Qu’est ce que la Franc-maçonnerie ? Comment sont perçus les francs-maçons par la société ? Comment les francs-maçons se perçoivent-ils eux-mêmes ? Comment les initiés réagissent-ils face aux idées qui circulent à leur encontre et tentent-ils de s’en défendre ? Depuis le début du XVIIIème siècle, la Franc-maçonnerie, Ordre initiatique qui a vu le jour sous sa forme moderne en Angleterre, fascine, interroge, inquiète… Les francs-maçons se disent "discrets", mais paradoxalement, cette discrétion est à l’origine de nombreuses publications et éveille la curiosité, suscite parfois la méfiance, la suspicion ou encore l’hostilité des "profanes". Cette étude a pour but de rendre compte des rapports entretenus entre la Franc-maçonnerie (et ses membres) et la société globale et les institutions qui la composent, et ce de l’apparition du "phénomène maçonnique" à nos jours. Elle en dégagera également les fondements, qu’ils soient exogènes à la Maçonnerie c'est-à-dire issus de déterminismes historico-culturels, ou endogènes c'est-à-dire inhérents à la Franc-maçonnerie, qualifiée parfois de "société dans la société". / What the Freemasonry? How are perceived the freemasons by the society? How do the freemasons perceive themselves ? How do freemasons react in front of ideas which circulate against them and do they try to defend themselves from it ? Since the beginning of the XVIIIth century, the Freemasonry, initiatory Order which was born under its modern shape in England, fascinates, questions, worries … The freemasons say themselves "discreet", but paradoxically, this discretion is at the origin of numerous publications and awakens the curiosity, the suspicion or the hostility in France. This study aims at reporting maintained connections between the Freemasonry (and his members) and the global society and the institutions which compose it, since the appearance of the " masonic phenomenon " in our days.

abstract: Recombinant protein expression is essential to biotechnology and molecular medicine, but facile methods for obtaining significant quantities of folded and functional protein in mammalian cell culture have been lacking. Here I describe a novel 37-nucleotide in vitro selected sequence that promotes unusually high transgene expression in a vaccinia driven cytoplasmic expression system. Vectors carrying this sequence in a monocistronic reporter plasmid produce >1,000-fold more protein than equivalent vectors with conventional vaccinia promoters. Initial mechanistic studies indicate that high protein expression results from dual activity that impacts both transcription and translation. I suggest that this motif represents a powerful new tool in vaccinia-based protein expression and vaccine development technology. / Dissertation/Thesis / M.S. Biochemistry 2012

Ribosomal protein genes are crucial for ribosome biogenesis. The ribosome itself is responsible for protein synthesis and hence cellular growth and development. Intertwining network of proteins in conjugation with cellular environment such as nutrition and growth factors collectively regulate expression of the ribosomal protein genes. DNA microarray analysis has implicated the role of 26S proteasome in transcriptional regulation of the ribosomal protein genes tying protein degradation to protein synthesis pathway. To determine the mechanism as to how the 26S proteasome promotes transcription of the ribosomal protein genes a series of experiments were performed. The results reveal that the 19S subcomplex of the 26S proteasome is recruited to the promoters of the ribosomal protein genes in a TOR (Target of Rapamycin)-dependent manner. TOR signals environmental cues and controls the expression of the ribosomal protein genes. Thus recruited 19S proteasome subcomplex promotes transcriptional initiation via facilitation of the recruitment of co-activator NuA4 (Nucleasome acetyltransferase of histone H4) complex to activator Rap1. NuA4 enhances PIC (Pre-initiation complex) formation at the core promoter, but it is not clearly understood how does it do so. Researches have identified two different forms of TBP: TAF (TBP associated factor)-dependent form of TBP and TAF-independent form of TBP. This work shows that impaired association of NuA4 interferes with TFIID recruitment, but recruits TAF-independent form of TBP to the core promoter. This recruitment of TBP is dependent on SAGA (Spt-Ada-Gcn5-acetyltransferase). Like ribosomal protein genes, antisense transcription is also enhanced by TAFs. However, it remains unknown whether NuA4 also promotes TAF-regulated antisense transcription. The results illustrate that like ribosomal protein genes, transcription of GAL10 antisense is also promoted by NuA4 HAT (histone acetyl transferase). NuA4 HAT is recruited to the 3’-end of the GAL10 coding sequence, acetylates histone H4 and promotes GAL10 antisense transcription. This work also reveals the roles of other chromatin regulatory factors in controlling antisense transcription. Collectively, these results significantly advance our current understanding of the regulatory mechanisms of ribosomal protein genes’ expression and antisense transcription. The ribosome and antisense are involved in virtually all the biological processes. Aberrant expression of the ribosomal protein genes and antisense transcripts are associated with numerous human disorders including cancers and cardiovascular diseases. Therefore, analyses of their regulatory processes provide valuable information toward understanding the etiology of numerous human diseases with potential therapeutic interventions.

