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Consumption of red wine with meals reduces the susceptibility of human plasma and low-density lipoprotein to lipid peroxidation.

The effect of consumption of red or white wine (11% alcohol) with meals on the propensity of plasma and low-density lipoprotein (LDL) to undergo lipid peroxidation was studied in 17 healthy men who were divided into two groups: 8 received 400 mL red wine/d for 2 wk, and 9 received a similar amount of white wine. Red wine consumption for 2 wk resulted in a 20% reduction in the propensity of plasma to undergo lipid peroxidation (in the presence of a free-radical-generating system) as determined by the thiobarbituric acid reactive substances (TBARS) assay. In parallel, red wine consumption reduced the propensity of the volunteers' LDL to undergo lipid peroxidation (in response to copper ions) as determined by a 46%, 72%, and 54% decrease in the content of TBARS, lipid peroxides, and conjugated dienes in LDL, respectively, as well as by a substantial prolongation of the lag phase required for the initiation of LDL oxidation. On the contrary, dietary consumption of white wine for 2 wk resulted in a 34% increase in plasma's propensity to undergo lipid peroxidation and also in a 41% increased propensity of the LDL to undergo lipid peroxidation. The antioxidant effect of dietary red wine on plasma lipid peroxidation was not secondary to changes in the plasma vitamin E or beta-carotene content but could be related to the elevation of polyphenol concentration in plasma and LDL. Thus, some phenolic substances that exist in red wine, but not in white wine, are absorbed, bind to plasma LDL, and may be responsible for the antioxidant properties of red wine.

Adult↗

Effect of tumor promoters on lipid peroxidation in mouse skin.

Products of lipid peroxidation were measured in mouse epidermis. These were shown to increase with advancing age. Conversely, a decline in these products was observed on treatment with 12-O-tetradecanoylphorbol-13-acetate (TPA), a potent tumor promoting agent. A decline in lipid peroxidation occurred within 4 h after application of 2 micrograms TPA and the maximum effect was seen at 22-24 h. A lesser active tumor promoter, phorbol dibenzoate; and ethyl phenylpropiolate, a purely hyperplastic agent, also lowered lipid peroxidation; while phorbol, a non-promoter, did not show any significant effect. Mezerein, a resiniferonal derivative with weak promoting activity but a potent stage II promoter, appeared to be more potent than TPA in lowering the basal levels of peroxidation. The TPA-induced decrease in lipid peroxidation could be prevented by fluocinolone acetonide, a potent antipromoting and antimitotic agent, but not by retinoic acid and tosylamino-2-phenylethylchlorimethyl-ketone which are relatively potent antipromoting agents but lack antimitotic activity, suggesting that the decrease of lipid peroxidation by tumor promoting agents is possibly related to their mitotic activity. Furthermore, skin papilloma and carcinoma contain lower levels of lipid peroxidation compared to epidermis from the same mice.

Age Factors↗

Preferential incorporation of docosahexaenoic acid into nonphosphorus lipids and phosphatidylethanolamine protects rats from dietary DHA-stimulated lipid peroxidation.

In a previous study, we found that dietary docosahexaenoic acid (DHA)-stimulated tissue lipid peroxide formation was suppressed to a lesser extent than expected from the peroxidizability index of tissue total lipids. This suppression was presumed to be potentiated by mechanisms other than the lipid peroxide-scavenging system. In this study, we focused primarily on the incorporation of DHA into tissue nonphosphorus lipids and phospholipid species. DHA and different levels of dietary vitamin E (VE; 7.5, 54, 134 and 402 mg/kg of diet) were fed to rats for 32 d. In rats with poor VE status, liver chemiluminescence intensity and kidney and testis thiobarbituric acid (TBA) values correlated with the tissue's peroxidizability index. In rats with normal VE nutriture, liver lipid peroxide formation was suppressed to a level below that expected from the peroxidizability index, likely because DHA was present in nonphosphorus lipids and utilized preferentially for phosphatidylethanolamine synthesis. In the kidney, differences in the TBA values were associated with differences in the peroxidizability index of total lipids, even in the DHA groups fed VE at higher than normal levels. This may be because the levels of lipid peroxide scavengers were lower than those of liver and because DHA was utilized preferentially for phosphatidylcholine synthesis. In testis, the lipid peroxide levels were not as high as expected from the peroxidizability index, even in rats fed a high DHA diet containing the normal level of VE. This may be because the testis was composed of a high proportion of (n-6) polyunsaturated fatty acids (PUFA), which are low in unsaturation, and thus the proportion of DHA was low. In addition, in testis, VE and ascorbic acid, which act as antioxidants, were retained at higher levels in rats with particularly poor and normal VE nutriture than those of liver and kidney. These results suggest that antioxidant protection against dietary DHA-stimulated lipid peroxidation below the extent expected from the peroxidizability index of tissue total lipids differed from tissue to tissue. The suppression was likely due to not only the lipid peroxide scavenging system but also preferential incorporation of DHA into nonphosphorus lipids and phosphatidylethanolamine, particularly in liver.

