978 resultados para TRYPSIN INHIBITORY-ACTIVITY
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Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)
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Lipases from oilseeds have a great potential for commercial exploration as industrial enzymes. Lipases are used mixed with surfactants in cleaning and other formulated products, and accordingly, both components must be compatible with each other. This work presents the results of the effects of anionic, cationic and nonionic surfactants, polyethylene glycol and urea on the activity and stability of a lipase extracted of oilseeds from Pachira aquatica. The enzyme was purified and the spectrophotometric assays were done using p-nitrophenyl acetate (p-NPA) as substrate pH 7.5 and 25 degrees C. The activity was significantly enhanced by the cationic surfactant CTAB. Bile salts increased the lipase activity in the tested concentration range, whereas anionic and nonionic surfactants showed an inhibitory effect. Aqueous solutions of PEG activated the lipase and maximum activation (161%) occurred in PEG 12,000. This effect on lipase that can be due to exposition of some hydrophobic residues located in the vicinity of the active site or aggregation.
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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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Mycobacterium tuberculosis is responsible for over 8 million cases of tuberculosis (TB) annually. Natural products may play important roles in the chemotherapy of TB. The immunological activity of Davilla elliptica chloroform extract (DECE) was evaluated in vitro by the determination of hydrogen peroxide (H2O2), nitric oxide (NO), and tumor necrosis factor-alpha (TNF-alpha) release in peritoneal macrophages cultures. DECE was also tested for its antimycobacterial activity against M. tuberculosis using the microplate alamar blue assay. DECE (50, 150, 250 µg/ml) stimulated the production of H2O2 (from 1,79 ± 0,23 to 7,27 ± 2,54; 15,02 ± 2,86; 20,5 ± 2,1 nmols) (means ± SD), NO (from 2,64 ± 1,02 to 25,59 ± 2,29; 26,68 ± 2,41; 29,45 ± 5,87 µmols) (means ± SD) and TNF-alpha (from 2,44 ± 1,46 to 30,37 ± 8,13; 38,68 ± 1,59; 41,6 ± 0,90 units/ml) (means ± SD) in a dose-dependent manner and also showed a promising antimycobacterial activity with a minimum inhibitory concentration of 62,5 µg/ml. This plant may have therapeutic potential in the immunological and microbiological control of TB.
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Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
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Com o objetivo de avaliar a atividade de células natural killer na paracoccidioidomicose experimental do hamster, 80 hamsters foram infectados por via intratesticular com Paracoccidioides brasiliensis e sacrificados após 24h, 48h, 96h, 1, 2, 4, 8 e 11 semanas de infecção. Como controle foram avaliados 40 hamsters normais, não infectados. Os animais foram submetidos ao estudo da atividade citotóxica de células NK pela técnica de single-cell assay e da resposta imune humoral pelas técnicas de imunodifusão dupla e Elisa. A produção do fator inibidor da migração de macrófagos em presença de Phytohemaglutinina e antígeno de P. brasiliensis e a histopatologia das lesões foram estudadas após 1,4, 8e 11 semanas de infecção. Os animais infectados, quando comparados aos controles, apresentaram níveis de atividade NK significativamente elevados durante as 4 primeiras semanas de infecção, havendo diminuição dessa atividade a partir da 8ª semana. Foi observada correlação inversa entre atividade NK e níveis de anticorpos específicos e, associação entre diminuição da atividade NK, depressão de resposta imune celular e aumento da extensão das lesões histopatológicas. Os resultados sugerem que após ativação inicial, as células NK são incapazes de controlar a disseminação do fungo pelos tecidos. A depressão da atividade NK na fase tardia da infecção parece estar relacionada com os distúrbios imunorregulatórios associados à paracoccidioidomicose.
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Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
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Gelatin microparticles containing propolis ethanolic extractive solution were prepared by spray-drying technique. Particle,, with regular morphology, mean diameter ranging of 2.27 mu m to 2.48 mu m, and good entrapment efficiency for propolis were obtained. The in vitro antimicrobial activity of microparticles was evaluated against microorganisms of oral importance (Enterococcus faecalis, Streptococcus salivarius, Streptococcus sanguinis, Streptococcus mitis, Streptococcus mutans, Streptococcus sobrinus, Candida albicans, and Lactobacillus casei). The utilized techniques were diffusion in agar and determination of minimum inhibitory concentration. The choice of the method to evaluate the antimicrobial activity of microparticles showed be very important. The microparticles displayed activity against all tested strains of similar way to the propolis, showing greater activity against the strains of E. salivarius, S. sanguinis, S. mitis, and C albicans.
