Rhodanese Rdl2 produces reactive sulfur species to scavenge hydroxyl radical and protect mitochondria

During aerobic respiration, mitochondria generate superoxide anion (O2&middot−), hydrogen peroxide (H2O2), and hydroxyl radical (HO·), and these reactive oxygen species (ROS) are detrimental to mitochondria. Mitochondrial damage is linked to a broad spectrum of pathologies such as Alzheimer's disease, hemochromatosis, and diabetes. Mitochondria contain several enzymes for rapidly removing superoxide anion and hydrogen peroxide, but how they antagonize HO· is elusive, representing a loophole in the anti-ROS system. Herein, we discovered that Rhodanese 2 (Rdl2) is critical for maintaining the functionality and integrity of mitochondria under sub-lethal ROS stress in Saccharomyces cerevisiae. Rdl2 converts stable sulfur species (thiosulfate and dialkyl polysulfide) to reactive sulfane sulfur including persulfide that protects mitochondrial DNA via scavenging HO·. Surprisingly, hydrogen sulfide (H2S) promotes HO· production through stimulating the Fenton reaction, leading to increased DNA damage. Our study may reveal an ex-ante mean for antagonizing HO·, patching the loophole of the anti-ROS system in mitochondria.

Functional study of Csrbohs in defence response against Xanthomonas citri ssp. citri

NADPH oxidases, encoded by rbohs (respiratory burst oxidase homologues), transfer electrons from NADPH to molecular oxygen (O2) to generate superoxide anion (O2•–), which is the first step in the formation of hydrogen peroxide (H2O2) in the plant–pathogen interaction system. In the present work, six citrus rbohs (Csrbohs) genes were identified in citrus, and their possible involvement in resistance to Xanthomonas citri ssp. citri (Xcc) was examined. Inoculation with Xcc promoted the H2O2 production and induced expression of the Csrbohs, especially CsrbohD. Results showed that CsrbohD was markedly induced in the resistant genotype kumquat ‘Luofu’ [Fortunella margarita (Lour.) Swingle] compared with grapefruit ‘Duncan’ [Citrus paradisi (Linn.) Macf.]. Virus-induced gene silencing (VIGS) of CsrbohD resulted in reduced resistance to Xcc in grapefruit, but not in kumquat. Compared with non-silenced plants, canker-like symptoms were observed earlier, and they were more extensive in the CsrbohD-silenced grapefruit. Silencing of CsrbohD also suppressed the Xcc induced reactive oxygen species (ROS) burst, and resulted in accumulation of more Xcc bacterial colonies. Taken together, these data indicate that CsrbohD promotes resistance to Xcc, especially in grapefruit.

Primary concept of nickel toxicity – an overview

Toxic metals, including excessive levels of essential metals tend to change biological structures and systems into either reversible or irreversible conformations, leading to the derangement of organ functions or ultimate death. Nickel, a known heavy metal is found at very low levels in the environment. Nickel is available in all soil types and meteorites and also erupts from volcanic emissions. In the environment, nickel is principally bound with oxygen or sulfur and forms oxides or sulfides in earth crust. The vast industrial use of nickel during its production, recycling and disposal has led to widespread environmental pollution. Nickel is discharged into the atmosphere either by nickel mining or by various industrial processes, such as power plants or incinerators, rubber and plastic industries, nickel-cadmium battery industries and electroplating industries. The extensive use of nickel in various industries or its occupational exposure is definitely a matter of serious impact on human health. Heavy metals like nickel can produce free radicals from diatomic molecule through the double step process and generate superoxide anion. Further, these superoxide anions come together with protons and facilitate dismutation to form hydrogen peroxide, which is the most important reason behind the nickel-induced pathophysiological changes in living systems. In this review, we address the acute, subchronic and chronic nickel toxicities in both human and experimental animals. We have also discussed nickel-induced genotoxicity, carcinogenicity, immunotoxicity and toxicity in various other metabolically active tissues. This review specifically highlighted nickel-induced oxidative stress and possible cell signaling mechanisms as well.

Generation of superoxide and hydrogen peroxide by side reactions of mitochondrial 2-oxoacid dehydrogenase complexes in isolation and in cells

Mitochondrial 2-oxoacid dehydrogenase complexes oxidize 2-oxoglutarate, pyruvate, branched-chain 2-oxoacids and 2-oxoadipate to the corresponding acyl-CoAs and reduce NAD+ to NADH. The isolated enzyme complexes generate superoxide anion radical or hydrogen peroxide in defined reactions by leaking electrons to oxygen. Studies using isolated mitochondria in media mimicking cytosol suggest that the 2-oxoacid dehydrogenase complexes contribute little to the production of superoxide or hydrogen peroxide relative to other mitochondrial sites at physiological steady states. However, the contributions may increase under pathological conditions, in accordance with the high maximum capacities of superoxide or hydrogen peroxide-generating reactions of the complexes, established in isolated mitochondria. We assess available data on the use of modulations of enzyme activity to infer superoxide or hydrogen peroxide production from particular 2-oxoacid dehydrogenase complexes in cells, and limitations of such methods to discriminate specific superoxide or hydrogen peroxide sources in vivo.

