992 resultados para 2-D electrophoresis


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Esta tesis versa sobre el an álisis de la forma de objetos 2D. En visión articial existen numerosos aspectos de los que se pueden extraer información. Uno de los más usados es la forma o el contorno de esos objetos. Esta característica visual de los objetos nos permite, mediante el procesamiento adecuado, extraer información de los objetos, analizar escenas, etc. No obstante el contorno o silueta de los objetos contiene información redundante. Este exceso de datos que no aporta nuevo conocimiento debe ser eliminado, con el objeto de agilizar el procesamiento posterior o de minimizar el tamaño de la representación de ese contorno, para su almacenamiento o transmisión. Esta reducción de datos debe realizarse sin que se produzca una pérdida de información importante para representación del contorno original. Se puede obtener una versión reducida de un contorno eliminando puntos intermedios y uniendo los puntos restantes mediante segmentos. Esta representación reducida de un contorno se conoce como aproximación poligonal. Estas aproximaciones poligonales de contornos representan, por tanto, una versión comprimida de la información original. El principal uso de las mismas es la reducción del volumen de información necesario para representar el contorno de un objeto. No obstante, en los últimos años estas aproximaciones han sido usadas para el reconocimiento de objetos. Para ello los algoritmos de aproximaci ón poligonal se han usado directamente para la extracci ón de los vectores de caracter ísticas empleados en la fase de aprendizaje. Las contribuciones realizadas por tanto en esta tesis se han centrado en diversos aspectos de las aproximaciones poligonales. En la primera contribución se han mejorado varios algoritmos de aproximaciones poligonales, mediante el uso de una fase de preprocesado que acelera estos algoritmos permitiendo incluso mejorar la calidad de las soluciones en un menor tiempo. En la segunda contribución se ha propuesto un nuevo algoritmo de aproximaciones poligonales que obtiene soluciones optimas en un menor espacio de tiempo que el resto de métodos que aparecen en la literatura. En la tercera contribución se ha propuesto un algoritmo de aproximaciones que es capaz de obtener la solución óptima en pocas iteraciones en la mayor parte de los casos. Por último, se ha propuesto una versi ón mejorada del algoritmo óptimo para obtener aproximaciones poligonales que soluciona otro problema de optimización alternativo.

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Descreve-se, no presente trabalho, os esforços envidados no sentido de criar uma solução informática generalista, para os problemas mais recorrentes do processo de produção de videojogos 20, baseados em sprites, a correr em plataformas móveis. O sistema desenvolvido é uma aplicação web que está inserida no paradigma cloud­computing, usufruindo, portanto, de todas as vantagens em termos de acessibilidade, segurança da informação e manutenção que este paradigma oferece actualmente. Além das questões funcionais, a aplicação é ainda explorada do ponto de vista da arquitetura da implementação, com vista a garantir um sistema com implementação escalável, adaptável e de fácil manutenção. Propõe-se ainda um algoritmo que foi desenvolvido para resolver o problema de obter uma distribuição espacial otimizada de várias áreas retangulares, sem sobreposições nem restrições a nível das dimensões, quer do arranjo final, quer das áreas arranjadas. ABSTRACT: This document describes the efforts taken to create a generic computing solution for the most recurrent problems found in the production of two dimensional, sprite­based videogames, running on mobile platforms. The developed system is a web application that fits within the scope of the recent cloud-computing paradigm and, therefore, enjoys all of its advantages in terms of data safety, accessibility and application maintainability. In addition, to the functional issues, the system is also studied in terms of its internal software architecture, since it was planned and implemented in the perspective of attaining an easy to maintain application, that is both scalable and adaptable. Furthermore, it is also proposed an algorithm that aims to find an optimized solution to the space distribution problem of several rectangular areas, with no overlapping and no dimensinal restrictions, neither on the final arrangement nor on the arranged areas.

