Cytochrome c-promoted cardiolipin oxidation generates singlet molecular oxygen

The interaction of cytochrome c (cyt c) with cardiolipin (CL) induces protein conformational changes that favor peroxidase activity. This process has been correlated with CL oxidation and the induction of cell death. Here we report evidence demonstrating the generation of singlet molecular oxygen [O-2((1)Delta(g))] by a cyt c-CL complex in a model membrane containing CL. The formation of singlet oxygen was directly evidenced by luminescence measurements at 1270 nm and by chemical trapping experiments. Singlet oxygen generation required cyt c-CL binding and occurred at pH values higher than 6, consistent with lipid-protein interactions involving fully deprotonated CL species and positively charged residues in the protein. Moreover, singlet oxygen formation was specifically observed for tetralinoleoyl CL species and was not observed with monounsaturated and saturated CL species. Our results show that there are at least two mechanisms leading to singlet oxygen formation: one with fast kinetics involving the generation of singlet oxygen directly from CL hydroperoxide decomposition and the other involving CL oxidation. The contribution of the first mechanism was clearly evidenced by the detection of labeled singlet oxygen [O-18(2)((1)Delta(g))] from liposomes supplemented with 18-oxygen-labeled CL hydroperoxides. However quantitative analysis showed that singlet oxygen yield from CL hydroperoxides was minor (<5%) and that most of the singlet oxygen is formed from the second mechanism. Based on these data and previous findings we propose a mechanism of singlet oxygen generation through reactions involving peroxyl radicals (Russell mechanism) and excited triplet carbonyl intermediates (energy transfer mechanism).

Electrocatalytic Activity of Pd, Pt, and Rh for the Electro-oxidation of Ethanol in Alkaline Electrolyte: An Online DEMS Study

The electro-oxidation of ethanol was investigated on electrodeposited layers of Pd, Pt, and Rh in alkaline electrolyte. The reaction products were monitored by experiments of online differential electrochemical mass spectrometry (DEMS). Potentiodynamic curves for the ethanol electro-oxidation catalyzed by these three different metal electrocatalysts showed similar onset potentials, but the highest Faradaic current peak was observed for the Pt electrocatalyst. Online DEMS experiments evidenced similar amounts of CO2 for the three different materials, but Pd presented the higher production of ethylacetate (acetic acid). This indicated that the electrochemical oxidation of ethanol on the Pd surface occurred to a higher extent. The formation of methane, which was observed for Pt and Rh, after potential excursions to lower potentials, was absent for Pd. On the basis of the obtained results, it was stated that, on Pt and Rh, the formation of CO2 occurs mainly via oxidation of CO and CH (x,ad) species formed after dissociative adsorption of ethanol or ethoxy species that takes place only at low potentials. This indicates that the dissociative adsorption of ethanol or ethoxy species is inhibited at higher potentials on Pt and Rh. On the other hand, on the Pd electrocatalyst, the reaction may occur via nondissociative adsorption of ethanol or ethoxy species at lower potentials, followed by oxidation to acetaldehyde and, after that, by a further oxidation step to acetic acid on the electrocatalyst surface. Additionally, in a parallel route, the acetaldehyde molecules adsorbed on the Pd surface can be deprotonated, yielding a reaction intermediate in which the carbon-carbon bond is less protected, and therefore, it can be dissociated on the Pd surface, producing CO2, after potential excursions to higher potentials.

Is the poly (methylene blue)-modified glassy carbon electrode an adequate electrode for the simple detection of thiols and amino acid-based molecules?

