Bioquímica e Biologia Molecular

URI permanente desta comunidadehttps://locus.ufv.br/handle/123456789/11837

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Data inicial: 2013Data final: 2013

Resultados da Pesquisa

Agora exibindo 1 - 10 de 21
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    Identification and expression levels of pig miRNAs in skeletal muscle
    (Livestock Science, 2013-06) Verardo, L. L.; Nascimento, C. S.; Silva, F. F.; Gasparino, E.; Toriyama, E.; Barbosa, A. R.; Costa, K. A.; Lopes, P. S.; Guimarães, S. E. F.; Périssé, I. V.
    MicroRNAs are a class of naturally occurring non-coding RNAs. Typically they are $22 nucleotides long and suppress translation of their targets genes. Several laboratories have attempted to identify miRNAs from pig muscle and the bioinformatics strategies using ESTs have proved to be successful for this aim. In this study we report an in silico identification of ncRNA in pig EST libraries focusing on novel pig miRNAs and further investigated the differential expression of pigs miRNAs (known and novel) by quantitative real-time PCR during preand postnatal stage from Commercial and local breed Piau pigs skeletal muscle tissue. We identified two miRNAs not yet described in pigs: hsa-miR-1207-5p and hsa-miR-665. Besides, we found 288 target genes for hsa-miR-1207-5p and 214 for hsa-miR-665; from them, four are muscle specific genes. Through expression analyses, differences were found between pre- and postnatal stages and genetics groups. The findings of miRNAs and their muscle-specific targets in pigs will be helpful for understanding the function and processing of this RNA class in the future. Besides, the miRNAs differentially expressed between Commercial and Piau breeds suggest that they can be used to uncover phenotypic differences across different genetic groups.
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    Concentração de proteína solúvel por bradford revela diferenças no metabolismo de plantas de ora-pro-nobis em diferentes doses de nitrogênio
    (Revista Brasileira de Agropecuária Sustentável, 2013-07) Guimarães, Dimitrius Santiago P. Simões Fróes; Souza, Maria Regina de Miranda; Hirano, Rafael T.; Pereira, Paulo Roberto Gomes; Baracat-Pereira, Maria Cristina
    O ora-pro-nobis (Pereskia aculeata Mill.) é um alimento rico em proteínas, muito utilizado no meio rural e com importância crescente na indústria de alimentos e farmacológica. O objetivo deste trabalho foi determinar a concentração de proteínas pelos métodos de Bradford e Kjeldahl, em folhas de plantas de ora-pro-nobis submetidas a diferentes doses de adubação nitrogenada, comparando estes métodos. As folhas das plantas de ora-pro-nobis adubadas com diferentes doses de N (0, 50, 100, 200 e 400 kg de N/ha) foram coletadas aos 423 dias após o plantio (DAP). Para o método de Bradford, as folhas foram trituradas com nitrogênio líquido e maceradas em Tris-HCl 50 mM, pH 7,0, o homogenato centrifugado e a proteína solúvel determinada no sobrenadante. Para avaliar o perfil proteico, as amostras dos diferentes tratamentos foram separadas por SDS-Tricina-PAGE 14%. O método de Kjeldahl tradicional foi realizado usando-se o fator de correção 6,25. Os resultados por ambos os métodos indicaram que houve alterações nas concentrações e composição de proteínas presentes em função da disponibilidade de N no solo. A proteína total por Kjeldahl aumentou até a dose de 100 kg de N/ha, e a proteína solúvel por Bradford aumentou nas doses de N entre 50 e 200 kg/ha. Pelo SDS-Tricina-PAGE, verificou-se aumento da intensidade das bandas consonante com o método de Bradford. Estes resultados sugerem que a avaliação de proteínas solúveis pelo método de Bradford permite detectar diferenças no metabolismo das plantas de ora-pro-nobis, expressando informações biológicas relevantes para estudos fisiológicos e nutricionais.
