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PubMed1978

The kinetics of methyl viologen oxidation and reduction by the hydrogenase from Clostridium pasteurianum.

A mechanism for the reduction and oxidation of methyl viologen by Clostridium pasteurianum hydrogenase (hydrogen:ferredoxin oxidoreductase, EC 1.12.7.1) is proposed. Double reciprocal plots for methyl viologen reduction and oxidation at pH values 7.0-9.85 are linear, and the plots for reduction and oxidation are intersecting. Such data are consistent with a mechanism in which the H2 and one methyl viologen bind (either in order or randomly) with subsequent reduction and release of the methyl viologen. A second methyl viologen then is bound, reduced and released. Comparison of the calculated Keq' with the Haldane expression in which both methyl viologens react at the same rate show a large difference. This difference indicates that the two methyl viologens react at different rates. Addition of oxidized electron carriers inhibits the hydrogen-deuterium exchange reaction (i.e., the exchange of protons between H2 and 2H2O). CO reversibly inhibits methyl viologen reduction and is competitive vs. H2. O2 acts as an irreversible inhibitor.

Carbon Monoxide↗
PubMed2006

Decavanadate interacts with microsomal NADH oxidation system and enhances cytochrome c reduction.

Oxidation of NADH with accompanying oxygen consumption (NADH:O(2) = 1:1) was observed in the combined presence of metavanadate (MV), decavanadate (DV) and microsomes. Oxygen consumption was negligible in the absence of MV, but NADH was oxidized and DV was reduced to a form of vanadyl-V(IV), colored blue like vanadyl sulfate but differed from it in having a 23-fold higher absorbance at 700 nm. DV can interact with the NADH oxidation system of microsomes as an electron acceptor, in addition to the known ferricyanide and cytochrome c. DV enhances rate of cytochrome c reduction significantly at microM concentrations. These studies indicate potential of DV as a redox intermediate.

Animals↗
PubMed1984

The effects of quinone analogues on cytochrome b6 reduction and oxidation in a reconstituted system.

The reconstituted system containing Photosystem I, plastocyanin and the cytochrome b6-f complex is used to study the effects of various quinone analogues on the redox behavior of cytochrome b6. The effects of DBMIB, DNP-INT and HQNO are compared in an attempt to discern the modes of action of these quinone analogues. Both DBMIB and DNP-INT are potent inhibitors of the plastocyanin reductase activity of the isolated cytochrome complex. However, while DBMIB abolished the oxidant-induced reduction of cytochrome b6, DNP-INT only inhibited about 25% of the net reduction. On the other hand, HQNO does not show any significant inhibition of plastocyanin reductase activity of the isolated cytochrome complex at concentrations up to 20 microM. An enhancement of the net amount of cytochrome b6 reduced is observed in the presence of HQNO. Both DNP-INT and HQNO inhibited the dark oxidation rate of cytochrome b6. The possible identity of the oxidant for cytochrome b6 is discussed. Plastoquinone is concluded to be the most likely candidate. DNP-INT is concluded to have at least two sites of inhibition in the cytochrome complex. The implications of these findings on quinone functions in the cytochrome b6-f complex are discussed.

Cytochrome b Group↗
PubMed2006

Gemfibrozil decreases atherosclerosis in experimental diabetes in association with a reduction in oxidative stress and inflammation.

AIMS/HYPOTHESIS: It is postulated that peroxisome proliferator-activated receptor alpha agonists confer cardiovascular benefits in diabetes, independently of their effects on lipid metabolism. We investigated putative mechanisms responsible for these anti-atherogenic effects in an in vivo model of diabetes-associated atherosclerosis. MATERIALS AND METHODS: Control and streptozotocin-induced diabetic apolipoprotein-deficient mice received gemfibrozil (100 mg kg(-1) day(-1)) or no treatment for 20 weeks. Aortic plaque deposition was assessed by Sudan IV staining and subsequent en face quantification. Superoxide production was measured using lucigenin-enhanced chemiluminescence. Markers of pathways including inflammation and oxidative stress were measured using real-time RT-PCR. RESULTS: No significant effect of gemfibrozil was observed on glycated haemoglobin, cholesterol or insulin in diabetic mice. Diabetes was associated with a three-fold increase in plaque area and a significant increase in NADPH-dependent superoxide compared with control mice. Gemfibrozil significantly attenuated plaque area and superoxide production in diabetic mice. In addition, gemfibrozil reduced the expression of the genes encoding the NADPH oxidase subunits p47phox, gp91phox and Rac-1. In addition, gemfibrozil reduced the expression of the genes encoding nuclear factor kappa B (NF-kappaB) subunit, p65, the NF-kappaB-dependent chemokine monocyte chemoattractant protein-1, and tissue factor. CONCLUSIONS/INTERPRETATIONS: This study demonstrates that gemfibrozil exerts anti-atherogenic actions, independently of changes in cholesterol and glucose metabolism. Such findings emphasise the possible usefulness of fibrates such as gemfibrozil in a setting of atherosclerosis even in the absence of dyslipidaemia.

