Electrochemical characterization of the paste carbon modified electrode with KSr2Ni0.75Nb4.25O15-delta solid in catalytic oxidation of the dipyrone

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

Beneficial effects of diclofenac therapy on serum lipids, oxidized low-density lipoprotein and antioxidant defenses in rats

To study the effects of diclofenac, a nonselective nonsteroidal anti-inflammatory drug (NSAID), on lipid profile, oxidized low-density-lipoprotein (Ox-LDL), serum antioxidant defenses and markers of oxidative stress, male Wistar rats were divided into two groups (n = 10): (C) receiving intramuscularly a single daily dose of saline (NaCl 0.9%), and (AI) receiving intramuscularly a single daily dose of 10 mg/kg diclofenac sodium (C14H10C12NNaO2). After 28 days diclofenac-treated rats had lower Ox-LDL, apoprotein B (apo-B), apo-B/LDL-cholesterol and lipid hydroperoxide than C. Total antioxidant substances and superoxide dismutase were increased in diclofenac-treated rats, while no significant changes were observed in catalase, glutathione peroxidase and nitric oxide. A perincubation test done to examine the possibility of mechanism-based activation showed that diclofenac had no effect on maximal superoxide dismutase velocity, but significantly reduced the Michaelis-Menten (K-M) constant, indicating that diclofenac induced SOD activation increasing substrate linkage affinity to the enzyme-catalytic site. In conclusion, diclofenac had beneficial effects decreasing Ox-LDL and improving antioxidant defense. It appears that the application of this agent may be feasible and beneficial for serum antioxidant protection, which certainly would bring new insights on dyslipidemia control. (C) 2008 Elsevier B.V. All rights reserved.

Production, purification and characterization of a minor form of xylanase from Aspergillus versicolor

A strain of Aspergillus versicolor produces a xylanolytic complex containing two components, the minor component being designated xylanase II. The highest production of xylanase II was observed in cultures grown for 5 days in 1% wheat bran as carbon source, at pH 6.5. Xylanase II was purified 28-fold by DEAE-Sephadex and HPLC GF-5 10 gel filtration. Xylanase II was a monomeric glycoprotein, exhibiting a molecular mass of 32 kDa with 14.1% of carbohydrate content. Optimal pH and temperature values for the enzyme activity were about 6.0-7.0 and 55 degreesC, respectively. Xylanase II thermoinactivation at 50degreesC showed a biphasic curve. The ions Hg2+, Cu2+ and the detergent SDS were strong inhibitors, while Mn2+ ions and dithiothreitol were stimulators of the enzyme activity. The enzyme was specific for xylans, showing higher specific activity on birchwood xylan. The Michaelis-Menten constant (K-m) for birchwood xylan was estimated to be 2.3 mg ml(-1) while maximal velocity (V-max) was 233.1 mumol mg(-1) min(-1) of protein. The hydrolysis of oat spell xylan released only xylooligosaccharides. Published by Elsevier Ltd.

Purification and properties of an acid beta-xylosidase from Penicillium sclerotiorum

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Estudo da oxidação dos sulfetos sintéticos molibdenita (MoS2) e covelita (CuS) por Acidithiobacillus ferrooxidans via respirometria celular

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

ALLOSTERIC MODULATION BY ATP, CALCIUM AND MAGNESIUM-IONS OF RAT OSSEOUS PLATE ALKALINE-PHOSPHATASE

Alkaline phosphatase from rat osseous plate is allosterically modulated by ATP, calcium and magnesium at pH 7.5. At pH 9.4, the hydrolysis of ATP and PNPP follows Michaelis-Menten kinetics with K0.5 values of 154 muM and 42 muM, respectively. However, at pH 7.5 both substrates exhibit more complex saturation curves, while only ATP exhibited site-site interactions. Ca2+-ATP and Mg2+-ATP were effective substrates for the enzyme, while the specific activity of the enzyme for the hydrolysis of ATP at pH 7.5 was 800-900 U/mg and was independent of the ion species. ATP, but not PNPP, was hydrolyzed slowly in the absence of metal ions with a specific activity of 140 U/mg. These data demonstrate that in vitro and at pH 7.5 rat osseous plate alkaline phosphatase is an active calcium or magnesium-activated ATPase.

Isolation of a new L-amino acid oxidase from Crotalus durissus cascavella venom

A novel L-amino acid oxidase (LAO) (Casca LAO) from Crotalus durissus cascavella venom was purified to a high degree of molecular homogeneity using a combination of molecular exclusion and ion-exchange chromatography system. The purified monomer of LAO presented a molecular mass of 68 kDa and pI estimated in 5.43, which were determined by two-dimensional electrophoresis. The 71st N-terminal amino acid sequence of the LAO from Crotalus durissus cascavella presented a high amino acid sequence similarities with other LAOs from Colloselasma rhosostoma, Crotalus adamanteus, Agkistrodon h. blomhoffi, Agkistrodon h. halys and Trimeresurus stejnegeri. LAO displayed a Michaelis-Menten behavior with a kilometer of 46.7 mu M and an optimum pH for enzymatic activity of 6.5. Casca LAO induced a dose-dependent platelet aggregation, which was abolished by catalase and inhibited by indomethacin and aspirin. These results suggest that the production of H2O2 is involved in subsequent activation of inflammatory enzymes, such as thromboxane. Casca LAO also inhibited the bacterial Growth of Gram-negative (Xanthomonas axonopodis pv passiflorae) and Gram-positive (S. mutans) strains. Electron microscopy assessments of both bacterial strains suggest that the hydrogen peroxide produced by LAO induce bacterial membrane rupture and consequently loss of cytoplasmatic content. This LAO exhibited a high antileishmanic activity against the promastigote of Leishmania amazonensis in vitro, its activity was dependent on the production of hydrogen peroxide, and the 50% inhibitory concentration was estimated in 2.39 mu g/ml. (C) 2005 Elsevier Ltd. All rights reserved.

