In dieser Dissertation wurden Verfahren für die konvergente Synthese von Peptide entwickelt. Für die erweiterte native chemische Ligation kamen bisher raumbeanspruchende Auxiliare zum Einsatz, die anspruchsvolle Ligationen behinderten. Zudem waren die drastischen säure-basierten Abspaltbedinungen mit vielen post-translationalen Modifikationen nicht vereinbar. In dieser Arbeit wurde ein neuartiges Auxiliargerüst mit geringerem Raumanspruch untersucht, dass eine erheblich mildere Abspaltung unter basischen Bedingungen ermöglichte. Dazu wurde ein kleiner Elektronenakzeptorsubsituent eingeführt, durch den nach der Ligation eine das Zielpeptid freisetzende Eliminierungsreaktion induziert werden konnte. Weiterhin konnte die Verwendung des kommerziell verfügbaren Penicillamins als Vorläufer in der Ligations-Entschwefelungs-Strategie demonstriert werden. Abschließend wurde die Ligation mit sequenz-internem Cystein untersucht. Hierbei zeigte sich, dass Peptide mit Cystein in einer bestimmten Position bevorzugt in thioester-basierten Kondensationen reagierten. / In this thesis approaches for convergent synthesis of peptides have been developed. In the extended native chemical ligation so far space demanding auxiliaries encumbered challenging condensations. Furthermore the required drastic acidolytic cleavage conditions were furthermore incompatible with many post-translational modifications. In the thesis a nouvelle less space demanding scaffold was scrutinized which allowed a milder basic cleavage. Therefore a small electron-accepting substituent was introduced that enabled the induction of an elimination reaction liberating the target peptide after the peptide ligation had taken place. Furthermore the applicability of the commercially available penicillamine as precursor of valine in the ligation-desulfurization strategy could be demonstrated. Finally the ligation with sequence internal cysteines was scrutinized. Herein it could be shown that certain peptides with cysteine in a distinctive position of the sequence preferable reacted in thioester-based condensation reactions.

La Lycie, région mystérieuse et fascinante, avec une identité mixte qui englobait des éléments locaux et des éléments gréco-perses, n’a pas encore une étude « générale » : pour cette raison, j’ai décidé de consacrer mon travail à son analyse. Cette thèse a été divisée en trois grandes parties : la première présente une analyse de l’évolution géographique des frontières régionales à partir du VIe siècle av. J.-C. jusqu’à la provincialisation romaine avec une perspective synchronique et diachronique. La deuxième décrit l’histoire régionale, en tenant naturellement compte des sources arrivées jusqu’à nous. La troisième contient l’édition des fragments des historiens auteurs de Lykiaka, c’est à dire les fragments des historiens de langue grecque, auteurs d’oeuvres monographiques sur cette région asiatique (Menecrate de Xanthos ; Policarme ; Léon d’Alabande ; les fragments de la « Constitution des Lyciens » ; Alexandre Polyhistor ; Capiton de Lycie ; Aristenète). Les trois parties de ce travail ne sont pas des sections indépendantes et séparées, mais interagissent et communiquent constamment les unes avec les autres : un ouvrage compliqué et composite, mais qui cherche de reproduire la complexité d’une région « de frontière ». / A general study lacks about Lycia, mysterious and fascinating region with a mixed identity that included local and Greco-Persian elements: for this reason, I decided to dedicate my PhD thesis to her analysis. This work has been divided into three parts: the first presents a study on the changing geography of regional borders from the sixth century B.C. until the creation of the Roman province, with a synchronic and diachronic perspective. The second describes the regional history. The third contains the fragments of the authors of Lykiaka, in other words the fragments of Greek historians, authors of monographic works on Lycia (Menecrates of Xanthos; Policarme; Léon of Alabanda; the fragments of the "Constitution of the Lycians"; Alexander Polyhistor; Capito of Lycia; Aristaenetus). The three parts of this work constantly interact and communicate each other: a complicated and composite work, but able to reproduce the complexity of a “ border” region.

