Showing papers on "Nitrite published in 1993"

Oxidation of nitric oxide in aqueous solution to nitrite but not nitrate: comparison with enzymatically formed nitric oxide from L-arginine.

Abstract: Nitric oxide (NO) in oxygen-containing aqueous solution has a short half-life that is often attributed to a rapid oxidation to both NO2- and NO3-. The chemical fate of NO in aqueous solution is often assumed to be the same as that in air, where NO is oxidized to NO2 followed by dimerization to N2O4. Water then reacts with N2O4 to form both NO2- and NO3-. We report here that NO in aqueous solution containing oxygen is oxidized primarily to NO2- with little or no formation of NO3-. In the presence of oxyhemoglobin or oxymyoglobin, however, NO and NO2- were oxidized completely to NO3-. Methemoglobin was inactive in this regard. The unpurified cytosolic fraction from rat cerebellum, which contains constitutive NO synthase activity, catalyzed the conversion of L-arginine primarily to NO3- (NO2-/NO3- ratio = 0.25). After chromatography on DEAE-Sephacel or affinity chromatography using 2',5'-ADP-Sepharose 4B, active fractions containing NO synthase activity catalyzed the conversion of L-arginine primarily to NO2- (NO2-/NO3- ratio = 5.6) or only to NO2-, respectively. Unpurified cytosol from activated rat alveolar macrophages catalyzed the conversion of L-arginine to NO2- without formation of NO3-. Addition of 30 microM oxyhemoglobin to all enzyme reaction mixtures resulted in the formation primarily of NO3- (NO2-/NO3- ratio = 0.09 to 0.20). Cyanide ion, which displaces NO2- from its binding sites on oxyhemoglobin, inhibited the formation of NO3-, thereby allowing NO2- to accumulate. These observations indicate clearly that the primary decomposition product of NO in aerobic aqueous solution is NO2- and that further oxidation to NO3- requires the presence of additional oxidizing species such as oxyhemoproteins.

Reactions of the bioregulatory agent nitric oxide in oxygenated aqueous media: determination of the kinetics for oxidation and nitrosation by intermediates generated in the NO/O2 reaction.

Abstract: The reaction kinetics of nitric oxide autoxidation in aerobic solutions were investigated by direct observation of the nitrite ion product and by trapping the strongly oxidizing and nitrosating intermediates formed in this reaction. The rate behavior observed for nitrite formation [rate = k3[O2][NO]2, k3 = (6 +/- 1.5) x 10(6) M-2 s-1 at 22 degrees C] was the same as found for oxidation of Fe(CN)6(4-) and of 2,2'-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS) and as for the nitrosation of sulfanilamide. There was a slight decrease in k3 to (3.5 +/- 0.7) x 10(6) M-2 s-1 at 37 degrees C. The second-order dependency for NO was observed at NO concentrations as low as 3 microM. The results of the competitive kinetics studies suggest that the key oxidizing intermediates, species which are both strong oxidants and nitrosating agents, are not one of those commonly proposed (NO2, N2O3, NO+, or O2NO-) but are one or more as yet uncharacterized NOx species.

The effects of nitrate, nitrite and N-nitroso compounds on human health: a review.

Abstract: The effects of nitrate, nitrite, and N-nitroso compounds on human health are reviewed. Special emphasis has been placed on the role of these compounds on infant methemoglobinemia and gastric cancer. The discussion on methemoglobinemia includes the source of nitrate or nitrite, diagnosis, treatment, prevention and the contributions of age, gastric pH, gastrointestinal illness, and ingestion of vitamin C to this illness. The maternal transfer of these compounds and the potential effect on fetal death and malformation are also described. The etiology and development of gastric cancer is reviewed as well as the roles of nitrate, nitrite, and N-nitroso compounds in this disease. Endogenous nitrosation and the experimental and epidemiologic evidence linking these compounds to gastric cancer is examined. Other sections include adult methemoglobinemia and acute toxicity, hypo- and hypertension, Balkan nephropathy, slowing of motor reflexes in children, nitrate esters dependence. Sources of nitrate, nitrite, and N-nitroso compounds are detailed. Future areas of research are given.

Interaction of myeloperoxidase with peroxynitrite. A comparison with lactoperoxidase, horseradish peroxidase and catalase.

