Le système spirocyclique [6.4] est un motif récurrent dans un certain nombre de produits naturels tels que les Lancifodilactones, les Micrandilactones ou les Rubriflordilactones. Ces structures polycycliques représentent un défi synthétique pour les chimistes organiciens puisqu’elles présentent au moins neuf centres stéréogènes dont plusieurs sont quaternaires. L’objectif principal de ce travail était de développer de nouvelles réactions métallo-catalysées, et de les utiliser comme étape clé afin d’obtenir rapidement et efficacement le squelette polycyclique de ces composés. La première partie de ces travaux a été consacrée au développement d’une nouvelle réaction domino pallado-catalysée donnant accès, selon les conditions employées, à différents cœurs polycycliques de nortriterpénoïdes issus d’une même famille, de manière totalement diastéréosélective. Cette séquence domino de trois réactions en un seul pot permet donc la synthèse unifiée de différents squelettes de produits naturels isolés du genre Schisandra, en une seule étape. Une approche au squelette tétracyclique ABCD de la Lancifodilactone F appliquant cette méthodologie pour la construction du motif spirolactone a été envisagée. Un cycle D hautement fonctionnalisé a ainsi été synthétisé de manière diastéréosélective via un réarrangement d’Ireland-Claisen et une réaction de métathèse cyclisante. Une réaction de cyclopropanation intramoléculaire et la réaction domino devraient permettre d’obtenir, par la suite, le squelette complet de la Lancifodilactone F. Dans une deuxième partie, deux approches aux coeurs CDEF et ACDE de la Rubriflordilactone A ont été développées autour d’une même réaction clé, c’est-à-dire une cycloaddition [2+2+2] d’un composé triynique permettant d’accéder en une seul étape au squelette tricyclique CDE. Le motif tétracyclique ACDE devrait être rapidement accessible après quelques aménagements de la voie de synthèse initiale. / The spiro [6.4] ring system is a recurring structural motif in numerous natural products such as Lancifodilactones, Micrandilactones and Rubriflordilactones. These polycyclic structures represent a synthetic challenge for organic chemists. Indeed, these molecules present at least nine stereogenic centers including several quaternary ones. The main goal of this work was to develop new metal-catalyzed reactions as key steps to obtain quickly and efficiently the polycyclic core of those natural products. The first part of these studies was dedicated to the development of a new palladium-catalyzed domino reaction leading to, depending on the conditions used, different polycyclic cores of nortriterpenoids coming from the same family, in a totally diastereoselective manner. This domino sequence of three reactions allowed the collective synthesis of various skeletons of natural products isolated from the Schisandra genus, in one pot. An approach to the tetracyclic core of Lancifodilactone F applying this methodology for the construction of the spirolactone moiety was then envisaged. A highly functionalized cycle D was synthetized in a diastereoselctive way via an Ireland-Claisen rearrangement and a ring closing metathesis reaction. Afterwards, an intramolecular cyclopropanation followed by the domino reaction should allow an access to the ABCD skeleton of Lancifodilactone F. In the last part, two approaches to the CDEF and ACDE cores of Rubriflordilactone A were developed around the same key reaction: a [2+2+2] cycloaddition of an acyclic triynic compound. The tricyclic skeleton CDE was reached at once in a single step. The ACDE tetracyclic moiety should be quickly accessible after few modifications of the initial strategy.

