874 resultados para eletroforese bidimensional


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En la hipótesis de trabajo del presente proyecto se considera la importancia del metabolismo de lípidos y proteínas en los insectos hematófagos, en particular en los vectores de la enfermedad de Chagas, para afrontar exitosamente la demanda energética de la reproducción. Las hembras de estas especies pueden ingerir una comida de sangre abundante en lípidos y proteínas, los que son modificados en el intestino para su utilización y posterior almacenamiento en estructuras organizadas en el tejido ovárico, sustentando así el rápido crecimiento de los ovocitos. Estos aspectos resultan críticos para el ciclo de vida del insecto y para el mantenimiento de la cadena epidemiológica de la enfermedad. En estas especies, recientemente hemos caracterizado a nivel bioquímico y celular la interacción entre lipoproteínas y tejidos [Fruttero y col., Insect Biochem. Mol. Biol. 39: 322-331 (2009); Fruttero y col. Biocel 33 (3): 260 (2009)] y las fases del ciclo reproductivo [Aguirre y col., J. Insect Physiol. 54: 393-402 (2008)]. No obstante, los factores que participan en su regulación son aún escasamente conocidos. En este contexto, el estudio propone emplear dos especies de triatominos con el objeto de: (1) caracterizar los factores involucrados en la formación y regulación de reservas nutricionales en los ovocitos; (2) analizar los eventos que participan en la regresión del tejido ovárico: atresia folicular y mecanismos de muerte celular. (3) evaluar el impacto de productos naturales (ureasas vegetales y péptidos derivados) en el desarrollo del tejido ovárico. Para la ejecución de los objetivos se llevarán a cabo ensayos in vivo e in vitro con trazadores fluorescentes, fraccionamiento subcelular, estudios de expresión de proteínas (mRNA y proteína), estudios histo-morfológicos, ultraestructurales e inmunocitoquímicos, microscopía láser confocalizada, ensayos de actividad enzimática, ELISA, western-blot, electroforesis bidimensional, espectrometria de masas en tándem, etc. También se evaluarán los mecanismos de muerte celular (apoptosis/autofagia) mediante microscopía electrónica, detección de apoptosis in situ (TUNEL), inmunofluorescencia, etc. Los resultados obtenidos permitirán un mejor conocimiento sobre la fisiología y bioquímica de estos vectores, los que resultan indispensables en el diseño de nuevas estrategias para su control. Debido a la carencia de un tratamiento específico para la enfermedad y a la falta de métodos preventivos (vacuna), el control del vector es una de las vías más importantes para reducir la incidencia de la enfermedad. Actualmente, la situación socio-económica que sufren amplios núcleos de nuestra población propicia condiciones de vida que facilitan la reproducción de los vectores y la transmisión vectorial del parásito. El estudio permitirá además explorar aspectos bioquímicos y celulares básicos, generando conocimientos que podrían ser extensivos a otros insectos de importancia económica en la ganadería y/o agricultura.

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Trabalho Final do Curso de Mestrado Integrado em Medicina, Faculdade de Medicina, Universidade de Lisboa, 2014

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Tese de mestrado, Medicina Legal e Ciências Forenses, Universidade de Lisboa, Faculdade de Medicina, 2016

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Dissertação para obtenção do grau de Mestre no Instituto Superior de Ciências da Saúde Egas Moniz

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O presente trabalho tem como fundamento a continuidade da investigação desenvolvida por Assis e Nascimento, cujo objetivo é a adaptação e validação para Portugal de uma escala de medição do cinismo organizacional, desenvolvida originalmente para o contexto americano. Não tendo sido conclusivo o estudo dos autores, estes deixaram pistas para investigações futuras, sendo sugerida a repetição do mesmo com novas amostras. Achou-se assim interessante desenvolver a pesquisa num contexto da Administração Pública indireta, concretamente num Instituto Público que resultou de um processo de fusão. O tema é pertinente, na medida em que a sua análise poderá dar relevo a variáveis passíveis de minimizar eventuais consequências menos positivas advindas do processo recentemente ocorrido com a fusão. Pode ainda fornecer pistas que permitam, por um lado, um contributo para a melhoria do desempenho e coesão organizacional e, por outro, colocar na agenda da investigação da Administração Pública um estudo atual que vise estudar esta temática tendo em conta justamente o universo próprio da Administração Pública. Apesar de este estudo não confirmar as conclusões da investigação em que se baseou, revela um novo foco, relativo ao contexto estudado, apresentando um constructo bidimensional em vez de unidimensional, como sugerido por Assis e Nascimento.

