Structure-activity relationships of hypervalent organochalcogenanes as inhibitors of cysteine cathepsins V and S

A new series of organotelluranes were synthesized and investigated, and the structure-activity relationships in cysteine proteases inhibition were determinated. It was possible to identify the relevance of structural components linked to the reactivity of these compounds as inhibitors. For example, dibromo-organotelluranes showed to be more reactive than dichloro-organotelluranes towards cysteine cathepsins V and S. Besides, no remarkable enantio-selectivity was verified. In general the achiral organotelluranes were more reactive than the chiral congeners against cysteine cathepsins V and S. A reactivity order for organochalcogenanes and cysteine cathepsins was proposed after the comparison of the inhibitory potencies of organotelluranes with the related organoselenanes. (C) 2011 Elsevier Ltd. All rights reserved.

Anti-Helicobacter pylori activity and immunostimulatory effect of extracts from Byrsonima crassa Nied. (Malpighiaceae)

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

Selective activity of butyrylcholinesterase in serum by a chemiluminescent assay

In a previous study, we showed that purified commercial esterase activity can be detected in a chemiluminescent assay based on the hydrolysis of 2-methyl-1-propenylbenzoate (MPB) to 2-methyl-1-propenol, which is subsequently oxidized by the horseradish peroxidase (HRP)-H2O2 system. The purpose of this study was to verify the applicability of this assay to human serum. The existence of an esterase activity capable of hydrolysing MPB is indicated by the fact that the MPB-scruin-HRP-H2O2 System consumes oxygen and emits light. Both signals were abolished by prior serum heat inactivation and were preserved when serum was stored at less than or equal to4 degreesC. Addition of aliesterase inhibitors, such as fluoride ion and trichlorfon or the cholinesterase inhibitor eserine, totally prevents light emission. The butyrylcholinesterase-specific substrate benzoylcholine causes a delay in both O-2 uptake and light emission, while the specific acetylcholinesterase substrate, acetyl-beta -methylcholine, had practically no effect. Purified butyrylcholinesterase, but not acetylcholinesterase, triggered light emission. The finding that butyryleholinesterase is responsible for the hydrolysis of MPB in serum should serve as the basis for the development of a specific chemiluminescent assay for this enzyme. Copyright (C) 2001 John Wiley & Sons, Ltd.

Ionically bound peroxidase from peach fruit

Soluble, ionically bound peroxidase (POD) and polyphenoloxidase (PPO) were extracted from the pulp of peach fruit during ripening at 20degreesC Ionically bound form was purified 6.1 -fold by DEAE-cellulose and Sephadex G-100 chromatography. The purified enzyme showed only one peak of activity on Sephadex G-100 and PAGE revealed that the enzyme was purified by the procedures adopted. The purified enzyme showed a molecular weight of 29000 Da, maximum activity at pH 5.0 and at 40degreesC the calculated apparent activation energy (Ea) for the reaction was 10.04 kcal/mol. The enzyme was heat-labile in the temperature range of 60 to 75degreesC with a fast inactivation at 75degreesC Measurement of residual activity showed a stabilizing effect of sucrose at various temperature/sugar concentrations (0, 10, 20 %, w/w), with an activation energy (Ea) for inactivation increasing with sucrose concentration from 0 to 20% (w/w). The Km and V-max values were 9.35 and 15.38 mM for O-dianisidine and H2O2, respectively. The bound enzyme was inhibited competitively by (.)ferulic, caffeic and protocatechuic acids with different values of Ki,. L-cysteine, p-coumaric and indolacetic acid and Fe++ also inhibited the enzyme but at a lower grade. N-ethylmaleimide and p-CMB were not effective to inhibit the enzyme demonstrating the non-essentiality of SH groups.

Peroxidase and phenylalanine ammonia-lyase activities, phenolic acid contents, and allelochemicals-inhibited root growth of soybean

The influence of the allelochemicals ferulic (FA) and vanillic (VA) acids on peroxidase (POD, EC 1.11.1.7) and phenylalanine ammonia-lyase (PAL, EC 4.3.1.5) activities and their relationships with phenolic acid (PhAs) contents and root growth of soybean (Glycine max (L.) Merr.) were examined. Three-day-old seedlings were cultivated in nutrient solution containing FA or VA (0.1 to 1 mM) for 48 h. Both compounds (at 0.5 and 1 mM) decreased root length (RL), fresh weight (FW) and dry weight (DW) and increased PhAs contents. At 0.5 and 1 mM, FA increased soluble POD activity (18% and 47%, respectively) and cell wall (CW)-bound POD activity (61% and 34%), while VA increased soluble POD activity (33% and 17%) but did not affect CW-bound POD activity. At I mM, FA increased (82%) while VA reduced (32%) PAL activities. The results are discussed on the basis of the role of these compounds on phenylpropanoid metabolism and root growth and suggest that the effects caused on POD and PAL activities are some of the many mechanisms by which allelochemicals influence plant growth.

