Related Reactive Species Research Articles

Mitochondrial Thiols in Antioxidant Protection and Redox Signaling: Distinct Roles for Glutathionylation and Other Thiol Modifications

The mitochondrial matrix contains much of the machinery at the heart of metabolism. This compartment is also exposed to a high and continual flux of superoxide, hydrogen peroxide, and related reactive species. To protect mitochondria from these sources of oxidative damage, there is an integrated set of thiol systems within the matrix comprising the thioredoxin/peroxiredoxin/methionine sulfoxide reductase pathways and the glutathione/glutathione peroxidase/glutathione-S-transferase/glutaredoxin pathways that in conjunction with protein thiols prevent much of this oxidative damage. In addition, the changes in the redox state of many components of these mitochondrial thiol systems may transduce and relay redox signals within and through the mitochondrial matrix to modulate the activity of biochemical processes. Here, mitochondrial thiol systems are reviewed, and areas of uncertainty are pointed out, focusing on recent developments in our understanding of their roles. The areas of particular focus are on the multiple, overlapping roles of mitochondrial thiols and on understanding how these thiols contribute to both antioxidant defenses and redox signaling. Recent technical progress in the identification and quantification of thiol modifications by redox proteomics means that many of the questions raised about the multiple roles of mitochondrial thiols can now be addressed.

Peroxynitrite—An ugly biofactor?

Cellular damage occurring under oxidative conditions has been ascribed mainly to the formation of peroxynitrite (ONOOH/ONOO(-)) that originates from the reaction of NO(*) with O(2) (*-). The detrimental effects of peroxynitrite are exacerbated by the reaction with CO(2) that leads to ONOOC(O)O(-), which further decays to the strong oxidant radicals NO(2) (*) and CO(3) (*-). The reaction with CO(2), however, may redirect peroxynitrite specificity. An excessive formation of peroxynitrite represents an important mechanism contributing to the DNA damage, the inactivation of metabolic enzymes, ionic pumps, and structural proteins, and the disruption of cell membranes. Because of its ability to oxidize biomolecules, peroxynitrite is implicated in an increasing list of diseases, including neurodegenerative and cardiovascular disorders, inflammation, pain, autoimmunity, cancer, and aging. However, peroxynitrite displays also protective activities: (i) at high concentrations, it shows anti-viral, anti-microbial, and anti-parasitic actions; and (ii) at low concentrations, it stimulates protective mechanisms in the cardiovascular, nervous, and respiratory systems. The detrimental effects of peroxynitrite and related reactive species are impaired by (pseudo-) enzymatic systems, mainly represented by heme-proteins (e.g., hemoglobin and myoglobin). Here, we report biochemical aspects of peroxynitrite actions being at the root of its biomedical effects.

Fungal responses to reactive oxygen species.

Reactive oxygen species (ROS) such as hydrogen peroxide, produced externally or during normal metabolism, can damage different cell components and usually trigger a counteracting antioxidant response. The fact that animals and humans utilize ROS and related nitrogen reactive species to prevent fungal infection has generated great interest in defining the components of the antioxidant response and studying their role as virulence determinants in fungi. Here we review the role of specific enzyme and non-enzyme mediated antioxidant mechanisms in virulence, as well as the signal transduction mechanisms that fungal cells use to perceive high ROS levels and induce gene expression. We focus on Schizosaccharomyces pombe antioxidant responses, which involve a prokaryotic-type multistep phosphorelay coupled to a stress-response MAP kinase pathway and an AP-1 type transcription factor, in relation to homologous mechanisms in Aspergillus nidulans and the human pathogen A. fumigatus. Compared to S. pombe and other unicellular fungi, filamentous fungi have additional mechanisms to handle ROS, such as the presence of a larger number of phosphorelay sensor kinases, antioxidant enzymes and secondary metabolites with antioxidant functions. In addition, filamentous fungi have enzymes like the NADPH oxidases, which regulate multicellular development through ROS production and therefore, offer a unique opportunity to study the interplay between ROS production, perception and detoxification, and the role of these processes in cell differentiation and pathogenesis.