Abstract: Polymorphonuclear neutrophils generate both nitric oxide and superoxide and these molecules can combine to form peroxynitrite. Neutrophils also contain myeloperoxidase which reacts with peroxynitrous acid (HOONO). On mixing myeloperoxidase with HOONO compound II was formed. Compound I could not be detected as an intermediate. The apparent second-order rate constant of formation of compound II was strongly pH-dependent (2.5×105 M−1· s−1 at pH 8.9 and 6.2×106 M−1· s−1 at pH 7.2). The pKa of this effect is 6.9 and it was concluded that the enzyme reacts with the protonated form of the peroxide, that is peroxynitrous acid, with a pH-independent second-order rate constant of 2.0×107 M−1· s−1 at 12°C. The interaction of HOONO with lactoperoxidase was studied for comparison. As was observed for myeloperoxidase, compound I could not be detected as an intermediate. The apparent second-order rate constant of compound II formation is pH-dependent and is 3.3×105 M−1· s−1 at pH 7.4 and 8.4×104 M−1· s−1 at pH 9.0. In contrast, horseradish peroxidase reacts with HOONO to form compound I, which is subsequently followed by the formation of compound II. The second-order rate constant for the formation of compound I is 3.2×106 M−1· s−1 and is pH-dependent, the pKa for this effect is 6.8. Catalase (up to 3 μM) does not affect the rate of decomposition of peroxynitrite and no compound I formation is observed. Since nitrite may be present in the peroxynitrite preparation and to discriminate between the reaction of the enzyme with nitrite or peroxynitrite, the effect of nitrite on myeloperoxidase was studied. The dissociation constant for the myeloperoxidase-nitrite complex is pH-dependent and has values of 580 μM at pH 6.0 and 55 mM at pH 8.5.

The effects of nitrate, nitrite, and N-nitroso compounds on animal health.

Abstract: The clinical signs of acute nitrate toxicity vary according to species. In general, ruminant animals develop methemoglobinemia while monogastric animals exhibit severe gastritis. Nitrate ingestion has also been linked to impairment of thyroid function, decreased feed consumption, and interference with vitamin A and E metabolism. Hematologic changes seen with chronic high nitrate exposure include both compensatory increases in red blood cells and anemia, along with increased neutrophils and eosinophils. Unlike nitrate, nitrite is capable of inducing methemoglobinemia in a wide range of species, ie cattle, sheep, swine, dogs, guinea pigs, rats, chickens and turkeys. In rats, chronic nitrite exposure causes pathologic changes in a variety of tissues, alterations in motor activity and brain electrical activity, and alters gastric mucosal absorption. Nitrite affects the metabolism of sulfonamide drugs in animals such as the pig, guinea pig, and rat. The N-nitroso compound dimethylnitrosamine causes toxic hepatosis in cattle, sheep, mink, and fox. Nitrosamines have been reported in cows milk and been found to pass into the milk of goats under experimental conditions.

Competition for limiting amounts of oxygen between Nitrosomonas europaea and Nitrobacter winogradskyi grown in mixed continuous cultures

Abstract: Chemolithotrophic nitrifying bacteria are dependent on the presence of oxygen for the oxidation of ammonium via nitrite to nitrate. The success of nitrification in oxygen-limited environments such as waterlogged soils, will largely depend on the oxygen sequestering abilities of both ammonium- and nitrite-oxidizing bacteria. In this paper the oxygen consumption kinetics of Nitrosomonas europaea and Nitrobacter winogradskyi serotype agilis were determined with cells grown in mixed culture in chemostats at different growth rates and oxygen tensions. Reduction of oxygen tension in the culture repressed the oxidation of nitrite before the oxidation of ammonium was affected and hence nitrite accumulated. K m values found were within the range of 1–15 and 22–166 μM O2 for the ammonium- and nitrite-oxidizing cells, respectively, always with the lowest values for the N. europaea cells. Reduction of the oxygen tension in the culture lowered the half saturation constant K m for oxygen of both species. On the other hand, the maximal oxygen consumption rates were reduced at lower oxygen levels especially at 0 kPa. The specific affinity for oxygen indicated by the V max/K m ratio, was higher for cells of N. europaea than for N. winogradskyi under all conditions studied. Possible consequences of the observed differences in specific affinities for oxygen of ammonium-and nitrite-oxidizing bacteria are discussed with respect to the behaviour of these organisms in oxygen-limited environments.

Prebiotic ammonia from reduction of nitrite by iron (II) on the early Earth.