Made available in DSpace on 2016-04-28T19:59:08Z (GMT). No. of bitstreams: 1 Riviane Medino da Rocha.pdf: 569620 bytes, checksum: 0b430ce5152574ef01c32b50a10fd358 (MD5) Previous issue date: 2007-10-16 / The objective of this dissertation is the understanding of the character in Estorvo, by Chico Buarque, a contemporary novel. Considering the plot form, the character is resulted by the mixture of author-narrator, living the conflict of the fragmentation of his individuality. Basic theories had been supports for the revision of the narrative and character process: Mikhail Bakhtin, Walter Benjamin, besides the concept of imitation of Aristoteles, Antonio Candido, Brait, Schwartz, Muecke, Coast Young chicken, Massi and others. It also was necessary, by a comparative methodology, to approach some characteristics of two modern workmanships of Franz Kafka - Metamorphosis of 1915 and the Process of 1925, in order to give support to the characterization of the contemporary character in Estorvo. The first chapter A personagem-narrador em figuração focuses the character in the novel text and the essential narrative elements. It is discussed the taking of conscience of his unfinished individuality, living the dilemma of the dilution of his capacities, unknown to himself. The second chapter O Intertexto em Estorvo , points out some parallels between the characters, kafkiana and buarquiana, with the objective to create a methodological intertextuality of definition about his psychological and metaphorical characteristics. It is described some divergences and convergences that helps to characterize the points of view of the narrator-character in fusing of fragment selves broken up by the proper story in Estorvo, in functional duplicity: in time and space. The third chapter, O Cronotopo em Estorvo , uses the psychological and metaphorical concept of chronoscope under the transforming action of the irony, and the contamination of the character by the ambiguous performance of the narrator in first person which gives the disfigurement to the ironic character, between two consciences: individual and public / O objetivo desta dissertação é a leitura da personagem em Estorvo, de Chico Buarque, romance contemporâneo. Em forma de enredo, a personagem é resultado da mixagem autornarrador, vivendo o conflito da fragmentação de sua individualidade. Teorias fundamentais foram suportes da revisão do conceito do processo narrativo e de personagem: Mikhail Bakhtin, Walter Benjamin, além do conceito de imitação de Aristóteles, Antonio Candido, Brait, Schwartz, Muecke, Costa Pinto, Massi e outros mais. Fez-se necessário, por uma metodologia comparativa, aproximar algumas características de duas obras modernas de Franz Kafka Metamorfose de 1915 e O Processo de 1925, de modo a dar suporte à caracterização da personagem contemporânea de Estorvo. O primeiro capítulo intitulado A personagem-narrador em figuração tem por foco a personagem no texto romanesco e seus elementos essenciais narrativos. Discutimos a tomada de consciência de sua individualidade inacabada, vivendo o dilema da diluição de suas capacidades, por ela própria desconhecidas. O segundo capítulo intitulado O Intertexto em Estorvo, traçamos alguns paralelos entre as personagens, kafkiana e buarqueana, com o objetivo de criar uma intertextualidade metodológica de definição das suas características psicológicas e metafóricas. Chegamos a descrever algumas divergências e convergências que nos auxiliaram a melhor caracterizar os pontos de vista do narrador-personagem em fusão de Eus fragmentados pelo próprio relato em Estorvo, em duplicidade funcional: em tempo e espaço. O terceiro capítulo, com título O Cronotopo em Estorvo, faz uso do conceito de cronotopo psicológico e metafórico sob a ação transformadora da ironia, e a contaminação da personagem pela performance ambígua de narrador em primeira pessoa. Característica essa que dá a desfiguração à personagem irônica, entre duas consciências: a individual e a pública