Introdução: Determinar critérios de indicação e contra-indicação para os tratamentos psicanalíticos é um ponto técnico crucial para a obtenção de melhora terapêutica. Entretanto, estes critérios costumam basear-se em especulações teóricas, não em investigações empíricas. Mesmo havendo discussão a respeito do que se considera melhora para o referencial psicanalítico, é relevante investigar se há diferença entre o perfil de pacientes que melhoram e não melhoram nas fases iniciais da psicoterapia psicanalítica, pois este dado tem o potencial de ser útil na indicação de tratamentos. Objetivos: O objetivo deste estudo foi identificar preditores de indicação/contra-indicação, concordância/não concordância em iniciar tratamento e melhora/não melhora inicial na psicoterapia psicanalítica de adultos. Método: Tratase de uma coorte realizada em ambulatório de saúde mental de Porto Alegre com 557 pacientes que buscaram atendimento em um período de 20 meses. Foram aplicados instrumentos para verificar qualidade de vida, sintomas e estilo defensivo dos pacientes na triagem e ao sexto mês de tratamento. Os pacientes foram alocados, com base na avaliação dos triadores e terapeutas e a partir de questionários autoaplicáveis, em: não indicação para psicoterapia psicanalítica, não concordância em iniciar tratamento, melhora/não melhora. Resultados: Primeiramente, constatou-se que os terapeutas que atendem neste ambulatório aderem, na amostra estudada, à técnica de psicoterapia psicanalítica em suas intervenções. O acompanhamento dos pacientes possibilitou encontrar que aqueles que buscam atendimento por queixas somáticas ou por queixas de atenção e com baixa escolaridade apresentaram menor probabilidade de receber indicação para psicoterapia psicanalítica; pacientes que percebem seus sintomas como muito intensos no momento da busca apresentaram maior probabilidade de receberem indicação para este tratamento. Uma vez indicados para psicoterapia psicanalítica, os preditores de não concordância em iniciar tratamento foram: baixa escolaridade, baixa renda familiar, diagnóstico de esquizofrenia, transtorno esquizotípico ou delirante, queixas depressivas e/ou nunca ter feito psicoterapia antes da busca atual. Os pacientes que permaneciam em tratamento ao final do sexto mês relataram melhor qualidade de vida, menor número de sintomas, redução na intensidade dos sintomas presentes e mudanças em alguns aspectos do seu estilo defensivo. Pacientes que buscaram tratamento por problemas de ansiedade apresentaram maior risco de não relatar melhora na qualidade de vida, sintomatologia e estilo defensivo nos seis primeiros meses de psicoterapia. Conclusões: Apesar de a psicoterapia psicanalítica ser um tratamento de longa duração, pacientes demonstraram melhora na qualidade de vida, nos sintomas apresentados e no estilo defensivo logo nos primeiros seis meses de atendimento. É relevante fomentar estudos que investiguem o papel da interação paciente-terapeuta na determinação da melhora do paciente e na indicação para psicoterapia psicanalítica, e não apenas características individuais do paciente. É necessário rever a técnica empregada durante as avaliações iniciais para aqueles pacientes com o perfil de não concordância em iniciar tratamento, considerando tanto a análise das resistências capazes de serem superadas durante este período inicial como a verificação de outros possíveis tratamentos mais bem indicados para estes