293 resultados para Cooking (Gelatin)
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Foram utilizados 78 cabritos de ambos os sexos e de cinco genótipos: raça Alpina, ½ Boer + ½ Alpina, ¾ Boer + ¼ Alpina, ½ Anglo-nubiana + ½ Alpina e tricross (½ Anglo-nubiana + ¼ Boer + ¼ Alpina), com peso médio inicial de 14,1 ± 2,5 kg. Objetivou-se avaliar os efeitos do genótipo, do sistema de terminação e do sexo sobre as características físico-químicas da carne. Os sistemas de terminação foram constituídos: ST1 - cabrito + mãe em pasto e ST2 - cabrito desmamado confinado. Os cabritos do ST1 foram mantidos em piquetes formados com Panicum maximum cv. Tanzânia e os do ST2 receberam dieta completa com 16% PB e 73% NDT. Os valores de pH, a* (teor de vermelho), Perda de Peso ao Cozimento (PPC) e porcentagem de Extrato Etéreo (EE) foram influenciados pelo genótipo. Os teores de vermelho (a*) e L* (luminosidade), PPC e porcentagens de umidade, proteína, EE e cinzas foram influenciados pelo sistema de terminação. O músculo longissimus dorsidos animais ½ BA apresentou as melhores características físico-químicas. Se a preferência do consumidor for por uma carne mais macia e com maior teor de gordura, as fêmeas são mais indicadas.
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Objetivou-se avaliar a qualidade da carne por meio da determinação da composição centesimal e das características físicas da paleta e do lombo de 20 cordeiros mestiços Ideal, não-castrados (NC) e castrados (C), submetidos a fotoperíodos curto, de 12 horas (FC), e longo, de 18 horas (FL). Os animais foram divididos ao acaso em esquema fatorial 2 x 2 (condição sexual x fotoperíodo), em quatro tratamentos (NC em FC; C em FC; NC em FL; e C em FL), com cinco repetições, sendo terminados em confinamento individual até que atingissem 37 kg de PV. A qualidade da carne foi determinada em amostras da paleta e do músculo Longissimus lumborum (LL). Os mais baixos teores de umidade e os mais altos de EE, tanto na paleta como no lombo, foram determinados na carne dos animais castrados. O fotoperíodo longo influenciou significativamente os teores de EE e proteína total do músculo LL. A capacidade de retenção de água foi afetada pelo FL, no qual foram encontrados percentuais mais baixos. Não houve efeito do fotoperíodo nem da condição sexual sobre a capacidade de absorção de água e a perda de peso por cozimento do músculo LL. Os valores de força de cisalhamento (FoCi) na carne de cordeiros NC e dos submetidos ao FL foram superiores aos da carne dos animais castrados ou expostos ao FC.
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Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
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Here, we evaluated collagen distribution and matrix metalloproteinases (MMPs) MMP-2 and MMP-9 activities in skeletal muscle of pacu (Piaractus mesopotamicus) during juvenile and adult growth phases. Muscle samples from juvenile and adult fishes were processed by histochemistry for collagen system fibers and for gelatin-zymography for MMP-2 and MMP-9 activities analysis. Picrosirius staining revealed a myosept, endomysium, and perimysium-like structures in both growth phases and muscle types, with increased areas of collagen fibers in adults, mainly in red muscle. Reticulin staining showed that reticular fibers in the endomysium-like structure were thinner and discontinuous in the red muscle fibers. The zymography revealed clear bands of the pro-MMP-9, active-MMP-9, intermediate-MMP-2, and active-MMP-2 forms in red and white muscle in both growth phases. MMP-2 activity was more intense in juvenile than adult muscle fibers. Comparing the red and white muscle types, MMP-2 activity was significantly higher in red muscle in adult phase only. The activity of MMP-9 forms was similar in juvenile red and white muscles and in the adult red muscle, without any activity in adult white muscle. In conclusion, our results show that, in pacu, the higher activities of MMP-2 and -9 are associated with the rapid muscle growth in juvenile age and in adult fish, these activities are related with a different red and white muscle physiology. This study may contribute to the understanding muscle growth mechanisms and may also contribute to analyse red and the white muscle parameters of firmness and softness, respectively, of the commercial product. Anat Rec, 292:387-395, 2009. (C) 2009 Wiley-Liss, Inc.
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Fresh cut cassava is an alternative to prolong roots commercialization period, adding value to the raw material and assisting the consumer's needs that seeks for products of easy preparation. The aim of this work was to evaluate the quality of fresh cut cassava subjected to gamma radiation. For processing cassava roots (IAC - 567-70) were used, after preparation were packed in expanded polystyrene (EPS) trays, covered by low density polyethylene (LDPE) plastic film, being subjected to gamma radiation in the doses 0 (control), 0.5; 1.0; 1.5; 2.0 KGy. Another treatment was also carried out in which roots were vacuum packed in nylon + polyethylene. Packed roots of all treatments were stored under refrigeration (5 +/- 1 degrees C) and evaluations were made after 0, 3, 6, 9 and 12 days. Evaluated analyses were: titrable acidity, pH, firmness, coloration, polyphenoloxidase activity, cooking time and roots hydration percentage. Evaluations just proceeded until the nine storage day, because after this period the roots already presented visually detectable physiologic and microbiological alterations. For titrable acidity, the vacuum packed roots had the lowest values, followed by the control packed in trays. For the other treatments, values of acidity increased and influenced the pH values. For the other appraised parameters there was no significant difference among treatments, but alterations were observed with the storage. The firmness decreased with the storage. Brightness values and yellow color component decreased with the storage and the b * (green) color values increased. Polyphenoloxidase activity was maximum in the day of elaboration of the product and it decreased with the storage. Cooking time and roots hydration percentage decreased with the roots storage.