O objetivo deste trabalho foi investigar o uso de NbCl5 como ácido de Lewis em reações de cicloadição. Foram estudadas algumas reações de cicloadição [2 + 2], Reações de Diels-Alder entre enonas e ciclopentadieno e Reações da aza-Diels-Alder com bases de Schiff, avaliando o efeito da temperatura e da concentração molar do NbCl5. A comparação dos rendimentos, dos produtos formados e do tempo de reação com NbCl5, também foi um ponto fundamental da pesquisa. As reações de cicloadição [2 + 2] foram realizadas entre ésteres propiólicos e diferentes tipos de alcenos (éteres enólicos de silício e alcenos alifáticos). Nas reações com os éteres enólicos de silício não foi verificada a formação dos respectivos adutos de ciclobuteno, pois o NbCl5 promove a quebra da ligação oxigênio-silício, não levando à formação do produto desejado. Nas reações com os alcenos alifáticos foi possível obter o respectivo aduto, porém com rendimentos menores que com outros ácidos de Lewis. Quanto às reações de Diels-Alder, foram investigadas as reações entre diferentes ciclo-enonas (dienófilos de baixa reatividade) com ciclopentadieno (dieno) na presença de NbCl5. Os resultados obtidos indicam que o NbCl5 é um bom ácido de Lewis para ativar reações de Diels-Alder, apresentando tempos reacionais menores e alta diastereosseletividade a temperaturas mais baixas, quando comparado com outros ácidos de Lewis. A possibilidade de efetuar reações do Diels-Alder a -78 0C é um dos aspectos de destaque neste trabalho, pois, além de demonstrar a forte ativação do sistema enona exercido pelo NbCl5, possibilita obter produtos com alta seletividade. Paralelamente aos estudos de sistemática reacional foram realizados estudos de elucidação estrutural completa de alguns dos compostos sintetizados, utilizando-se diversas técnicas de RMN (uni e bidimensionais), e o auxílio de cálculos teóricos. Nas reações de aza-Diels-Alder entre bases de Schiff e di-hidropirano, o NbCl5 se mostrou um ótimo catalisador para a síntese de derivados de piranoquinolinas. Estes derivados formam uma importante classe de produtos naturais que apresentam ampla atividade biológica. As reações foram conduzidas com baixas concentrações de nióbio e em tempos relativamente curtos, obtendo-se rendimentos variando de 72 a 96 %. Outro fator a se destacar é a alta diastereosseletividade encontrada nestas reações. Também foram realizados estudos da atividade tripanocida dos derivados de piranoquinolinas preparados através das reações de aza-Diels-Alder catalisadas por NbCl5. / The aim of this work was to investigate the use of NbCl5 as Lewis acid in cycloadittion reactions. We have studied [2 + 2] cycloaddition reactions, Diels-Alder reactions between enonas and cyclopentadiene and aza-Diels-Alder reactions with Schiff bases. The effects of the temperature and of the molar concentration of NbCl5 were also evaluated. Comparasion of reaction yields, obtained products as well as reation time with NbCl5 were also a key point on this work. The [2 + 2] cycloaddition reactions were performed using propiolic ester and different types of alkenes (silyl enol ethers and aliphatic alkenes). In the reactions with silyl enol ethers, formation of the corresponding cyclobutene aductts was not verified, since the NbCl5 promotes the rupture of oxygen-silicon bonds, and the desired product is not obtained. In the reactions with aliphatic alkenes, it was possible to obtain the adduct, however, in lower yields as compared to those obtained with others Lewis acids. Regarding the Diels-Alder reactions, we have investigated reactions using different cycloenones (dienophiles of low reactivity) with cyclopentadiene (diene) with NbCl5. The obtained results indicate that NbCl5 is a good Lewis acid to activate these Diels-Alder reactions, resulting in shorter reaction times and higher diastereoselectivity at lower temperatures than other Lewis acids. The possibility of carrying out Diels-Alder reaction at -78 0C is another remarkable aspect of this work. Besides demonstrating the strong activation of the enone system by NbCl5, it opens the possibility of obtaining high stereoselectivity. We have also performed studies of complete structural elucidation of some compounds by using different NMR techniques (uni and bidimensional), with the help of theoretical calculations. In the aza-Diels-Alder reactions between Schiff bases and dihydropyran, the NbCl5 was an excellent catalyst for the synthesis of pyranoquinoline derivatives. These derivatives are an importante class of natural products that exhibit wide range of biological activity. The reactions were carried out at low concentration of niobium and in relatively short times, resulting in yields varying from 72 to 96 %. Another aspect that should be remarked is the high diastereoselectivity found in these reactions. We have also carried out studies of tripanocydal activity of pyranoquinoline derivatives prepared through aza-Diels-Alder reactions catalyzed by NbCl5.