Animals↗

Moderate wine consumption protects against hydrogen peroxide-induced DNA damage.

We have tested the hypothesis that moderate wine drinking can protect somatic cells against the DNA-damaging effect of hydrogen peroxide which is an endogenous source of reactive oxygen metabolites. In this preliminary investigation, four male volunteers were placed on a plant-polyphenol-free (PPF) diet to ensure that the wine provided was the only main source of plant phenolic compounds. After 48 h on the PPF diet the volunteers were required to consume 300 ml of red or white wine and blood samples collected 1, 3, 8 and 24 h post-consumption while still on a PPF diet. Plasma was isolated from the blood samples and stored frozen for subsequent assays. In the subsequent assays, fresh lymphocytes from each donor were incubated in their corresponding plasma from the various intervention time-points for 30 min. The capacity of the plasma to prevent damage to DNA in lymphocytes by hydrogen peroxide was assessed using the cytokinesis-block micronucleus technique. The data from this preliminary investigation indicated that there was a strong inhibition (>70%) of hydrogen peroxide-induced micronucleated cells by the plasma samples from the blood collected 1 h after consumption of wine as compared to plasma samples from blood immediately before the consumption of wine. This protective effect was apparent for both red and white wine although statistical significance (P = 0.0068) was achieved only in the white wine intervention. A higher degree of statistical significance (P = 0.0008) was achieved when the data for samples following the consumption of red and white wine were combined. There was no difference in the hydrogen-peroxide-induced micronucleated cell frequency when comparing results immediately before starting on the PPF diet, before consumption of wine, 8 h after or 24 h after wine consumption. The hydrogen peroxide-induced micronucleated cell frequency in cells incubated with plasma from blood collected 3 h after wine consumption was intermediate to that observed for plasma after 1 and 8 h after wine intake. The protective effect of plasma against DNA damage cannot be readily explained by the red wine content of phenolic compounds because results for red wine were similar to those for white wine even though white wine had a much lower level of total polyphenols. A possible explanation could be that alcohol, glycerol and ascorbate in wine together with specific wine phenolic compounds that are also equally present in red and white wine (e.g. hydroxycinnamates) may have contributed to the observed protection of nuclear material from hydrogen peroxide-derived reactive oxygen metabolites. This explanation is supported by data from in vitro experiments showing that incubation of lymphocytes either with alcohol or wine stripped of phenolic compounds resulted in a statistically significant (P < 0.05) dose-related reduction (up to 87% reduction) in hydrogen peroxide-induced micronucleated cell frequency.

Adult↗

The effectiveness of Salmonella strains TA100, TA102 and TA104 for detecting mutagenicity of some aldehydes and peroxides.