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A presença de sementes em frutos, dependendo da espécie, pode ser dispensável ou até mesmo indesejável ao consumo e à culinária, como seria o caso, por exemplo, de algumas pimentas. Este trabalho teve como objetivo avaliar o efeito da aplicação de ácido giberélico (GA3) na produção e nas características de frutos partenocárpicos em plantas de pimenta escabeche (Capsicum baccatum L.). A semeadura foi realizada em bandejas de poliestireno expandido e o plantio das mudas em estrutura plástica com irrigação por gotejamento. O delineamento experimental foi em blocos casualizados com quatro tratamentos (0; 700; 1.400 e 2.800 mg L-1 de GA3). Foram feitas duas aplicações de GA3: a primeira quando as plantas estavam em plena floração e a segunda aos 10 dias após a primeira aplicação. Observou-se que o GA3 favoreceu a produção de frutos partenocárpicos (94,6% com 2.100 mg L-1 de GA3), diminuiu a produção de frutos (87,1 g fruto/planta com 2.250 mg L-1 de GA3), aumentou a altura da planta (168,0 cm com 2.450 mg L-1 de GA3), porém, prejudicou as características comerciais dos frutos. É possível produzir frutos partenocárpicos com a aplicação de GA3 em pimenta escabeche, porém esta técnica não foi agronomicamente viável.
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BjussuMP-II is an acidic low molecular weight metalloprotease (Mr similar to 24,000 and pI similar to 6.5), isolated from Bothrops jararacussu snake venom. The chromatographic profile in RP-HPLC and its N-terminal sequence confirmed its high purity level. Its complete cDNA was obtained by RT-PCR and the 615 bp codified for a mature protein of 205 amino acid residues. The multiple alignment of its deduced amino acid sequence and those of other snake venom metalloproteases showed a high structural similarity, mainly among class P-I proteases. The molecular modeling analysis of BjussuMP-II showed also conserved structural features with other SVMPs. BjussuMP-II did not induce hemorrhage, myotoxicity and lethality, but displayed dose-dependent proteolytic activity on fibrinogen, collagen, fibrin, casein and gelatin, keeping stable at different pHs, temperatures and presence of several divalent ions. BjussuMP-II did not show any clotting or anticoagulant activity on human citrated plasma, in contrast to its inhibitory effects on platelet aggregation. The aspects broached, in this work, provide data on the relationship between structure and function, in order to better understand the effects elicited by snake venom metalloproteases. (c) 2007 Elsevier B.V. All rights reserved.
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Results from our laboratory revealed propolis activity on Giardia trophozoites proliferation. Since therapeutic agents can inhibit the activity of proteases related to relevant biologic and physiologic processes of parasites, this study was undertaken to characterise the proteolytic activity of excretory/secretory products (ESP) of trophozoites treated with propolis. ESP was obtained from culture supernatants of trophozoites exposed to 250 and 500 mu g mL(-1) of propolis. ESP were tested in sodium dodecyl sulfate-polyacrylamide gel electrophoresis for the protein profiles and the protease activity was assayed in gelatin-containing gels. Synthetic inhibitors were used to characterise the protease classes. Treated and non-treated ESP showed a similar protein and hydrolysis pattern. A simple pattern of protein composed by five evident bands of approximately 167, 132, 79, 61 and 51 kDa was found, and the zymograms comprised hydrolysis zones distributed from > 170 to 23 kDa. No inhibition was seen on protease activity of propolis-treated trophozoites, whose hydrolysis pattern was similar to control. One may conclude that both ESP degraded gelatin and the activity was predominantly due to cysteine proteases. Although propolis had no effect on the proteolytic activity, further studies could identify the active constituents responsible for propolis antigiardial activity and their mechanisms of action.
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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 present investigation was undertaken to identify and characterize trophozoite proteases of five axenic strains of Giardia duodenalis isolated in Brazil and the reference strain Portland 1 isolated in the United States. Trophozoite cell lysates of each strain were analysed for the pattern of proteins and for proteolytic activity. Samples were tested in SDS-polyacrylamide gel electrophoresis for the protein profiles, and the detection of proteases in cell lysates was performed using substrate gel electrophoresis [gelatin, collagen, bovine serum albumin (BSA) and haemoglobin] and azocasein assays. Indeed, synthetic inhibitors were included in the assays to characterize the protease classes. Differences on the hydrolysis patterns of protein substrates were observed in relation to the substrate composition as much as the Giardia trophozoite strain. The substrate-containing gels revealed hydrolysis bands with molecular masses ranging from > 97 to 20-15 kDa, and most zones were common to the five strains. However, some pronounced differences could be detected in the BTU-11 pattern. Azocasein was also degraded; however, depending on the lysate assayed, the degree of substrate degradation was variable. It was observed that inhibitory effects are substrate-dependent since the activity was predominantly due to cysteine proteases against gelatin, collagen, BSA and azocasein substrates and due to serine against haemoglobin. The presence of aspartic protease and aminopeptidase activity in the lysates was also indicated.