casos. / Introduction: To determine criteria of indication and contraindication for psychoanalytic treatments is a crucial technical point for obtaining therapeutic improvement. However, these criteria are usually based on theoretical speculation, not on empirical investigations. Even though there is debate about how to define improvement within the psychoanalytic approach, it is important to investigate whether there are differences between the profile of patients who improve and those who do not improve during early stages of psychoanalytic psychotherapy, because this information have potential to be useful when indicating treatments. Objectives: The aim of this study was to identify predictors of indication/contraindication, agreement/disagreement in starting treatment and improvement/no improvement during initial stages of psychoanalytic psychotherapy of adults. Methods: This was a cohort study conducted at an outpatient mental health clinic of Porto Alegre (Southern Brazil) with 557 patients who sought treatment in a period of 20 months. Instruments were administered to check quality of life, symptoms and defensive style of patients at the intake interview and at the sixth month of treatment. Patients were allocated, based on the evaluation of their interviewers and therapists and from selfadministered questionnaires, into: not indicated for psychoanalytic psychotherapy, not concordant to initiate treatment, improved / did not improve after six months of treatment. Results: First, we found that a sample of therapists who attend at this clinic adhere to the psychoanalytic psychotherapy technique in their interventions. The follow-up analyzes showed that patients who seek care because of somatic or attention complaints or complaints and with low educational level were less likely to be indicated for psychoanalytic psychotherapy; patients who perceive their symptoms as very intense at the time of the search were more likely to be indicated to this treatment. Once referred to psychoanalytic psychotherapy, the predictors of non-agreement in starting treatment were: low educational level, low family income, diagnosis of schizophrenia, schizotypal or delusional disorder, depressive symptoms, and/or never having undergone psychotherapy before. Patients who remained on treatment at the end of the sixth month reported better quality of life, fewer symptoms, reduction in the intensity of symptoms, and changes in some aspects of defensive style. Patients seeking treatment for anxiety disorders had higher risk of not reporting improvement in quality of life, symptoms and defensive style in the first six months of psychotherapy. Conclusions: Although psychoanalytic psychotherapy is considered a long-term treatment, patients demonstrated improvement in quality of life, symptoms and defensive style during the first six months of treatment. It is important to encourage studies to investigate the role of the patient-therapist interaction in the determination of patient improvement and indication/contraindication for psychoanalytic psychotherapy, not just individual characteristics of the patient. It is necessary to review the technique used during the initial assessments for those patients with the profile of non-agreement to start treatment, considering both the analysis of resistances capable to be overcome during this initial period as checking for other possible treatments better indicated for these cases.