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    Differentially expressed proteins during an incompatible interaction between common bean and the fungus Pseudocercospora griseola
    (Molecular Breeding, 2013-07-30) Borges, Leandro Luiz; Santana, Fernanda Abreu; Castro, Isabel Samila Lima; Arruda, Klever Márcio Antunes; Ramos, Humberto Josué de Oliveira; Moreira, Maurilio Alves; Barros, Everaldo Gonçalves de
    The common bean (Phaseolus vulgaris L.) is the main source of protein and an important source of minerals in several countries around the world. Angular leaf spot, caused by the fungus Pseudocercospora griseola, is one of the major diseases of the common bean. In this work, we used two-dimensional gel electrophoresis and mass spectrometry to analyze alterations in the proteome of common bean leaves challenged with an incompatible race of P. griseola. Twenty-three differentially expressed proteins were detected in leaves of cultivar AND 277 collected at 12, 24 and 48 h after inoculation. The proteins were digested with trypsin and submitted to MALDI-TOF/TOF and MicrOTOF-Q electrospray mass spectrometry. Nineteen of them were identified upon MS/MS fragmentation. Most of these proteins are involved with amino acid metabolism, terpenoid metabolism, phenylpropanoid biosynthesis, antioxidant systems, vitamin and cofactor metabolism, plant–pathogen interaction, carbohydrate metabolism, photosynthesis, or genetic information processing, showing that the interaction in this pathosystem affects different genes from various metabolic pathways and processes.
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    The nematophagous fungus Monacrosporium thaumasium and its nematicidal activity on Angiostrongylus vasorum
    (Revista Iberoamericana de Micología, 2013-09-24) Soares, Filippe Elias de Freitas; Braga, Fabio Ribeiro; Araújo, Jackson Victor de; Lima, Walter dos Santos; Queiroz, José Humberto de
    The dog acts as a reservoir and environmental disseminator of potentially zoonotic parasites. The objective of this work was to study the fungus Monacrosporium thaumasium regarding its nematicidal potential in laboratory trials and its proteolytic profile. The in vitro test was carried out through two assays (A and B). In assay A, conidia of the fungus N34a were added in positive coprocultures for Angiostrongylus vasorum. In assay B, crude extract (treated group) and distilled water (control group) were added to coprocultures. Next, the proteolytic profile of crude extract of the nematophagous fungus M. thaumasium (NF34a) was revealed by performing a zymogram. There was a reduction (p < 0.01) in the averages of larvae recovered from the treated groups (conidia and crude extract) in relation to control groups. The zymogram suggested that the nematophagous fungus M. thaumasium produces a protease of approximately 40 kDa. The results of this work confirm that the conidia as well as the crude extract of the fungus M. thaumasium may be used to control A. vasorum L1. The proteolytic profile suggested the presence of one protease (Mt1) of approximately 40 kDa that in the future may be used in biological control of L1 of this nematode.
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    Nematicidal activity of three novel extracellular proteases of the nematophagous fungus Monacrosporium sinense
    (Parasitology Research, 2013-01-31) Soares, Filippe E. F.; Braga, Fabio R.; Araújo, Jackson V.; Geniêr, Hugo L. A.; Gouveia, Angélica S.; Queiroz, José H.
    Extracellular proteases are an important virulence factor for the nematophagous fungi Monacrosporium. The objective of this study was to optimize, purify, partially characterize, and to evaluate the nematicidal activity of the proteases produced by the nematophagous fungus Monacrosporium sinense (SF53) by solid-state fermentation. Wheat bran was used as substrate for protease production. The variables moisture, pH, incubation time, temperature, glucose, yeast extract, and the number of conidia were tested for their influences on protease production by SF53. To determine the optimal level of the selected variables the central composite design was applied. The crude extract obtained was purified in two steps, an ion exchange chromatography and a gel excision. SDS-PAGE and zymogram were performed for analysis of the purification process. Proteolytic activity was also tested at different pHs and temperatures. In the in vitro assay, the nematicidal activity of the three proteases was evaluated. pH and incubation time showed a significant effect (p < 0.05) on production of protease. The highest value of activity was 38.0 (U/ml) under the conditions of pH 5.0 and incubation time of 211 h. SF53 produced three different proteases (Ms1, Ms2, and Ms3) which were directly purified from the zymogram. Ms1, Ms2, and Ms3 showed the following percentage of reduction (p < 0.05) on the number of Panagrellus redivivus compared to control after 24 h: 76.8, 68.1, and 92.1 %. This is the first report of the use of proteases of the isolate SF53 on a phytonematode, which may be a research tool in future works.
