Threefoil is a small engineered protein of 141 amino acids which is a member of the beta-trefoil superfamily, with three-fold symmetry and high thermal and kinetic stability. Its primary sequence was designed based on a predicted beta-trefoil glycosidase from the halophilic Archaeon Haloarcula marismortui. Threefoil predominantly forms inclusion bodies when over-expressed in Escherichia coli at 37??C, with little to no protein soluble in the cytoplasm. Nevertheless, Threefoil is capable of adopting a native beta-trefoil structure when refolded from solubilized inclusion bodies. The focus of this thesis is on characterization of the folding and stability of Threefoil through thermodynamic and kinetic experiments for wild-type Threefoil, in addition to sugar- and metal-binding studies and characterization of Threefoil inclusion bodies. Various Threefoil mutants, designed to increase protein stability, are also characterized to probe the origins of, as well as to give insight into, the mechanism of inclusion body formation. The thermodynamic and kinetic stability of wild-type Threefoil was studied using spectral probes, mainly fluorescence, circular dichroism (CD) and dynamic light scattering (DLS). The major observed spectral changes in kinetic and thermodynamic experiments can be fit to a 2-state transition between the folded state and a denatured state containing extensive residual secondary structure. At high protein concentrations, the folding of wild-type Threefoil is complicated by protein misfolding and aggregation. As Threefoil is remarkably resistant to denaturation even at high concentrations of urea and guanidine hydrochloride (GuHCl), studies were also conducted in guanidine isothiocyanate (GuSCN), which is a much stronger denaturant than urea and GuHCl. Remarkably, the time that is required for Threefoil samples to reach equilibrium in renaturation curves is approximately 100 days, while equilibrium by denaturation in the stronger denaturant, GuSCN, requires more than two years. The expression levels of Threefoil mutants A62V, Q78I, D85P and D93P were also studied. None of the four mutants studied exhibited any pronounced increase in solubility compared to wild-type when expressed in E. coli.

No description available.

Orientador: Arlindo Rebechi Júnior / Banca: Marcelo Magalhães Bulhões / banca: Pedro Plaza Pinto / Resumo: Esta dissertação analisa o trabalho do crítico de cinema Ely Azeredo, a partir de um corpus de vinte e sete artigos sobre o cinema brasileiro publicados entre 1956 a 1973, nos periódicos cariocas Tribuna da Imprensa e Jornal do Brasil. Este recorte temporal foi delimitado levando em consideração as profundas transformações e manifestações fílmicas inovadoras que a cinematografia do país apresentou nesse período. Nessa fase, surge o Cinema Novo, influenciado pela Neorrealismo Italiano e pela Nouvelle Vague Francesa, cujo objetivo era realizar uma leitura a partir do seus filmes dos problemas sócio-políticos do Brasil. Paralelamente, o Cinema Marginal surge como contraponto ao Cinema Novo, alegando que o cinema nacional só poderia criar obras legitimamente brasileiras, a partir do momento em que aceitasse sua posição de subdesenvolvido, trabalhando todos os elementos do "dejeto industrial" que era despejado na cultura brasileira. Por fim, tem-se o advento das comédias eróticas, que seriam rotuladas, no futuro, como "pornochanchadas", pois o gênero possuía caráter popular ao combinar a comédia de costumes e o erotismo. Durante esse período, o Brasil passou por um golpe militar, que depôs o então Presidente João Goulart, e, consequentemente, instaurou uma ditadura no país, que perdurou por 21 anos (1964-1985), fazendo com que a produção cinematográfica brasileira fosse afetada, direta ou indiretamente por esse cenário político. Com o propósito de investigar a construção de sentid... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: This dissertation analyzes the work of the film critic Ely Azeredo, based on a corpus of twenty -seven articles on Brazilian cinema published between 1956 and 1973, in the Rio's Tribuna da Imprensa and Jornal do Brasil periodicals. This temporal cut was delimited taking into account the profound transformations and innovative filmic manifestations that the cinematography of the country presented in that period. In this phase, Cinema Novo appears, influenced by Italian Neorealism and by the French Nouvelle Vague, whose objective was to make a reading from his films of the socio-political problems of Brazil. At the same time, Marginal Cinema emerges as a counterpoint to Cinema Novo, claiming that national cinema could only create legitimately Brazilian works, from the moment it accepted its underdeveloped position, working all elements of the "industrial plot" that was dumped in the culture Brazilian. Finally, one has the advent of erotic comedies, which would be labeled, in the future, as "pornochanchadas", since the genre had a popular character when combining comedy of customs and eroticism. During this period, Brazil underwent a military coup, which deposed the then President João Goulart, and, consequently, established a dictatorship in the country, which lasted for 21 years (1964-1985), causing Brazilian cinematographic production to be affected, directly or indirectly through this political scenario. In order to investigate the construction of meaning in a specific media production, represented by the critics of Ely Azeredo, the methodology adopted for the analysis of the corpus was the French Line Discourse Analysis / Mestre

