931 resultados para Protein incubation time


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A antracnose, causada por Colletotrichum dematium var. truncata, é a principal doença da soja que afeta a fase inicial de formação das vagens, podendo causar a morte das plântulas. Cercospora kikuchii é o fungo causador da doença mancha púrpura nas sementes de soja, responsável por severas reduções no rendimento e na qualidade das sementes. O objetivo deste trabalho foi determinar o tempo mais apropriado para infecção das sementes de soja por C. dematium var. truncata e C. kikuchii, para posterior avaliação dos danos causados pelo fungo na germinação. Tais fungos foram cultivados em meio BDA. As sementes de soja cultivar MSOY 6101 foram colocadas sobre meio contendo o patógeno C. dematium var. truncata por 0 (testemunha), 4, 16, 24, 32 e 40h. As sementes da cv. CD 208 foram inoculadas com C. kikuchii pelos mesmos períodos citados, com adição de 48 e 56h. Após os respectivos tempos de contato, as sementes foram submetidas ao teste de sanidade (papel de filtro), com duas variações, sem e com assepsia superficial (hipoclorito de sódio 1% por três minutos). Determinado o tempo mais adequado de infecção, outras sementes foram infectadas pelo patógeno e, posteriormente, foram realizados testes de germinação em papel e areia com uma mistura de sementes sadias (colocadas sobre o meio BDA) e sementes inoculadas, resultando em lotes com 0, 20, 40, 60, 80 e 100% de sementes infectadas. O tempo de incubação de 40 e 56 horas para C. dematium var truncata e C. Kikuchii, respectivamente são suficientes para obtenção da totalidade das sementes infectadas. C. Kikuchii não prejudica a germinação das sementes.

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O objetivo deste trabalho foi avaliar o efeito da aplicação superficial de calcário em plantio direto, na produtividade e nas características tecnológicas dos grãos de cultivares de feijão. O trabalho foi desenvolvido durante dois anos agrícolas, em Latossolo Vermelho distrófico, em condições de sequeiro. O delineamento experimental utilizado foi o de blocos casualizados em esquema de parcelas subdivididas, com quatro repetições. As parcelas foram representadas por cultivares de feijão (Carioca, IAC Carioca Eté, Pérola, IAPAR 81 e Campeão 2) e as subparcelas formadas pelas doses de calcário dolomítico (sem aplicação de calcário; 1,8; 3,6 e 5,4 t ha-1). As cultivares de feijão tiveram desenvolvimento produtivo distinto em função das doses de calcário aplicadas superficialmente no sistema plantio direto. A cultivar Campeão 2 foi mais produtiva nos dois anos de experimentação e a cultivar IAPAR 81 respondeu linearmente ao aumento das doses de calcário no segundo ano agrícola. Houve efeito da interação dos fatores cultivares x doses de calcário para o tempo de cozimento, nos dois anos de experimentação, com aumento dessa característica na cultivar IAPAR 81, em função do incremento das doses de calcário. Comportamento semelhante foi obtido pela cultivar Pérola, porém apenas no segundo ano agrícola. O teor de proteína bruta variou de 210 a 230 g kg-1 e a relação de hidratação ficou próxima de 2,0 não sendo influenciados pelas doses de calcário e pelas cultivares de feijão.

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Foram realizados três experimentos procurando avaliar a utilização de dois marcadores internos (FDN e FDA indigestíveis), obtidos por meio da incubação in vitro e in situ, e de um marcador externo (óxido crômico) para estimativa da produção fecal e do fluxo da digesta duodenal em bovinos. Para análise dos dados, adotou-se o delineamento em blocos inteiramente ao acaso, sendo os tratamentos distribuídos em esquema fatorial, constituindo cinco marcadores e três volumosos. Os teores de FDN e FDA indigestíveis mostraram-se variáveis para cada volumosos, independentemente da metodologia utilizada (in vitro ou in situ), indicando que possivelmente a incubação por 144 horas não reproduz a fração indigestível total. As estimativas de produção fecal e de fluxo da digesta duodenal, obtidas por intermédio dos marcadores avaliados, apresentaram comportamento bastante diferenciado de acordo com cada volumoso. Os marcadores internos (FDN e FDA indigestíveis) podem ser utilizados como preditores dos parâmetros avaliados, desde que alguns cuidados sejam tomados na sua determinação.

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Trypanosoma cruzi proteins from epimastigote membranes, herein referred as antigens, have been used for the construction of an amperometric immunosensor for serological diagnosis of Chagas' disease. The proteins used had a molecular mass ranging from 30 to 100 kDa. The gold electrode was treated with cysteamine and glutaraldehyde prior to antigen immobilization. Antibodies present in the serum of patients with Chagas' disease were captured by the immobilized antigens and the affinity interaction was monitored by chronoamperometry at a potential of -400 mV (versus Ag pseudo-reference electrode) using peroxidase-labeled IgG conjugate and hydrogen peroxide, iodide substrate. The incubation time to allow maximum antigen-antibody and antibody-peroxidase-labeled IgG interactions was 20 min with a reactivity threshold at -0.104 mu A. (c) 2004 Elsevier B.V. All rights reserved.