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    Proteolytic activity of gut bacteria isolated from the velvet bean caterpillar Anticarsia gemmatalis
    (Journal of Comparative Physiology B, 2013-02-08) Pilon, F. M.; Visôtto, L. E.; Guedes, R. N. C.; Oliveira, M. G. A.
    The development of proteinase inhibitors as potential insect control agents has been constrained by insect adaptation to these compounds. The velvet bean caterpillar (Anticarsia gemmatalis) is a key soybean pest species that is well-adapted to proteinase inhibitors, particularly serine-proteinase inhibitors, which are abundant in the caterpillar host. The expression of diverse proteolytic enzymes by gut symbionts may allow the velvet bean caterpillar to circumvent proteinase inhibitors produced by the host plant. In this study, we characterized the proteolytic activity of the four nonpathogenic species of gut bacteria isolated from the velvet bean caterpillar—Bacillus cereus, Enterococcus gallinarum, Enterococcus mundtii and Staphylococcus xylosus. Two proteinase substrates, N-a-benzoyl- L -Arg-p-nitroanilide ( L -BApNA) and N-a-p-tosyl- L -Arg methyl ester ( L -TAME) and five proteinase inhibitors [aprotinin, E-64, ethylenediamine tetraacetic acid (EDTA), pepstatin and N-a-tosyl- L -lysine chloromethyl ketone (TLCK)] as well as CaCl 2 , pH and temperature profiles were used to characterize the expressed proteolytic activity of these bacterial strains in vitro. Kinetic parameters for proteolytic activity were also estimated. The results of these experiments indicated that serine- and cysteine-proteinase activities were expressed by all four gut bacteria symbionts of the velvet bean caterpillar. The cysteine- and serine-proteinase activities of these gut symbionts were distinct and different from that of gut proteinases of the caterpillar itself. This finding provides support for the potential involvement of gut symbionts in the mitigation of the negative effects of serine-proteinase inhibitors in the velvet bean caterpillar.
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    Efeito do consumo da aveia e farinha da casca de maracujá sobre a glicemia e lipemia em um grupo de voluntários
    (Revista de Ciências Farmacêuticas Básica e Aplicada, 2013-04-13) Oliveira, Marcos Rodrigo; Miranda, Gilson Silva; Rennó, Luciana Navajas; Machado, Bruno Brandão; Silva, Juliana Lopes; Pinto, Rogério
    O objetivo do presente artigo foi analisar os efeitos do consumo da aveia em flocos e da farinha da casca de maracujá sobre a glicemia e a lipemia em um grupo de voluntários, com hábitos sedentários. Foi realizado um estudo clínico com 28 pessoas na cidade de Viçosa, MG, no ano de 2010, com idade variando entre 30 a 60 anos, onde essas foram divididas de forma aleatória em dois grupos de 14 pessoas. O grupo 1 recebeu 30 g de aveia em flocos enquanto o grupo 2, 30 g de farinha da casca de maracujá, durante 60 dias. Para verificação dos resultados bioquímicos, foram realizadas três coletas de sangue: a primeira, no início do estudo (tempo zero), a segunda, após 30 (t30) dias; e a terceira, após 60 dias (t60). Com o uso da aveia em flocos, observou-se que a concentração de HDL aumentou e a concentração sérica de glicose diminuiu significativamente, porém as variáveis peso, colesterol total e LDL não diferiram estatisticamente. Com a ingestão da farinha da casca do maracujá, houve diminuição significativa das concentrações de glicose e de colesterol total, contudo, as variáveis peso, HDL e LDL não diferiram estatisticamente. Os produtos utilizados foram eficazes na redução da concentração sérica de glicose e houve também melhora no perfil lipídico.