Orientador: Igor Rossoni / Banca: Suzi Frankl Sperber / Banca: Evelina de Carvalho Sá Hoisel / Banca: José Carlos Zamboni / Banca: Jeane Mari Sant'Ana Spera / Resumo: Magma é o volume de poemas que marcou a estréia literária de João Guimarães Rosa, tendo no entanto permanecido inédito por mais de sessenta anos e sido até o momento relegado à marginalidade pela maior parte da crítica. O presente trabalho tem como objetivo contribuir para o resgate desse livro, bem como levantar, no discurso do mesmo, elementos úteis para o reconhecimento da sua inserção ideológica no conjunto da obra do autor. Com esse intento, procede-se à análise do universo simbológico plasmado por seus textos componentes, através do que se consegue identificar em Magma a inquietação anagógica que é o cerne da literatura rosiana. / Abstract: Magma is the poems volume that set out the literary debut of João Guimarães Rosa, although have stayed inedited over sixty years and until now have been discarded by the majority of the critique. The aim of the present work is to contribute to the ransom of that book and to find in its discourse able factors to its insertion into the whole author production. With that purpose, is accomplished the analysis of the symbological universe shaped by its component texts, with what is possible to identify in Magma the anagogical inquietud that is the pith of the Guimarães Rosa literature. / Doutor

Introdução: Determinar critérios de indicação e contra-indicação para os tratamentos psicanalíticos é um ponto técnico crucial para a obtenção de melhora terapêutica. Entretanto, estes critérios costumam basear-se em especulações teóricas, não em investigações empíricas. Mesmo havendo discussão a respeito do que se considera melhora para o referencial psicanalítico, é relevante investigar se há diferença entre o perfil de pacientes que melhoram e não melhoram nas fases iniciais da psicoterapia psicanalítica, pois este dado tem o potencial de ser útil na indicação de tratamentos. Objetivos: O objetivo deste estudo foi identificar preditores de indicação/contra-indicação, concordância/não concordância em iniciar tratamento e melhora/não melhora inicial na psicoterapia psicanalítica de adultos. Método: Tratase de uma coorte realizada em ambulatório de saúde mental de Porto Alegre com 557 pacientes que buscaram atendimento em um período de 20 meses. Foram aplicados instrumentos para verificar qualidade de vida, sintomas e estilo defensivo dos pacientes na triagem e ao sexto mês de tratamento. Os pacientes foram alocados, com base na avaliação dos triadores e terapeutas e a partir de questionários autoaplicáveis, em: não indicação para psicoterapia psicanalítica, não concordância em iniciar tratamento, melhora/não melhora. Resultados: Primeiramente, constatou-se que os terapeutas que atendem neste ambulatório aderem, na amostra estudada, à técnica de psicoterapia psicanalítica em suas intervenções. O acompanhamento dos pacientes possibilitou encontrar que aqueles que buscam atendimento por queixas somáticas ou por queixas de atenção e com baixa escolaridade apresentaram menor probabilidade de receber indicação para psicoterapia psicanalítica; pacientes que percebem seus sintomas como muito intensos no momento da busca apresentaram maior probabilidade de receberem indicação para este tratamento. Uma vez indicados para psicoterapia psicanalítica, os preditores de não concordância em iniciar tratamento foram: baixa escolaridade, baixa renda familiar, diagnóstico de esquizofrenia, transtorno esquizotípico ou delirante, queixas depressivas e/ou nunca ter feito psicoterapia antes da busca atual. Os pacientes que permaneciam em tratamento ao final do sexto mês relataram melhor qualidade de vida, menor número de sintomas, redução na intensidade dos sintomas presentes e mudanças em alguns aspectos do seu estilo defensivo. Pacientes que buscaram tratamento por problemas de ansiedade apresentaram maior risco de não relatar melhora na qualidade de vida, sintomatologia e estilo defensivo nos seis primeiros meses de psicoterapia. Conclusões: Apesar de a psicoterapia psicanalítica ser um tratamento de longa duração, pacientes demonstraram melhora na qualidade de vida, nos sintomas apresentados e no estilo defensivo logo nos primeiros seis meses de atendimento. É relevante fomentar estudos que investiguem o papel da interação paciente-terapeuta na determinação da melhora do paciente e na indicação para psicoterapia psicanalítica, e não apenas características individuais do paciente. É necessário rever a técnica empregada durante as avaliações iniciais para aqueles pacientes com o perfil de não concordância em iniciar tratamento, considerando tanto a análise das resistências capazes de serem superadas durante este período inicial como a verificação de outros possíveis tratamentos mais bem indicados para estes casos. / Introduction: To determine criteria of indication and contraindication for psychoanalytic treatments is a crucial technical point for obtaining therapeutic improvement. However, these criteria are