Embora o romance Em busca do tempo perdido seja incontestavelmente uma obra inacabada, não se trata, entretanto, de uma obra incompleta. Seu fechamento circular, promovido pelo diálogo entre o primeiro e último volumes, foi tema de grande parte da crítica proustiana. Com relação à sua composição, seu processo escritural passou por diversas mudanças e a construção, equiparada à execução de uma catedral, poderia igualmente ser caracterizada pela colagem, montagem ou costura dos fragmentos textuais esboçados nos setenta e cinco cadernos de rascunho. A busca pela unidade em meio a essa profusão de textos levou o escritor à atividade incessante de releitura e reescritura e, consequentemente, ao inacabamento da obra. O trabalho que ora apresentamos tem por objetivo o estudo dessa composição, a partir da leitura e análise dos cadernos 53 e 55, ambos consagrados à elaboração da história de Albertine. / Although the novel In Search of Lost Time is certainly unfinished, it is not an incomplete work. Its round ending, promoted by the dialogue between the first and last volumes, was the subject of much Proustian criticism. With respect to its composition, its writing process has gone through many changes and the construction, equivalent to the execution of a cathedral, could also be characterized by the process of montage or the stitching of textual fragments contained in Prousts seventy-five exercise books. The search for unity amongst this profusion of texts has led the writer to the ceaseless activity of rereading and rewriting and thus to the incompleteness of the work. The analysis presented here is aimed at studying this composition, having the reading and the analysis of exercise books 53 and 55, both related to the elaboration of the story of Albertine, as a starting point.

Recent years have seen the emergence of Mycobacterium abscessus, a highly drug-resistant non-tuberculous mycobacterium, which causes life-threatening infections in people with chronic lung conditions like cystic fibrosis. This opportunistic pathogen is refractory to treatment with standard anti-tuberculosis drugs and most currently available antibiotics, often resulting in accelerated lung function decline. This project aims to use a structure-guided fragment-based drug discovery approach to develop effective drugs to treat M. abscessus infections. During the early stage of the project, three bacterial targets were identified, based on analysis of the structural proteome of M. abscessus and prior knowledge of M. tuberculosis drug targets, followed by gene knockout studies to determine target essentiality for bacterial survival. The three targets from M. abscessus were then cloned, expressed and purified and suitable crystallization conditions were identified leading to the determination of high resolution structures. Further, a large number of starting fragments that hit the three target proteins were determined, using a combination of biophysical screening methods and by defining crystal structures of the complexes. For target 3, PPAT (Phosphopantethiene adenylyl transferase), a chemical linking of two fragments followed by iterative fragment elaboration was carried out to obtain two compounds with low micromolar affinities in vitro. However, these compounds afforded only low inhibitory activity on M. abscessus whole cell. All starting fragments of target 2, PurC (SAICAR synthase), occupied the ATP indole pocket. Efforts were then made to identify further fragment hits by screening diverse libraries. Sub-structure searches of these initial fragment hits and virtual screening helped to identify potential analogues amenable to further medicinal chemistry intervention. While fragment hits of target 1, TrmD (tRNA-(N1G37) methyl transferase), were prioritized, whereby two chemical series were developed using fragment growing and merging approaches. Iterative fragment elaboration cycle, aided by crystallography, biophysical and biochemical assays led to the development of several potential lead candidates having low nano-molar range of in vitro affinities. Two such compounds afforded moderate inhibition of M. abscessus and stronger inhibition of M. tuberculosis and S. aureus cultures. Further chemical modifications of these compounds as well as others are now being done, to optimize cellular and in vivo activities, to be ultimately presented as early stage clinical candidates.