Antioxidant Potential of Black Tea Against 7,12-Dimethylbenz(a)anthracene- Induced Oxidative Stress in Swiss Albino Mice

Oxygen free radicals and related reactive species have been implicated in the etiology of many diseases, such as atherosclerosis, neurodegenerative disorders, and cancer. Antioxidant enzymes exist in cells to protect against the effects of these free radicals and other oxygen-derived species, which are produced during the oxidative stress. Tea (Camellia sinensis) is the most commonly consumed beverage worldwide. Both green and black tea are known to possess many pharmacological properties, including antioxidant, antipyretic, antibacterial, and antineoplastic effects. In the present study, the preventive effects of black tea extract (BTE) was evaluated in Swiss albino mice against 7,12-dimethyl benz(a)anthracene (DMBA)-induced oxidative stress. The animals were given 0.5%, 1%, and 1.5% BTE as the sole source of drinking solution for 1 week prior to the administration of DMBA, which was given orally as a single dose of 50 mg/kg body weight. At the end of the study period, the liver, kidney, and prostate tissues were dissected out for the determination of antioxidant enzyme levels (catalase, superoxide dismutase, glutathione reductase, glutathione-S-transferase), and lipid peroxidation. A dose-dependent protective effect of BTE against DMBA-induced depletion in enzymes activity was observed in all three tissues examined. Similarly, a significant dose-dependent inhibition of the lipid peroxidation caused by DMBA was observed in the BTE-administered animals in all three tissues examined. Our results revealed that BTE provides protection against oxidative damage induced by xenobiotics.

Control of plasma chemistry for preparing highly stabilized amorphous silicon at high growth rate

Abstract Recent progress in the growth process of hydrogenated amorphous silicon (a-Si:H) from a silane glow-discharge plasma is reviewed, being focused on the improvement of photo-induced degradation properties in a-Si:H deposited at high growth rate. Contribution of higher-silane related reactive species (HSRS) during film growth is suggested as a key event to increase the degree of photo-induced degradation in the resulting a-Si:H through an incorporation of excess Si–H 2 bonds in the network as far as the substrate temperature is kept constant. The contribution ratio of HSRS to film growth is derived using a couple of gas-phase reaction-rate equations, which shows strong dependence on electron temperature, hydrogen-dilution ratio, gas-flow rate, and gas temperature in the plasma. Various methods are adopted to reduce the contribution ratio of HSRS during film growth and the validity of theoretical approach is confirmed experimentally. Finally, stabilized conversion efficiency of 8.2% is demonstrated in the a-Si:H-based single junction solar cell with high deposition rate of 2.0 nm/s.

Prostaglandins and Nitric Oxide Mediate Superoxide-induced Myocardial Contractile Dysfunction in Isolated Rat Hearts

Oxygen-derived free radicals have been implicated in the pathogenesis of myocardial injury. We therefore investigated the pathophysiology of myocardial injury induced in isolated rat hearts by perfusion with superoxide radical generated by reacting 2.5 mmol/l purine, 0.03 U/ml xanthine oxidase and 300 U/ml catalase. Perfusion with superoxide significantly (P<0.05) increased left ventricular end-diastolic pressure within 15 to 20 min. During the same time period, heart rate and left-ventricular developed pressure significantly declined to 44.6±8.2% and 31.0±4.9% of control, respectively. Superoxide perfusion also significantly increased production of prostaglandins, nitric oxide (detected as nitrites) and peroxynitrite (detected immunohistochemically as nitrotyrosine). NG-nitro-l -arginine (100 μ mol/l), a nitric oxide synthase inhibitor, attenuated superoxide-induced generation of peroxynitrite, increased synthesis of prostacyclin, and partially blocked myocardial dysfunction, as did 1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one (30 μ mol/l), a selective inhibitor of soluble guanylate cyclase, and ONO-3708 (10 μ mol/l), a selective thromboxane A2receptor antagonist. In contrast, nitroglycerin (4 μ mol/l) and sodium nitroprusside (1 μ mol/l) each exacerbated the superoxide-induced myocardial dysfunction. These results suggest that nitric oxide and related reactive species contribute to myocardial injury induced by superoxide. Moreover, they suggest that oxidative stress can be delayed or inhibited by reducing levels of nitric oxide, by inhibiting soluble guanylate cyclase, and by blocking thromboxane/prostaglandin receptors.