The aim of my PhD Thesis was to explore the potential of helically chiral N-heterocyclic carbene (NHC) ligands in asymmetric catalysis. Helicenes and helicene-like molecules are inherently chiral. Their application in this field has been rather limited. To date, only a few examples of enantiopure helically chiral NHCs have been described in the literature. Using a well-established method based on the diastereoselective metal catalysed [2+2+2] cycloisomerisation of centrally chiral triynes as the key step, I have synthesised a series of optically pure 2H-pyran based penta- and hexahelicenes bearing an amino group on the terminal benzene ring. The triynes were prepared by a sequence of Sonogashira and Mitsunobu coupling reactions using the commercially available (S)-but-3-yn-2-ol as the source of chirality. The resulting aminooxa[5]- and aminooxa[6]helicenes were then converted into the corresponding 1,3-disubstituted imidazolium salts, from which, upon deprotonation, the helically chiral N- heterocyclic carbenes were generated. To evaluate the performance of the new helically chiral ligands, the enantioselective Ni0 - catalysed [2+2+2] intramolecular cycloisomerisation of prochiral triynes to nonracemic dibenzohelicenes was chosen as a model reaction. All the synthesised imidazolium salts provided,...

Bien que le Diabète de Type 2 soit parfois considéré comme évitable, il est généralement irréversible et les traitements actuels sont communément inefficaces pour stopper la progression inexorable de la maladie vers un mauvais contrôle glycémique et des complications dégénératives micro- et macrovasculaires. Le DT2 est caractérisé par une altération de la sécrétion d’insuline par les cellules beta pancréatiques, combinée à une absence de réponse des organes cibles à l’insuline (insulino-résistance) incluant le foie, le tissu adipeux et le muscle squelettique. Malgré 40 ans de recherches intensives, les mécanismes moléculaires sous-jacents sont toujours largement débattus à ce jour. Le DT2 est une maladie à forte composante génétique, comme l’ont montré les études de jumeaux concordants en général pour le DT2. Au moins 2% des patients atteints de DT2 ont un diabète d’origine monogénique, survenant en général pendant l’enfance ou l’adolescence. L’une de ces formes survient spécifiquement à la naissance (diabète néonatal) et 50% de ses étiologies génétiques sont connus à ce jour. Les formes communes de DT2 survenant chez l’adulte sont polygéniques, liées à l’interaction entre des variants génétiques à effet modeste et faible pénétrance, et l’environnement ; en outre, des mécanismes « épigénétiques » seraient aussi en cause. L’identification des déterminants génétiques du DT2 a été dopée par les études d’association pangénomique (GWAS pour Genome Wide Association Studies) grâce à la mise au point de puces à ADN, qui permettent d’analyser conjointement plusieurs centaines de milliers de polymorphismes nucléotidiques simples (SNPs pour Single Nucleotide Polymorphisms) chez plusieurs centaines de milliers d’individus. Les GWAS ont permis d’identifier à ce jour une trentaine de gènes associées au risque de DT2. Le diagnostic du DT2 repose sur la mesure d’un trait continu, la glycémie à jeun. A partir de 1,25g/l, un individu est considéré diabétique. La glycémie à jeun, même chez des personnes non diabétiques, est fortement héritable, c’est-à-dire que ce paramètre métabolique est sous contrôle génétique. Nous avons ainsi utilisé la méthode des GWAS pour rechercher des loci régulant la glycémie dans des populations générales comme la population française DESIR. Nous avons mis en évidence la contribution du gène MTNR1B (codant le récepteur 2 de la mélatonine) dans la sécrétion d’insuline et la régulation de l’homéostasie glycémique. La mélatonine est l’hormone clef des rythmes circadiens de l’organisme, et il a été rapporté dans plusieurs études que la perturbation de ces rythmes est nuisible à l’homéostasie glycémique. Nous avons pu ensuite préciser quelles étaient les séquences d’ADN ayant un effet dans ce contrôle glycémique. Nous avons ainsi mis en évidence des mutations non-synonymes rares qui annihilaient