Angiotensins↗
PubMed2003

Red blood cells upregulate cytoprotective proteins and the labile iron pool in dividing human T cells despite a reduction in oxidative stress.

We have recently reported that red blood cells (RBC) promote T cell growth and survival by inhibiting activation-induced T cell death. In the present study, we have examined parameters of oxidative stress and intracellular iron in activated T cells and correlated these data with the expression of ferritin, heme oxygenase-1 (HO-1), and the transferrin receptor CD71. T cells growing in the presence of RBC had reduced levels of reactive oxygen species (ROS) and oxidatively modified proteins, suggesting that RBC efficiently counteracted ROS production on the activated T cells. Flow cytometry and immunodetection demonstrated that T cells dividing in the presence of RBC had increased levels of intracellular ferritin rich in L-subunits and HO-1 along with a downmodulation in CD71 expression. Finally, using the fluorescent iron indicator calcein and flow cytometry analysis, we were able to show that a relative amount of the labile iron pool (LIP) was upregulated in T cells growing in the presence of RBC. These findings are consistent with a typical response to iron overload. However, neither heme compounds nor ferric iron reproduced the levels of expansion and survival of T cells induced by intact RBC. Altogether, these data suggest that RBC inhibit apoptosis of activated T cells by a combination of ROS scavenging and upregulation of cytoprotective proteins such as ferritin and HO-1, which may counteract a possible toxic effect of the increased intracellular free iron.

Antigens, CD↗
PubMed2006

Efficiency of apples, strawberries, and tomatoes for reduction of oxidative stress in pigs as a model for humans.

OBJECTIVE: The present study investigated the potency of apples, strawberries, and tomatoes in decreasing oxidative stress in pigs as a model for humans. METHODS: Forty-eight growing pigs were individually penned in metabolic cages and divided into six groups. All groups received isocaloric daily rations composed of a basal diet supplemented with starch, linseed oil, linseed oil and apples, linseed oil and strawberries, linseed oil and tomatoes, or linseed oil, apples, strawberries and tomatoes. Oxidative stress was evaluated at the beginning and at the end of the 22-d experimental period by measuring the degree of DNA damage in mononuclear blood cells, the concentration of malondialdehyde (MDA) in plasma, the MDA excretion rate, the concentration of tocopherols in plasma, plasma total antioxidant status, and erythrocyte glutathione peroxidase concentration. RESULTS: The results confirmed that the linseed oil diet significantly increased MDA formation in the body and DNA damage to mononuclear blood cells in comparison with the starch diet. Although the MDA concentration in plasma was significantly decreased in all fruit groups, urine MDA excretion rate was decreased only in the linseed oil/apple and linseed oil/apple/strawberry/tomato groups. Supplementation with fruit significantly decreased DNA damage, was more pronounced in the linseed oil/apple and linseed oil/apple/strawberry/tomato groups. Significantly lower glutathione peroxidase concentrations were observed only in the linseed oil/strawberry group. CONCLUSIONS: Our findings support the hypothesis that supplementation with apples, strawberries, or tomatoes effectively decreases oxidative stress by decreasing MDA formation in the body and by protecting mononuclear blood cells against increased DNA damage. This effect was particularly pronounced in the group supplemented with a fruit mixture; among the single fruit supplements, the most beneficial effect was obtained with apples.

Analysis of Variance↗
PubMed1997

Mitochondrial oxygen radical formation during reductive and oxidative stress to intact hepatocytes.