Antioxidant regulation of genes encoding enzymes that detoxify xenobiotics and carcinogens

Antioxidants are substances that delay or prevent the oxidation of cellular oxidizable substrates. The various antioxidants exert their effect by scavenging superoxide or by activating a battery of detoxifying/defensive proteins. In this chapter, we have focused on the mechanisms by which antioxidants induce gene expression. Many xenobiotics (e.g., beta-naphthoflavone) activate genes similar to those activated by antioxidants. The promoters of these genes contain a common cis-element, termed the antioxidant response element (ARE), which contains two TRE (TPA response element) or TRE-like elements followed by GC box. Mutational studies have identified GTGAC***GC as the core of the ARE sequence. Many transcription factors, including Nrf, Jun, Fos, Fra, Maf, YABP, ARE-BP1, Ah (aromatic hydrocarbon) receptor, and estrogen receptor bind to the ARE from the various genes. Among these factors, Nrf-Jun heterodimers positively regulate ARE-mediated expression and induction of genes in response to antioxidants and xenobiotics. This Nrf-Jun heterodimerization and binding to the ARE requires unknown cytosolic factors. The mechanism of signal transduction from antioxidants and xenobiotics includes several steps: (1) Antioxidants and xenobiotics undergo metabolism to generate superoxide and related reactive species, leading to the generation of a signal to activate expression of detoxifying/defensive genes. (2) The generation of superoxide and related reactive species is followed by activation of yet to be identified cytosolic factors by unknown mechanism(s). (3). Activated cytosolic factors catalyze modification of Nrf and/or Jun proteins, which bind to the ARE in promoters of the various detoxifying/defensive genes. (4) The transcription of genes encoding detoxifying/defensive proteins is increased. The unknown cytosolic factors are significant molecules because they represent the oxidative sensors within the cells. Identification of the cytosolic factors will be of considerable importance in the field of antioxidants and gene regulation research. Future studies will also be required to completely understand the molecular mechanism of signal transduction from antioxidants and xenobiotics to Nrf-Jun. In addition to the Nrf-Jun pathway, mammalian cells also contain other pathways that activate gene expression in response to oxidative stress. These include NF-KB-, HIF-1-, Mac-1-, and SRF-mediated pathways. It is expected that collectively these pathways increase transcription of more than four dozen genes to protect cells against oxidative stress.

New functions for the ancient globin family: bacterial responses to nitric oxide and nitrosative stress.

Globin-like oxygen-binding proteins occur in bacteria, yeasts and other fungi, and protozoa. The simplest contain protohaem as sole prosthetic group, but show considerable variation in their similarity to the classical animal globins and plant globins. Flavohaemoglobins comprise a haem domain homologous to classical globins and a ferredoxin-NADP+ reductase (FNR)-like domain that converts the globin into an NAD(P)H-oxidizing protein with diverse reductase activities. In Escherichia coli, the prototype flavohaemoglobin (Hmp) is clearly involved in responses to nitric oxide (NO) and nitrosative stress: (i) the structural gene hmp is upregulated by NO and nitrosating agents; (ii) purified Hmp binds NO avidly, but also converts it to nitrate (aerobically) or nitrous oxide (anaerobically); (iii) hmp mutants are hypersensitive to NO and nitrosative stresses. Here, we review recent advances in E. coli and the growing number of microbes in which globins are known, draw particular attention to the essential chemistry of NO and related reactive species and their interactions with globins, and suggest that microbial globins have additional functions unrelated to 'NO' stresses.