Simultaneous effects of ferulic and vanillic acids on peroxidase and phenylalanine ammonia-lyase in soybean (Glycine max) roots

Simultaneous effects of ferulic (FA) and vanillic (VA) acids on peroxidase (POD, EC 1.11.1.7) and phenylalanine ammonia-lyase (PAL, EC 4.3.1.5) activities on soybean (Glycine max (L.) MERR.) root growth were analyzed. Three-day-old seedlings were cultivated in nutrient solution containing FA or VA (0.5 mM; 1.0 mM or equimolar mixtures) for 48 h. Acting alone, both compounds (at 0.5 or 1.0 mM) decreased root length (RL), fresh weight (FW), dry weight (DW) and increased soluble POD and cell wall (CW)-bound POD activities. At 1.0 mM, FA increased (but VA decreased) the PAL activity. Acting simultaneously, the effects of the allelochemical interaction were lower than the sum of the effects of each compound tested separately, suggesting antagonism.

Efeito do cobre na atividade da enzima pirogalol peroxidase em plantas de Myriophyllum aquaticum cultivadas em solução nutritiva

Myriophyllum aquaticum é uma planta perene, herbácea, que pode se desenvolver totalmente submersa ou com a porção terminal dos ramos acima da superfície da água. É também considerada uma planta daninha que possui elevado potencial de colonização, o qual, dependendo da densidade populacional, pode causar aumento no teor de matéria orgânica e redução de oxigênio na água, comprometendo a qualidade da água e seus usos múltiplos. O objetivo do presente trabalho foi verificar a influência do cobre na atividade da pirogalol peroxidase de plantas de M. aquaticum submetidas à solução nutritiva contendo concentrações de cobre de 1,2; 11,2; 21,2; 31,2; e 41,2 µg L-1. O experimento foi conduzido em um delineamento experimental inteiramente casualizado, com quatro repetições e cinco tratamentos, aos quais as plantas foram submetidas durante 21 dias. Aos 81 dias após a instalação das mudas em solução nutritiva contendo os diferentes níveis de cobre, as folhas foram colhidas a partir do ápice da planta até o final do ramo, que não estavam em contato com a solução. Esse material fresco foi envolvido por plástico transparente e papel-alumínio e, a seguir, congelado em nitrogênio líquido, sendo armazenado em freezer a -20 ºC até o momento da determinação da atividade da enzima pirogalol peroxidase. A atividade da enzima foi progressiva com o aumento das doses de cobre. As plantas cultivadas com 40 µg L-1 de Cu2+ após três semanas, com base em avaliação visual, apresentaram redução no desenvolvimento.

Pro-oxidant activity of apocynin radical

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

Antioxidant activity of indigo and its preventive effect against ethanol-induced DNA damage in rat gastric mucosa

Ethanol-induced oxidative damage is commonly associated with the generation of reactive oxygen molecules, leading to oxidative stress. Considering that antioxidant activity is an important mechanism of action involved in cytoprotection, the aim of this work was to evaluate the antioxidant properties of the alkaloid indigo (1) (2 mg/kg, p. o.), obtained from the leaves of Indigofera truxillensis Kunth (Fabaceae), on rat gastric mucosa submitted to ethanol-induced (100%, 1 mL, p.o.) gastric ulcer. Enzymatic assays and DNA fragmentation analysis were performed. When ethanol was administered to the control group, the sulfhydryl content (SH) and the glutathione peroxidase (GPx) activity decreased by 41% and 50%, respectively; in contrast, superoxide dismutase (SOD) and glutathione reductase (GR) activities increased by 56% and 67%, respectively. Additionally, myeloperoxidase (MPO) activity, a marker for free radical generation caused by polymorphonuclear neutrophil (PMN) tissue infiltration, also increased 4.5-fold after ethanol treatment. Rat gastric mucosa exposed to ethanol showed DNA fragmentation. Indigo alkaloid pretreatment protected rats from ethanol-induced gastric lesions. This effect was determined by the ulcerative lesion area (ULA), indicating an inhibition of around 80% at 2 mg/kg. This alkaloid also diminished GPx activity, which was higher than that observed with ethanol alone. However, this effect was counterbalanced by increased GR activity. Indigo was unable to restore alterations in SOD activity promoted by ethanol. After indigo pretreatment, SH levels and MPO activity remained normal and gastric mucosa DNA damage caused by ethanol was also partially prevented by indigo. These results suggest that the gastroprotective mechanisms of indigo include non-enzymatic antioxidant effects and the inhibition of PMN infiltration which, in combination, partially protect the gastric mucosa against ethanol-induced DNA damage.