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Purpose: A number of proteome studies have been performed recently to identify pheromone-related protein expression and their molecular function using genetically modified rodents' urine. However, no such studies have used Indian commensal rodents; interestingly, in a previous investigation, we confirmed the presence of volatile molecules in commensal rodents urine and these molecules seem to be actively involved in pheromonal communication. Therefore, the present study aims to identify the major urinary protein [MUP] present in commensal rat urine, which will help us to understand the protein's expression pattern and intrinsic properties among the rodents globally. Experimental Design: Initially, the total urinary proteins were separated by 1-D and 2-D electrophoresis and then subsequently analyzed by Matrix Assisted Laser Desorption Ionization-Time of Flight and Mass Spectrometer (MALDI-TOF/MS). Furthermore, they were then fragmented with the aid of a Tandem Mass Spectrometer (TOF/TOF) and the identified sequences aligned and confirmed using similarity with the deduced primary structures of members of the lipocalin superfamily.Results: The SDS-PAGE protein profiles showed distinct proteins with molecular masses of 15, 22.4, 25, 28, 42, 50, 55, 68, and 91 kDa. Of these proteins, the 22.4 kDa protein was considered to be target candidate. When 2D electrophoresis was carried out, about similar to 50 spots were detected with different masses and various pI ranges. The 22.4 kDa protein was found to have a pI of about 4.9. This 22.4 kDa protein spot was digested and subjected to mass spectrometry; it was identified as rat MUP. The fragmented peptides from the rat MUP at 935, 1026, 1192, and 1303 m/z were further fragmented with the aid of MS/MS and generated de novo sequence and this confirmed this protein to be the MUP present in the urine of commensal rats.Conclusion: The present investigation confirms the presence of MUP with a molecular mass of 22.4 kDa in the urine of commensal rats. This protein may be involved in the binding of volatile molecules and opens up a discussion about how volatile and non-volatile molecules in the commensal rats' urine may contribute chemo-communication.

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叶绿素突变泛指能导致叶绿素代谢失调的核基因或叶绿体基因突变。发生叶绿素突变的植物个体普遍表现为叶色的变化,目前已报道的多数叶绿素突变体为人工诱变产物。叶绿素缺失突变导致的叶结素代谢缺陷实际上反映了叶绿体发育过程的缺陷,研究叶绿素突变更重要的意义是在于阐明叶绿体发育过程。 本研究所用材料1103s是一类特殊的叶绿素突变体,为籼性光敏核不育水稻(Oryza sativa L.)8902s群体中发现的自发突变体。该突本所具有的失绿特性为特定温度条件下才表现出来的瞬时性状,在环境温度恢复后,失绿组织可复发。遗传分析表明该突变由隐性核基因控制。本文不1103s所具有的温度敏感和失绿复绿特性,在亚细胞水平和生理化水平进行了详细的探讨。 叶绿素含量的检测表明,诱导后表现失绿的叶片组织内叶绿素含量明显降低,叶绿素a/b比值升高,原脱植基叶绿素含量低于绿色组织。失绿组织中的这种原脱植基叶绿素在失绿组织中含量的减少是由两方面因素造成的,其一是叶绿素合成过程中原脱植基叶绿素合成之前的某一步过程反应受阻;其二是原脱植基叶绿素向叶绿素转化的过程是正常进行的。 对1103s叶绿体内部的超微结构观察表明:1.控制叶绿素缺失性状的是一多效基因。该基因在特定温度条件下表达时,不仅影响到叶绿体的发育,也对细胞质中的其他细胞器产生重要的影响,其结果是细胞质中的高度有序的内膜系统被大量形状不规则的泡状结构所取代。放大后发现,这类泡状结构由(1)线粒体(2)功能未知的泡状结构I,其内部含颗粒状物质泡结构被膜内还有数层不连续的膜残片(3)功能未知的泡状结构II,其内部含大颗粒状物质。2.该突变体表达失绿和复绿过程中,叶绿体内部膜结构的变化伴随叶绿素含量的变化也有退脂和恢复的过程,但与已报道的其他突体有两个明显的不同:首先,在退化细胞的叶绿体内未观察到前片层体的存在。前片层体是叶绿体发育过程中黄化体阶段常见的非常明显的特殊结构,在电镜下为有规律的晶格状结构。已有研究表明,前片层体的形成与原脱植基叶绿素的积累有密切关系。与组培白化苗中检测到的结果不同,失绿组织中原脱植基叶绿素的含量不但没有积累,反而少于绿色组织中的含量,而造成该突变体在失绿过程中质体内无前片层体形成。其次,1103s在叶绿体退化过程中类囊体膜的变化不同于其化温敏的转绿型叶绿素突变体,尤其是在失绿过程中,其类囊体膜不是以直接解体的方式减少而是以单类囊体膜紧靠为主要特征。 对野生型 8902s与1103s类囊体膜结构的冰冻蚀刻分析表明,1103s失绿叶片上的失绿组织和绿色组织中,EFs面的大颗粒结构均异常。其异常之处表现在每个颗粒明显解离成两个亚单位(上面观),而在野生型8902s中则无上述现象出现。亚单位的解离程度在失绿组织中更明显。有间接证据研究表明,EFs面上的大颗粒代表PS II。如果该推论正确,那么失绿叶片的失绿组织和绿色组织中,PS II都可能是异常的。 另外,通过对失绿组织和绿色组织全叶蛋白双向电泳图谱的比较,得到了一个特异缺失的叶蛋白组分,该蛋白的分子量为51kd。此蛋白在失绿叶片上的失绿组织和绿色组织之间存在组织差异性。通过对该蛋白在不同温度处理和不同遗传背景下的变化规律分析,发现该蛋白是一存在于许多水稻品系叶片中的高含量组分,此蛋白表达本身不受变温诱导过程的影响,而是受另一感温过程的调控。初步分析表明该蛋白为一失绿相关蛋白。 综上所述,1103s所具有的失绿和复绿特性是核基因多效表达的结果,有一感温过程调控下游蛋白表达的复杂过程。此外,该突变特性很可能与PS II的结构异常有关。