Orientador: Wagner André Pedro / Banca: Paulo de Marco Junior / Banca: Luiz Dino Vizotto / Resumo: A região Noroeste do estado de São Paulo, bem como microregiões adjacentes, carece de informações concisas sobre a composição floristica e faunística, em especial à avifauna. Esta condição aliada ao alto grau de degradação ambiental apresentado favorece a perda incontestável de biodiversidade. A grande maioria dos remanescentes florestais do interior paulista são representados por pequenos trechos de mata, isolados na maior parte daz vezes por matrizes inóspitas como cultivo de cana-de-açúcar e pastagens. As formações florestais representadas pela Floresta Estacional tipo Semidecidual cobrem a maior parte do interior do estado de São Paulo e, portanto refletem um quadro sazonal relativamente acentuado com dois períodos bastante distintos de seca e chuva. Esta condição sazonal interfere em vários aspectos da historia de vida da fauna inserida na região, especialmente das aves e artrópodes. Neste sentido, procuro-se aqui descrever a estrutura da comunidade de aves em um remanescente florestal de pouco mais de 2.400 ha, com inferências ao grau de endemismo de algumas espécies, grau de conservação e ameaça, habitats e dependência florestal. Ainda, em segundo momento, teve-se por objetivo analisar e avaliar os efeitos da sazonalidade e disponibilidade de presas invertebradas para a comunidade como um todo, mas em especial aos insetívoros. Ao todo foram xi registradas 171 espécies na área de estudo, sendo 95 no remanescente florestal e 76 em áreas de entorno que englobaram ambientes antropizados e aquáticos. Sete espécies foram consideradas endêmicas da Mata Atlântica ou Cerrado, 22 apresentando algum grau de ameaça e 28 espécies dependentes da floresta. Os resultados para as relações sazonais demostraram-se diferentes para as diferentes guildas, sendo que frugívoros tiveram suas populações oscilando mais do que insetívoros e onívoros entre... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: The Northwest region of São Paulo, as well as adjacent microregions, lacks concise information on the floristic composition and wildlife, especially the avifauna. This condition coupled with the high degree of environmental degradation presented incostestável favors the loss of biodiversity. Most of the interior of forest remnants are represented by small patches of forest, mostly isolated daz times inhospitable matrix as the cultivation of sugar cane and pasture. The forest types represented by seasonal forest type Semidecidual cover most of the state of Sao Paulo, and thus reflect a relatively strong seasonal table with two very distinct periods of drought and rain. This seasonal condition interferes with many aspects of life history fauna inserted into the region, especially birds and arthropods. In this sense, I try here to describe the community structure of birds in a forest remnant of just over 2,400 ha, with inferences to the degree of endemism of some species, conservation status and threat, and dependence on forest habitats. Still, in the second time it was taken to analyze and evaluate the effects of seasonality and availability of invertebrate prey for the community as a whole, but especially the insectivores. Altogether 171 species were recorded in the study area, 95 and 76 in the remaining forest in the surrounding areas have included aquatic and anthropogenic environments. Seven species are endemic to the Atlantic Forest and Cerrado, 22 showing some degree of threat and 28 species of forest-dependent. The xiii results demonstrated for seasonal relationships are different for different guilds, and their populations ranging frugivores had more than insectivores and omnivores between the dry and rainy seasons. The species richness was significantly correlated with invertebrate abundance of foliage. The breeding season of birds recorded in the remaining forest... (Complete abstract click electronic access below) / Mestre

