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Snake venom proteomes have long been investigated to explore a multitude of biologically active components that are used for prey capture and defense, and are involved in the pathological effects observed upon mammalian envenomation. Glycosylation is a major protein post-translational modification in venoms and contributes to the diversification of proteomes. We have shown that Bothrops venoms are markedly defined by their content of glycoproteins, and that most N-glycan structures of eight Bothrops venoms contain sialic acid, while bisected N-acetylglucosamine was identified in Bothrops cotiara venom. To further investigate the mechanisms involved in the generation of different venoms by related snakes, here the glycoproteomes of nine Bothrops venoms (Bothrops atrox, B. cotiara, Bothrops erythromelas, Bothrops fonsecai, B. insularis, Bothrops jararaca, Bothrops jararacussu, Bothrops moojeni and Bothrops neuwiedi) were comparatively analyzed by enrichment with three lectins of different specificities, recognizing bisecting N-acetylglucosamine- and sialic acid-containing glycoproteins, and mass spectrometry. The lectin capture strategy generated venom fractions enriched with several glycoproteins, including metalloprotease, serine protease, and L- amino acid oxidase, in addition to various types of low abundant enzymes. The different contents of lectin-enriched proteins underscore novel aspects of the variability of the glycoprotein subproteomes of Bothrops venoms and point to the role of distinct types of glycan chains in generating different venoms by closely related snake species.

Histone variants play a crucial role in chromatin structure organization and gene expression. Trypanosomatids have an unusual H2B variant (H2B.V) that is known to dimerize with the variant H2A.Z generating unstable nucleosomes. Previously, we found that H2B.V protein is enriched in tissue-derived trypomastigote (TCT) life forms, a nonreplicative stage ofTrypanosoma cruzi, suggesting that this variant may contribute to the differences in chromatin structure and global transcription rates observed among parasite life forms. Here, we performed the first genome-wide profiling of histone localization inT.cruziusing epimastigotes and TCT life forms, and we found that H2B.V was preferentially located at the edges of divergent transcriptional strand switch regions, which encompass putative transcriptional start regions; at some tDNA loci; and between the conserved and disrupted genome compartments, mainly at trans-sialidase, mucin and MASP genes. Remarkably, the chromatin of TCT forms was depleted of H2B.V-enriched peaks in comparison to epimastigote forms. Interactome assays indicated that H2B.V associated specifically with H2A.Z, bromodomain factor 2, nucleolar proteins and a histone chaperone, among others. Parasites expressing reduced H2B.V levels were associated with higher rates of parasite differentiation and mammalian cell infectivity. Taken together, H2B.V demarcates critical genomic regions and associates with regulatory chromatin proteins, suggesting a scenario wherein local chromatin structures associated with parasite differentiation and invasion are regulated during the parasite life cycle.

Este documento apresenta uma dissertação sobre o desenvolvimento de um sistema de integração de dados para geração de estruturas tridimensionais a partir de vídeo 3D. O trabalho envolve a extensão de um sistema de vídeo 3D em tempo real proposto recentemente. Esse sistema, constituído por projetor e câmera, obtém imagens de profundidade de objetos por meio da projeção de slides com um padrão de faixas coloridas. Tal procedimento permite a obtenção, em tempo real, tanto do modelo 2,5 D dos objetos quanto da textura dos mesmos, segundo uma técnica denominada luz estruturada. Os dados são capturados a uma taxa de 30 quadros por segundo e possuem alta qualidade: resoluções de 640 x 480 pixeis para a textura e de 90 x 240 pontos (em média) para a geometria. A extensão que essa dissertação propõe visa obter o modelo tridimensional dos objetos presentes em uma cena por meio do registro dos dados (textura e geometria) dos diversos quadros amostrados. Assim, o presente trabalho é um passo intermediário de um projeto maior, no qual pretende-se fazer a reconstrução dos modelos por completo, bastando para isso apenas algumas imagens obtidas a partir de diferentes pontos de observação. Tal reconstrução deverá diminuir a incidência de pontos de oclusão (bastante comuns nos resultados originais) de modo a permitir a adaptação de todo o sistema para objetos móveis e deformáveis, uma vez que, no estado atual, o sistema é robusto apenas para objetos estáticos e rígidos. Até onde pudemos averiguar, nenhuma técnica já foi aplicada com este propósito. Este texto descreve o trabalho já desenvolvido, o qual consiste em um método para detecção, rastreamento e casamento espacial de componentes conexas presentes em um vídeo 3D. A informação de imagem do vídeo (textura) é combinada com posições tridimensionais (geometria) a fim de alinhar partes de superfícies que são vistas em quadros subseqüentes. Esta é uma questão chave no vídeo 3D, a qual pode ser explorada em diversas aplicações tais como compressão, integração geométrica e reconstrução de cenas, dentre outras. A abordagem que adotamos consiste na detecção de características salientes no espaço do mundo, provendo um alinhamento de geometria mais completo. O processo de registro é feito segundo a aplicação do algoritmo ICP---Iterative Closest Point---introduzido por Besl e McKay em 1992. Resultados experimentais bem sucedidos corroborando nosso método são apresentados. This document presents a MSc thesis focused on the development of a data integration system to generate tridimensional structures from 3D video. The work involves the extension of a recently proposed real time 3D video system. This system, composed by a video camera and a projector, obtains range images of recorded objects using slide projection of a coloured stripe pattern. This procedure allows capturing, in real time, objects´ texture and 2,5 D model, at the same time, by a technique called structured light. The data are acquired at 30 frames per second, being of high quality: the resolutions are 640 x 480 pixels and 90 x 240 points (in average), respectively. The extension that this thesis proposes aims at obtaining the tridimensional model of the objects present in a scene through data matching (texture and geometry) of various sampled frames. Thus, the current work is an intermediary step of a larger project with the intent of achieving a complete reconstruction from only a few images obtained from different viewpoints. Such reconstruction will reduce the incidence of occlusion points (very common on the original results) such that it should be possible to adapt the whole system to moving and deformable objects (In the current state, the system is robust only to static and rigid objects.). To the best of our knowledge, there is no method that has fully solved this problem. This text describes the developed work, which consists of a method to perform detection, tracking and spatial matching of connected components present in a 3D video. The video image information (texture) is combined with tridimensional sites (geometry) in order to align surface portions seen on subsequent frames. This is a key step in the 3D video that may be explored in several applications such as compression, geometric integration and scene reconstruction, to name but a few. Our approach consists of detecting salient features in both image and world spaces, for further alignment of texture and geometry. The matching process is accomplished by the application of the ICP---Iterative Closest Point---algorithm, introduced by Besl and McKay in 1992. Succesful experimental results corroborating our method are shown.