virus type 1 (HSV 1) affects the majority of the world population. HSV 1 causes various deleterious symptoms with the most common being facial mucosal lesions usually named cold sores. The virus can also contribute to more serious effects such as corneal blindness and neurological problems. The virus is permanently residing in the host body. Despite the existence of several treatments against HSV 1 symptoms, no drug is able to eliminate the virus. In order to improve knowledge of the viral cycle of HSV 1, this project focuses on the transport of the virus in the host cell. During this project we collect data to detail the modus operandi used by HSV 1 to leave cellular compartments such as the nucleus and the TGN. The different experimentations achieved during this PhD allowed the publication of three articles, including one selected as worthy of note by the editors of “Journal of virology” and a fourth article that has been submitted. Firstly, an in vitro assay that reproduces the exit of HSV 1 virus from nuclei was established via the isolation of nuclei from infected cells. We found that, as in intact cells, capsids escaped the isolated nuclei in the in vitro assay by budding through the inner nuclear membrane, accumulated as enveloped capsids between the two nuclear membranes, and were released in cytoplasm exclusively as unenveloped capsids. These observations support the de-envelopment / re-envelopment model of transport. Secondly, to identify viral players implicated in the nuclear egress of HSV 1, viral proteins associated with nuclear released capsids were investigated. HSV 1 has a multilayered morphology that includes a DNA genome, a capsid, a tegument and an envelope. The tegument represents viral proteins added sequentially on the viral particle. The sequential order and intracellular compartments where the tegument is added are the subject of intense research. The in vitro assay was used to investigate this tegumentation process. The acquired data suggest a sequential process that involved the acquisition of viral proteins UL36, UL37, ICP0, ICP8, UL41, UL42, US3 and possibly ICP4 on capsids released by the nucleus. Thirdly, to obtain information regarding another process of egress of HSV 1 from a membranous cellular organelle, the egress of HSV 1 from the TGN was also studied. The study revealed the implication of the cellular protein kinase D (PKD) in HSV 1 post-TGN transport. The involvement of this kinase, known to regulate the transport of small cargos, highlights the post TGN trafficking of both small and large entities (such as HSV 1) by a common machinery, in sharp contrast to earlier steps of transport. This indicates that the TGN is not only a sorting station but also a meeting point where different intracellular routes can meet. All these outcomes contribute to a better understanding of the complex maturation process of HSV 1 that could lead to the development of better tools to fight the virus. Results acquired concerning HSV 1 could also be applied to other viruses. Besides their relevance in the virology field, findings provided by this project also supply new details about cellular biology concerning intracellular transport.

Ma thèse s'inscrit dans l'étude des modifications du globule rouge humain induites par P. falciparum. Ces modifications qui représentent une remarquable adaptation du parasite à un environnement plus complexe qu'il n'y paraît au premier abord et expliquent sa persistance chez l'Homme sont détaillées dans une revue et un chapitre de livre dont je suis co-auteur. Mes travaux de recherche ont porté sur la caractérisation fonctionnelle des structures de Maurer, un compartiment membranaire exporté par le parasite dans le globule rouge parasitaire et directement lié à la physiopathologie du paludisme grave. J'ai contribué à la caractérisation fonctionnelle de nouvelles protéines de ces structures, codées par trois familles multigéniques sub-télomériques en cluster avec la famille Pfmc-2tm, et présentant de façon étonnante un fort degré de conservation (article 1). La diminution d'expression de ces gènes, obtenue par titration d'un facteur transcriptionnel, entraine un défaut de libération des mérozoïtes. Mon deuxième projet porte sur l'identification des modalités d'export de la protéine transmembranaire résidente des structures de Maurer PfSBP1. Mes travaux montrent que PfSBP1 est exportée sous forme soluble dans le cytoplasme érythrocytaire, en interaction avec le complexe chaperon parasitaire PfTCP1 (article 2). / Plasmodium falciparum causes the most severe forms of human malaria, a pathology associated with the erythrocytic asexual stages of the parasite. My work focused on the remodeling of the infected erythrocytes induced by P. falciparum and detailed in a review and a book chapter that I co-authored. These modifications illustrate a remarkable adaptation of P. falciparum resulting in its persistence in humans. My PhD thesis was dedicated to the functional characterization of Maurer's clefts, a membrane compartment transposed by the parasite in the cytoplasm of its host cell, and central to the export of virulence factors to the host cell surface. I have conducted two projects and contributed first to the functional characterization of novel exported protein encoded by three highly conserved multigene sub-telomeric families in cluster with the Pfmc-2tm family. Down regulation of these gene families by promoter titration impacted the release of infectious merozoites from the host cell (annex 1). My second project was dedicated to the identification of the modality of export of the resident and Maurer's clefts transmembrane protein PfSBP1. I have shown that PfSBP1 is exported as a soluble protein in the host cell cytoplasm in interaction with the parasite Thermosome complex protein 1 (PfTCP1) chaperone complex (annex 2).