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O objeto de estudo deste projeto de mestrado centra-se na análise e desenvolvimento de personagens na animação, mais especificamente, nas problemáticas associadas à união entre o desenho bidimensional e objectos tridimensionais. Esta união entre a animação 2D e 3D surge de um contexto um tanto recente, como forma de tentar facilitar e melhorar as animações, procurando, em simultâneo, ultrapassar os conhecimentos técnicos da animação para inovar o cinema em live action e em animação. O objetivo desta investigação, em paralelo com a construção de uma curta-metragem e da sua análise, é compreender a harmonização e as dificuldades associadas à união de personagens 2D e 3D. Este trabalho procura tambem analisar os estudos já elaborados nesta materia de aprioximação do 3D ao desenho, contribuindo para o seu desenvolvimento, e compreender de forma mais aprofundada a construção de personagens numa animação.

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The method (algorithm BIDIMS) of multivariate objects display to bidimensional structure in which the sum of differences of objects properties and their nearest neighbors is minimal is being described. The basic regularities on the set of objects at this ordering become evident. Besides, such structures (tables) have high inductive opportunities: many latent properties of objects may be predicted on their coordinates in this table. Opportunities of a method are illustrated on an example of bidimentional ordering of chemical elements. The table received in result practically coincides with the periodic Mendeleev table.

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2000 Mathematics Subject Classification: 60J80.

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The sulfated polysaccharides (SP) from the edible red seaweed Gracilaria birdiae were obtained using five different condition extraction (GB1: Water; GB1p: Water/proteolysis; GB1s: Water/sonication; GB1sp: Water/sonication/proteolysis; GB2s: NaOH/sonication; GB2sp: NaOH/sonication/proteolysis. The yield (g) increased in the following order GB2sp>GB1sp>GB1p>GB2s>GB1s>GB1. However, the amount of SP extracted increased in different way GB2sp>GB1p>GB1>GB1sp>GB1s>GB2s. Infrared and electrophoresis analysis showed that all conditions extracted the same SP. In addition, monosaccharide composition showed that ultrasound promotes the extraction of other polysaccharides than SP. In the prothrombin time (PT) test, which evaluates the extrinsic coagulation pathway, none of the samples showed anticoagulant activity. While in the activated partial thromboplastin time (aPTT) test, which evaluates the intrinsic coagulation pathway, all samples showed anticoagulant activity, except GB2s. The aPTT activity decreased in the order of GB1sp>GB2sp>GB1p>GB1>GB1s>GB2s. Total capacity antioxidant (TCA) of the SP was also affected by condition extraction, since GB2s and GB1 showed lower activity in comparison to the other conditions. In conclusion, the conditions of SP extraction influence their biological activities and chemical composition. The data showed NaOH/sonication/proteolysis was the best condition to extract anticoagulant and antioxidant SPs from Gracilaria birdiae.

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Data Visualization is widely used to facilitate the comprehension of information and find relationships between data. One of the most widely used techniques for multivariate data (4 or more variables) visualization is the 2D scatterplot. This technique associates each data item to a visual mark in the following way: two variables are mapped to Cartesian coordinates so that a visual mark can be placed on the Cartesian plane; the others variables are mapped gradually to visual properties of the mark, such as size, color, shape, among others. As the number of variables to be visualized increases, the amount of visual properties associated to the mark increases as well. As a result, the complexity of the final visualization is higher. However, increasing the complexity of the visualization does not necessarily implies a better visualization and, sometimes, it provides an inverse situation, producing a visually polluted and confusing visualization—this problem is called visual properties overload. This work aims to investigate whether it is possible to work around the overload of the visual channel and improve insight about multivariate data visualized through a modification in the 2D scatterplot technique. In this modification, we map the variables from data items to multisensoriy marks. These marks are composed not only by visual properties, but haptic properties, such as vibration, viscosity and elastic resistance, as well. We believed that this approach could ease the insight process, through the transposition of properties from the visual channel to the haptic channel. The hypothesis was verified through experiments, in which we have analyzed (a) the accuracy of the answers; (b) response time; and (c) the grade of personal satisfaction with the proposed approach. However, the hypothesis was not validated. The results suggest that there is an equivalence between the investigated visual and haptic properties in all analyzed aspects, though in strictly numeric terms the multisensory visualization achieved better results in response time and personal satisfaction.