usually based on theoretical speculation, not on empirical investigations. Even though there is debate about how to define improvement within the psychoanalytic approach, it is important to investigate whether there are differences between the profile of patients who improve and those who do not improve during early stages of psychoanalytic psychotherapy, because this information have potential to be useful when indicating treatments. Objectives: The aim of this study was to identify predictors of indication/contraindication, agreement/disagreement in starting treatment and improvement/no improvement during initial stages of psychoanalytic psychotherapy of adults. Methods: This was a cohort study conducted at an outpatient mental health clinic of Porto Alegre (Southern Brazil) with 557 patients who sought treatment in a period of 20 months. Instruments were administered to check quality of life, symptoms and defensive style of patients at the intake interview and at the sixth month of treatment. Patients were allocated, based on the evaluation of their interviewers and therapists and from selfadministered questionnaires, into: not indicated for psychoanalytic psychotherapy, not concordant to initiate treatment, improved / did not improve after six months of treatment. Results: First, we found that a sample of therapists who attend at this clinic adhere to the psychoanalytic psychotherapy technique in their interventions. The follow-up analyzes showed that patients who seek care because of somatic or attention complaints or complaints and with low educational level were less likely to be indicated for psychoanalytic psychotherapy; patients who perceive their symptoms as very intense at the time of the search were more likely to be indicated to this treatment. Once referred to psychoanalytic psychotherapy, the predictors of non-agreement in starting treatment were: low educational level, low family income, diagnosis of schizophrenia, schizotypal or delusional disorder, depressive symptoms, and/or never having undergone psychotherapy before. Patients who remained on treatment at the end of the sixth month reported better quality of life, fewer symptoms, reduction in the intensity of symptoms, and changes in some aspects of defensive style. Patients seeking treatment for anxiety disorders had higher risk of not reporting improvement in quality of life, symptoms and defensive style in the first six months of psychotherapy. Conclusions: Although psychoanalytic psychotherapy is considered a long-term treatment, patients demonstrated improvement in quality of life, symptoms and defensive style during the first six months of treatment. It is important to encourage studies to investigate the role of the patient-therapist interaction in the determination of patient improvement and indication/contraindication for psychoanalytic psychotherapy, not just individual characteristics of the patient. It is necessary to review the technique used during the initial assessments for those patients with the profile of non-agreement to start treatment, considering both the analysis of resistances capable to be overcome during this initial period as checking for other possible treatments better indicated for these cases.

Introdução: Determinar critérios de indicação e contra-indicação para os tratamentos psicanalíticos é um ponto técnico crucial para a obtenção de melhora terapêutica. Entretanto, estes critérios costumam basear-se em especulações teóricas, não em investigações empíricas. Mesmo havendo discussão a respeito do que se considera melhora para o referencial psicanalítico, é relevante investigar se há diferença entre o perfil de pacientes que melhoram e não melhoram nas fases iniciais da psicoterapia psicanalítica, pois este dado tem o potencial de ser útil na indicação de tratamentos. Objetivos: O objetivo deste estudo foi identificar preditores de indicação/contra-indicação, concordância/não concordância em iniciar tratamento e melhora/não melhora inicial na psicoterapia psicanalítica de adultos. Método: Tratase de uma coorte realizada em ambulatório de saúde mental de Porto Alegre com 557 pacientes que buscaram atendimento em um período de 20 meses. Foram aplicados instrumentos para verificar qualidade de vida, sintomas e estilo defensivo dos pacientes na triagem e ao sexto mês de tratamento. Os pacientes foram alocados, com base na avaliação dos triadores e terapeutas e a partir de questionários autoaplicáveis, em: não indicação para psicoterapia psicanalítica, não concordância em iniciar tratamento, melhora/não melhora. Resultados: Primeiramente, constatou-se que os terapeutas que atendem neste ambulatório aderem, na amostra estudada, à técnica de psicoterapia psicanalítica em suas intervenções. O acompanhamento dos pacientes possibilitou encontrar que aqueles que buscam atendimento por queixas somáticas ou por queixas de atenção e com baixa escolaridade apresentaram menor probabilidade de receber indicação para psicoterapia psicanalítica; pacientes que percebem seus sintomas como muito intensos no