Oxidative Stress in Brain Ischemia

Brain ischemia initiates a complex cascade of metabolic events, several of which involve the generation of nitrogen and oxygen free radicals. These free radicals and related reactive chemical species mediate much of damage that occurs after transient brain ischemia, and in the penumbral region of infarcts caused by permanent ischemia. Nitric oxide, a water- and lipid-soluble free radical, is generated by the action of nitric oxide synthases. Ischemia causes a surge in nitric oxide synthase 1 (NOS 1) activity in neurons and, possibly, glia, increased NOS 3 activity in vascular endothelium, and later an increase in NOS 2 activity in a range of cells including infiltrating neutrophils and macrophages, activated microglia and astrocytes. The effects of ischemia on the activity of NOS 1, a Ca2+-dependent enzyme, are thought to be secondary to reversal of glutamate reuptake at synapses, activation of NMDA receptors, and resulting elevation of intracellular Ca2+. The up-regulation of NOS 2 activity is mediated by transcriptional inducers. In the context of brain ischemia, the activity of NOS 1 and NOS 2 is broadly deleterious, and their inhibition or inactivation is neuroprotective. However, the production of nitric oxide in blood vessels by NOS 3, which, like NOS 1, is Ca2+-dependent, causes vasodilatation and improves blood flow in the penumbral region of brain infarcts. In addition to causing the synthesis of nitric oxide, brain ischemia leads to the generation of superoxide, through the action of nitric oxide synthases, xanthine oxidase, leakage from the mitochondrial electron transport chain, and other mechanisms. Nitric oxide and superoxide are themselves highly reactive but can also combine to form a highly toxic anion, peroxynitrite. The toxicity of the free radicals and peroxynitrite results from their modification of macromolecules, especially DNA, and from the resulting induction of apoptotic and necrotic pathways. The mode of cell death that prevails probably depends on the severity and precise nature of the ischemic injury. Recent studies have emphasized the role of peroxynitrite in causing single-strand breaks in DNA, which activate the DNA repair protein poly(ADP-ribose) polymerase (PARP). This catalyzes the cleavage and thereby the consumption of NAD+, the source of energy for many vital cellular processes. Over-activation of PARP, with resulting depletion of NAD+, has been shown to make a major contribution to brain damage after transient focal ischemia in experimental animals. Neuronal accumulation of poly(ADP-ribose), the end-product of PARP activity has been demonstrated after brain ischemia in man. Several therapeutic strategies have been used to try to prevent oxidative damage and its consequences after brain ischemia in man. Although some of the drugs used in early studies were ineffective or had unacceptable side effects, other trials with antioxidant drugs have proven highly encouraging. The findings in recent animal studies are likely to lead to a range of further pharmacological strategies to limit brain injury in stroke patients.

Simulating transport and removal of xylene during remediation of a sandy aquifer

Xylene, originating from a spill, is present both as a nonaqueous-phase liquid (NAPL) at residual saturation near the water table, and as a dissolved groundwater component contaminating a sandy aquifer beneath an abandoned refinery. Three remediation wells are in operation on the site to prevent further xylene migration in the groundwater. Field observations indicate that microbially-mediated xylene degradation and oxygen and nitrate reduction occur in the aquifer. To realistically simulate dissolved xylene migration at this site, a three-dimensional numerical flow and transport model incorporating biochemical multispecies interactions and xylene dissolution from the NAPL has been developed. In the calibration process the variable contact area between the NAPL and groundwater and the vertical transverse dispersivity were identified as crucial parameters controlling the fate of xylene. The simultaneous modeling of a whole set of related reactive species made it also possible to quantify the observed biodegradation. Results indicate that it contributes in the same order of magnitude to total xylene removal than does extraction by the wells. The calibrated model will be used to assist in the design of an in situ bioremediation scheme, where biodegradation in the aquifer is enhanced by injection of an electron acceptor.

Interactions of the B3 cluster with H atoms and H2 molecules

A b initio electronic structure calculations on several σ- and three-center bonded B3H and B3H2 structures that correlate with the ground and first excited states of B3 (plus H or H2 ) have been carried out using correlation-consistent polarized valence double-zeta basis sets and complete-active-space self-consistent-field treatments of electron correlation. Geometries, electronic energies, and local harmonic vibrational frequencies were determined for two locally stable structures of B3H and three stable and one metastable structures of B3H2 as well as for transition states connecting such structures. Reaction energies for several processes and a barrier for the B3+H2→B3H2 reaction have also been calculated. A picture of the B3 reactivity is given in terms of two coupled potential energy surfaces and their avoided crossings. The relevance of our findings to future experimental work and similarities with the chemistry of boranes and related reactive species are emphasized.