No description available.

Le développement d'une résistance à l'action de l'insuline est fréquemment observée au cours de l'obésité et est à l'origine du diabète de type 2. L'amélioration du signal insulinique au sein de la cellule adipeuse (i.e adipocyte) est une stratégie thérapeutique intéressante en vue d'améliorer la sensibilité à l'insuline systémique de patients pré-diabétiques et diabétiques. Dans cette optique, l'inhibition de la lipase hormono-sensible (LHS) adipocytaire (enzyme responsable de la libération d'acides gras par le tissu adipeux) protège de l'insulino-résistance. Les mécanismes impliqués dans l'amélioration de la sensibilité à l'insuline n'étaient cependant pas encore connus. Mon travail de thèse a donc été axé sur l'identification des mécanismes liant l'inhibition de la LHS et l'amélioration de la sensibilité à l'insuline. Dans des adipocytes humains, l'invalidation de la LHS augmente le transport du glucose, la voie de la lipogenèse de novo (synthèse d'acides gras à partir du glucose) et le signal insulinique. Parmi les enzymes lipogéniques, l'élongase des acides gras à longue chaîne ELOVL6 voit son expression très fortement induite in vitro et in vivo lors d'une déficience pour la LHS, conduisant à un enrichissement en oléate des phospholopides et des triglycérides. L'invalidation d'ELOVL6 dans des adipocytes humains a permis de démontrer le rôle clé de l'élongase dans la médiation des effets bénéfiques de l'invalidation de la LHS. In vivo, une déficience pour ELOVL6 conduit également à une détérioration du signal insulinique dans le tissu adipeux. Dans des études cliniques, l'expression d'ELOVL6 s'effondre au cours de l'insulino-résistance et est restaurée après une chirurgie bariatrique. L'enrichissement en oléate dans les phospholipides par ELOVL6 est responsable des effets protecteurs de l'inhibition de la LHS sur le signal insulinique. Des adipocytes surexprimant ELOVL6 présentent d'ailleurs une augmentation de la fluidité membranaire associée à une amélioration du signal insulinique. Dans le foie, ELOVL6 est une cible du facteur de transcription ChREBP. ChREBP adipocytaire, notamment l'isoforme ChREBPß, a récemment été mis en évidence pour son rôle déterminant sur la sensibilité à l'insuline systémique. Chez l'Homme, nous avons observé in vitro et in vivo d'étroites corrélations positives entre les expressions adipocytaires de ChREBPß et d'ELOVL6. L'inhibition simultanée de la LHS et ChREBP réduit fortement l'expression d'ELOVL6 et annule l'enrichissement en oléate des phospholipides ainsi que l'amélioration du signal insulinique. De manière particulièrement intéressante, nous avons mis en évidence qu'une interaction physique entre la LHS et ChREBP inhibe la translocation nucléaire du facteur et son activité transcriptionnelle. L'activité catalytique de la LHS n'est pas requise pour assurer l'interaction. Nous avons aussi montré que cette interaction est spécifique de la LHS et est restreinte à l'adipocyte. En conclusion, notre travail a mis au jour une nouvelle voie déterminante pour la sensibilité à l'insuline adipocytaire, liant la LHS au facteur de transcription ChREBP et à sa cible, l'élongase des acides gras ELOVL6. L'enrichissement consécutif en oléate des phospholipides conduit à une augmentation de la fluidité membranaire et à une amélioration du signal insulinique. L'inhibition de l'interaction entre la LHS et ChREBP dans le tissu adipeux pourrait donc être bénéfique dans la prise en charge de l'insulino-résistance associée à l'obésité. / Insulin resistance is a feature frequently associated to obesity and an early defect in the development of type 2 diabetes. Improvement of fat cell insulin signaling may favor recovery of whole body systemic insulin sensitivity in pre-diabetic and diabetic states. In this context, inhibition of hormone-sensitive lipase (HSL) in adipocytes (an enzyme responsible for fatty acid release by adipose tissue) was demonstrated to be protective against insulin resistance. However, the mechanisms remained unclear. Consequently, my PhD work aimed at understanding the mechanisms linking HSL inhibition and improvement of insulin sensitivity. In human adipocytes, HSL gene silencing increased glucose transport, de novo lipogenesis and insulin signaling. Among de novo lipogenesis enzymes, ELOVL6 was preferentially induced in vitro and in vivo during HSL partial deficiency, resulting in enrichment of phospholipids and triglycerides in oleic acid. ELOVL6 gene silencing in human adipocytes provided the direct demonstration of the role of the enzyme in the beneficial effect of HSL inhibition. Fat cell insulin signaling was also impaired in adipose tissue of Elovl6 null mice. In clinical studies, ELOVL6 expression was blunted in insulin resistant states and restored after bariatric surgery. ELOVL6-mediated oleic acid enrichment of phospholipids was responsible for the positive effect of HSL inhibition on insulin signaling. FRAP studies revealed an increase in plasma membrane fluidity and insulin signaling in adipocytes overexpressing ELOVL6. In the liver, ELOVL6 is a target of ChREBP. Adipose ChREBP, notably the constitutively active isoform ChREBPß, recently emerged as a major determinant of systemic insulin action on glucose metabolism. In humans, we observed in several in vitro models and in vivo studies a strong positive association between adipose ChREBPß and ELOVL6. Dual HSL-ChREBP inhibition blunted adipose ELOVL6 expression in vivo and in vitro and mirrored ELOVL6 gene silencing on fatty acid profile and insulin signaling. Importantly, we found that physical interaction between HSL and ChREBP impairs ChREBP translocation into the nucleus and its transcriptional activity. A naturally short form of HSL devoid of catalytic activity retained the capacity to bind ChREBP. We also demonstrated that ChREBP-HSL interaction was specific of the lipase and restricted to adipocyte. To conclude, our work identifies a novel pathway critical for optimal insulin signaling in fat cells which links the neutral lipase HSL to the glucose-responsive transcription factor ChREBP and its target gene, the fatty acid elongase, ELOVL6. ELOVL6-mediated oleate enrichment in phospholipids increases membrane fluidity and improves insulin signaling. Inhibition of HSL/ChREBP interaction in adipose tissue may be beneficial in the treatment of obesity-associated insulin resistance.