Several aldehydes and peroxides were tested for mutagenicity using Salmonella typhimurium tester strains TA97a, TA100, TA102 and TA104, in the presence and absence of Aroclor-induced liver S9 mix from F344 rats and B6C3F1 mice, in either preincubation or vapour phase protocols. Some chemicals were tested in additional Salmonella strains. Benzaldehyde, butyraldehyde, benzoyl peroxide, 4-chlorobenzaldehyde, isobutyraldehyde, propionaldehyde and veratraldehyde were non-mutagenic. Acetaldehyde and dicumyl peroxide gave inconsistent results and furfural gave equivocal responses in TA100 and TA104. Cumene hydroperoxide, formaldehyde and glutaraldehyde were mutagenic in TA100, TA102 and TA104. trans-Cinnamaldehyde exhibited a weak mutagenic response in TA100 with mouse liver S9 only. 2,4,5-Trimethoxybenzaldehyde was mutagenic only in strain TA1538 with rat liver S9. With the exception of butanone peroxide, which was mutagenic only in TA104, all chemicals mutagenic in strains TA102 and/or TA104 were also mutagenic in TA100. The data do not, therefore, support the preferential use of strains TA102 and TA104 for screening aldehydes and peroxides for mutagenicity. For a number of these chemicals the advantages of using TA102 or TA104 was in the increased responses compared with those obtained with TA100. Two of the four peroxides were mutagenic and one of these was mutagenic only with TA104. This suggests that strains TA102 and TA104 be used if peroxides are not mutagenic in TA100 or TA97.

Aldehydes↗

Effect of aluminum ion on Fe(2+)-induced lipid peroxidation in phospholipid liposomes under acidic conditions.

The effects of Al3+ on Fe(2+)-induced lipid peroxidation in phospholipid liposomes consisting of phosphatidylcholine (PC) and phosphatidylserine (PS) were examined under acidic conditions. The stimulatory effect of Al3+ on Fe(2+)-induced lipid peroxidation in the liposomes showed a biphasic response against pH variation, and the maximum stimulation was observed around pH 6.0. In addition, it was found that the stimulatory effect of Al3+ on the lipid peroxidation was dependent on the proportion of PS in the liposomes. On the other hand, the lipid peroxidation in PC liposomes was not stimulated by the addition of Al3+. From these findings, it is suggested that the Al3+ effect on Fe(2+)-induced lipid peroxidation under acidic conditions is largely dependent on the phospholipid composition. Trivalent cations such as Tb3+ and Ga3+ also stimulated Fe(2+)-induced lipid peroxidation in PC/PS liposomes under acidic conditions, but divalent cations (Zn2+ and Mn2+) showed no stimulatory effect. The extents of Fe2+ disappearance and Fe3+ formation during the reaction were enhanced by the addition of Al3+ or Ga2+, but Tb3+ had no effect on Fe2+ disappearance. The results with 1,6-diphenyl-1,3,5-hexatriene (DPH) showed that the fluorescence anisotropy of DPH-labeled PC/PS liposomes under acidic conditions was increased by the addition of Al3+. Furthermore, there is a relation between the extents of the fluorescence anisotropy of the complex and TBARS production. In contrast, the fluorescence anisotropy of DPH molecules embedded in PC liposomes was not changed by the addition of Al3+. Based on these results, a possible mechanism of the stimulatory effect of Al3+ on Fe(2+)-induced lipid peroxidation under acidic conditions is discussed.

Aluminum↗

Role of apoplastic ascorbate and hydrogen peroxide in the control of cell growth in pine hypocotyls.

The growth cessation of plant axis has been related with the formation of diphenyl bridges among the pectic components of the cell wall caused by the action of apoplastic peroxidases using hydrogen peroxide as electron acceptor. The formation of diphenyl bridges is prevented by the presence of ascorbate in the apoplastic fluid which acts as a hydrogen peroxide scavenger. The current work focuses on the role of the apoplastic ascorbate and hydrogen peroxide in the cell growth. The addition of hydrogen peroxide caused an inhibition of the auxin-induced growth as well as a significant decrease in the cell wall creep induced by acid-pH solutions. The hydrogen peroxide content in apoplastic fluid increased with the hypocotyl age and along the hypocotyl axis of 10-day-old pine seedlings, as the growth capacity decreased. On the other hand, the ascorbate content in the apoplastic fluid decreased with the hypocotyl age and along the hypocotyl axis of 10-day-old seedlings. A very significant correlation between the hydrogen peroxide apoplastic level and the growth rate as well as between the ascorbate/hydrogen peroxide molar ratio and the growth rate of hypocotyls have been found suggesting that the redox state is the main factor controlling the cell wall stiffening mechanism and thus growth in pine hypocotyls.