momento da busca apresentaram maior probabilidade de receberem indicação para este tratamento. Uma vez indicados para psicoterapia psicanalítica, os preditores de não concordância em iniciar tratamento foram: baixa escolaridade, baixa renda familiar, diagnóstico de esquizofrenia, transtorno esquizotípico ou delirante, queixas depressivas e/ou nunca ter feito psicoterapia antes da busca atual. Os pacientes que permaneciam em tratamento ao final do sexto mês relataram melhor qualidade de vida, menor número de sintomas, redução na intensidade dos sintomas presentes e mudanças em alguns aspectos do seu estilo defensivo. Pacientes que buscaram tratamento por problemas de ansiedade apresentaram maior risco de não relatar melhora na qualidade de vida, sintomatologia e estilo defensivo nos seis primeiros meses de psicoterapia. Conclusões: Apesar de a psicoterapia psicanalítica ser um tratamento de longa duração, pacientes demonstraram melhora na qualidade de vida, nos sintomas apresentados e no estilo defensivo logo nos primeiros seis meses de atendimento. É relevante fomentar estudos que investiguem o papel da interação paciente-terapeuta na determinação da melhora do paciente e na indicação para psicoterapia psicanalítica, e não apenas características individuais do paciente. É necessário rever a técnica empregada durante as avaliações iniciais para aqueles pacientes com o perfil de não concordância em iniciar tratamento, considerando tanto a análise das resistências capazes de serem superadas durante este período inicial como a verificação de outros possíveis tratamentos mais bem indicados para estes casos. / Introduction: To determine criteria of indication and contraindication for psychoanalytic treatments is a crucial technical point for obtaining therapeutic improvement. However, these criteria are usually based on theoretical speculation, not on empirical investigations. Even though there is debate about how to define improvement within the psychoanalytic approach, it is important to investigate whether there are differences between the profile of patients who improve and those who do not improve during early stages of psychoanalytic psychotherapy, because this information have potential to be useful when indicating treatments. Objectives: The aim of this study was to identify predictors of indication/contraindication, agreement/disagreement in starting treatment and improvement/no improvement during initial stages of psychoanalytic psychotherapy of adults. Methods: This was a cohort study conducted at an outpatient mental health clinic of Porto Alegre (Southern Brazil) with 557 patients who sought treatment in a period of 20 months. Instruments were administered to check quality of life, symptoms and defensive style of patients at the intake interview and at the sixth month of treatment. Patients were allocated, based on the evaluation of their interviewers and therapists and from selfadministered questionnaires, into: not indicated for psychoanalytic psychotherapy, not concordant to initiate treatment, improved / did not improve after six months of treatment. Results: First, we found that a sample of therapists who attend at this clinic adhere to the psychoanalytic psychotherapy technique in their interventions. The follow-up analyzes showed that patients who seek care because of somatic or attention complaints or complaints and with low educational level were less likely to be indicated for psychoanalytic psychotherapy; patients who perceive their symptoms as very intense at the time of the search were more likely to be indicated to this treatment. Once referred to psychoanalytic psychotherapy, the predictors of non-agreement in starting treatment were: low educational level, low family income, diagnosis of schizophrenia, schizotypal or delusional disorder, depressive symptoms, and/or never having undergone psychotherapy before. Patients who remained on treatment at the end of the sixth month reported better quality of life, fewer symptoms, reduction in the intensity of symptoms, and changes in some aspects of defensive style. Patients seeking treatment for anxiety disorders had higher risk of not reporting improvement in quality of life, symptoms and defensive style in the first six months of psychotherapy. Conclusions: Although psychoanalytic psychotherapy is considered a long-term treatment, patients demonstrated improvement in quality of life, symptoms and defensive style during the first six months of treatment. It is important to encourage studies to investigate the role of the patient-therapist interaction in the determination of patient improvement and indication/contraindication for psychoanalytic psychotherapy, not just individual characteristics of the patient. It is necessary to review the technique used during the initial assessments for those patients with the profile of non-agreement to start treatment, considering both the analysis of resistances capable to be overcome during this initial period as checking for other possible treatments better indicated for these cases.