After simple respiratory inhibition, glycolytic substrates prevent cell death by providing an alternate source of cellular ATP. When mitochondrial uncoupling ensues, the uncoupler-stimulated mitochondrial ATPase hydrolyzes ATP formed by glycolysis and protection is lost. Electron transfer components abnormally reduced by respiratory inhibition, especially ubisemiquinone, react directly with oxygen to form toxic radicals. Mitochondria also generate reactive oxygen species after exposure to oxidant chemicals. A consequence is onset of the mitochondrial permeability transition, which leads to uncoupling, cellular ATP depletion and loss of viability. Thus, mitochondria are both a source and a target of toxic oxygen radicals in cell injury.

Animals↗
PubMed2006

Up regulation of IL-6 by ischemic preconditioning in normal and fatty rat livers: association with reduction of oxidative stress.

We analyzed the role of IL-6 in the protection that ischemic preconditioning (IP) exerts against hepatic ischemia reperfusion-mediated (I/R) oxidative damage, particularly in fatty livers. IP-related IL-6 up-regulation during reperfusion in steatotic and non-steatotic livers was correlated with reduced indices of liver damage, as also demonstrated by pharmacological modulation of IL-6. IP activated NF-kB and HSF during ischemia (Isc), whereas AP-1 activity was unaffected. IP blunted the activation of STAT3 and stress-responsive genes, such as NF-kB, AP-1 and heme oxygenase (HO-1) during reperfusion. The role of reduced oxidative stress in hepatoprotection of fatty livers was further demonstrated by the fact that: (i) IP prevented the decrease of glutathione levels and the increase of lipid peroxidation; (ii) the anti-oxidant GSH-ester prevented lipid peroxidation and necrosis. In conclusion, IP modulates the activity of transcription factors and triggers IL-6 production; this may prevent hepatic I/R damage in a oxidative stress-dependent way, particularly in fatty livers.

Animals↗
PubMed2006

Reduction of oxidative stress and modulation of autoantibodies against modified low-density lipoprotein after rosuvastatin therapy.

AIMS: To examine the effect of 24 weeks' rosuvastatin treatment on oxidative stress and changes in immune response to oxidized low-density lipoprotein (LDL). METHODS: This was an open-label study of patients in Austria receiving 10 or 40 mg rosuvastatin daily alternately during 12 and 24 weeks. Circulating concentrations of antibodies to malondialdehyde-oxidized LDL (MDA-LDL), both IgG and IgM type, to copper-oxidized LDL (Cu-OxLDL-IgG), concentrations of oxidized LDL complexed to IgG (OxLDL-IC) and markers of oxidative stress and systemic inflammation in subjects with plasma LDL cholesterol concentrations between 130 mg dl-1 and 250 mg dl-1 and triglycerides<or=400 mg dl-1 were determined. RESULTS: During statin therapy, plasma endogenous peroxides (POX-ACT) concentrations and peroxidase activity were significantly decreased, associated with a modest increase in total antioxidant capacity (TAC). Antibody titres to MDA-LDL-IgM, Cu-OxLDL-IgG and OxLDL-IC decreased, whereas MDA-LDL-IgG concentrations were increased after therapy. These changes were dose- and LDL-independent. POX-ACT concentrations were significantly positively correlated with inflammation markers before and after therapy and inversely with high-density lipoprotein-cholesterol concentrations after therapy. CONCLUSION: This study provides in vivo evidence that rosuvastatin significantly reduces oxidative stress and has immunomodulatory properties in a dose- and LDL-independent manner.

Antioxidants↗
PubMed2005

The influence of chokeberry juice supplementation on the reduction of oxidative stress resulting from an incremental rowing ergometer exercise.

The aim of the study was to investigate the influence of an increased intake of anthocyanins, contained in chokeberry juice, on the redox parameters in rowers performing a physical exercise during a 1-month training camp. The athletes were randomly assigned to receive 150 mL of chokeberry juice daily, containing 23 mg/100 mL of anthocyanins (supplemented group), or placebo (control group). Before and after the supplementation period, the subjects performed an incremental rowing exercise test. Blood samples were taken from the antecubital vein before each exercise test, 1 min after the test, and following a 24-h recovery period. After the supplementation period, TBARS concentrations in the samples collected 1 min after the exercise test and following a 24-h recovery period were significantly lower in the subjects receiving chokeberry juice than in the control group. In the supplemented group, glutathione peroxidase activity was lower in the samples collected 1 min after the exercise test, and superoxide dismutase activity was lower in the samples taken following a 24-h recovery, as compared to the subjects receiving placebo. These findings indicate that an increased intake of anthocyanins limits the exercise-induced oxidative damage to red blood cells, most probably by enhancing the endogenous antioxidant defense system.