Aging↗

Glutathione correlates with lipid peroxidation in liver mitochondria of triiodothyronine-injected hypophysectomized rats.

The temporal relationship of changes in state 3 respiration, lipid peroxidation, and glutathione (GSH) content was investigated in liver mitochondria of hypophysectomized rats after an injection of 3,3',5-triiodo-L-thyronine (T3). Lipid peroxidation induced by ADP/Fe3+/NADPH was determined by the amount of malondialdehyde formed. Hypophysectomy decreased respiration and lipid peroxidation (from 19.88 +/- 3.04 to 14.19 +/- 1.14 nmol malondialdehyde.mg protein-1.10 min-1) but increased GSH content (from 7.06 +/- 2.08 to 12.46 +/- 3.58 nmol/mg protein). Daily injections of a low dose (5 micrograms/100 g) of T3 for 7 days restored the parameters. Time course (up to 96 h) of these changes was followed after one injection of a moderate (100 micrograms/100 g) and high (1000 micrograms/100 g) dose of the hormone. Respiration showed a significant increase at 24 h and declined slightly at 96 h. There was a slow loss of respiratory control ratio after 24 h. Lipid peroxidation remained unchanged at 24 h and showed a gradual increase, becoming significantly higher at 72-96 h depending on the hormone dosage. Changes in GSH content followed a time course similar to that of lipid peroxidation except that it showed a decrease instead of an increase. There was a high degree of inverse linear correlation between lipid peroxidation and GSH (correlation coefficient = 0.95). Because GSH is required for detoxification of hydroperoxides generated by the respiratory chain, it is suggested that lipid peroxidation may play a major role in the modulation of intramitochondrial GSH.

Animals↗

Mechanism of peroxide-induced cellular injury in cultured adult cardiac myocytes.

Reactive oxygen species contribute to the tissue injury seen after reperfusion of ischemic myocardium. We propose that toxicity originates from the effect that mitochondrial peroxide metabolism has on substrate entry into oxidative pathways. To support our contention, cultured adult rat cardiomyocytes were incubated with physiological concentrations of peroxide. The cellular extract and incubation medium were analyzed for adenine nucleotides and purines by reverse-phase high-pressure liquid chromatography. Cellular glutathione efflux was determined by enzymatic analysis of the incubation medium. Pyruvate dehydrogenase (PDH) activity was determined in the cultured myocytes as well as in freshly isolated cardiac mitochondria using [1-C14]pyruvate. Extracellular glutathione rose 3.3-fold in response to small doses of peroxide (approximately 108 nmol/mg protein). Likewise, small quantities of peroxide reduced total cellular adenine nucleotides to 50-60% of control values with only a modest (0.95-0.91) reduction in energy charge [ATP + 1/2 ADP)/(ATP + ADP + AMP]. Peroxide-treated myocytes selectively release inosine and adenosine, as only these two purine degradation products were detected in the incubation medium. The most dramatic response was a peroxide dose-dependent inhibition of PDH activity in cultured myocytes as well as freshly isolated mitochondria; just 65 and 30 nmol peroxide/mg protein induced a 50% reduction in cellular and mitochondrial PDH activity, respectively. In conclusion, physiological quantities of peroxide potently inhibit PDH in cultured cardiomyocytes and isolated cardiac mitochondria. PDH inhibition blocks the aerobic oxidation of glucose and inhibits the oxidative phosphorylation of ADP, which in turn leads to cellular adenine nucleotide degradation.

Adenine Nucleotides↗

Early hydrogen peroxide-induced pulmonary endothelial cell dysfunction: detection and prevention.