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Arginine kinase (AK) was previously reported as a phosphagen-ATP phosphotransferase found in invertebrates. In this study, an 1184 bp cDNA was cloned and sequenced. It contained an open reading frame of 1068 bp that coded for 356 deduced amino acids of AK in Fenneropenaeus chinensis. The calculated molecular mass of AK is 40129.73 Da and pI is 5.92. The predicted protein showed a high level of identity to known AK in invertebrates and creatine kinase from vertebrates, which belong to a conserved family of ATP:guanidino phospho-transferases. In addition, AK protein in plasma of F. chinensis was identified using two-dimensional electrophoresis (2DE) and electrospray ionization mass spectrometry (ESI-MS) according to the calculated molecular mass and pI. AK was significantly decreased in the plasma of F. chinensis at 45 min and recovered at 3 It after laminarin injection as confirmed by 2DE and ESI-MS. The results showed that AK was one of the most significantly changed proteins on two-dimensional gel in the plasma proteins of F. chinensis at 45 min and 3 It after simulation. (c) 2005 Elsevier Ltd. All rights reserved.

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Chromobacterium violaceum is a free-living bacillus, Gram-negative commonly found in water and sand of tropical and subtropical regions. One of its main characteristic it's the ability to produce the purple pigment named violacein, that shows countless biological activities. In 2003, the genome of this organism was totally sequenced and revealed important informations about the physiology of this bacteria. However, few post-genomics studies had been accomplished. This work evaluated the protein profile of C. violaceum cultivated in LB medium at 28ºC that allowed the identification and characterization of proteins related to a possible secretion system that wasn't identified and characterized yet in C. violaceum, to the quorum sensing system, to regulatory process of transcription and translation, stress adaptation and biotechnological potential. Moreover, the response of the bacteria to UVC radiation was evaluated. The comparison of the protein profile, analyzed through 2-D electrophoresis, of the control group versus the treatment group allowed the identification of 52 proteins that arose after stress induction. The obtained results enable the elaboration of a stress response pathway in C. violaceum generated by the UVC light. This pathway, that seems to be a general stress response, involves the expression of proteins related to cellular division, purine and pirimidine metabolism, heat chock or chaperones, energy supply, regulation of biofilm formation, transport, regulation of lytic cycle of bacteriophages, besides proteins that show undefined function. Despite the response present similarities with the classic SOS response of E. coli, we still cannot assert that C. violaceum shows a SOS-like response, mainly due to the absence of characterization of a LexA-like protein in this organism

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Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

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Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

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Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

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Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

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The venom proteome of Daboia russelli siamensis, a snake of medical importance in several Asian countries, was analysed by 2-D electrophoresis, subsequent MS/MS and enzymatic assays. The proteome comprises toxins from six protein families: serine proteinases, metalloproteinases, phospholipases A(2), L-amino acid oxidases, vascular endothelial growth factors and C-type lectin-like proteins. The venom toxin composition correlates with the clinical manifestation of the Russell's viper bite and explains pathological effects of the venom such as coagulopathy, oedema, hypotensive, necrotic and tissue damaging effects. The vast majority of toxins are potentially involved in coagulopathy and neurotoxic effects. The predominant venom components are proteinases capable of activating blood coagulation factors and promoting a rapid clotting of the blood, and neurotoxic phospholipase A(2)s. The analysis of the venom protein composition provides a catalogue of secreted toxins. The proteome of D. r. siamensis exhibits a lower level of toxin diversity than the proteomes of other viperid snakes. In comparison to the venoms of Vipera ammodytes ammodytes and Vipera ammodytes meridionalis, the venom from D. r. siamensis showed quantitative differences in the proteolytic, phospholipase A2, L-amino acid oxidase and alkaline phosphatase activities. (c) 2009 Published by Elsevier B.V.