This paper describes the preparation, characterization, and use of poly (methylene blue) (PMB)-modified glassy carbon electrodes (GCE) (GCE-PMB) in the detection of the thiols L-cysteine (L-CySH) and N-acetyl cysteine (Acy), and the herbicide glyphosate (GLYP) in pH 5.3 aqueous solution. The polymer film prepared by electropolymerization showed different characteristics such as robustness, stability, and redox properties satisfactorily. The surface coverage concentration (Gamma) of PMB was found to be 7.90 x 10(-9) - mol cm(-2). Moreover, we observed strong adhesion of the polymer film to the electrode surface. The results using GCE-PMB as a sensor indicated that this modified electrode exhibited electrocatalytic activity toward the detection of thiols and glyphosate in 0.1 mol L-1 KO (pH 5.3). Meanwhile, strong adsorption of the analytes on the GCE-PMB electrodes was also observed. Otherwise, using a low concentration (1 x 10(-4) mol L-1) of L-cysteine and N-acetyl cysteine and 8.9 x 10(-6) mol L-1 of glyphosate, separately, it was possible to observe a well-defined electrochemical response, thus providing an opportunity to further understand the applicability of PMB as a sensor for amino acid-based molecules. (C) 2012 Elsevier B.V. All rights reserved.

The dual pathway in action: decoupling parallel routes for CO2 production during the oscillatory electro-oxidation of methanol

As in the case of most small organic molecules, the electro-oxidation of methanol to CO2 is believed to proceed through a so-called dual pathway mechanism. The direct pathway proceeds via reactive intermediates such as formaldehyde or formic acid, whereas the indirect pathway occurs in parallel, and proceeds via the formation of adsorbed carbon monoxide (COad). Despite the extensive literature on the electro-oxidation of methanol, no study to date distinguished the production of CO2 from direct and indirect pathways. Working under, far-from-equilibrium, oscillatory conditions, we were able to decouple, for the first time, the direct and indirect pathways that lead to CO2 during the oscillatory electro-oxidation of methanol on platinum. The CO2 production was followed by differential electrochemical mass spectrometry and the individual contributions of parallel pathways were identified by a combination of experiments and numerical simulations. We believe that our report opens some perspectives, particularly as a methodology to be used to identify the role played by surface modifiers in the relative weight of both pathways-a key issue to the effective development of catalysts for low temperature fuel cells.

Cerebral White Matter Oxidation and Nitrosylation in Young Rodents With Kaolin-Induced Hydrocephalus

Hydrocephalus is associated with reduced blood flow in periventricular white matter. To investigate hypoxic and oxidative damage in the brains of rats with hydrocephalus, kaolin was injected into the cisterna magna of newborn 7- and 21-day-old Sprague-Dawley rats, and ventricle size was assessed by magnetic resonance imaging at 7, 21, and 42 days of age. In-situ evidence of hypoxia in periventricular capillaries and glial cells was shown by pimonidazole hydrochloride binding. Biochemical assay of thiobarbituric acid reaction and immunohistochemical detection of malondialdehyde and 4-hydroxy-2-nonenal indicated the presence of lipid peroxidation in white matter. Biochemical assay of nitrite indicated increased nitric oxide production. Nitrotyrosine immunohistochemistry showed nitrosylated proteins in white matter reactive microglia and astrocytes. Activities of the antioxidant enzymes catalase and glutathione peroxidase were not increased, and altered hypoxia-inducible factor 1 alpha was not detected by quantitative reverse transcription-polymerase chain reaction. Cerebral vascular endothelial growth factor expression determined by quantitative reverse transcription-polymerase chain reaction and enzyme-linked immunosorbent assay was not changed, but vascular endothelial growth factor immunoreactivity was increased in reactive astrocytes of hydrocephalic white matter. To determine if nitric oxide synthase is involved in the pathogenesis, we induced hydrocephalus in 7-day-old wild-type and neuronal nitric oxide synthase-deficient mice. At 7 days, the wild-type and mutant mice exhibited equally severe ventriculomegaly and no behavioral differences, although increased glial fibrillary acidic protein was less in the mutant mice. We conclude that hypoxia, via peroxidation and nitrosylation, contributes to brain changes in young rodents with hydrocephalus and that compensatory mechanisms are negligible.