la voie de signalisation de la mélatonine et qui sont associées à un risque élevé de DT2. Par ailleurs, via le consortium international MAGIC, nous avons contribué à l’identification de 16 marqueurs génétiques de la glycémie à jeun, de cinq marqueurs de la glycémie après charge de glucose, et de 10 loci intervenant dans la variance de l’hémoglobine glyquée (HbA1c, qui est un marqueur communément utilisé pour évaluer le contrôle glycémique d’un diabète traité). Ce faisant, nous avons montré que si le gène HK1 codant l’hexokinase 1 est le facteur génétique ayant le plus d’effet sur les valeurs d’HbA1c, il n’intervient pas sur les mécanismes physiologiques de la régulation de la glycémie. C’est uniquement de par les effets des variants d’HK1 sur la fonction des globules rouges (et sur le risque d’anémie) que ce gène modifie indirectement l’HbA1c. / Type 2 Diabetes is a major health care problem responsible for early morbidities and mortality. T2D prevalence inexorably increases due to dramatic changes in our way of life. T2D is preventable but no generally curable and present medications fail to prevent the worsening of glucose control and the development of complications. T2D is a systemic disease characterized by both insulin secretion defects and by insulin resistance at the levels of several tissues. Despite more than 40 years of research the aetiologies of T2D are still elusive. T2D is a multifactorial disease with a significant genetic component. However, T2D is a polygenic disorder with the effects of multiple DNA variants having a modest effect and a weak penetrance interaction with environmental factors. The identification of T2D susceptibility genes has been transformed by Genome Wide Association Studies (GWAS) which allow the analysis of hundred of thousands Single Nucleotide Polymorphisms (SNPs) in thousands of samples. Case/control GWAS have identified about 30 loci/genes, so far. However, these loci only explain a small part of T2D inheritance. T2D is defined by the measurement of the continuous trait “glycemia”. A fasting plasma glucose (FPG) higher than 7mM is considered to be abnormal. FPG is highly genetically determined and we have reanalyzed our GWAS data obtained in non diabetic general populations to identify genes that control (normal) glucose values. We first found that SNPs at MTNR1B (encoding the melatonin 2 receptor) locus regulate both FPG and insulin secretion. Melatonin is the hormone of darkness that plays a major role in circadian rhythms. The alteration of these physiological rhythms has been shown to impair glucose homeostasis. Subsequently we have screened and functionally analysed the coding part of MTNR1B in thousands of T2D cases and controls for rare lack of function mutations that strongly increase the risk for T2D. In addition via our contribution to the international consortium MAGIC we have identified 16 genetic markers modulating FPG, five controlling blood glucose after oral glucose load, and ten involved in the variance of glycated haemoglobin (HbA1c, a clinical marker of glucose control in treated diabetic patients). In this respect, we have demonstrated that if HK1 (encoding hexokinase 1) is the most potent gene controlling HbA1c, it was not at all involved in the physiology of glucose homeostasis. Instead, variant at HK1 locus act on red cell function, increase risk for anemia, and only indirectly perturb HbA1c measurement. This study shows that GWAS findings don’t mean causality and it is always mandatory to question the physiological validity of any association found through GWAS before implying a gene as causal. In this respect, the MAGIC consortium showed that among all the genes that regulate HbA1c, a large proportion is indeed directly related to red cell function. In conclusion, during my PhD, I have contributed to achieve a map of frequent SNPs regulating the major glucose control quantitative traits that define T2D. I also showed that rare DNA variants with a stronger biological impact also contribute to T2D risk. Although still of limited value for the prediction of T2D, GWAS have proven extremely useful to make progress in T2D physiology.