Triton X-100 solubilized bone matrix-induced alkaline phosphatase

1. 1. Solubilized and membrane-bound alkaline phosphatase showed Michaelis-Menten behavior in a wide range of different substrate concentrations. 2. 2. Membrane-bound alkaline phosphatase has a molecular weight of 130,000 and its minimum active configuration comprises two identical subunits of about 65,000. 3. 3. The two forms of the enzyme behave similarly with respect to NaCl, urea and guanidine HCl. 4. 4. Catalytic groups have pK values of about 8.5 and 9.7 for both membrane-bound and solubilized enzyme. © 1987.

Use of sorghum seed tissue as a biocatalyst in a stirred reactor for oxalic acid determination

The enzyme oxalate oxidase, E.C. 1.2.3.4 from Sorghum vulgare seeds (variety BR303) was used to develop a new sensor for oxalate determination without any purification. The sorghum seeds were conditioned in a 0.10 mol I-1 KCl solution. Then, these seeds were put in a stirring bar type enzymic reactor and coupled with an electrode for CO2. This device was introduced into a cell containing 10.0 ml of a 0.10 mol I-1 KCl solution saturated with oxygen. This sensor showed a linear response between 1.0 and 4.0 × 10-3 mol I-1 with a slope of 30 mV per decade of oxalate concentration at 25.0°C. The sensor was stable for one month or 200 determinations. The response time was about 60 s. The Michaelis-Menten constant determined for this enzyme was 1.5 × 10-3 mol I-1.

Estudo da dissolução oxidativa microbiológica de uma complexa amostra mineral contendo pirita (FeS2), pirrotita (Fe 1-XS) e molibdenita (MoS2)

This work aims to study the oxidation of a complex molybdenite mineral which contains pyrite and pyrrotite, by Acidithiobacillus ferrooxidans. This study was performed by respirometric essays and bioleaching in shake flasks. Respirometric essays yielded the kinetics of mineral oxidation. The findings showed that sulfide oxidation followed classical Michaelis-Menten kinetics. Bioleaching in shake flasks allowed evaluation of chemical and mineralogical changes resulting from sulfide oxidation. The results demonstrated that pyrrotite and pyrite were completely oxidized in A. ferrooxidans cultures whereas molybdenite was not consumed. These data indicated that molybdenite was the most recalcitrant sulfide in the sample.

Electrogravimetric Analysis by Quartz-Crystal Microbalance on the Consumption of the Neurotransmitter Acetylcholine by Acetylcholinesterase

Electrogravimetric analysis was performed on the consumption of the neurotransmitter Acetylcholine (ACh) by Acetylcholinesterase (AChE) in situ and in real time. Michaelis-Menten assumption was achieved by using an enzyme micro-reactor in which the total enzyme was anchored in a quartz crystal microbalance chip (QCM-chip) with a strategically engineered self-assembled monolayer (SAM) of alkanethiols, which can prevent diffusion-controlled or spatially restricted kinetics. The real-time frequency changes indicated the rate of the products formation from enzymatic reaction. The QCM-chip was tested showing that it could demonstrate AChE inhibition by physostigmine. © 2013 Copyright Taylor and Francis Group, LLC.

Inbreeding depression on production and reproduction traits of buffaloes from Brazil

The objective of this study was to evaluate the influence of inbreeding depression on traits of buffaloes from Brazil. Specifically, the traits studied were body weight at 205 and 365 days of age, average daily gain from birth to 205 days (ADG_205), average daily gain between 205 and 365 days (ADG205_365) in Mediterranean buffaloes, and milk yield, lactation length, age of first calving and calving intervals in Murrah buffaloes. Inbreeding effects on the traits were determined by fitting four regression models (linear, quadratic, exponential and Michaelis-Menten) about the errors generated by the animal model. The linear model was only significant (P<0.05) for growth traits (exception of ADG205_365). The exponential and Michaelis-Menten models were significant (P<0.01) for all the studied traits while the quadratic model was not significant (P>0.05) for any of the traits. Weight at 205 and 365 days of age decreased 0.25kg and 0.39kg per 1% of increase in inbreeding, respectively. The inbred animals (F=0.25) produced less milk than non-inbred individuals: 50.4kg of milk. Moreover, calving interval increased 0.164 days per 1% of increase in inbreeding. Interestingly, inbreeding had a positive effect on age at first calving and lactation length, decreasing age of first calving and increasing lactation length. © 2012 Japanese Society of Animal Science.

Estudo da oxidação de covelita (CuS) e molibdenita (MoS2) sintéticas por Acidithiobacillus ferrooxidans

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Indução da expressão e caracterização de uma fosfatase ácida ligada à membrana produzida por Burkholderia sp

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Desenvolvimento e aplicação de sensor biomimético descartável para detecção seletiva de hidroquinona

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)