OBJECTIVES: To determine a) whether hydrogen peroxide-induced, early lung endothelial cell dysfunction can be detected in an isolated, perfused, rat lung model; and b) whether the organic phosphothioate N-(2-mercaptoethyl)-1,3-propanediamine, which protects cells in culture against hydrogen peroxide-mediated damage, can exert the same protection in this model. DESIGN: Intervention study; before-after trial. SETTING: Research laboratory. MODEL: Isolated, perfused, rat lung model. INTERVENTION: Continuous hydrogen peroxide infusion at increasing concentrations and infusion times, preceded or not by a N-(2-mercaptoethyl)-1,3-propanediamine infusion. MEASUREMENTS AND MAIN RESULTS: Early pulmonary endothelial cell alterations, assessed by the lung extraction (% extraction) of 123I-metaiodobenzylguanidine. Permeability edema by % extraction of 125I-human serum albumin and the lung dry-to-wet weight ratio. Control experiments: % extraction-123I-metaiodobenzylguanidine: 21.7 +/- 3.8% (n = 7). With increasing concentrations of hydrogen peroxide (0.025, 0.125, 0.5, and 2 mmol), % extraction-123I-metaiodobenzylguanidine was progressively depressed (n = 28, ANOVA, p < .05), significantly decreased from controls at 2 mmol (10.2 +/- 5.0%, n = 7, p < .05). When the 2-mmol hydrogen peroxide infusion was preceded by the N-(2-mercaptoethyl)-1,3-propanediamine (2 mmol) infusion, % extraction-123I-metaiodobenzylguanidine (19.9 +/- 2.9%, n = 5) was not significantly different from controls (n = 7) and was significantly greater than after the 2-mmol hydrogen peroxide infusion alone (8.7 +/- 7.4%, p < .05, n = 8). In all experiments, % extraction of human serum albumin ratio and dry-to-wet weight ratio were not significantly different from that of controls. CONCLUSIONS: a) Hydrogen peroxide-induced lung endothelial cell dysfunction was detected at an early stage, before any permeability defect appeared; b) N-(2-mercaptoethyl)-1,3-propanediamine protected against such damage.

Amifostine↗

Plasma hydrogen peroxide production in hypertensives and normotensive subjects at genetic risk of hypertension.

BACKGROUND: Oxygen free radicals may play roles in hypertension both in arteriolar constriction and in formation of lesions. OBJECTIVE: To quantify free radical production in blood plasma of genetic hypertensives. DESIGN: Hydrogen peroxide levels were measured, because it is one of the most stable reactive oxygen species. METHODS: An electrode technique was used to determine plasma hydrogen peroxide levels after blockade of endogenous catalase with sodium azide. This method was validated by an independent spectrophotometric technique. RESULTS: Members of the essential hypertensive group (n=21) had higher plasma hydrogen peroxide levels (3.16+/-0.14 versus 2.50+/-0.16 micromol/l, P=0.005) than did members of the normotensive group (n=29). Furthermore, within the normotensive group, those with a family history of hypertension (n=15) exhibited higher hydrogen peroxide levels (2.83+/-0.27 versus 2.14< or =0.13 micromol/l, P=0.03) than did those without such a family history (n=14). Plasma hydrogen peroxide levels in these 50 subjects were correlated to their mean arterial pressures (r=0.54, P < 0.001). When hypertensives were grouped with normotensives without a family history of hypertension, the correlation improved (r=0.70, P< 0.001). Statistical analysis (two-way analysis of variance) revealed that a family history of hypertension was a better predictor of plasma hydrogen peroxide production than was blood pressure status (P=0.003 versus P=0.093). Further investigations showed that superoxide is produced in plasma and that one of its sources is xanthine oxidase. CONCLUSIONS: Hydrogen peroxide is produced in blood plasma and elevation of its level could constitute a pathogenic factor in vascular organ damage attendant upon systemic hypertension.

Adult↗

Mixed tocopherols have a stronger inhibitory effect on lipid peroxidation than alpha-tocopherol alone.

Intake of vitamin E with food (mixed tocopherols) has been found to counteract the development of atherosclerotic cardiovascular disease, whereas intake of large amounts of pure alpha-tocopherol has shown only a slight or no effect in clinical studies. This study was designed to investigate the effects of alpha-tocopherol alone and a mixed tocopherol preparation (gamma-, delta-, and alpha-tocopherol) on hydrogen peroxide-induced lipid peroxidation in human erythrocytes. Erythrocytes were incubated with different concentrations of alpha-tocopherol or mixed tocopherols and then exposed to hydrogen peroxide. Tocopherol levels and malondialdehyde-thiobarbituric acid-reactive substances were determined by high-performance liquid chromatography and fatty acids by gas chromatography. Incubation of erythrocytes with tocopherols (30-120 microM) increased the tocopherol level in a concentration-dependent manner. The uptake of gamma- and delta-tocopherols was much higher than that of alpha-tocopherol. Hydrogen peroxide strongly increased lipid peroxidation and decreased polyunsaturated fatty acids in erythrocytes. Both alpha-tocopherol and the tocopherol mixture protected the cells from lipid peroxidation, the mixture being much more potent than alpha-tocopherol alone. This study indicates that a mixture of tocopherols has a stronger inhibitory effect on lipid peroxidation induced in human erythrocytes than alpha-tocopherol alone, due to higher uptake of gamma- and delta-tocopherol in the cells.