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Background: The hypothalamus plays a pivotal role in numerous mechanisms highly relevant to the maintenance of body homeostasis, such as the control of food intake and energy expenditure. Impairment of these mechanisms has been associated with the metabolic disturbances involved in the pathogenesis of obesity. Since rodent species constitute important models for metabolism studies and the rat hypothalamus is poorly characterized by proteomic strategies, we performed experiments aimed at constructing a two-dimensional gel electrophoresis (2-DE) profile of rat hypothalamus proteins. Results: As a first step, we established the best conditions for tissue collection and protein extraction, quantification and separation. The extraction buffer composition selected for proteome characterization of rat hypothalamus was urea 7 M, thiourea 2 M, CHAPS 4%, Triton X-100 0.5%, followed by a precipitation step with chloroform/methanol. Two-dimensional (2-D) gels of hypothalamic extracts from four-month-old rats were analyzed; the protein spots were digested and identified by using tandem mass spectrometry and database query using the protein search engine MASCOT. Eighty-six hypothalamic proteins were identified, the majority of which were classified as participating in metabolic processes, consistent with the finding of a large number of proteins with catalytic activity. Genes encoding proteins identified in this study have been related to obesity development. Conclusion: The present results indicate that the 2-DE technique will be useful for nutritional studies focusing on hypothalamic proteins. The data presented herein will serve as a reference database for studies testing the effects of dietary manipulations on hypothalamic proteome. We trust that these experiments will lead to important knowledge on protein targets of nutritional variables potentially able to affect the complex central nervous system control of energy homeostasis.

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Chromobacterium violaceum is a free-living bacillus, Gram-negative commonly found in water and sand of tropical and subtropical regions. One of its main characteristic it's the ability to produce the purple pigment named violacein, that shows countless biological activities. In 2003, the genome of this organism was totally sequenced and revealed important informations about the physiology of this bacteria. However, few post-genomics studies had been accomplished. This work evaluated the protein profile of C. violaceum cultivated in LB medium at 28ºC that allowed the identification and characterization of proteins related to a possible secretion system that wasn't identified and characterized yet in C. violaceum, to the quorum sensing system, to regulatory process of transcription and translation, stress adaptation and biotechnological potential. Moreover, the response of the bacteria to UVC radiation was evaluated. The comparison of the protein profile, analyzed through 2-D electrophoresis, of the control group versus the treatment group allowed the identification of 52 proteins that arose after stress induction. The obtained results enable the elaboration of a stress response pathway in C. violaceum generated by the UVC light. This pathway, that seems to be a general stress response, involves the expression of proteins related to cellular division, purine and pirimidine metabolism, heat chock or chaperones, energy supply, regulation of biofilm formation, transport, regulation of lytic cycle of bacteriophages, besides proteins that show undefined function. Despite the response present similarities with the classic SOS response of E. coli, we still cannot assert that C. violaceum shows a SOS-like response, mainly due to the absence of characterization of a LexA-like protein in this organism

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The spherical Lindquist type polyoxometalate, Mo6O192-, has been used as a noncoordinating anionic template for the construction of novel three-dimensional lanthanide-aromatic monocarboxylate dimer supramolecular networks [Ln(2)(DNBA)(4)(DMF)(8)][Mo6O19] (Ln = La 1, Ce 2, and Eu 3, DNBA = 3,5-dinitrobenzoate, DMF = dimethylformamide). The title compounds are characterized by elemental analyses, IR, and single-crystal X-ray diffractions. X-ray diffraction experiments reveal that two Ln(III) ions are bridged by four 3,5-dinitrobenzoate anions as asymmetrically bridging ligands, leading to dimeric cores, [Ln(2)(DNBA)(4)(DMF)(8)](2+); [Ln(2)(DNBA)(4)(DMF)(8)](2+) groups are joined together by pi-pi stacking interactions between the aromatic groups to form a two-dimensional grid-like network; the 2-D supramolecular layers are further extended into 3-D supramolecular networks with 1-D box-like channels by hydrogen-bonding interactions, in which hexamolybdate polyanions reside. The compounds represent the first examples of 3-D carboxylate-bridged lanthanide dimer supramolecular "host" networks formed by pi-pi stacking and hydrogen-bonding interactions encapsulating noncoordinating "guest" polyoxoanion species. The fluorescent activity of compound 3 is reported.