Chemical Selectivity during the Electro-Oxidation of Ethanol on Unsupported Pt Nanoparticles

In this paper we present results on the electro-oxidation of ethanol on unsupported (carbon free) platinum nanoparticles, considering the effects of the alcohol concentration. The case of the so-called dual pathway mechanism during the electro-oxidation of ethanol showed to be influenced by the surface coverage of adsorbed carbon monoxide (COad) at unsupported platinum. The influences of adsorbed intermediates were followed by in situ infrared spectroscopy (FTIR) and by electrochemical experiments. Unsupported platinum showed that the reaction leads to the formation of CO2 and acetic acid as main products at low concentrations of ethanol (0.01 to 0.1 mol L-1). At least in this case of 0.01 mol L-1 ethanol, most formation of CO2 occurred via COad (indirect pathway). At higher concentration of ethanol, however, most CO2 was formed via a reactive intermediate such as acetaldehyde (direct pathway). In addition, in this higher concentration of ethanol, the acetic acid was produced via formation of adsorbed acetaldehyde (via acetate) at higher overpotentials. In case of the acetic acid formation, a dual pathway was identified during the electro-oxidation of ethanol at low alcohol concentrations, whereas a parallel pathway occurred without the formation of adsorbed acetate intermediates at low overpotentials. (C) 2012 The Electrochemical Society. [DOI: 10.1149/2.101203jes] All rights reserved.

Effect of natural antioxidant combinations on lipid oxidation in cooked chicken meat during refrigerated storage

The effect of combinations of sage, oregano and honey on lipid oxidation in cooked chicken meat during refrigeration at 4 degrees C for 96 h was determined. Chicken samples (thigh and breast) were then separated into five groups; control; butylated hydroxytoluene; oregano + sage; oregano + sage + 5%honey and oregano + sage + 10%honey. Quantitative measurements of thiobarbituric acid reactive substances, conjugated dienes, hexanal, fatty acids, cholesterol and cholesterol oxides were used as indicators of lipid oxidation. Acceptability and preference were also evaluated. The effectiveness of the natural antioxidants for reducing the velocity of lipid oxidation in cooked chicken thigh and breast was demonstrated after 48 and 96 h of refrigeration at 4 degrees C. The treatments that presented the lowest hexanal values after 96 h of refrigeration were oregano + sage + 5%honey and oregano + sage + 10%honey. Only traces of free cholesterol oxides were found (25-OH, 7-k, 7 alpha-OH and 7 beta-OH). The natural antioxidants protected cooked chicken meat from oxidation processes and resulted in great acceptability. (C) 2012 Elsevier Ltd. All rights reserved.

Platinum nanoparticles incorporated in silsesquioxane for use in LbL films for the simultaneous detection of dopamine and ascorbic acid

We describe the preparation of platinum nanoparticles (PtNPs) using the 3-n-propylpyridinium silsesquioxane chloride (SiPy+Cl-) as a nanoreactor and stabilizer. The formation of PtNPs was monitored by UV-Vis spectroscopy by measuring the decrease in the intensity of the band at 375 nm, which is attributed to the electronic absorption of PtCl62- ions. TEM images of Pt-SiPy+Cl- nanohybrid indicated an average size of 3-40 nm for PtNPs. The Pt-SiPy+Cl- was used as a polycation in the preparation of layer-by-layer films (LbL) on a glass substrate coated with fluorine-doped tin oxide (FTO) alternating with the polyanion poly(vinyl sulfonic acid) (PVS). The films were electrochemically tested in sulfuric acid to confirm the deposition of Pt-SiPy+Cl- onto the LbL films, observing the adsorption and desorption of hydrogen (E-pa = 0.1 V) and by the redox process of formation for PtO with E-pa = 1.3 V and E-pc = 0.65 V. FTIR and Raman spectra confirmed the presence of the PVS and Pt-SiPy+Cl- in the LbL films. A linear increase in the absorbance in the UV-Vis spectra of the Pt-SiPy+Cl- at 258 nm (pi -> pi* transition of the pyridine groups) with a number of Pt-SiPy+Cl-/PVS or PVS/SiPy+Cl- bilayers (R = 0.992) was observed. These LbL films were tested for the determination of dopamine (DA) in the presence of ascorbic acid (AA) with a detection limit (DL) on the order of 2.6 x 10(-6) mol L-1 and a quantification limit (QL) of 8.6 x 10(-6) mol L-1. The films exhibited a good repeatability and reproducibility, providing a potential difference of 550 mV for the oxidation of DA with AA interferent.