Tobacco Smoke Induces Ventricular Remodeling Associated with an Increase in NADPH Oxidase Activity

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Influence of stationary and bioreactor cultivation on Lentinula edodes (Berk) pegler lignocellulolitic activity

This work aimed to study the stationary and periodically mixed culture of L. edodes to the production of lignocellulolitic enzymes activity. LE 95/17, LE 96/22 and Leax strains were incubated in 25 g of eucalyptus sawdust substrate in Erlenmeyer flasks in stationary culture at 25 degrees C and in a bioreactor with four complete rotations daily at 25 degrees C and 3% CO2. The samples were collected at 8, 11, 14, 17 and 20 days after the incubation. Oxidative and hydrolytic enzymes analyses were performed. Lignin peroxidase enzyme was not found in the lignolytic systernfor LE 95/17, LE 96/22 and Leax strains in the different incubation methods. The use of bioreactor could be a practicable system to induce the laccase activity for L22 and Leax and MnP activity for L17 and L22. The activity of the hydrolytic enzymes was higher in the stationary system in comparison to periodically mixed system in the bioreactor.

PRODUCTION of FREE RADICALS and CATALASE ACTIVITY DURING ACUTE EXERCISE TRAINING IN YOUNG MEN

Reactive oxygen species (ROS) are constantly produced by cells that promote cellular oxidative damage and are neutralized by an antioxidant system including superoxide dismutase, glutathione, peroxidase and catalase. Male volunteers were exercised for 20 minutes, three days (60, 70 and 80% of maximum heart rate). Catalase activity and plasma malondialdehyde concentration were measured. The mean age of the volunteers was 25 +/- 7 years, with body mass index 2 of 24.03 +/- 4.32 kg/m(2). Acute exercise training produced an increase of malondialdehyde concentration that was exercise intensity-dependent in young volunteers. However, catalase activity shows a great variability at baseline and the percentual of reduction was exercise intensity-independent in this particular population. Therefore, our study shows that acute cycling exercise promotes an increase of oxidative stress that was exercise intensity-dependent in young volunteers. Furthermore, the antioxidant system measured by catalase activity was effective to counterbalance the ROS production showing a saturation behavior at an intensity of 70% of maximum heart rate.

Ligninolytic activity from newly isolated basidiomycete strains and effect of these enzymes on the azo dye orange II decolourisation

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

Physiological quality and enzymatic activity of crambe seeds after the accelerated aging test

O crambe é uma cultura promissora para produção de biodiesel, principalmente pelo alto conteúdo de óleo de suas sementes. No entanto, não há metodologias estabelecidas para avaliar a qualidade fisiológica das sementes desta espécie e lotes não podem ser comparados, especialmente por testes de vigor, como o de envelhecimento acelerado. O objetivo da presente pesquisa foi avaliar o efeito da alta temperatura e do período de exposição durante o teste de envelhecimento acelerado na qualidade fisiológica e atividade enzimática de sementes de crambe. Dois lotes de sementes de crambe, cultivar Brilhante, foram analisados por meio dos testes de teor de água, massa de 1.000 sementes, germinação, primeira contagem, condutividade elétrica, atividade enzimática (peroxidase e superóxido dismutase) e comprimento de plântulas. As avaliações foram conduzidas antes e após o envelhecimento acelerado, que foram testadas diferentes temperaturas (38, 40 e 42ºC) e períodos de exposição (24, 48 e 72h). O delineamento experimental foi o inteiramente casualizado com quatro repetições. Os dados foram submetidos à análise de variância e as médias foram comparadas pelo teste de Tukey (p < 0.05). O teste de Dunnet (p < 0.05) foi utilizado para comparar os valores da testemunha (antes do envelhecimento acelerado) com cada valor médio individualmente. O teste de correlação linear simples também foi aplicado. Conclui-se que a interação temperatura x período de exposição afeta a qualidade fisiológica das sementes e a atividade enzimática e que as melhores condições durante o teste de envelhecimento acelerado são dependentes do genótipo.

Callus cell culture of Pothomorphe umbellata (L.) under stress condition leads to high content of peroxidase enzyme

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