Este trabalho diz respeito à diversidade genética de Myrciaria floribunda (West ex Willdenow) Berg (cambuí), uma espécie arbórea de sub-bosque, em paisagem fragmentada da Serra da Mantiqueira, MG. A diversidade genética foi estudada a partir dos dados de análises eletroforéticas de isoenzimas. Foram coletadas folhas de indivíduos jovens em duas populações dentro de um fragmento controle (5810,27 ha) e em dois fragmentos menores (18 e 10 ha), sendo quarenta plantas por fragmento. Através da análise de sete locos isoenzimáticos foi avaliada o número médio de alelos por loco (Â), número médio de alelos por loco polimórfico (Âp), porcentagem de locos polimórficos ( $ P ), heterozigosidade média esperada ( $ He ), heterozigosidade observada ( $ Ho ) e freqüências alélicas, utilizando-se alelos de baixa freqüência. A heterozigosidade observada para os quatro fragmentos foi elevada (entre 0,288 a 0,386). A variação genética intrapopulacional, avaliada pela riqueza alélica foi menor nos fragmentos menores que nas populações contida no fragmento controle. Alguns alelos encontrados em menor freqüência no fragmento controle tiveram suas freqüências reduzidas nos fragmentos menores, sendo que um alelo foi perdido nos fragmentos menores, provavelmente devido à redução do tamanho populacional durante a fragmentação, ou à deriva genética após a fragmentação ou a amostragem realizada. O índice de diversidade gênica de Nei (heterozigosidade esperada) não foi muito diferente entre as quatro populações, apesar de mostrar uma tendência à diminuição nos fragmentos menores. Os indivíduos jovens analisados apresentaram valores de f ˆ = 0,0864, sugerindo que estas populações tem vindo de cruzamentos panmíticos, provavelmente anteriores ao processo de fragmentação. Estas quatro populações apresentaram pequena diferenciação genética entre elas ( p q ˆ = 0,0555), o número estimado de migrantes foi relativamente alto (Nm= 4,25). Este trabalho desta forma não foi capaz de detectar o efeito da fragmentação na endogamia desta espécie. / This research is about effects of forest fragmentation on the genetic diversity of Myrciaria floribunda (West ex Willdenow) Berg (cambuí), a tropical tree species from the Atlantic Forest (Mantiqueira hills), State of Minas Gerais. This work is about genetic diversity studies of de Myrciaria floribunda (cambuí), a late successional species from fragmented landscapes of the Mantiqueira hills. Genetic diversity was assessed using electrophoresis of isozymes. Leaves from juveniles trees were collected from four populations, two within a control fragment (5810,27 ha) and from two other small fragments (18 e 10 ha). Forty plants were sampled per populations. Seven loci were studied regarding the average number of alleles per locus (Â), the average number of alleles per polymorphic loci (Âp), percentage of polymorphic loci ( $ P ), expected ( $ He ) and observed heretozigosity ( $ Ho ), and allelic frequency, using alleles of low frequency. The observed heterozigosity was high for all fragments (from 0,288 to 0,386). Within population genetic diversity, measured by allelic richness was lower in small fragments compared with the large fragment. Low frequency alleles found in the large fragment showed an even lower frequencies in the two small fragments. One allele was lost in the small fragments, probably due to a reduction in population size because of the fragmentation process, and probably due to genetic drift or sampling strategy. Nei’s diversity index (expected heterozigosity) did not differ among populations, although lower in the small fragments. The result for f ˆ = 0,0864 indicated that the current populations probably were originated from panmmitic populations, established before the fragmentation process. Little genetic differentiation was detected among populations ( p q ˆ = 0,0555), however the number of migrants was considerably high (Nm= 4,25). Therefore this work was not able to detect the effects of endogamy due to the forest fragmentation on the studied species.

La méthode de conception de médicaments à partir de molécules « fragments » (connue sous le nom de « Fragment-Based Drug Design ») a été proposée au milieu des années 90, et a depuis été reconnue comme une alternative tangible aux techniques plus classiques de recherche de médicaments telles que le criblage à haut débit par exemple. La méthode des fragments consiste à cribler un petit nombre (< 10000) de composés organiques de faible poids moléculaire (< 300 Da) afin de détecter ceux qui se lient à la cible (protéine ou acides nucléiques). Du fait de leur faible complexité, les fragments présentent une affinité faible pour la cible, et la détection s'effectue généralement grâce à une technique biophysique (en particulier, résonance