Adult↗
PubMed2004

Reduction of oxidative stress marker in lung fluid of preterm infants after administration of intra-tracheal liposomal glutathione.

BACKGROUND: Low levels of glutathione are associated with subsequent chronic lung disease in preterm infants. Incorporation of glutathione into liposomes offers a method of increasing levels with a prolonged half-life compared with direct inhalation. OBJECTIVES: The aim of this study was to examine the clinical feasibility of administering a single dose of liposomal glutathione and its effectiveness at raising glutathione at 12 and 24 h after treatment. METHODS: Fourteen ventilated preterm infants from the Regional Neonatal Intensive Care Unit at Liverpool Women's Hospital received 1 mg/kg or 10 mg/kg liposomal glutathione intra-tracheally and bronchoalveolar lavage fluid was collected prior to treatment, 12 and 24 h after dosing for glutathione and malondialdehyde estimation. RESULTS: Mean glutathione was initially 12.2 micromol/l, increasing to 52.8 micromol/l at 12 h (p = 0.006). Mean malondialdehyde was initially 265.6 nmol/l decreasing to 11.2 nmol/l at 12 h (p = 0.018). CONCLUSIONS: Intra-tracheal liposomal glutathione instillation offers a feasible method of raising pulmonary glutathione in preterm infants and shows biochemical antioxidant effects.

Biomarkers↗
PubMed2004

Electrocatalytic reductions of nitrite, nitric oxide, and nitrous oxide by thermophilic cytochrome P450 CYP119 in film-modified electrodes and an analytical comparison of its catalytic activities with myoglobin.

Previous investigations of nitrite and nitric oxide reduction by myoglobin in surfactant film modified electrodes characterized several distinct steps in the denitrification pathway, including isolation of a nitroxyl adduct similar to that proposed in the P450nor catalytic cycle. To investigate the effect of the axial ligand on these biomimetic reductions, we report here a comparison of the electrocatalytic activity of myoglobin (Mb) with a thermophilic cytochrome P450 CYP119. Electrocatalytic nitrite reduction by CYP119 is very similar to that by Mb: two catalytic waves at analogous potentials are observed, the first corresponding to the reduction of nitric oxide, the second to the production of ammonia. CYP119 is a much more selective catalyst, giving almost exclusively ammonia during the initial half-hour of reductive electrolysis of nitrite. More careful investigations of specific steps in the catalytic cycle show comparable rates of nitrite dehydration and almost identical potentials and lifetimes for ferrous nitroxyl intermediate (Fe(II)-NO(-)) in CYP119 and Mb. The catalytic efficiency of nitric oxide reduction is reduced for CYP119 as compared to Mb, attributable to both a lower affinity of the protein for NO and a decreased rate of N-N coupling. Isotopic labeling studies show ammonia incorporation into nitrous oxide produced during nitrite reduction, as has been termed co-denitrification for certain bacterial and fungal nitrite reductases. Mb has a much higher co-denitrification activity than CYP119. Conversely, CYP119 is shown to be slightly more efficient at the two-electron reduction of N(2)O to N(2). These results suggest that thiolate ligation does not significantly alter the catalytic reactivity, but the dramatic difference in product distribution may suggest an important role for protein stability in the selectivity of biocatalysts.

Ammonia↗
PubMed2005

Sequential electrolytic oxidation and reduction of aqueous phase energetic compounds.

Contamination of soils and groundwater with energetic compounds has been documented at many former ammunition manufacturing plants and ranges. Recent research at Colorado State University (CSU) has demonstrated the potential utility of electrolytic degradation of organic compounds using an electrolytic permeable reactive barrier (e-barrier). In principle, an electrolytic approach to degrade aqueous energetic compounds such as hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX) or 2,4,6-trinitrotoluene (TNT) can overcome limitations of management strategies that involve solely oxidation or reduction, through sequential oxidation-reduction or reduction-oxidation. The objective of this proof-of-concept research was to evaluate transformation of aqueous phase RDX and TNT in flow-through electrolytic reactors. Laboratory experiments were conducted using six identical column reactors containing porous media and expanded titanium-mixed-metal-oxide electrodes. Three columns tested TNT transformation and three tested RDXtransformation. Electrode sequence was varied between columns and one column for each contaminant acted as a no-voltage control. Over 97% of TNT and 93% of RDX was transformed in the reactors under sequential oxidation-reduction. Significant accumulation of known degradation intermediates was not observed under sequential oxidation-reduction. Removal of approximately 90% of TNT and 40% of RDX was observed under sequential reduction-oxidation. Power requirements on the order of 3 W/m2 were measured during the experiment. This suggests that an in-situ electrolytic approach may be cost-practical for managing groundwater contaminated with explosive compounds.