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Este estudo de caráter qualitativo, com base em metodologia pesquisa – ação, procurou descrever características funcionais da estratégia de ensino „Imitating Art”, construída com base em métodos de dramatização, solução de problemas e tempestade cerebral e alicerçada no processo de ensino de arte do conhecer, apreciar e fazer arte. Participaram da pesquisa 96 alunos do ensino em saúde com idade média de 21 anos. Com o objetivo de desenhar as características funcionais da estratégia, a mesma foi planejada, descrita, modificada e avaliada em 4 etapas: experimentação, metodização, adequação e replicação. Cada etapa foi verificada a partir de avaliação observacional pelos docentes envolvidos e análise de conteúdo sobre as respostas dos alunos participantes, em instrumento de avaliação próprio. Da fase experimental participaram 14 alunos dos quais 100 % aprovaram a estratégia. A análise de conteúdo mostrou Identificação de ferramentas cognitivas e afetivas (30,6%), características da atividade (22,2%), e mediação de trabalho em grupo (16,7%). A observação identificou ativo trabalho atitudinal em uma dinâmica de grupo criativa. Na fase de metodização participaram 31 alunos, dos quais 96,9% aprovaram a estratégia. Na análise de conteúdo, as ideias mais presentes foram, dificuldades encontradas pelos participantes (28%), identificação de ferramentas cognitivas e afetivas (17,6%), Implicação da estratégia no cotidiano profissional (13,2%) e mediação do trabalho em grupo (13,2%). Na fase de Adequação, participaram 32 alunos, dos quais 90,7% demonstraram aprovação da estratégia. A avaliação dos participantes evidenciou como mais importante a mediação do trabalho em grupo (29,6%), identificação de ferramentas cognitivas e afetivas (21,3%) e características da atividade (19,7%). A fase de replicação teve 19 alunos, dos quais 100% aprovaram a estratégia e cuja avaliação demonstrou principalmente dificuldades encontradas pelo participante (27,8%), mediação do trabalho em grupo (25%) e Habilidade criativa da atividade (22,2%). A avaliação bidimensional mostrou que se constitui como uma estratégia de abordagem ativa e colaborativa, podendo ser usada como instrumento linear de diagnóstico e incremento de competências atitudinais tais como criatividade e trabalho em grupo.

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Chondroitin sulfate (CS) is a naturally glycosaminoglycan found in the extracellular matrix of connective tissues and it may be extracted and purified those tissues. CS is involved in various biological functions, which may be related to the having structural variability, despite the simplicity of the linear chain structure from this molecule. Researches in biotechnology and pharmaceutical field with wastes from aquaculture has been developed in Brazil. In recent decades, tilapia (Oreochromis niloticus), native fish from Africa, has been one of the most cultivated species in various regions of the world, including Brazil. The tilapia farming is a cost-effective activity, however, it generates large amount of wastes that are discarded by producers. It is understood that waste from tilapia can be used in research as a source of molecules with important biotechnological applications, which also helps in reducing environmental impacts and promote the development of an ecofriendly activity. Thus, nile tilapia viscera were subjected to proteolysis, then the glycosaminoglycans were complexed with ion exchange resin (Lewatit), it was fractionated with increasing volumes of acetone and purified by ion exchange chromatography DEAE-Sephacel. Further, the fraction was analyzed by agarose gel electrophoresis and nuclear magnetic resonance (NMR). The electrophoretic profile of the compound together the analysis of 1H NMR spectra and the HSQC correlation allow to affirm that the compound corresponds to a molecule like chondroitin sulfate. MTT assay was used to assess cell viability in the presence of CS tilapia isolated and showed that the compound is not cytotoxic to normal cells such as cells from the mouse embryo fibroblast (3T3). Then, this compound was tested for the ability to reduce the influx of leukocytes in model of acute peritonitis (in vivo) induced by sodium thioglycolate. In this context, it was done total and differential leukocytes counting in the blood and peritoneal fluid collected respectively from vena cava and the peritoneal cavity of the animals subjected to the experiment. The chondroitin sulfate for the first time isolated from tilapia (CST ) was able to reduce the migration of leukocytes to the peritoneal cavity of inflamed mice until 80.4 per cent at a dose 10µg/kg. The results also show that there was a significant reduction (p<0.001) of the population of polymorphonuclear leukocytes from peritoneal cavity in the three tested doses (0.1µg/kg; 1µg/kg and 10µg/kg) when it was compared to the positive control (just thioglycolate). Therefore, since the CST structure and mechanism of action has been completely elucidated, this compound may have potential for therapeutic use in inflammatory diseases