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The binding selectivity of the M(phen)(edda) (M = Cu, Co, Ni, Zn; phen = 1,10-phenanthroline, edda = ethylenediaminediacetic acid) complexes towards ds(CG)(6), ds(AT)(6) and ds(CGCGAATTCGCG) B-form oligonucleotide duplexes were studied by CD spectroscopy and molecular modeling. The binding mode is intercalation and there is selectivity towards AT-sequence and stacking preference for A/A parallel or diagonal adjacent base steps in their intercalation. The nucleolytic properties of these complexes were investigated and the factors affecting the extent of cleavage were determined to be: concentration of complex, the nature of metal(11) ion, type of buffer, pH of buffer, incubation time, incubation temperature, and the presence of hydrogen peroxide or ascorbic acid as exogenous reagents. The fluorescence property of these complexes and its origin were also investigated. The crystal structure of the Zn(phen)(edda) complex is reported in which the zinc atom displays a distorted trans-N4O2 octahedral geometry; the crystal packing features double layers of complex molecules held together by extensive hydrogen bonding that inter-digitate with adjacent double layers via pi...pi interactions between 1,10-phenanthroline residues. The structure is compared with that of the recently described copper(II) analogue and, with the latter, included in molecular modeling. (C) 2008 Elsevier B.V. All rights reserved.

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Salmonella enterica serovar Typhi is the causative agent of typhoid fever in humans, and the use of antibiotics is essential for controlling this infection; however, the excessive use of antibiotics may select resistant strains. Propolis is a honeybee product and its antimicrobial activity has been intensively investigated. Thus, the objective of this study was to investigate a possible synergism between propolis (collected in Brazil and Bulgaria) and antibiotics acting on the ribosome (chloramphenicol, tetracycline and neomycin) against Salmonella Typhi in vitro. The synergism was investigated by using 1/2 and 1/4 of the minimum inhibitory concentration for propolis and these antimicrobial agents, evaluating the number of viable cells according to the incubation time. Brazilian propolis showed a bacteriostatic action against S. Typhi, while Bulgarian propolis showed a bactericidal activity and a synergistic effect with the three antibiotics. Variations in the biological assays might be due to the differences in their chemical compositions. Based on the results, one may conclude that Bulgarian propolis showed an important antibacterial action, as well as a synergistic effect with antibiotics acting on the ribosome, which points out a possible therapeutic strategy evaluating the use of propolis preparations for the treatment of Salmonella Typhi infection.

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Propolis is a honeybee product that has been used since ancient times because of its therapeutic effects. It can be used in the development of alternative therapies for the treatment of many diseases, and because propolis shows antibacterial action, this work was carried out in order to investigate a possible synergism between propolis and antibiotics acting on DNA (ciprofloxacin and norfloxacin) and on the metabolism (cotrimoxazole) against Salmonella Typhi. Propolis samples collected in Brazil and Bulgaria were compared in these assays, and the synergism was investigated by using 1/2 and 1/4 of the minimal inhibitory concentration for propolis and antibiotics, evaluating the number of viable cells according to the incubation time. Brazilian and Bulgarian propolis showed antibacterial activity, but no synergistic effects with the three tested antibiotics were seen. Previous works by our laboratory have revealed that propolis has synergistic effects with antibiotics, acting on the bacterial wall and ribosome, but it does not seem to interact with antibiotics acting on DNA or folic acid, and only a bacteriostatic action was seen in these assay conditions.

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Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

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We review the information currently available on the reproduction of the Amphisbaenia and provide original data on the reproductive biology of four Neotropical species: Amphisbaena alba; A. mertensi; Cercolophia roberti, and Leposternon infraorbitale. In total, we compiled data for 22 species: 17 Amphisbaenidae, 1 Rhineuridae, 3 Bipedidae, and 1 Trogonophidae. The majority of the species were oviparous with the exceptions of Loveridgea ionidesii, Monopeltis anchietae, M. capensis, and Trogonophis wiegmanni. Viviparity was interpreted as a derived trait that evolved independently for at least 3 times within the Amphisbaenia. In most species, reproduction is synchronized with the hot and rainy season and seems to vary with latitude. Although Amphisbaenia eggs have been found in ant nests, it remains disputable whether this is an obligatory or even a preferable location for egg-laying. Incubation time in A. mertensii lasts 59 days and this is the first report encompassing egg-laying to hatching for any Amphisbaenia species. Nonetheless, a two months incubation period seems to be the common rule for oviparous Amphisbaenia. The general pattern of reproductive output in Amphisbaenia is characterized by a low number of eggs/embryos per clutch whose individual size is comparatively large in relation to adult body size. Eggs are markedly elongated on the long axis and arranged in-line within the abdominal cavity possibly to prevent/diminish biomechanic drawbacks of egg bearing. Hatchlings of A. mertensi possess an egg-tooth implanted at the upper jaw, exhibit positive geotropism, and display defensive behaviors known to be present in adults. Our review shows that our current knowledge of Amphisbaenia reproduction is fragmentary, often based on the examination of small samples, and heavily dependent on the publication of anedoctal observations. Future publications on this subject are encouraged.