O presente estudo objetivou avaliar o efeito da dexmedetomidina, associada ou não à bupivacaína, na raquianestesia de fêmeas felinas submetidas à ovariohisterectomia. Foram utilizadas 34 gatas, jovens e adultas, sem raça definida, saudáveis submetidas a anestesia inalatória com isofluorano e aos seguintes tratamentos, após distribuição aleatória: grupo bupivacaína (GB) - raquianestesia com a bupivacaína isolada (0,5 mg/kg), grupo dexmedetomidina (GD) - raquianestesia com dexmedetomidina (1 mcg/kg) e grupo de dexmedetomidina bupivacaína (GDB) - raquianestesia com dexmedetomidina (1 mcg/kg) e bupivacaína (0,5 mg/kg). Após a indução da anestesia e manutenção com isofluorano, os animais foram posicionados em decúbito lateral direito para punção subaracnoide realizada no espaço lombossacro com agulha espinhal 25G. Os animais foram mantidos em decúbito dorsal até o final do procedimento cirúrgico e os atributos fisiológicos foram avaliados no período pré, trans e por 3 horas no período pós-operatório. Nenhum animal apresentou arritmia ou hipotensão arterial. O GDB apresentou redução significativa da frequência cardíaca e incremento pressórico quando comparado ao GB (p<0,01). Não houve diferença significativa no consumo de fentanil e no requerimento de isofluorano entre os grupos durante o procedimento cirúrgico. Na dose e diluição empregadas, a bupivacaína não determinou bloqueio motor significativo. As associações utilizadas promoveram analgesia adequada no período pós-operatório. O GDB apresentou maior grau de sedação durante parte da recuperação da anestesia (90min) (p<0,05), sem aumento no tempo de extubação. A adição da dexmedetomidina à bupivacaína na raquianestesia não aumentou o bloqueio motor e sensitivo; entretanto aumentou o grau de sedação dos animais promovendo melhor qualidade na recuperação anestésica sem deflagrar complicações cardiorrespiratória ou neurológica. / The present study aimed to evaluate the effect of dexmedetomidine, associated or not to bupivacaine, on spinal anesthesia in female felines submitted to ovariohysterectomy. Thirty four mixed breed healthy cats, young and adult, underwent inhalation anesthesia with isoflurane and the following treatments, after random distribution: bupivacaine group (BG) - spinal anesthesia with bupivacaine alone (0.5 mg/kg), dexmedetomidine group (DG) - spinal anesthesia with dexmedetomidine (1 mcg/kg). and dexmedetomidine/bupivacaine group (DBG) - spinal anesthesia with dexmedetomidine (1 mcg/kg) and bupivacaine (0.5 mg/kg). After anesthetic induction and maintenance with isoflurane, the animals were positioned in the right lateral recumbency for subarachnoid puncture performed in the lumbosacral space with a 25G spinal needle. The animals were kept in dorsal recumbency until the end of the surgical procedure and the physiological parameters were assessed in the pre, trans and 3 hours postoperative period. No animal presented arrhythmia or arterial hypotension. DBG presented a significant reduction in heart rate and pressure increase when compared to BG (p <0.01). There was no significant difference in fentanyl consumption and in the isoflurane requirement between groups during the surgical procedure. At the dose and dilution used, bupivacaine did not determine significant motor blockage. The associations used promoted adequate analgesia in the postoperative period. DBG had a higher degree of sedation during part of the anesthetic recovery (90min) (p <0.05), without an increase in extubation time. The addition of dexmedetomidine to bupivacaine in spinal anesthesia did not increase motor and sensory blockage; however, it increased the sedation level of the animals, promoting better quality of anesthetic recovery without triggering cardiorespiratory or neurological complications.