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The understanding of the occurrence and flow of groundwater in the subsurface is of fundamental importance in the exploitation of water, just like knowledge of all associated hydrogeological context. These factors are primarily controlled by geometry of a certain pore system, given the nature of sedimentary aquifers. Thus, the microstructural characterization, as the interconnectivity of the system, it is essential to know the macro properties porosity and permeability of reservoir rock, in which can be done on a statistical characterization by twodimensional analysis. The latter is being held on a computing platform, using image thin sections of reservoir rock, allowing the prediction of the properties effective porosity and hydraulic conductivity. For Barreiras Aquifer to obtain such parameters derived primarily from the interpretation of tests of aquifers, a practice that usually involves a fairly complex logistics in terms of equipment and personnel required in addition to high cost of operation. Thus, the analysis and digital image processing is presented as an alternative tool for the characterization of hydraulic parameters, showing up as a practical and inexpensive method. This methodology is based on a flowchart work involving sampling, preparation of thin sections and their respective images, segmentation and geometric characterization, three-dimensional reconstruction and flow simulation. In this research, computational image analysis of thin sections of rocks has shown that aquifer storage coefficients ranging from 0,035 to 0,12 with an average of 0,076, while its hydrogeological substrate (associated with the top of the carbonate sequence outcropping not region) presents effective porosities of the order of 2%. For the transport regime, it is evidenced that the methodology presents results below of those found in the bibliographic data relating to hydraulic conductivity, mean values of 1,04 x10-6 m/s, with fluctuations between 2,94 x10-6 m/s and 3,61x10-8 m/s, probably due to the larger scale study and the heterogeneity of the medium studied.

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In this work we have investigated some aspects of the two-dimensional flow of a viscous Newtonian fluid through a disordered porous medium modeled by a random fractal system similar to the Sierpinski carpet. This fractal is formed by obstacles of various sizes, whose distribution function follows a power law. They are randomly disposed in a rectangular channel. The velocity field and other details of fluid dynamics are obtained by solving numerically of the Navier-Stokes and continuity equations at the pore level, where occurs actually the flow of fluids in porous media. The results of numerical simulations allowed us to analyze the distribution of shear stresses developed in the solid-fluid interfaces, and find algebraic relations between the viscous forces or of friction with the geometric parameters of the model, including its fractal dimension. Based on the numerical results, we proposed scaling relations involving the relevant parameters of the phenomenon, allowing quantifying the fractions of these forces with respect to size classes of obstacles. Finally, it was also possible to make inferences about the fluctuations in the form of the distribution of viscous stresses developed on the surface of obstacles.

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The Waltheria genus belonging to the Sterculiaceae family, it is reported as a prolific source of flavonoids and quinolone alkaloids, substances of great interest due to several associated biological activities. This work describes a novel phytochemical study from Waltheria ferruginea, aiming to contribute to the chemical knowledge of this specie and the isolation of substances with biological potential. For the phytochemical study were used chromatography techniques on silica gel and molecular exclusion in Sephadex LH-20.The structural elucidation of the isolated compounds was performed through spectrometric techniques 1H and 13C NMR, including uni and bidimensional pulse sequences, and comparison with data from literature. Five substances were isolated, namely: the flavonids kaempferol-3-O-β-(6''-cumaroil)-glucopyranoside (F1) and kaempferol -3 -O- β - glucopyranoside (F2), both analyzes with pharmacological properties, the flavonol quercetin-3-O-β-glucopyranoside (F3 ) pure and in the epimeric mixture α (F3') and (F3), the terpenegeranyl - geranyl (G1) and the 12-hydroxi-octadecanoic acid, all no previous reported in the literature.