Electrodes↗
PubMed2001

Enzymatic and non-enzymatic reduction of brucine N-oxide by aldehyde oxidase and catalase.

1. Brucine N-oxide was reduced by aldehyde oxidase in rabbit liver cytosol in the presence of an electron donor, 2-hydroxypyrimidine, under anaerobic conditions. The flavoprotein purified from rabbit liver exhibited significant reductase activity in the presence of electron donors. 2. Brucine N-oxide was also reduced by rabbit liver cytosol and blood in the presence of both a reduced pyridine nucleotide and FAD under anaerobic conditions. The N-oxide reductase activities were inhibited by carbon monoxide and air. However, these activities were not abolished whe n liver cytosol and blood were boiled. Rabbit erythrocytes exhibited the reductase activity, but not plasma. 3. When liver cytosol or blood was separated by DEAE-cellulose column chromatography, the fractions with the reducing activity in the presence of both NADH and FAD also showed catalase activity. 4. Catalase catalysed the brucine N-oxide reduction in the presence of both NAD(P)H and FAD. Hematin also exhibited the reductase activity in the presence of both NAD(P)H and FAD. Photochemically reduced FAD was effective in the reduction instead of NAD(P)H and FAD. 5. Bricine N-oxide reduction proceeds via two routes in liver cytosol and blood. One is enzymatic reduction by aldehyde oxidase; the other is non-enzymatic reduction catalysed by the haem group of catalase in the presence of reduced flavin.

Aldehyde Oxidase↗
PubMed1981

Cytochrome b oxidation and reduction reactions in the ubiquinone-cytochrome b/c2 oxidoreductase from Rhodopseudomonas sphaeroides.

1. The kinetics of cytochrome b reduction and oxidation in the ubiquinone-cytochrome b/c2 oxidoreductase of chromatophores from Rhodopseudomonas sphaeroides Ga have been measured both in the presence and absence of antimycin, after subtraction of contributions due to absorption changes from cytochrome c2, the oxidized bacteriochlorophyll dimer of the reaction center, and a red shift of the antenna bacteriochlorophyll. 2. A small red shift of the antenna bacteriochlorophyll band centered at 589 nm has been identified and found to be kinetically similar to the carotenoid bandshift. 3. Antimycin inhibits the oxidation of ferrocytochrome b under all conditions; it also stimulates the amount of single flash activated cytochrome b reductions 3- to 4-fold under certain if not all conditions. 4. A maximum of approximately 0.6 cytochrome b-560 (Em(7) = 50 mV, n = 1, previously cytochrome b50) hemes per reaction center are reduced following activating flashes. This ratio suggests that there is one cytochrome b-560 heme functional per ubiquinone-cytochrome b/c2 oxidoreductase. 5. Under the experimental conditions used here, only cytochrome b-560 is observed functional in cyclic electron transfer. 6. We describe the existence of three distinct states of reduction of the ubiquinone-cytochrome b/c2 oxidoreductase which can be established before activation, and result in markedly different reaction sequences involving cytochrome b after the flash activation. Poising such that the special ubiquinone (Qz) is reduced and cytochrome b-560 is oxidized yields the conditions for optimal flash activated electron transfer rates through the ubiquinone-cytochrome b/c2 oxidoreductase. However when the ambient redox state is lowered to reduce cytochrome b-560 or raised to oxidize Qz, single turnover flash induced electron transfer through the ubiquinone-cytochrome b/c2 oxidoreductase appears impeded; the points of the impediment are tentatively identified with the electron transfer step from the reduced secondary quinone (QII) of the reaction center to ferricytochrome b-560 and from the ferrocytochrome b-560 to oxidized Qz, respectively.

Bacteriochlorophylls↗