A família Potamotrygonidae constitui um grupo monofilético único dentre todos os elasmobrânquios por ser exclusivo de água doce e endêmico aos rios América do Sul. Apesar de seu monofiletismo, apresenta grandes problemas taxonômicos, uma vez que a grande variação nos padrões de coloração de disco e a ausência de um grande número de caracteres diagnósticos adequados proporcionam muitos erros e equívocos quanto a identificação de seus membros, principalmente pela presença de diversos padrões intermediários de coloração. Potamotrygon scobina Garman 1913, espécie cuja ampla distribuição e considerável variação morfológica geraram incertezas quanto a conspicuidade taxonômica do grupo, foi estudada morfologicamente de modo aprofundado, visando delimitar o status taxonômico dessa espécie e ajudar na resolução da problemática envolvida na taxonomia da família Potamotrygonidae. Espécimes cujas características pigmentares e corporais se assemelhavam à Potamotrygon scobina foram estudados e ao fim se obteve quatro novas espécies proximamente relacionadas à original. O refinamento das áreas de distribuição de cada uma das espécies, auxiliado da descrição taxonômica destas, auxiliaram na elaboração de uma hipótese de relacionamento entre estas espécies. Esta hipótese fora então comparada com dados geológicos da região amazônica e elaboraram-se inferências quanto à dinâmica biogeográfica tanto da família quanto das regiões amazônicas, sob um ponto de vista de um grupo de organismos nunca antes apresentado / The family Potamotrygonidae constitutes a monophyletic group unique among all elasmobranch in being exclusive to freshwater and endemic to South America. Despite its supraspecific taxa monophyletic status, some major taxonomic problems exists, as many species present great variation in colour patterns; the absence of a large number of suitable diagnostic characters yield various errors and misconceptions regarding the identification of its members, mainly regarding the presence of several intermediate patterns of dorsal pigmentation. Potamotrygon scobina Garman 1913, a species whose wide distribution and morphological variation generated considerable uncertainties about its taxonomic conspicuity, was morphologically studied in depth in order to delimit its taxonomic status of this species and assist in the resolution of the problems involved in the taxonomy of the family Potamotrygonidae. Specimens whose pigmentary characteristics and body resembled that ofPotamotrygon scobina were studied and four new species closely related to P. scobina were discovered. The refinement of the areas of distribution of each species, and the taxonomic description of these, assisted in the development of a hypothesis of relationship between these species. This result was then compared with geological data of the Amazon region in order to draw inferences as to the dynamics of both the family and biogeographical regions of the Amazon, from a point of view of a group of organisms never before studied in this way

O Estado de São Paulo encontra-se em uma situação epidemiológica da raiva humana, canina e felina que permite a declaração de área livre de raiva humana transmitida por cães (AgV-2). Entretanto, a vigilância precisa ser avaliada e capaz de detectar a doença com velocidade e sensibilidade. O objetivo do presente estudo foi avaliar a situação epidemiológica da raiva, o seu sistema de vigilância e propor uma reestruturação com base em vigilância baseada em risco. A situação epidemiológica foi avaliada de maneira descritiva. A avaliação foi realizada com o guia de avaliação de sistemas de vigilância do Centro de Controle de Doenças dos Estados Unidos. A proposta de reestruturação foi realizada com árvores de cenários da sensibilidade do sistema e uma proposição, com cálculos de probabilidade, de amostragem para maximizar tal sensibilidade. A situação epidemiológica mostrou que não houve casos de raiva humana entre 2003 e 2013 e que os casos em cães e gatos são esporádicos, dispersos no tempo e no espaço e ocasionados pelas AgV-3 e AgV-4, não sendo registrados casos pela AgV-2 também há mais de dez anos. Para cães e gatos, a vigilância foi dependente de estrutura específica dos serviços locais para enviar amostras de cérebro para os laboratórios (baixa simplicidade). Os veterinários privados tiveram baixa participação e percepção sobre o sistema (baixa aceitabilidade). A área sem vigilância em cães cresceu 30% entre 2003 e 2013 (baixa representatividade). Recebendo cães com sintomas associados à raiva, 39% dos serviços locais e 72% dos veterinários privados não suspeitaram de raiva em 2012 (baixa sensibilidade). O tempo entre o início dos sintomas e o resultado laboratorial dos cães suspeitos, em 2012 e 2013, foi, em média, 25 dias (baixa oportunidade). Os indicadores mostraram que o sistema tem baixa capacidade de detectar a doença precocemente. Se os esforços de amostragem para vigilância em cães forem direcionados aos animais com sintomas neurológicos (mesmo os inespecíficos de raiva), em ação conjunta entre médicos veterinários privados e serviços locais de controle de zoonoses, a sensibilidade do sistema chegaria ao máximo, utilizando-se da estrutura laboratorial de diagnóstico já existente. / The State of São Paulo is in an epidemiological situation of human, canine and feline rabies allowing a declaration of free area of human rabies transmitted by dogs (AgV-2). However, the surveillance needs to be evaluated and be able to detect the disease with speed and sensitivity. The aim of this study was to evaluate the epidemiological situation of rabies and the surveillance system and propose a restructure on a risk-based surveillance strategy. The epidemiological situation was descriptively assessed. The evaluation was performed according to the evaluation guidelines for surveillance systems in public health from the US Center for Disease Control (CDC). The proposed restructure was performed by building scenario trees of the system sensitivity and a proposition, with probability calculations, of sampling to maximize this sensitivity. The evaluation of the epidemiological situation showed no cases of human rabies between 2003 and 2013, cases in dogs and cats were sporadic, dispersed in time and space and occasioned by AgV-3 and AgV-4, cases not reported by AgV-2 for over ten years. For dogs and cats, surveillance was dependent on the specific structure of the local services to send brain samples to laboratories (low simplicity). Private veterinarians had low participation and perception of the system (low acceptability). The area unattended in dogs grew 30% between 2003 and 2013 (low representativeness). Getting dogs with symptoms associated with rabies, 39% of local and 72% of private veterinarians not suspected of rabies in 2012 (low sensitivity). The time between onset of symptoms and laboratory results from dogs suspected in 2012 and 2013 was on average 25 days (low timeliness). The indicators have shown that the system has low ability to disease detect early. If the sampling efforts for surveillance in dogs were directed to animals with neurological symptoms (even the nonspecific of rabies), a joint action between private veterinarians and local services of zoonoses controls, the system sensitivity could reach the its maximum, using the structure of existing laboratory diagnosis.

Thesis advisor: Marc L. Snapper / Intramolecular [2+2] photocycloadditions have been studied: (i) Previous studies in our group have demonstrated that there is a regiochemical dependence of intramolecular [2+2] photocycloadditions on the solvent employed. In order to understand the generality of hydrogen bonding effects of photochemical reactions of β-hydroxyl enones, we examined the photochemical outcome of enones with both acyclic and cyclic olefin tethers. (ii) An application of an intramolecular [2+2] photocycloaddition/ fragmentation strategy to form 5-8-5 ring systems has been demonstrated toward the synthesis of the natural product cycloaraneosene. / Thesis (PhD) — Boston College, 2008. / Submitted to: Boston College. Graduate School of Arts and Sciences. / Discipline: Chemistry.

Situar a pequena peça romântica para piano no contexto histórico e cultural do século XIX, e dela extrair propostas no campo da didática, é o objetivo desse trabalho. Para isso, procurou-se buscar não só a origem, mas justificar a preferência da pequena forma por parte dos compositores, em detrimento de formas maiores, já consagradas e aceitas universalmente. Fatores como o aperfeiçoamento do piano, estrutura, características e titulação das peças são abordados, assim como o movimento nacionalista, que tão bem delas se utilizou como veículo de expressão. A obra Macchiette op.2, coletânea de Henrique Oswald, serviu de exemplo vivo para ilustrar e justificar o que foi proposto: peças descritivas, intimistas, danças, revelam não só as tendências da época, mas as características, o estilo do compositor, que, através delas e de maneira particular, original, subjetiva, expressa suas emoções e se deixa conhecer. / The objective of this paper is to place this small romantic piano piece in the historical and cultural setting of the XIX century and extract from it some new ideas for teaching. For this reason, we tried to get at its very origins and justify the preference for the small form favored by the composers rather than larger, already consecrated and universally accepted forms. Factors such as perfecting the piano as well as structure, characteristics and studying the titles of the pieces were looked at, just as the nationalist movement that at the time used them for expression. Macchiette op.2, compiled by Henrique Oswald, is a lively example to illustrate and justify the purpose: descriptive, intimistic pieces, dances, revealing not only trends of their time, but the features and style of the composer who, in a very particular, original and subjective way, used them to express his feelings and make himself known.

