904 resultados para Lysine-rich peptides


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

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O presente trabalho objetivou determinar o tempo mínimo necessário à realização do teste de germinação para sementes de B. brizantha cv. Marandu, o método de superação de dormência e a condição de temperatura que proporcionem a maior germinação no menor tempo. Numa primeira etapa, realizou-se o teste de germinação em trinta lotes de sementes, sob duas condições de temperaturas (15-35ºC e 20-35ºC), avaliadas em conjunto com três métodos para a superação de dormência (H2SO4, KNO3 e Controle), constituindo seis tratamentos. Realizaram-se contagens diárias da germinação para a determinação da data mais apropriada para o término do teste. Na segunda etapa, realizaram-se testes de germinação em oito lotes de diferentes níveis de vigor, utilizando-se os mesmos seis tratamentos e com encerramento do teste nas datas definidas na primeira etapa. Conclui-se que, para o teste de germinação de B. brizantha, a escarificação com H2SO4 e a temperatura de 20-35ºC são tratamentos que resultam na maior germinação em um menor tempo, possibilitando o encerramento do teste aos 11 dias após a semeadura.

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Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

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Enzymatic synthesis of peptides using proteases has attracted a great deal of attention in recent years. One key challenge in peptide synthesis is to find supports for protease immobilization capable of working in aqueous medium at high performance, producing watersoluble oligopeptides. At present, few reports have been described using this strategy. Therefore, the aim of this thesis was to immobilize proteases applying different methods (Immobilization by covalent bound, entrapment onto polymeric gels of PVA and immobilization on glycidil metacrylate magnetic nanoparticles) in order to produce water-soluble oligopeptides derived from lysine. Three different proteases were used: trypsin, α-chymotrypsin and bromelain. According to immobilization strategies associated to the type of protease employed, trypsin-resin systems showed the best performance in terms of hydrolytic activity and oligopeptides synthesis. Hydrolytic activities of the free and immobilized enzymes were determined spectrophotometrically based on the absorbance change at 660 nm at 25 °C (Casein method). Calculations of oligolysine yield and average degree of polymerization (DPavg) were monitored by 1H-NMR analysis. Trypsin was covalently immobilized onto four different resins (Amberzyme, Eupergit C, Eupergit CM and Grace 192). Maximum yield of bound protein was 92 mg/g, 82 mg/g and 60 mg/g support for each resin respectively. The effectiveness of these systems (Trypsin-resins) was evaluated by hydrolysis of casein and synthesis of water-soluble oligolysine. Most systems were capable of catalyzing oligopeptide synthesis in aqueous medium, albeit at different efficiencies, namely: 40, 37 and 35% for Amberzyme, Eupergit C and Eupergit CM, respectively, in comparison with free enzyme. These systems produced oligomers in only 1 hour with DPavg higher than free enzyme. Among these systems, the Eupergit C-Trypsin system showed greater efficiency than others in terms of hydrolytic activity and thermal stability. However, this did not occur for oligolysine synthesis. Trypsin-Amberzyme proved to be more successful in oligopeptide synthesis, and exhibited excellent reusability, since it retained 90% of its initial hydrolytic and synthetic activity after 7 reuses. Trypsin hydrophobic interactions with Amberzyme support are responsible for protecting against strong enzyme conformational changes in the medium. In addition, the high concentration of oxirane groups on the surface promoted multi-covalent linking and, consequently, prevented the immobilized enzyme from leaching. The aforementioned results suggest that immobilized Trypsin on the supports evaluated can be efficiently used for oligopeptides synthesis in aqueous media

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Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

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I thank to my advisor, João Marcos, for the intellectual support and patience that devoted me along graduate years. With his friendship, his ability to see problems of the better point of view and his love in to make Logic, he became a great inspiration for me. I thank to my committee members: Claudia Nalon, Elaine Pimentel and Benjamin Bedregal. These make a rigorous lecture of my work and give me valuable suggestions to make it better. I am grateful to the Post-Graduate Program in Systems and Computation that accepted me as student and provided to me the propitious environment to develop my research. I thank also to the CAPES for a 21 months fellowship. Thanks to my research group, LoLITA (Logic, Language, Information, Theory and Applications). In this group I have the opportunity to make some friends. Someone of them I knew in my early classes, they are: Sanderson, Haniel and Carol Blasio. Others I knew during the course, among them I’d like to cite: Patrick, Claudio, Flaulles and Ronildo. I thank to Severino Linhares and Maria Linhares who gently hosted me at your home in my first months in Natal. This couple jointly with my colleagues of student flat Fernado, Donátila and Aline are my nuclear family in Natal. I thank my fiancée Luclécia for her precious a ective support and to understand my absence at home during my master. I thank also my parents Manoel and Zenilda, my siblings Alexandre, Paulo and Paula.Without their confidence and encouragement I wouldn’t achieve success in this journey. If you want the hits, be prepared for the misses Carl Yastrzemski

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Objectives: To analyze the healing of autogenous onlay bone grafts in three different situations, focusing on the interface area.Material and methods: Sixteen rabbits underwent autogenous bone graft surgeries in the calvaria. The block bone grafts were positioned in three different situations: direct contact between bone graft and receptor bed, graft interposed by particulate bone, and graft interposed by platelet-rich plasma (PRP). After 7, 15, 30, and 60 days, the specimens were retrieved for histological and morphometric evaluation.Results: All groups healed uneventfully and presented incorporation of the grafts after 30 days. A slightly more evident new bone formation could be observed in the PRP group in the first analyzed period, and an earlier maturation of bone in the last period, although no statistically significant differences were achieved.Conclusion: the use of additional material between the bone graft and the receptor bed when using the onlay technique must be carefully considered, taking into account the size of the reconstruction and the cost/benefit relation. The addition of PRP in between autogenous bone blocks and the receptor bed did not confer significant benefit for the new bone formation and healing on the calvaria of bone of rabbits.

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Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

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Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

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The purpose of this study was to analyze histologically the influence of platelet-rich plasma (PRP) coagulated with two different activators on bone healing in surgically created critical-size defects (CSD) in rat calvaria.Forty-eight rats were divided into three groups: C, PRP-C and PRP-T. An 8 mm diameter CSD was created in the calvarium of each animal. In group C, the defect was filled by a blood clot only. In groups PRP-C and PRP-T, the defect was filled with PRP activated with either calcium chloride or thromboplastin solution, respectively. Each group was divided into two subgroups (n = 8 per subgroup) and killed at either 4 or 12 weeks postoperatively. Histologic and histometric analyses were performed. The amount of new bone formed was calculated as a percentage of the total area of the original defect. Percentage data were transformed into arccosine for statistical analysis (analysis of variance, Tukey's post hoc test, p < 0.05).No defect completely regenerated with bone. Group PRP-C had a statistically greater amount of bone formation than groups C and PRP-T at both time points of analysis. No statistically significant differences were observed between groups C and PRP-T.It can be concluded that the type of activator used to initiate PRP clot formation influences its biological effect on bone healing in CSD in rat calvaria.