Submitted by Caroline Falcao (caroline.rfalcao@ufpe.br) on 2017-04-10T17:54:57Z No. of bitstreams: 2 license_rdf: 1232 bytes, checksum: 66e71c371cc565284e70f40736c94386 (MD5) 2012-Tese-TamaraLima.pdf: 15100477 bytes, checksum: 79a87ae32652a2bd9c0dae960dc0a539 (MD5) / Made available in DSpace on 2017-04-10T17:54:57Z (GMT). No. of bitstreams: 2 license_rdf: 1232 bytes, checksum: 66e71c371cc565284e70f40736c94386 (MD5) 2012-Tese-TamaraLima.pdf: 15100477 bytes, checksum: 79a87ae32652a2bd9c0dae960dc0a539 (MD5) Previous issue date: 2012-09-21 / Em eucariotos,a proteína de ligação à cauda poli-A (PABP) se liga especificamente a seqüência de poli-adenosinas na extremidade 3’ dos mRNAs, atuando em múltiplas funções associadas ao metabolismo destes, incluindo biogênese, processamento, transporte núcleo-citoplasmático e degradação. A PABP também participa na síntese proteíca, via interações diretas com fatores de tradução, como o fator de iniciação eIF4G, parceiro da proteína de ligação ao capeIF4E. O objetivo desta Tese foi dar continuidade a caracterização de três homólogos da PABP identificados em Leishmania sp.(PABPs1,2 e 3), protozoários patogênicos de características biológicas diferenciadas. Em estudos prévios estas proteínas se mostraram abundantes e de expressão simultânea e a PABP1 se mostrou representada por isoformas associadas a sua fosforilação. Neste trabalho, foi observado que as três proteínas possuem localização citoplasmática, porém sob condições de inibição da transcrição, mas não de tradução ou processamento de mRNAs, as PABPs 2 e 3 migram para o núcleo. Esse comportamento está de acordo com ensaios de interação proteína-proteina, onde foi observada uma associação direta entre as PABPs 2 e 3, independente de RNA. A expressão de isoformas fosforiladas da PABP1 foi então analisada e observou-se que esta era afetada pela inibição dos processos de síntese de mRNAs. Investigando sua participação no processo de tradução, foram confirmadas interações específicas entre a PABP1 e um homólogo de eIF4G in vivo e interações inéditas entre esta e homólogos de eIF4E. As três proteínas se mostraram capazes de se associar com os polissomas de células em fase de crescimento exponencial, mas não de fase estacionária. Ao longo do trabalho foram mapeadosmotivos envolvidos com as interações observadas identificando características novas não descritas. Por fim, ensaios de co-imuno precipitação seguido de análise protéica por espectrometria de massa e de RNAs por RT-PCR confirmaram a presença de parceiros protéicos diferenciados para cada homólogo e mRNAs alvo para as PABPs 2 e 3 cuja tradução está associada a fase S do ciclo celular. Nossos resultados mostram que estas proteínas apresentam funções divergentes e reforça um papel mais geral da PABP1 na tradução. / In eukaryotes, the poly-A binding protein (PABP) binds specifically to a poly-adenosines sequence present in the 3' end of the mRNAs where performes multiple functions associated with their metabolism, including biogenesis, processing, nucleocytoplasmic transport, and degradation. The PABP also participates in protein synthesis, via direct interactions with translation factors such as the initiation factor eIF4G, partner of the cap binding protein eIF4E. The aim of this Thesis was to continue the characterization of the three PABP homologues identified in Leishmaniasp. (PABPs 1, 2 and 3), a pathogenic protozoan with differentiated biological characteristics. Previous studies have shown that these proteins are abundant and simultaneously expressed and that PABP1 is characterized by multiples isoformes associated to phosphorylation. In this study, it was observed that all three proteins have cytoplasmic localization, but under transcription inhibition conditions but not translation or mRNAs processing, the PABPs 2 and 3 migrate to the nucleus. This behavior is in agreement with a protein-protein interaction observed between PABPs 2 and 3, and this interaction isindependent of RNA. The expression of the phosphorylated isoforms of PABP1 was also analyzed and it was found that some of those isoforms were affected by the inhibition of the processessing of mRNAs sinthesis. When investigated their participation in thetranslation process were observed specific interactions between PABP1 and a homolog of eIF4G in vivoand a novel interaction between PABP 1 and the homologues of eIF4E. Throughout this study the motifs involved in the interactios mentioned above were mapped and new motifs not yet described were observed. All three proteins have been shown to be associated with polysomes in cells in in exponential phase, but not in stationary phase. Finally, co-immunoprecipitation assays followed by protein analysis by mass spectrometry and of RNAs by RT-PCR confirmed the presence of different protein partners and mRNAs targets to the PABPs 2 and 3 whose translation is associated with the S phase of cell cycle. Our results show that these proteins have different functions and reinforces a more general role of PABP1 in translation.