Exploiting the enantioselectivity of Baeyer-Villiger monooxygenases via boron oxidation

The enantioselective carbon-boron bond oxidation of several chiral boron-containing compounds by Baeyer-Villiger monooxygenases was evaluated. PAMO and M446G PAMO conveniently oxidized 1-phenylethyl boronate into the corresponding 1-(phenyl)ethanol (ee = 82-91%). Cyclopropyl boronic esters were also oxidized but with no enantioselectivity. beta-Boryl carboxylic esters were not oxidized by any BVMOs. (C) 2012 Elsevier Ltd. All rights reserved.

New Approaches for the Ethanol Oxidation Reaction of Pt/C on Carbon Cloth Using ATR-FTIR

This work describes the study of the ethanol oxidation reaction of a Pt/C Etek electrocatalyst that was supported on different substrates, such as gold, glassy carbon and carbon cloth treated with PTFE. In the ethanol oxidation reaction, the activity varies with the substrate, as well as the pathways for ethanol oxidation, as studied by an ATR-FTIR in situ setup using the carbon cloth as the electrocatalyst support. The electrocatalyst Pt/C supported on gold starts acetaldehyde production from ethanol oxidation at an onset potential of 0.1 V less than that observed for the same process on Teflon-treated carbon cloth. The Pt/C supported on the carbon cloth starts its CO2 production for the same oxidation process at 0.2 V less than on the Pt/C supported on gold substrate. The differences in catalytic activity for the ethanol oxidation reaction depend not only on the electrocatalyst but also on various electrode factors, such as the substrate, the roughness of the electrode and the charge transfer resistance.

Assessment of a UASB reactor for the removal of sulfate from acid mine water

A bench-scale Upflow Anaerobic Sludge Blanket (UASB) reactor was used to study the treatment of acid mine drainage through the biological reduction of sulfate. The reactor was fed with acid mine drainage collected at the Osamu Utsumi uranium mine (Caldas, MG, Brazil) and supplemented with ethanol as an external carbon source. Anaerobic granular sludge originating from a reactor treating poultry slaughterhouse wastewater was used as the inoculum. The reactor's performance was studied according to variations in the chemical oxygen demand (COD)/SO42- ratio, influent dilution and liquid-phase recirculation. The digestion of a dilution of the acid mine drainage resulted in a 46.3% removal of the sulfate and an increase in the effluent pH (COD/SO42- = 0.67). An increase in the COD/SO42- ratio to 1.0 resulted in an 85.6% sulfate reduction. The reduction of sulfate through complete oxidation of the ethanol was the predominant path in the reactor, although the removal of COD was not greater than 68% in any of the operational stages. The replenishment of the liquid phase with tap water positively affected the reactor, whereas the recirculation of treated effluent caused disequilibrium and decreased efficiency. (C) 2012 Elsevier Ltd. All rights reserved.

The influence of the Pt crystalline surface orientation on the glycerol electro-oxidation in acidic media

We investigated the electrochemical oxidation of glycerol on low-index Pt single crystals in acidic media (H2SO4 and HClO4) by cyclic voltammetry and Fourier Transform Infrared (FTIR) spectroscopy and we verified that this is a surface sensitive reaction. Pt(100) and Pt(110) surface structures favor the breaking of the C-C-C bond at low potentials (say 0.05 V), as seen by the formation of CO, one of the adsorbed residues of the glycerol dissociation, which poisons these surfaces even at high potentials. Pt(111) surface structure does not favor the C-C-C bond breaking at potentials as low as 0.05 V. However, Pt(111) is less poisoned by residues of glycerol dissociation and, for this reason, it is more active for glycerol oxidation than Pt(100) and Pt(110) at low potentials. Carbonyl containing compounds and CO2 were detected as reaction products of the glycerol oxidation on all investigated single-crystal Pt surfaces. The ratio between CO2 and carbonyl containing compounds is clearly much higher for Pt(100) and Pt(110) than for Pt(111). (C) 2012 Elsevier Ltd. All rights reserved.