magnétique nucléaire (RMN), cristallographie aux rayons X, résonance plasmonique de surface). Les fragments « hits » sont ensuite modifiés par addition de nouvelles fonctions chimiques, ou par liaison de deux fragments, afin d'élaborer, étape par étape, une molécule capable d'établir des interactions plus nombreuses avec la cible, et d'améliorer ainsi l'affinité. Comparée aux méthodes classiques de criblage haut débit, la méthode des fragments offre divers avantages, notamment une meilleure exploration de l'espace chimique, une meilleure efficacité de liaison des molécules « hits », et une plus grande facilité d'optimisation des hits en molécules plus affines. Dans le cadre de ce projet de thèse, plusieurs aspects de la méthode des fragments ont été abordés : dans une première partie, nous étudions un cas concret d'application de la méthode des fragments à la recherche d'un inhibiteur de la peroxiredoxine 5 humaine, en utilisant la RMN comme outil de criblage des fragments ainsi que comme outil d'étude des interactions protéine-fragment. La découverte d'un inhibiteur de cette enzyme représente une avancée importante, qui devrait permettre de mieux comprendre son fonctionnement. Les autres parties de ce projet de thèse abordent des aspects plus méthodologiques de la méthode des fragments : les fragments conservent-ils leur site de liaison, leur efficacité de liaison et leur mode d'interaction au cours de leur élaboration en inhibiteur ? Les fragments peuvent-ils être spécifiques d'une protéine ? D'un site de liaison particulier ? Ces questions, rarement traitées, sont pourtant essentielles à la compréhension du comportement des molécules fragments, et sont abordées d'une part en défragmentant plusieurs inhibiteurs de la protéine Bcl-xL et en étudiant par RMN le comportement de ces fragments vis-à-vis de la protéine en termes d'affinité et de site de liaison, d'autre part en réalisant le criblage par RMN d'une série de fragments sur cinq protéines différentes (peroxiredoxine 5 humaine, sérum albumine humaine et trois protéines homologues de la famille Bcl-2). De manière générale, ce projet de thèse vise à étudier des aspects peu abordés de la méthode des fragments et à proposer des pistes permettant de mieux comprendre le comportement des fragments vis-à-vis de leur cible, au cours du criblage initial comme lors de leur optimisation / Fragment-Based Drug Design (FBDD) has been proposed in 1996 and has since been recognized as a tangible alternative to the more classical drug discovery methods such as High-Throuput Screening. FBDD consists of screening a small number (< 10 000) of low-molecular weight (< 300 Da) compounds and detect those that bind to the target (protein or nucleic acids). Because of their low complexity, fragment molecules usually display low affinities for their target, hence detecting fragment-protein interactions is mostly achieved using a biophysical technique (mostly Nuclear Magnetic Resonance (NMR), X-ray crystallography or Surface Plasmon Resonance). “Hit” fragments are then modified by addition of chemical substituents, or linked together, so as to elaborate a more complex molecule, forming more interactions with the target and hence displaying an improved affinity. As compared to the more classical High Throughput Screening method, fragment screening provides several advantages, including a better exploration of chemical space, highly ligand-efficient hits and easier optimization of the hits into more affine molecules. In this PhD project, several aspects of FDBB have been addressed : first, FBDD approaches were applied to the research of an inhibitor of the human peroxiredoxin 5 protein, using NMR not only as a screening method but also for the characterization of the protein-fragment interactions. The discovery of an inhibitor against this enzyme would allow to better understand its function. Next, methodological aspects of the FBDD method were addressed : Do fragments conserve their binding site, binding efficiency and mode of interaction upon optimization? Can the fragments display specificity towards a given target? Towards a given binding site? These issues, rarely studied, are yet essential to the understanding of the behavior of fragment molecules, and will be addressed firstly by defragmentating several Bcl-xL inhibitors into fragments and studying their behavior towards the protein in terms of a_nity and binding mode, secondly by screening a set of fragments against five different proteins (human peroxiredoxin 5, human serum albumin and three homologous proteins of the Bcl-2 family of proteins). More generally, this PhD project aims at studying less characterized aspects of the fragment methodology and proposing answers to better understand the behavior of fragment molecules towards their targets, both in the initial screening step and then during their optimization