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We have examined by gelatin-SDS-PAGE the protease activity in cell lysates of Giardia duodenalis trophozoites of two axenic strains isolated in Brazil from a symptomatic patient (BTU-11) and an asymptomatic carrier (BTU-10), and the reference strain Portland 1 (P1). The proteolysis band patterns showed differences among strains isolated from asymptomatic and symptomatic individuals. The lysate of the strain BTU-10, showed only five hydrolysis bands, while a greater number of bands (10-11 bands) was seen in strains BTU-11 and P1. The protease activity in all lysates was inhibited by cysteine (E-64 and iodoacetamide) and serine proteases (TPCK and TLCK) inhibitors, but not by PMSF and EDTA. In general, the results revealed protease activities in G. duodenalis trophozoites of Brazilian axenic strains and the predominance of cysteine proteinases. It should be stressed the inter-strain difference in hydrolysis band patterns observed between strains isolated from symptomatic patients and the strain obtained from an asymptomatic carrier.
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O presente estudo consiste em uma caracterização preliminar da atividade proteolítica de frações de proteínas purificadas a partir de lisados de trofozoítos de cepa isolada e axenizada no Brasil. Frações obtidas por cromatografia líquida (FPLC) foram analisadas quanto ao perfil eletroforético em géis de poliacrilamida (SDS-PAGE) e a atividade proteolítica foi avaliada em géis contendo gelatina como substrato. A caracterização das enzimas foi realizada a partir da análise do efeito de inibidores sintéticos de cisteína-proteases (E-64, IAA), serina-proteases (PMSF), serina e cisteína-proteases (TPCK, TLCK, elastatinal), metalo-proteases (EDTA) e aspartil proteases (pepstatina) sobre a degradação do substrato. Entre 30 frações eluídas, bandas de proteínas foram observadas em oito delas, entretanto, atividade proteolítica foi detectada apenas nas frações 23, 24, 25 e 26. O perfil eletroforético das proteínas revelou poucas bandas distribuídas na faixa de 45 a 18 kDa. Os zimogramas revelaram zonas de proteólise na faixa de aproximadamente 62 a 35 kDa, entretanto destacaram-se as bandas de hidrólise de 62, 55, 53, 50, 46 e 40 kDa. Nos ensaios de inibição, a proteólise foi marcantemente inibida por E-64, TPCK, TLCK e elastatinal. Redução discreta da proteólise foi observada com IAA e PMSF, enquanto que EDTA e pepstatina não promoveram alteração dos perfis de hidrólise. Estas observações são relevantes, especialmente se considerarmos que para elucidar o envolvimento das proteases na relação parasita-hospedeiro, a purificação dessas moléculas é um requisito importante.
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One of the major questions concerning Giardia is the understanding of pathophysiological processes associated with small intestine abnormalities. There are evidences that Giardia trophozoites contain and/or release proteolytic enzymes that may be implicated in the host intestinal epithelium. The present investigation was undertaken to examine the protease activity in excretory/secretory (E/S) products of Giardia duodenalis trophozoites of an axenic Brazilian strain (BTU-11) and the reference strain Portland 1 (P1). E/S products from trophozoites of each strain in conditioned medium were tested with sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) for the protein profiles, and the protease activity was analyzed using substrate-impregnated SDS-PAGE (gelatin and collagen) and hemoglobin assay. The proteases characterization was based on inhibition assays including synthetic inhibitors. Electrophoresis analysis of E/S products revealed a banding pattern composed by few bands (4 to 6 bands) in the migration region of 123 to 28 kDa. Proteolytic products were detected in the conditioned medium by trophozoites of both assayed strains. In the gels containing copolymerized gelatin and collagen, E/S products promoted substrate degradation and the most evident proteolysis zones were distributed in the migration regions of 77 to 18 kDa and 145 to 18 kDa, respectively, in the patterns of gelatinolytic and collagenolytic activities. Degradation of hemoglobin was also observed, and the pattern of hydrolysis was similar in both E/S products assayed. Inhibitor assays showed that the main proteolytic activity in both E/S products is due to cysteine proteases, although the presence of serine proteases was also indicated. Degradation of substrates including collagen and hemoglobin could lead us to speculate different functions of Giardia excreted/secreted proteases in vivo, but to confirm this possibility and to elucidate its implication on host-parasite interactions, further experiments applying protocols for the purification of proteases are necessary. Even so, our observations are relevant and hold the perspective for the understanding about protease activity in Giardia trophozoites of axenic strain isolated in an endemic area.