Abstract: Theories for the origin of life require the availability of reduced (or 'fixed') nitrogen-containing compounds, in particular ammonia. In reducing atmospheres, such compounds are readily formed by electrical discharges, but geochemical evidence suggests that the early Earth had a non-reducing atmosphere, in which discharges would have instead produced NO. This would have been converted into nitric and nitrous acids and delivered to the early oceans as acid rain. It is known, however, that Fe(II) was present in the early oceans at much higher concentrations than are found today, and thus the oxidation of Fe(II) to Fe(III) provides a possible means for reducing nitrites and nitrates to ammonia. Here we explore this possibility in a series of experiments which mimic a broad range of prebiotic seawater conditions (the actual conditions on the early Earth remain poorly constrained). We find that the reduction by Fe(II) of nitrites and nitrates to ammonia could have been a significant source of reduced nitrogen on the early Earth, provided that the ocean pH exceeded 7.3 and is favoured for temperatures greater than about 25 degrees C.

Formation and decay of peroxynitric acid : a pulse radiolysis study

Abstract: Peroxynitrous acid and peroxynitrite anion have been studied using pulse radiolysis of nitrite and nitrate solutions. The formation rate constant is determined to be k(OH+NO 2 )=(4.5±1.0)×10 9 M -1 s -1 , and the rate constant for the OH radical reaction with nitrite is determined to be k(OH+NO 2 - )= (6.0±1.0)×10 9 M -1 s -1 . In nitrate solutions, the competing reaction between OH and NO 3 2- is found to have a rate constant of k(OH+NO 3 2- ) =(3.0±1.0)×10 9 M -1 s -1

The periplasmic nitrite reductase of Thauera selenatis may catalyze the reduction of selenite to elemental selenium

Abstract: Thauera selenatis grows anaerobically with selenate, nitrate or nitrite as the terminal electron acceptor; use of selenite as an electron acceptor does not support growth. When grown with selenate, the product was selenite; very little of the selenite was further reduced to elemental selenium. When grown in the presence of both selenate and nitrate both electron acceptors were reduced concomitantly; selenite formed during selenate respiration was further reduced to elemental selenium. Mutants lacking the periplasmic nitrite reductase activity were unable to reduce either nitrite or selenite. Mutants possessing higher activity of nitrite reductase than the wild-type, reduced nitrite and selenite more rapidly than the wild-type. Apparently, the nitrite reductase (or a component of the nitrite respiratory system) is involved in catalyzing the reduction of selenite to elemental selenium while also reducing nitrite. While periplasmic cytochrome C551 may be a component of the nitrite respiratory system, the level of this cytochrome was essentially the same in mutant and wild-type cells grown under two different growth conditions (i.e. with either selenate or selenate plus nitrate as the terminal electron acceptors). The ability of certain other denitrifying and nitrate respiring bacteria to reduce selenite will also be described.

Feedback Regulation of Nitrate Influx in Barley Roots by Nitrate, Nitrite, and Ammonium

Abstract: The short-lived radiotracer 13N was used to study feedback regulation of nitrate influx through the inducible high-affinity transport system of barley (Hordeum vulgare L. cv Steptoe) roots. Both wild-type plants and the mutant line Az12:Az70 (genotype nar1a;nar7w), which is deficient in the NADH-specific and NAD(P)H-bispecific nitrate reductases (R.L. Warner, R.C. Huffaker [1989] Plant Physiol 91: 947–953) showed strong feedback inhibition of nitrate influx within approximately 5 d of exposure to 100 fmu]M nitrate. The result with the mutant, in which the flux of nitrogen into reduced products is greatly reduced, indicated that nitrate itself was capable of exercising feedback regulation upon its own influx. This conclusion was supported by the observation that feedback in wild-type plants occurred in both the presence and absence of L-methionine sulfoximine, an inhibitor of ammonium assimilation. Nitrite and ammonium were also found to be capable of exerting feedback inhibition upon nitrate influx, although it was not determined whether these ions themselves or subsequent metabolites were responsible for the effect. It is suggested that feed-back regulation of nitrate influx is potentially mediated through several nitrogen pools, including that of nitrate itself.

Biodegradation of 2-nitrotoluene by Pseudomonas sp. strain JS42.

Abstract: A strain of Pseudomonas sp. was isolated from nitrobenzene-contaminated soil and groundwater on 2-nitrotoluene as the sole source of carbon, energy, and nitrogen. Bacterial cells growing on 2-nitrotoluene released nitrite into the growth medium. The isolate also grew on 3-methylcatechol, 4-methylcatechol, and catechol. 2-Nitrotoluene, 3-methylcatechol, and catechol stimulated oxygen consumption by intact cells regardless of the growth substrate. Crude extracts from the isolate contained catechol 2,3-dioxygenase and 2-hydroxy-6-oxohepta-2,4-dienoate hydrolase activity. The results suggest that 2-nitrotoluene is subject to initial attack by a dioxygenase enzyme that forms 3-methylcatechol with concomitant release of nitrite. The 3-methylcatechol is subsequently degraded via the meta ring fission pathway.