INTRODUÇÃO: A incidência de tuberculose (TB) em Santos (SP) situa-se em 73/100.000 habitantes-ano. A prevalência média de coinfecção TB/HIV é de 16%, taxas de cura e abandono de tratamento, entre casos novos são, respectivamente, 71% e 12%. Tais indicadores sugerem elevado risco para TB multidroga resistente (TBMR) no município, com incidência estimada em 1,9/100.000 habitantes-ano. OBJETIVO: Descrever e analisar o perfil de sensibilidade às drogas (TS) de primeira e segunda linha de tratamento entre pacientes com TB pulmonar (TBP), estimar a incidência de TBMR e a proporção de TB extensivamente resistente (TBXR); analisar aspectos moleculares, epidemiológicos e institucionais dos casos de TBP resistentes em Santos (SP). MÉTODOS: Estudo descritivo de uma coorte de pacientes de TBP, com início de tratamento ou retratamento entre 01 de janeiro de 2011 a 31 de dezembro de 2012. Definiu-se como caso de TBP, indivíduos com 15 anos ou mais, de ambos os sexos, residentes no município de Santos, com manifestações clínicas compatíveis com TBP e confirmação por cultura com isolamento de Mycobacterium tuberculosis. As variáveis de interesse para o estudo foram as características sociodemográficas, história atual e pregressa de TB, aspectos relativos ao tratamento, co-morbidades, ao diagnóstico e resistência a drogas. Para as análises comparativas entre proporções foram usados os testes qui-quadrado de Pearson e o Exato de Fisher e para variáveis contínuas o teste T de Student ou o de Kruskal - Wallis. Os perfis genéticos dos isoladas resistentes a ao menos uma droga foram obtidos pela técnica RLFP e analisados pelo programa Bionumerics versão 5.0 (Applied Maths - Bélgica). A descrição da distribuição espacial da TB resistentes e clusters foram feitas mediante a inserção dos casos no mapa de Santos, por endereço de residência, segundo o índice de vulnerabilidade social. RESULTADOS: Dos 263 casos de TBP selecionados, 68,4% (180/263) eram do sexo masculino, a mediana da idade foi de 38 anos, 8,7% (23/263) eram diabéticos; 20,4% (42/206) dos casos novos apresentavam ao menos um fator de risco para TBMR; destacando-se entre estes casos 10,7% (22/206) de confecção HIV/TB; 47,3% (123/260) tiveram tratamento supervisionado, 14,7% (91/617) dos contatos foram examinados, 18,6% (49/263) foram hospitalizados durante o tratamento, perfazendo uma média de 145,4 dias por paciente. Entre os casos resistentes a ao menos uma droga, a resistência à isoniazida foi 8,4% (22/263) e à rifampicina 3,8% (10/263) dos casos. A TBMR primária foi encontrada em 1,9% (4/206) dos casos e destes 25,0% (1/4) eram TBXR. A incidência média anual de TBMR foi de 0,57/100,000 habitantes. Dos 25 isolados resistentes ao menos uma droga, submetidos à RFLP, 12 (48,0%) foram agrupados em seis grupos genéticos, com dois pacientes em cada grupo. CONCLUSÕES: A elevada proporção TBMR primária, com um caso de TBXR enfatizam a necessidade de universalizar a cultura e TS, ampliar a cobertura do tratamento supervisionado, a investigação rotineira dos contatos e o monitoramento da resistência a drogas. O fortalecimento da vigilância da resistência às drogas é indispensável para o contínuo aperfeiçoamento do Programa de Controle da TB, especialmente em regiões de elevada carga da doença / INTRODUCTION: The incidence of tuberculosis (TB) in Santos (SP) is located around 73 / 100,000-year, approximately double that found on average in the country. The average prevalence of TB / HIV is 16% cure rates and treatment dropout among new cases are, respectively, 71% and 12%. Such indicators suggest high risk for multidrug-resistant TB (MR-TB) in the city, with the incidence estimated at 1.9 / 100,000-year. OBJECTIVE: To describe and analyze the sensitivity to drugs of first and second line treatment of patients with pulmonary TB (PTB) to estimate the incidence of MR-TB and extensively drugresistant