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

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A new lipase from seeds of Pachira aquatica was purified to homogeneity by SDS-PAGE obtaining an enzyme with a molecular weight of approximately 55 kDa. The purified lipase exhibited maximum activity at 40 degrees C and pH 8.0, for an incubation time of 90 min. Concerning temperature stability, at the range from 4 to 50 degrees C, it retained approximately 47% of its original activity for 3 h. The enzyme activity increased in the presence of Ca(++) and Mg(++), but was inhibited by Hg(++), Mn(++), Zn(++), Al(+++) and various oxidizing and reducing agents. The lipase was highly stable in the presence of organic solvents, and its activity was stimulated by methanol. The values of K(m) and V(max) were 1.65 mM and 37.3 mu mol mL(-1) min(-1), respectively, using p-nitrophenylacetate as substrate. The enzyme showed preference for esters of long-chain fatty acids, but demonstrated significant activity against a wide range of substrates.

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Fractional factorial design and factorial with center point design were applied to the development of an amperometric biosensor for the detection of the hepatitis C virus. Biomolecules were immobilized by adsorption on graphite electrodes modified with siloxane-poly(propyleneoxide) hybrid matrix prepared using the sol-gel method. Several parameters were optimized, such as the streptavidin concentration at 0.01 mg mL(-1) and 1.0% bovine serum albumin, the incubation time of the electrodes in the complementary DNA solution for 30 minutes and a 1: 1500 dilution of the avidin-peroxidase conjugate, among others. The application of chemometric studies has been efficient, since the best conditions have been established with a restricted number of experiments, indicating the influence of different factors on the system.

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LASSBio-767 [(-)-3-O-acetyl-spectaline] and LASSBio-822 [(-)-3-O-tert-Boc-spectaline] were recently described as cholinesterase inhibitors derived from the natural piperidine alkaloid (-)-spectaline, obtained from the flowers of Senna spectabilis (Fabaccae). We investigated their mechanism of inhibition of acetylcholinesterase and their efficacy in reversing scopolamine-induced amnesia. Competition assays with the substrate acetylthiocholine showed a concentration-dependent reduction in rat brain cholinesterase V-max without changes in apparent K-m. The kinetic data for LASSBio-767 and LASSBio-822 were best fit by a model of simple linear noncompetitive inhibition with K-i of 6.1 mu M and 7.5 mu M, respectively. A dilution assay showed a fast and complete reversal of inhibition, independent of incubation time. Simulated docking of the compounds into the catalytic gorge of Torpedo acetylcholinesterase showed interactions with the peripheral anionic site, but not with the catalytic triad. Anti-amnestic effects in mice were assessed in a step-down passive avoidance test and in the Morris water maze 30 min after injection of scopolamine (1 mg/kg i.p.). Saline, LASSBio-767, or LASSBio-822 was administered 15 min before scopolamine. Both compounds reversed the scopolamine-induced reduction in step-down latency at 0.1 mg/kg i.p. LASSBio-767 reversed scopolamine-induced changes in water maze escape latency at 1 mg/kg i.p. or p.o., while its cholinergic side effects were absent or mild up to 30 mg/kg i.p. (LD50 above 100 mg/kg i.p.). Thus, the (-)-spectaline derivatives are potent cholinergic agents in vivo, with a unique profile combining noncompetitive cholinesterase inhibition and CNS selectivity, with few peripheral side effects. (C) 2007 Elsevier B.V. All rights reserved.

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Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

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Este estudo teve a finalidade de fornecer dados morfológicos de ovos de D. renale e do desenvolvimento de larvas de primeiro estádio em ovos mantidos em diferentes temperaturas. Os ovos foram obtidos por centrífugação da urina de cães parasitados e colocados em placas de Petri em estufa BOD, durante 90 dias. O experimento consistiu de três tratamentos (GI - 15 ºC, GII - 20 ºC e GIII - 26 ºC) com cinco repetições cada. Os ovos apresentaram tamanho médio de 67,23 x 42,78 µm, e o tempo médio de incubação foi inversamente proporcional à temperatura de incubação e as larvas apresentaram motilidade por aproximadamente uma semana após sua formação.