A norma brasileira que trata de Saídas de Emergência, a ABNT NBR 9077:2001, pressupõe que o perfil da população de alunos em edifícios escolares, do ensino fundamental à universidade, é homogêneo, e que, portanto, não há eventuais particularidades inerentes às diferentes faixas etárias no uso das saídas de emergência. Esta pesquisa partiu da hipótese de que o perfil da população e, em particular, o de crianças do ciclo básico do ensino fundamental, pode ser diferente do adulto e, por consequência, demandar um dimensionamento específico para as saídas de emergência. Assim, realizou-se a coleta e análise de dados relativos à velocidade de caminhamento e aspectos comportamentais de crianças no movimento de descida de escadas, além de buscar correlações entre as velocidades e os dados antropométricos. Participaram da pesquisa 783 alunos do ciclo básico do ensino fundamental, oriundos de três escolas de ensino gratuito, com idade entre 6 e 10 anos. Foram testadas duas técnicas para a coleta dos dados: filmagens obtidas por meio de sistema fechado de televisão (CFTV) e sistema de identificação por rádio frequência (RFID), sendo que essa última não atendeu às expectativas e necessidades da pesquisa. As velocidades obtidas por processamento de dados de filmagens foram comparadas com as adotadas nos softwares de simulação de abandono \"FDS+EVAC\" e \"Simulex\". Constatou-se que, para os lances das escadas, as velocidades das crianças obtidas se assemelham as de referência de adultos (0,45 m/s até 1,05 m/s). Já para a velocidades obtidas nos patamares, houve grande dispersão, em relação aos valores de referência para adultos e crianças. Verificou-se, também, que o trajeto mais utilizado nos patamares foi o interno (mais curto) e que o emprego de métodos simplificados para estimar esses trajetos pode resultar em dados de velocidade imprecisos. Não foi constatada nenhuma correlação significativa entre as medidas antropométricas (IMC e bideltóide) e as velocidades dos alunos em lances ou patamares de escadas. Devido a dispersão dos resultados obtidos, não é possível a confirmação da hipótese inicial desta pesquisa. / In the Brazilian Technical Standard concerning means of egress (ABNT NBR 9077. 2001) it is presumed that the profile of students\' population in educational buildings, from elementary school to college, is homogeneous. Therefore, it does not consider the eventual particularities intrinsic to specific age groups during the use of egress systems. Assuming that the profile of elementary school children may be different from adults\', the criteria applicable to a variety of means of egress components could be more specific according to the occupant\'s characteristics. Thus, data regarding walking speeds and behavioral aspects of children in descending movement of stairs were collected and analyzed. Anthropometric data (weight, height and bideltoid measurement) from each child were also collected in order to seek correlations between these dimensions and the walking speeds. 783 elementary students from 3 different public schools, ages varying from 6 to 10 years old, participated in this research. Two techniques were tested for collecting data: recorded images from closed-circuit television (CCTV) and signals from radio-frequency identification (RFID); however, the last one was discarded due to technical difficulties. When compared to the speeds adopted by the egress simulation softwares \"FDS+EVAC\" and \"Simulex\", it was noticed that children\'s travel speeds in stairs are actually quite similar to the ones adopted for adults (0,45 m/s to 1,05 m/s), contrasting with the common sense that infants are slower than grown-ups. As for the walking speeds on landings, a wide dispersion in relation to parameters determined for adults and children was found. It was also noticed that most infants chose the shortest path (internal) on landings and that designation of a simplified average trajectory can lead to inaccurate travel speeds. Regarding the anthropometric measurements and walking speeds, no direct correlation was found. Due to the significant dispersion of the results, the initial hypothesis of this research could not be confirmed.