Dissertação (mestrado) - Universidade Federal de Santa Catarina. Centro Socio-Economico / Made available in DSpace on 2012-10-16T07:37:06Z (GMT). No. of bitstreams: 0Bitstream added on 2016-01-08T19:08:15Z : No. of bitstreams: 1 93361.pdf: 6963239 bytes, checksum: 2f8d7f21eda8210fa865fc38d7b92393 (MD5) / A constante transformação das estratégias organizacionais, tornou-se fundamental à manutenção da competitividade, devido às crescentes mudanças no ambiente de negócio. No entanto, para que as mudanças das estratégias sejam efetivas, a correta percepção e avaliação das influências externas é necessária. A maneira como ambiente é percebido torna-se um elo crítico no ajuste da organização com o ambiente, evidenciando-se a figura do agente da escolha estratégica. Na presente dissertação analisou-se empiricamente as mudanças estratégicas de uma organização do ramo agroindustrial do estado de Santa Catarina, de acordo com o modelo da empresa no setor proposto por Child e Smith (1987). A pesquisa foi desenvolvida na forma de um estudo de caso longitudinal. Os dados foram obtidos através de entrevistas semi-estruturadas, dando-se ênfase na análise aos dados referentes aos últimos vinte anos da empresa. Os resultados reforçaram a necessidade das pesquisas, neste campo de investigação, abordarem aspectos relativos à história da organização, ao contexto onde ela está inserida e ao processo pelo qual as mudanças ocorrem. No estudo de caso detectaram-se ações reativas e proativas da organização às indicações do ambiente. Obedecendo em ambos caso, porém, a padrões particulares de ação e reação ligados, principalmente, a questões relacionadas com a memória da organização, a sua cultura, aos papéis de liderança e aos aspectos ligados a sua estrutura de poder.