The rising demand for powerful oncolytic virotherapy agents has led to the identification of Maraba virus, one of the most potent oncolytic viruses from Rhabdoviridae family which displays high selectivity for killing malignant cells and low cytotoxicity in normal cells. Although the virus is readied to be used for clinical trials, the interactions between the virus and the host cells is still unclear. Using a newly developed interferon-sensitive mutant Maraba virus (MG1), we have identified two key regulators of global translation (4E-BP1 and eIF2α) responsible for the inhibition of protein synthesis in the infected cells. Despite the translational arrest upon viral stress, we showed an up-regulation of anti-apoptotic Bcl-xL protein that provides a survival benefit for the host cell, yet facilitates effective viral propagation. Given the fact that eIF5B canonically regulates 60S ribosome subunit end joining, and is able to replace the role of eIF2 in delivering initiator tRNA to the 40S ribosome subunit upon the phosphorylation of eIF2α, we have tested whether eIF5B mediates the translation of target mRNAs during MG1 infection. Our results show that the inhibition of eIF5B significantly down-regulates the level of Bcl-xL steady-state mRNA, thus indirectly attenuates viral propagation.

This thesis explores the archetypal theme of initiation in selected texts by Joseph Conrad. The Introduction first surveys critical attention to initiatory motifs in Conrad with the objective of demonstrating the need for an approach to the topic informed by a more formal and theorized understanding of initiation. It then offers a prima facie case for the centrality of the idea of initiation in Conrad's oeuvre, based on references culled from a range of the author's writings. Chapter One seeks to contextualise initiation by providing a history of anthropological research into and theorisations of the rite, proceeding to a description of its typical structure and functions. A detailed account is given of the most widely accepted model of initiation, Arnold van Gennep's tripartite schema. Moving on to Conrad's writing, Chapter Two draws on both his fiction and more personal writings in order to provide a provisional account of the writer's own understanding of initiation and its importance, and to offer some explanation of why Conrad should have been prompted to accord the motif such prominence in his work. Conrad's presentation and (impliedly) his understanding of initiation was never entirely consistent and underwent some change in the course of his writing career. The critical assessment of "Typhoon" in Chapter Three depicts Conrad's more optimistic conception of initiation as a rite benefitting both society, by promoting solidarity, and the individual, by advancing self-knowledge. Chapter Four introduces, via analyses of the novellas "Youth" and "The Shadow Line", that variation on the motif of initiation which is more typical of its manifestation in Conrad: the failure of individuals to complete their cycles of initiation. Chapter Five identifies those characteristics of initiation which appear to be determinative in the representations of incomplete initiation in Conrad's work. Initiation seems to play out approximately seven paradoxes; the impact of some of these is examined through analysis of the initiatory ordeals of the main protagonists in The Secret Agent. Integral to this discussion is an attempt to demonstrate the vital role which initiation plays in the healthy maintenance not only of social order but also of faith and life itself. The Conclusion summarises the more important findings of the study and indicates some directions for further, related research.