TB (TBXR), describe molecular and institutional aspects, spatial distribution, epidemiological PTB resistant cases in the city of Santos (SP). METHODS: A descriptive study of a cohort of patients with PTB residing in the city who started treatment or retreatment in the period January 2011 to December 31, 2012. The case definition PTB individuals 15 years or more, both sexes, living in the city of Santos (SP), who present clinical manifestations compatible with PTB and whose confirmation was made by culture with isolation of M. tuberculosis. The variables of interest for the study were: bacteriological / laboratory socio-demographic characteristics, current and previous history of TB, aspects related to treatment, and comorbidities. For comparative analyzes of proportions the chi-squared tests and Fisher\'s exact were used for continuous variables and the Student t test or the Kruskal - Wallis. The genetic profiles of isolates resistant to at least one drug were obtained by RFLP (length polymorphism restriction fragment) and analyzed using version BioNumerics 5.0 (Applied Maths - Belgium) software. The description of the spatial distribution of resistant TB and the clusters was made by inserting the cases in Santos map, by address of residence, which was according to the index of social vulnerability. RESULTS: Of the 263 cases of PTB selected, 68.4% (180/263) were male, th median age was 38 years, 8.7% (23/263) were diabetes; 20.4% (42/206) of new cases had at least one risk factor for MR-TB, especially 10.7% (22/206) of making HIV / TB; 47.3% (123/260) underwent supervised treatment, 14.7% (91/617) of the contacts were examined, 18.6% (49/263) were hospitalized during treatment, totaling 7127 days of hospitalization with a mean 145.4 days per patient. Among the cases resistant to at least one drug resistance to isoniazid 8.4% (22/263) and rifampin 3.8% (10/263) of the cases was found. The primary MR-TB was found in 1.9% (4/206) of MR-TB cases and of these 25.0% (1/4) were TBXR. The average annual incidence of MDR-TB was 0.57/100,000 inhabitants. Of the 25 isolates resistant least one drug, subjected to molecular characterization of IS6110, 12 (48.0%) were grouped in six clusters, with each group including two isolates. CONCLUSIONS: A high proportion of primary MR-TB, including a case of TBXR emphasizes the need to universalize culture and TS, expand the coverage of supervised treatment, routine investigation of contacts and monitoring of drug resistance. The strengthening of the surveillance of drug resistance is essential for continuous improvement of the TB Control Program, especially in regions of high disease burden

<p>BACKGROUND: Abetalipoproteinemia (ABL) is a rare autosomal recessive disorder caused by mutations in the gene coding for microsomal triglyceride transfer protein (MTTP).</p><p>AIM: To characterize the genetic basis of ABL in two unrelated patients.</p><p>RESULTS: In the first patient, the substitution c.1911C>T in exon 12 of the <em>MTTP</em> gene, resulting in the protein substitution p.P552L, was discovered using mutation screening. The parents are heterozygous and the proband is a homozygous carrier of this substitution. Using restriction fragment length polymorphism (RFLP), 100 control subjects were analyzed and none carried the substitution indicating that it is a novel <em>MTTP </em>mutation. Sequencing of the other ABL patient showed that the proband carried a homozygous single base insertion, at position  c.2342IVS16+2-3insT, located at the donor splice-site of intron 16 resulting in skipping of exon 16 and truncation of the protein. The proband's mother is heterozygous for the insertion while the father does not carry the insertion. Multiplex ligation-dependent probe amplification (MLPA) did not identify any deletion encompassing exon 16 in the proband, father or mother. Nonpaternity was excluded using polymorphic markers from several chromosomes. Haplotype analysis using markers spanning chromosome 4 revealed  heterodisomy (two homologous chromosomes) of 4p and the distal part of 4q, and isodisomy (duplication of one chromosome) of 4q12-4q26.</p><p>CONCLUSION: These data show that the cause of ABL in one of the patients is a missense mutation, p.P552L, while the cause of ABL in the other patient is due to uniparental disomy, probably resulting from non-disjunstion in meiosis I.</p>