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In this study, we compared the anti-leishmanial activity of three crotalic venoms (Crotalus durissus terrificus-Cdt, Crotalus durissus cascavella-Cdca, and Crotalus durissus collilineatus-Cdcol). Different concentrations of each venom incubated with Leishmania (Leishmania) amazonensis promastigotes were used. Cdt venom exhibited a higher anti-leishmanial activity (Inhibitory concentration-IC50-value of 4.70 +/- 1.72 mu g/ml) in comparison with that of Cdca venom (IC50 value of 9.41 +/- 1.21 mu g/ml), while Cdcol venom increased parasite numbers in 50% at a concentration of 44.30 +/- 2.18 mu g/ml. In addition, this venom showed a low anti-leishmanial activity in higher concentrations (IC50 value of 281.00 +/- 9.50 mu g/ml). The main fractions of Cdca venom were isolated and assayed under similar conditions used for assessing crude venom. The most active fractions were gyroxin and crotamine that had IC50 values of 3.80 +/- 0.52 mu g/ml and 19.95 +/- 4.21 mu g/ml, respectively. Convulxin also inhibited parasite growth rate, although this effect was not dose-dependent. Crotoxin was the least effective fraction with an IC50 value of 99.80 +/- 2.21 mu g/ml. None of the protein fractions presented cytotoxic effects against J774 cells in culture. In vivo assays using BALB/c mice revealed that crotoxin and crotamine were the main toxic fractions. In conclusion, C. durissus cascavella venom has three main fractions with anti-leishmanial activity. These results open new possibilities to find proteins that might be used as possible agents against cutaneous leishmaniasis.
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The leukotoxic activity of 31 Actinobacillus actinomycetemcomitans isolates from Brazilian periodontal patients [nine from Localized Juvenile Periodontitis (LJP) patients, 22 from patients with AIDS-associated Necrotizing Ulcerative Periodontitis (AIDS/NUP)], and from the reference strain A. actinomycetemcomitans ATCC43718, were analyses for their cytotoxicity on human monocytes. A cytotoxicity inhibitory assay of the isolate P35 and the reference strain ATCC 43718 with sera from ten LJP patients and ten healthy subjects was also performed and leukotoxin reactivity was evaluated with serum from rabbits immune to leukotoxin from A. actinomycetemcomitans ATCC 43718. The cytotoxicity results were not statistically different among groups of A. actinomycetemcomitans isolates from LJP and AIDS/NUP patients, but the individual analysis of each isolate showed two isolates (P24 and P35) from LJP patients with high leukotoxic activity (P<0.05). Also, a high leucotoxic inhibitory effect with LJP patients' sera compared with healthy subjects with sonic extract from isolate P35 (P<0.05) and the reactivity of rabbit antiserum to leukotoxin were observed. Both leukotoxic and non-leukotoxic activity is more frequent in PJL than AIDS/NUP patients. Even though A. actinomycetemcomitans exhibits leukotoxic activity, there is an immune response to the leucotoxin in LJP patients. (C) 2000 Academic Press.
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Rat osseous plate alkaline phosphatase is a metalloenzyme with two binding sites for Zn2+ (sites I and III) and one for Mg2+ (site II). This enzyme is stimulated synergistically by Zn2+ and Mg2+ (Ciancaglini et al., 1992) and also by Mn2+ (Leone et al., 1995) and Co2+ (Ciancaglini et al., 1995). This study was aimed to investigate the modulation of enzyme activity by Ca2+. In the absence of Zn2+ and Mg2+, Ca2+ had no effects on the activity of Chelex-treated, Polidocanol-solubilized enzyme. However, in the presence of 10 mu M MgCl2, increasing concentration of Ca2+ were inhibitory, suggesting the displacement of Mg2+ from the magnesium-reconstituted enzyme. For calcium-reconstituted enzyme, Zn2+ concentrations Zip to 0.1 mu M were stimulatory, increasing specific activity from 130 U/mg to about 240 U/mg with a K-0.5 = 8.5 nM. Above 0.1 mu M Zn2+ exerted a strong inhibitory effect and concentrations of Ca2+ up to I mM were not enough to counteract this inhibition, indicating that Ca2+ was easily displaced by Zn2+. At fixed concentrations of Ca2+, increasing concentrations of Mg2+ increased the enzyme specific activity from 472 U/mg to about 547 U/mg, but K-0.5 values were significantly affected (from 4.4 mu M to 38.0 mu M). The synergistic effects observed for the activity of Ca2+ plus magnesium-reconstituted enzyme, suggested that these two ions bind to the different sites. A model to explain the effect of Ca2+ on the activity of the enzyme is presented. (C) 1997 Elsevier B.V.
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This report compares the in vitro activity of three cephalosporins (cephalothin, cefoxitin and ceftriaxone) against 57 Staphylococcus aureus strains isolated from cows with clinical mastitis on the basis of the minimal inhibitory (MIC) and minimal bactericidal concentrations (MBC). The majority of the S aureus strains showed resistance to cefoxitin and ceftriaxone and sensitivity to cephalothin. The highest MICs and MBCs were found for cefoxitin and ceftriaxone. Antimicrobial tolerance (MBC/MIC greater-than-or-equal-to 32:1) was observed in relation to cephalothin and ceftriaxone. The data suggest that these cephalosporins may not be effective for the treatment of staphylococcal bovine mastitis. The precise definition of their antimicrobial efficacies requires more detailed in vitro and in vivo studies.