Heparan sulfate (HS) is a class of linear, sulfated polysaccharides located on cell surface, secretory granules, and in extracellular matrices found in all animal organ systems. It consists of alternately repeating disaccharide units, expressed in animal species ranging from hydra to higher vertebrates including humans. HS binds and mediates the biological activities of over 300 proteins, including growth factors, enzymes, chemokines, cytokines, adhesion and structural proteins, lipoproteins and amyloid proteins. The binding events largely depend on the fine structure - the arrangement of sulfate groups and other variations - on HS chains. With the activated electron dissociation (ExD) high-resolution tandem mass spectrometry technique, researchers acquire rich structural information about the HS molecule. Using this technique, covalent bonds of the HS oligosaccharide ions are dissociated in the mass spectrometer. However, this information is complex, owing to the large number of product ions, and contains a degree of ambiguity due to the overlapping of product ion masses and lability of sulfate groups; as a result, there is a serious barrier to manual interpretation of the spectra. The interpretation of such data creates a serious bottleneck to the understanding of the biological roles of HS. In order to solve this problem, I designed HS-SEQ - the first HS sequencing algorithm using high-resolution tandem mass spectrometry. HS-SEQ allows rapid and confident sequencing of HS chains from millions of candidate structures and I validated its performance using multiple known pure standards. In many cases, HS oligosaccharides exist as mixtures of sulfation positional isomers. I therefore designed MULTI-HS-SEQ, an extended version of HS-SEQ targeting spectra coming from more than one HS sequence. I also developed several pre-processing and post-processing modules to support the automatic identification of HS structure. These methods and tools demonstrated the capacity for large-scale HS sequencing, which should contribute to clarifying the rich information encoded by HS chains as well as developing tailored HS drugs to target a wide spectrum of diseases.

Submitted by PPGH null (poshisto@ufba.br) on 2017-06-19T13:04:51Z No. of bitstreams: 1 LUANA MOURA QUADROS.pdf: 2118408 bytes, checksum: a974163719827e4b37376c934aa5b8ab (MD5) / Approved for entry into archive by Uillis de Assis Santos (uillis.assis@ufba.br) on 2017-06-26T18:00:06Z (GMT) No. of bitstreams: 1 LUANA MOURA QUADROS.pdf: 2118408 bytes, checksum: a974163719827e4b37376c934aa5b8ab (MD5) / Made available in DSpace on 2017-06-26T18:00:06Z (GMT). No. of bitstreams: 1 LUANA MOURA QUADROS.pdf: 2118408 bytes, checksum: a974163719827e4b37376c934aa5b8ab (MD5) / CAPES / Uma das expressões do cotidiano de Salvador nos anos trinta do século XX era a carestia de vida, que incidia diretamente sobre os gêneros de primeira necessidade, principalmente, nos alimentos básicos do consumo da população como carne verde, farinha, charque, feijão, pão, banha, bacalhau e café. As dificuldades de sobrevivência que se acentuavam com a elevação dos preços propiciavam uma atmosfera de tensão entre a população e comerciantes, que levavam as acusações de especuladores e gananciosos. Em 1937, com o golpe do Estado Novo, a carestia de vida ganhava contornos diferentes não só pela nova conformação política, mas pelas formas de intervenção na política econômica que visavam o controle do abastecimento. Para controlar os ânimos de revoltosos em potencial, em 1938 Getúlio Vargas definira os crimes contra a economia popular, impondo aos transgressores julgamento pelo Tribunal de Segurança Nacional. Com a guerra decretada às potências do Eixo em 1942, aumentava-se a vigilância sobre a produção de gêneros agropecuários e sua circulação, mas na contrapartida acentuava-se o clima de tensão e os preços disparavam. Uma confluência de fatores que caracteriza as condições de sobrevivência da população da capital baiana no período de 1937 a 1945. / One of the expressions of the daily day in Salvador in the 30s was the life high cost that ocurred directly over the foodstuff of first necessity, mainly, in the basics food of population such as green meat, farina, charque, beans, bread, banha, codfish and coffee. The surviving dificulties that become worst due to prices increases propitiates a tension ambiance among the people and traders, that took the accusations of speculators and greed. In 1937, with the Estado Novo coup, the life high cost got new different shapes not only for the new political conformation, but for the shapes of intervention in the economic politics that aims the supllies control. To control the moods of revolteds in potential, in 1938 Getúlio Vargas established the crimes against the popular economy, imposing judgement to the offenders by the Tribunal de Segurança Nacional. With the war decreted to the Eixo powers in 1942, the vigilance in the agro foodstuff increased and its circulation, but due that, the tension ambiance got worse and the prices ran up. A confluence of factors that characterize the conditions of surviving of the population of the bahian capital in the period of 1937 to 1945.