Increased production of superoxide anion contributes to dysfunction of the arteriovenous fistula

Vascular access dysfunction causes morbidity in hemodialysis patients. This study examined the generation and pathobiological significance of superoxide anion in a rat femoral arteriovenous fistula (AVF). One week after AVF creation, there was increased production of superoxide anion accompanied by decreased total superoxide dismutase (SOD) and Cu/Zn SOD activities and induction of the redox-sensitive gene heme oxygenase-1. Immunohistochemical studies of nitrotyrosine formation demonstrated that peroxynitrite, a product of superoxide anion and nitric oxide, was present in increased amounts in endothelial and smooth muscle cells in the AVF. Because uncoupled NOS isoforms generate superoxide anion, and NOS coupling requires tetrahydrobiopterin (BH4) as a cofactor, we assessed NOS uncoupling by determining the ratio of BH4 to dihydrobiopterin (BH2); the BH4-to-BH2 ratio was markedly attenuated in the AVF. Because Src is a vasculopathic signaling species upstream and downstream of superoxide anion, such expression was evaluated; expression of Src and phosphorylated Src was both markedly increased in the AVF. Expression of NADPH oxidase (NOX) 1, NOX2, NOX4, cyclooxygenase (COX) 1, COX2, p47phox, and p67phox was all unchanged, as assessed by Western analyses, thereby suggesting that these proteins may not be involved in increased production of superoxide anion. Finally, administration of tempol, a superoxide anion scavenger, decreased neointima formation in the juxta-anastomotic venous segment and improved AVF blood flow. We conclude that the AVF exhibits increased superoxide anion generation that may reflect the combined effects of decreased scavenging by SOD and increased generation by uncoupled NOS, and that enhanced superoxide anion production promotes juxta-anastomotic stenosis and impairs AVF function.

Hyperhomocysteinemia increases arterial permeability and stiffness in mice

We have reported that hyperhomocysteinemia (HHcy) evoked by folate depletion increases arterial permeability and stiffness in rats and that low folate without HHcy increases arterial permeability in mice. In this study, we hypothesized that HHcy independently increases arterial permeability and stiffness in mice. C57BL/6J mice that received rodent chow and water [control (Con), n = 12] or water supplemented with 0.5% l-methionine (HHcy, n = 12) for 18 ± 3 wk had plasma homocysteine concentrations of 8 ± 1 and 41 ± 1 μM, respectively ( P < 0.05), and similar liver folate (∼12 ± 2 μg folate/g liver). Carotid arterial permeability, assessed as dextran accumulation using quantitative fluorescence microscopy, was greater in HHcy (3.95 ± 0.4 ng·min−1·cm−2) versus Con (2.87 ± 0.41 ng·min−1·cm−2) mice ( P < 0.05). Stress versus strain curves generated using an elastigraph indicated that 1) maximal stress (N/mm2), 2) physiological stiffness (low-strain Young's modulus, mN/mm), and 3) maximal stiffness (high-strain Young's modulus, N/mm) were higher ( P < 0.05) in aortas from HHcy versus Con mice. Thus, chronic HHcy increases arterial permeability and stiffness. Carotid arterial permeability also was assessed in age-matched C57BL/6J mice before and after incubation with 1) xanthine (0.4 mg/ml)/xanthine oxidase (0.2 mg/ml; X/XO) to generate superoxide anion (O2−) or 50 μM dl-homocysteine in the presence of 2) vehicle, 3) 300 μM diethylamine-NONOate (DEANO; a nitric oxide donor), or 4) 10−3M 4,5-dihydroxy-1,3-benzene disulfonic acid (tiron; a nonenzymatic intracellular O2−scavenger). Compared with preincubation values, X/XO and dl-homocysteine increased ( P < 0.05) permeability by 66 ± 11% and 123 ± 8%, respectively. dl-Homocysteine-induced increases in dextran accumulation were blunted ( P < 0.05) by simultaneous incubation with DEANO or tiron. Thus, acute HHcy increases arterial permeability by generating O2−to an extent whereby nitric oxide bioavailability is reduced.

Superoxide Anion and Hydroxyl Radical Release by Collagen-induced Platelet Aggregation - Role of Arachidonic Acid Metabolism

SummaryPrevious study demonstrated that platelets undergoing anoxia-reoxygenation generate superoxide anion (O2 −) and hydroxyl radical (OH°) which in turn contribute to activate arachidonic acid (AA) metabolism. However it has not been clarified if oxygen free radicals (OFRs) are also generated when platelets are aggregated by common agonists. We used two probes, i.e. lucigenin and salicylic acid (SA), to measure platelet release of O2 − and OH°, respectively. Among the agonists used, such as ADP, thrombin and collagen, the release of O2 − and OH° was observed mainly when platelets were stimulated with collagen. Such release was inhibited in platelets pre-treated by aspirin suggesting that AA metabolism was the main source of O2 − and OH° formation. To further analyze this relationship, O2 and OH° formation was measured if other oxidant species, namely O2 − and OH°, contribute to the during AA-stimulated platelet aggregation (PA); we observed that O − and OH° release were dependent upon AA concentration. Furthermore, we found that the incubation of platelets with AACOCF3, a potent inhibitor of cytosolic phospholipase A2, inhibited collagen-induced platelet O− and OH° release. The incubation of platelets with salicylic acid or ascorbic acid, which blunt OH° and O2 − respectively, inhibited both collagen-induced platelet aggregation and AA-release. This study demonstrated that collagen-induced platelet aggregation is associated with O2 − and OH° formation, which is dependent upon AA release and analyzed if O2 − and OH° are released during aggregation induced by metabolism.