Adult↗

Cyclosporine nephrotoxicity: attenuation by an antioxidant-inhibitor of lipid peroxidation in vitro and in vivo.

The exact mechanisms by which cyclosporine (CsA) causes renal injury is not known. The possibility that reactive oxygen species (ROS) may play a role, since CsA induces renal microsomal lipid peroxidation and reduces glutathione levels, was investigated by examining whether administration of an antioxidant attenuates CsA-induced nephrotoxicity. One of three groups of uninephrectomized rats received vehicles, another CsA 25 mg/kg/day and the third, CsA plus antioxidant lazaroid (U-74389 G) 20 mg/kg/day, for 8 weeks. CsA-induced functional and structural derangements were accompanied by a significant induction of renal cortical lipid peroxidation (thiobarbituric acid reactive substances and conjugated diene). Administration of lazaroid significantly suppressed CsA-induced lipid peroxidation and provided significant functional and structural protection. That lazaroid affords renal functional protection against CsA in the rat was again demonstrable in a crossover study. To examine the relation between CsA nephrotoxicity and lipid peroxidation, cell culture studies were undertaken. CsA induced renal epithelial (LLC-PK1) cell injury (LDH release) as well as lipid peroxidation and degradation (prelabeled 3H arachidonic acid release). Lazaroid prevented CsA-induced cell injury and limited lipid alterations. These new findings--that an antioxidant inhibitor of lipid peroxidation limits CsA-induced renal toxicity in vitro and in vivo--suggest a pathogenic role for ROS-mediated lipid peroxidation in CsA-induced renal toxicity. Antioxidant therapy may minimize CsA-induced renal toxicity in humans.

Animals↗

The use of hydrogen peroxide for achieving dermal hemostasis after burn excision in a patient with platelet dysfunction.

As a topical hemostatic agent, hydrogen peroxide (3%) has been applied to tangential excisions of burn patients. Hydrogen peroxide provides an adjunct to topical epinephrine for hemostasis and clinically has been most useful in patients with known platelet dysfunction. Hydrogen peroxide should be irrigated from the wound bed before the placement of skin grafting because of its potential to inhibit wound healing. In our experience, hydrogen peroxide (3%) soaks have not resulted in complications related to corrosive damage, oxygen gas formation, or lipid peroxidation that are associated with high concentrations of hydrogen peroxide (30%). We also have experienced excellent skin graft take at the wound bed after using hydrogen peroxide.

Aged, 80 and over↗

Inducibility of the response of yeast cells to peroxide stress.

Exponential phase cells of the yeast, Saccharomyces cerevisiae when treated with a non-lethal concentration of hydrogen peroxide (H2O2; 0.2mM) for 60 min adapted to become resistant to the lethal effects of a higher dose of H2O2 (2mM). From studies using cycloheximide to inhibit protein synthesis it appears that protein synthesis is required for maximal induction of resistance but that some degree of protection from the lethal effects of peroxide can be acquired in the absence of protein synthesis. Treatment of cells with 50 micrograms cycloheximide ml-1 alone lead to them acquiring some protection from peroxide. Cells subjected to heat shock became more resistant to 2mM-H2O2; however, peroxide pretreatment did not confer thermotolerance. L-[35S]Methionine labelling of cells subjected to 0.2 mM-H2O2 stress showed that synthesis of at least ten polypeptides was induced by peroxide treatment. Some of these were also induced in cells subjected to heat shock (23 to 37 degrees C shift) but the synthesis of at least four polypeptides (45, 39.5, 38 and 24 kDa) was unique to peroxide-stressed cells. Resistance to peroxide was also inducible in an isogenic petite and an isogenic strain with a mutation in the HAP1 gene, indicating that the adaptive response does not require functional mitochondria.