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    Use of sorghum straw (Sorghum bicolor) for second generation ethanol production: pretreatment and enzymatic hydrolysis
    (Química Nova, 2013-04-04) Tavares, Gabriella Peterlini; Cardoso, Wilton Soares; Tardin, Flávio Dessaune; Queiroz, Paula Viana; Mota, Samuel Sampaio; Kasuya, Maria Catarina Megumi; Queiroz, José Humberto de
    Agronomic biomass yields of forage sorghum BRS 655 presented similar results to other energy crops, producing 9 to 12.6 tons/ha (dry mass) of sorghum straw. The objective of this study was to evaluate the lignocellulosic part of this cultivar in terms of its potential in the different unit processes in the production of cellulosic ethanol, measuring the effects of pretreatment and enzymatic hydrolysis. Three types of pre-treatments for two reaction times were conducted to evaluate the characteristics of the pulp for subsequent saccharification. The pulp pretreated by alkali, and by acid followed by delignification, attained hydrolysis rates of over 90%.
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    Concentração de proteína solúvel por Bradford releva diferenças no metabolismo de plantas de Ora-pro-nobis em diferentes doses de nitrogênio
    (Revista Brasileira de Agropecuária Sustentável, 2013-07-18) Guimarães, Dimitrius Santiago P. Simões Fróes; Souza, Maria Regina de Miranda; Hirano, Rafael T.; Pereira, Paulo Roberto Gomes; Baracat-Pereira, Maria Cristina
    O ora-pro-nobis (Pereskia aculeata Mill.) é um alimento rico em proteínas, muito utilizado no meio rural e com importância crescente na indústria de alimentos e farmacológica. O objetivo deste trabalho foi determinar a concentração de proteínas pelos métodos de Bradford e Kjeldahl, em folhas de plantas de ora-pro-nobis submetidas a diferentes doses de adubação nitrogenada, comparando estes métodos. As folhas das plantas de ora-pro-nobis adubadas com diferentes doses de N (0, 50, 100, 200 e 400 kg de N/ha) foram coletadas aos 423 dias após o plantio (DAP). Para o método de Bradford, as folhas foram trituradas com nitrogênio líquido e maceradas em Tris-HCl 50 mM, pH 7,0, o homogenato centrifugado e a proteína solúvel determinada no sobrenadante. Para avaliar o perfil proteico, as amostras dos diferentes tratamentos foram separadas por SDS-Tricina-PAGE 14%. O método de Kjeldahl tradicional foi realizado usando-se o fator de correção 6,25. Os resultados por ambos os métodos indicaram que houve alterações nas concentrações e composição de proteínas presentes em função da disponibilidade de N no solo. A proteína total por Kjeldahl aumentou até a dose de 100 kg de N/ha, e a proteína solúvel por Bradford aumentou nas doses de N entre 50 e 200 kg/ha. Pelo SDS-Tricina-PAGE, verificou-se aumento da intensidade das bandas consonante com o método de Bradford. Estes resultados sugerem que a avaliação de proteínas solúveis pelo método de Bradford permite detectar diferenças no metabolismo das plantas de ora-pro-nobis, expressando informações biológicas relevantes para estudos fisiológicos e nutricionais.
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    Optimization of Endoglucanase and Xylanase activities from fusarium verticillioides for simultaneous saccharification and fermentation of sugarcane bagasse
    (Applied Biochemistry and Biotechnology, 2013-10-30) Almeida, Maíra N. de; Guimarães, Valéria M.; Falkoski, Daniel L.; Paes, Guilherme B. T.; Ribeiro Jr., José Ivo; Visser, Evan M.; Alfenas, Rafael F.; Pereira, Olinto L.; Rezende, Sebastião T. de
    Enzymatic hydrolysis is an important but expensive step in the production of ethanol from biomass. Thus, the production of efficient enzymatic cocktails is of great interest for this biotechnological application. The production of endoglucanase and xylanase activites from F. verticillioides were optimized in a factorial design (25) followed by a CCDR design. Endoglucanase and xylanase activities increased from 2.8 to 8.0 U/mL and from 13.4 to 114 U/mL, respectively. The optimal pH and temperature were determined for endoglucanase (5.6, 80 °C), cellobiase (5.6, 60 °C), FPase (6.0, 55 °C) and xylanase (7.0, 50 °C). The optimized crude extract was applied in saccharification and fermentation of sugarcane bagasse from which 9.7 g/L of ethanol was produced at an ethanol/biomass yield of 0.19.