A Tuberculose é uma das doenças infecciosas mais antiga e bem descrita. Entretanto, ainda permanece como um dos principais problemas de saúde pública a ser enfrentado em âmbito global. A implantação de novas estratégias no controle da Tuberculose requer uma melhor compreensão dos mecanismos que sucedem a fagocitose das micobactérias por macrófagos. Após a fagocitose, as micobactérias dão início a um conjunto de ações para sobreviverem e se replicarem no ambiente intracelular, entre as quais a provável interferência no processo de morte celular. Estudos mostram que M. tuberculosis pode apresentar habilidade de interferir no mecanismo de morte celular. Essa habilidade se tornou um desafio a ser estudado devido às implicações que isso deve ter na patogênese da doença. O nosso estudo teve por objetivo analisar a expressão de genes anti-apoptóticos (bcl-2, bcl-x e mcl-1) e pro-apoptóticos (bak, bax e bid) por PCR em tempo Real em macrófagos humanos derivados de células THP-1 após diferenciação induzida por PMA. Além disso, analisar a porcentagem de fragmentação de DNA nesses macrófagos, utilizando a citometria de fluxo, pois a fragmentação internucleossômica do DNA é uma das características apresentadas por células apoptóticas. Para as infecções foram utilizados isolados clínicos de M. tuberculosis com perfil de suscetibilidade a fármacos diferentes e a cepa padrão H37Rv (ATCC). Os dados de expressão foram analisados pela diferença de entre os isolados clínicos sensíveis, resistentes a três dos fármacos utilizados no tratamento da tuberculose humana e a cepa padrão H37Rv, utilizando-se o método de 2-ΔΔCT. Para comparar os resultados de expressão gênica, bem como a porcentagem de fragmentação de DNA, nos macrófagos infectados com os diferentes isolados clínicos, foram utilizados análise de variância (ANOVA) e o teste de comparação múltipla de Tukey. Os resultados sugerem, que os isolados clínicos resistentes a INH, RIF e EMB utilizados no nosso estudo, bem como a cepa padrão H37Rv (ATCC), não induzem mecanismos anti-apoptóticos para evadir da resposta imune. A ocorrência de fragmentação de DNA nos macrófagos infectados é um indicativo de morte por apoptose ou pyroptose. Além disso, o tempo de infecção éum fator importante e, com certeza, infecções com tempos maiores poderiam induzir ainda mais a morte dos macrófagos infectados. / Tuberculsis, an ancient infection disease, continues to thrive. Although well described, it remains a world health problem to overcome. The development and application of new strategies to control Tuberculosis requires a better understanding of mechanisms involved in mycobacteria-macrophages interaction. Following phagocytosis, mycobacteria initiates a variety of actions to survive and multiply themselves inside macrophages. According to researches, mycobacteria might interfere in the cell death mechanism. This ability became a challenge to be studied due to its implications in the pathogenesis of the disease. The aim of this study was to analyze the gene expression of anti-apoptotic (bcl-2, bcl-x e mcl-1) and pro-apoptotic genes (bak, bax e bid) in PMA-treated THP-1 cells by Real Time qPCR. Moreover, the percentage of macrophage DNA fragmentation was assessed by flow citometry because internucleosomal DNA fragmentation is characteristic of apoptotic and pyroptotic cell death. The infection was carried out using clinical isolates of M. tuberculosis resistent to multiple drugs, drug susceptibility and the M. tuberculosis H37Rv strain. The difference in the expression profile among clinical isolates, susceptible and resistant to three drugs used in the TB treatment, and the M. tuberculosis H37Rv were evaluated with the method 2-ΔΔCT. In order to compare gene expression patterns as well as the percentage of DNA fragmentation in macrophages infected with different clinical isolates, it was used analysis of variance (ANOVA) and Tukey`s multiple comparison test. The results suggest that M. tuberculosis H37Rv and the clinical isolates presenting higher drug resistant profile might induce programmed cell death in macrophages after 24-h infection. This was observed in the gene expression pattern and also in the macrophage DNA fragmentation profile, which indentifies apoptosis or pyroptosis. Therefore, it is suggested these clinical isolates and M. tuberculosis H37Rv do not present anti-apoptotic mechanisms to evade immune response. Moreover, the infection time is an important factor and, definitely, infections for long time could induce increase death of the infectados macrophages.