Submitted by Pós-Com Pós-Com (pos-com@ufba.br) on 2013-09-11T14:40:27Z No. of bitstreams: 1 José Umbelino Brasil.pdf: 1336159 bytes, checksum: ebd994101951dc8825a310bc27af20ef (MD5) / Approved for entry into archive by Alda Lima da Silva(sivalda@ufba.br) on 2013-09-12T18:19:31Z (GMT) No. of bitstreams: 1 José Umbelino Brasil.pdf: 1336159 bytes, checksum: ebd994101951dc8825a310bc27af20ef (MD5) / Made available in DSpace on 2013-09-12T18:19:31Z (GMT). No. of bitstreams: 1 José Umbelino Brasil.pdf: 1336159 bytes, checksum: ebd994101951dc8825a310bc27af20ef (MD5) / AS CRÍTICAS DO JOVEM GLAUBER – BAHIA 1956-1963 expõe a face menos conhecida de Glauber Rocha, a de crítico cinematográfico. A proposição deste trabalho é a de interagir com a expressão conceitual do pensamento glauberiano, buscando compreender de que maneira o autor construiu a sua fundamentação teórica sobre o cinema. Nesse percurso, a tese examina e analisa parte das críticas dedicadas ao cinema americano e brasileiro, verificando como os seus conteúdos serviram na formulação da invenção do movimento Cinema Novo, alicerçaram a política endógena de autor e foram transformados em processo de criação de uma poética própria.

Dissertação (mestrado) - Universidade Federal de Santa Catarina, Centro de Ciências Jurídicas, Programa de Pós-Graduação em Direito, Florianópolis, 2009. / Made available in DSpace on 2012-10-24T07:45:51Z (GMT). No. of bitstreams: 1 269346.pdf: 1286228 bytes, checksum: 756cb33f0efac86e19161898c63116b8 (MD5) / Essa dissertação versa sobre o Tribunal de Segurança Nacional, criado no Brasil em 1936 para julgar casos de crimes políticos, sendo extinto em 1945. O objetivo principal consiste em esclarecer alguns pontos sobre o Tribunal de Segurança Nacional através da documentação encontrada # na forma de debates em torno da sua criação, da legislação que o compôs e da produção doutrinária que a ele se seguiu # e desse modo juntar elementos suficientes para propor-se uma definição do que o Tribunal de Segurança Nacional deveria ter sido e quais argumentos jurídicos contribuíram para a forma como foi criado e posteriormente modificado. O trabalho divide-se basicamente em duas partes, cada uma disposta em dois capítulos: a primeira parte trata do Tribunal de Segurança Nacional, buscando analisar os principais elementos que contribuíram para a sua formação, suas modificações e sua extinção, em 1945. A segunda parte aborda as matérias colocadas sob sua competência, agrupadas em crimes contra a segurança do Estado e em crimes contra a economia popular. / This dissertation refers to the National Security Court, created in Brazil in 1936 to judge cases of political crimes, extinct in 1945. The major purpose is to clarify some points about the National Security Court through documentation found # in form of discussion about it's creation, the legislation that composed it and the doutrinary production that followed it # and by this means gatter sufficient elements to propose a definition of what the National Security Court should have been and wich legal arguments concur to the form it was created and posteriorly modified. The study is basically divided in two parts, each one arranged in two chapters: the first part concerns the National Security Court, analyzing the main elements that did contribute to its formation, modifications and extinction, in 1945. The second part refers to the items putted on the competence of the Court, grouped in crimes against the State security and crimes against popular economy.