Adaptation, Physiological↗

The effect of hydrogen peroxide on spores of Clostridium bifermentans.

The effect of hydrogen peroxide on the germination, colony formation and structure of spores of Clostridium bifermentans was examined. Treatment with 0.35 M-hydrogen peroxide increased the germination rate at 25 degrees C but increasing the temperature or concentration of hydrogen peroxide decreased both the germination rate and colony formation. The presence of Cu2+ increased the lethal effect of hydrogen peroxide on colony formation as much as 3000-fold. Pre-incubation of spores with Cu2+ before treatment with hydrogen peroxide produced a similar increase, but this could be eliminated by washing the spores with dilute spores--apparently from the coat--and treatment with dithiothreitol, which also removes spore-coat protein, increased the lethal effect of hydrogen peroxide 500-fold, suggesting that spore-coat protein has a protective effect against hydrogen peroxide.

Clostridium↗

Peroxide sensitivity of cold-shocked Salmonella typhimurium and Escherichia coli and its relationship to minimal medium recovery.

Cold-shocked Salmonella typhimurium displayed minimal medium recovery (MMR), viable counts on M9 minimal agar being much higher than those on tryptone soya yeast extract agar (TSYA). The addition of catalase to TSYA restored counts to the level found on M9 agar. Peroxide concentrations between 12 and 30 mumol/l were measured in TSYB but none was detected in M9 medium. Cold-shocked cells were sensitive to reagent hydrogen peroxide at a concentration similar to that found in TSYB. The minimal medium recovery phenomenon of cold-shocked cells is thus a manifestation of peroxide sensitivity. Changing the composition of growth media affected both cellular catalase activity and the magnitude of the MMR effect but the two properties were not directly related. Factors additional to cellular catalase activity must therefore affect susceptibility to peroxide following cold shock. Mutational loss of catalase, exonuclease III or recA-dependent DNA repair functions all increased the sensitivity of cold-shocked Escherichia coli to the inhibitory effects of peroxide present in rich medium. The peroxide resistant fraction of a cold-shocked population of Salm. typhimurium (i.e. those cells able to grow on TSYA) was more resistant to gamma radiation than the population as a whole. Cold shock thus sensitizes cells to more than one form of oxidative stress. Prior exposure of growing cells to 30 mumol/l hydrogen peroxide abolished their sensitivity to rich medium following cold shock implying that Salm. typhimurium contains an inducible system protecting against oxidative stress.

Catalase↗

Activation of the redox sensor Pap1 by hydrogen peroxide requires modulation of the intracellular oxidant concentration.

The transcription factor Pap1 and the MAP kinase Sty1 are key regulators of hydrogen peroxide-induced responses in Schizosaccharomyces pombe. Pap1 can be activated quickly at low, but not high, hydrogen peroxide concentrations. The MAP kinase Sty1 has been reported to participate in Pap1 activation by the oxidant. Here, we provide biochemical and genetic evidence for the in vivo formation of a hydrogen peroxide-induced disulphide bond in Pap1, which precedes the rapid and reversible nuclear accumulation of the transcription factor. We show that activation of the Sty1 cascade before the oxidative insult, or overexpression of the Sty1-regulated genes ctt1 (encoding catalase) or gpx1 (encoding glutathione peroxidase), can accelerate Pap1 entry even at high doses of hydrogen peroxide. In fact, the lack of Sty1 impedes Pap1 nuclear localization, but only at high doses of the oxidant. We propose that, whereas low doses of hydrogen peroxide lead directly to Pap1 oxidation-activation, high concentrations of the oxidant initially activate the Sty1 pathway, with the consequent increase in scavenging enzymes, which in turn helps to decompose the excess of hydrogen peroxide and achieve an appropriate concentration for the subsequent activation of Pap1. Our results also suggest that activation of Sty1 at high doses of hydrogen peroxide may also be required to trigger other antioxidant activities such as those reverting the overoxidation of cysteine residues at the Pap1 pathway.

Active Transport, Cell Nucleus↗