Effects of exogenous calcium or oxalic acid on Pinus taeda treatment with the white-rot fungus Ceriporiopsis subvermispora

Pinus taeda wood chips were treated with the biopulping fungus Ceriporiopsis subvermispora in calcium-or oxalic acid-amended cultures. The secretion of hydrolytic and oxidative enzymes was inhibited only in the cultures having the highest concentration of calcium (1400 mg kg(-1) wood). Calcium decreased the availability of free oxalic acid, inhibited fungal growth, and reduced lignin mineralization and transformations. Oxalic acid amendment in the cultures was found not to affect the lignin mineralization and transformations; however, it did inhibit the depolymerization reactions detectable in the residual lignin that was retained in the biotreated wood. C. subvermispora presented catabolic activity for oxalic acid in the cultures amended with 1660 mg acid kg(-1) wood, whereas oxalic acid was synthesized when it was amended at low amounts or initially absent in the cultures. These data suggest one ideal ratio of oxalic acid in C. subvermispora cultures and indicate that its exogenous addition does not necessarily accompany the further degradation of lignin. Crown Copyright (C) 2012 Published by Elsevier Ltd. All rights reserved.

Gamma Radiation Induced Oxidation and Tocopherols Decrease in In-Shell, Peeled and Blanched Peanuts

In-shell, peeled and blanched peanut samples were characterized in relation to proximate composition and fatty acid profile. No difference was found in relation to its proximate composition. The three major fatty acids were palmitic acid, oleic acid, and linoleic acid. In order to investigate irradiation and storage effects, peanut samples were submitted to doses of 0.0, 5.0, 7.5 or 10.0 kGy, stored for six months at room temperature and monitored every three months. Peanuts responded differently to irradiation, particularly with regards to tocopherol contents, primary and secondary oxidation products and oil stability index. Induction periods and tocopherol contents were negatively correlated with irradiation doses and decreased moderately during storage. alpha-Tocopherol was the most gamma radiation sensitive and peeled samples were the most affected. A positive correlation was found among tocopherol contents and the induction period of the oils extracted from irradiated samples. Gamma radiation and storage time increased oxidation compounds production. If gamma radiation is considered an alternative for industrial scale peanut conservation, in-shell samples are the best feedstock. For the best of our knowledge this is the first article with such results; this way it may be helpful as basis for future studies on gamma radiation of in-shell crops.

Metabolic engineering of beta-oxidation in Penicillium chrysogenum for improved semi-synthetic cephalosporin biosynthesis

Industrial production of semi-synthetic cephalosporins by Penicillium chrysogenum requires supplementation of the growth media with the side-chain precursor adipic acid. In glucose-limited chemostat cultures of P. chrysogenum, up to 88% of the consumed adipic acid was not recovered in cephalosporinrelated products, but used as an additional carbon and energy source for growth. This low efficiency of side-chain precursor incorporation provides an economic incentive for studying and engineering the metabolism of adipic acid in P. cluysogenum. Chemostat-based transcriptome analysis in the presence and absence of adipic acid confirmed that adipic acid metabolism in this fungus occurs via beta-oxidation. A set of 52 adipate-responsive genes included six putative genes for acyl-CoA oxidases and dehydrogenases, enzymes responsible for the first step of beta-oxidation. Subcellular localization of the differentially expressed acyl-CoA oxidases and dehydrogenases revealed that the oxidases were exclusively targeted to peroxisomes, while the dehydrogenases were found either in peroxisomes or in mitochondria. Deletion of the genes encoding the peroxisomal acyl-CoA oxidase Pc20g01800 and the mitochondrial acyl-CoA dehydrogenase Pc20g07920 resulted in a 1.6- and 3.7-fold increase in the production of the semi-synthetic cephalosporin intermediate adipoyl-6-APA, respectively. The deletion strains also showed reduced adipate consumption compared to the reference strain, indicating that engineering of the first step of beta-oxidation successfully redirected a larger fraction of adipic acid towards cephalosporin biosynthesis. (C) 2012 Elsevier Inc. All rights reserved.