Catalytical removal of nitrate and nitrite from drinking water: 1. Screening for hydrogenation catalysts and influence of reaction conditions on activity and selectivity

Abstract: The rising concentrations of nitrate in ground water make it necessary to use processes to remove nitrate from drinking water. Nitrate is removed at present either by physicochemical or biological ...

Effect of Medium Composition on the Denitrification of Nitrate by Paracoccus denitrificans.

Abstract: The course of denitrification of nitrate in static cultures of Paracoccus denitrificans was studied. Reduction of nitrate to gaseous nitrogen without accumulation of nitrite because of parallel and balanced activities of nitrate and nitrite reductases was observed in nutrient broth. In minimal liquid cultures supplemented with either methanol, acetate, or ethanol as a sole carbon source, substantial amounts of nitrite (up to 70%) accumulated. The reduction in nitrite concentration began just after the transformation of nitrate to nitrite was completed. The addition of some growth factors to minimal media shortened the bacterial biomass doubling time. A correlation coefficient of 0.71 between the doubling time and the amount of accumulated nitrite in cultures was found. My results indicated that the type of denitrification carried out by P. denitrificans is not stable and depends on the nutritional composition of the culture medium.

Reduction of perchlorate by an anaerobic enrichment culture

Abstract: A mixed bacterial culture capable of reducing perchlorate stoichiometrically to chloride under naerobic conditions was enriched from municipal digester sludge. The reduction of 10 mM perchlorate resulted in oxidation of the medium and cessation of perchlorate reduction. The activity was recovered on addition of a reducing agent. Addition of air to the culture during perchlorate reduction immediately terminated the process and aeration for 12 h permanently destroyed the ability of the culture to reduce perchlorate. The culture also reduced nitrite, nitrate, chlorite, chlorate and sulfate. The presence of 10 mM nitrite or chlorite completely inhibited perchlorate reduction, whereas the same concentration of chlorate decreased the reduction rate. Nitrate or sulfate did not affect perchlorate reduction. Chlorate and chlorite, suspected intermediates in the reduction of perchlorate to chloride, were not detected in any cultures during reduction of perchlorate.

The purification of a cd1-type nitrite reductase from, and the absence of a copper-type nitrite reductase from, the aerobic denitrifier Thiosphaera pantotropha; the role of pseudoazurin as an electron donor.

Abstract: Thiosphaera pantotropha has been reported to contain a copper-type nitrite reductase on the basis that the copper chelator diethyldithiocarbamate inhibited the overall process of denitrification. It is now shown that nitrous oxide reduction is 100% inhibited by 10 mM diethyldithiocarbamate or 100 microM azide. We also found that both these inhibitors partially inhibited nitrite reduction in this organism. We purified the nitrite reductase of T. pantotropha and found that it was of the cytochrome cd1 type, contrary to the published report of it being a copper-type nitrite reductase. This is of importance since T. pantotropha is capable of aerobic nitrite reduction. The only detectable nitrite reductase in anaerobically or aerobically grown cells is the cd1 type. We also purified a small copper-containing protein, pseudoazurin. Pseudoazurin was found to be capable of donating electrons to the cd1-type nitrite reductase in vitro, and its copper centre was chelated by diethyldithiocarbamate. Since nitrite reduction is partially inhibited by diethyldithiocarbamate, it is thought that pseudoazurin is an electron donor to nitrite reductase in vivo.

Oxidation of aqueous sulfite ion by nitrogen dioxide

Abstract: The reaction of nitrogen dioxide with aqueous sulfite solutions, in the presence and in the absence of oxygen, has been studied using Raman spectroscopy to determine product concentrations. The products observed include nitrite, sulfate, and dithionate ions. The reaction appears to initially produce nitrite ion and sulfite radical: NO 2 +SO 3 2- →NO 2 - +SO 3 .- . The sulfite radical can undergo either recombination or reaction with oxygen to form SO 5 .-

Tetrahydrobiopterin is required for cytokine-induced nitric oxide production in a murine macrophage cell line (RAW 264).

Abstract: The murine macrophage cell line RAW 264 constitutively synthesizes tetrahydrobiopterin (BH4), the cofactor required for the hydroxylation of the aromatic amino acids and for the production of nitric oxide. Stimulation of the cells with interferon-gamma and lipopolysaccharide induced the production of nitric oxide and increased BH4 levels further. When the cells were stimulated in the presence of 2,4-diamino-6-hydroxypyrimidine (DAHP), an inhibitor of BH4 biosynthesis, biopterin levels decreased by 90% within 6 hr, whereas nitrite production was essentially unaffected. Pretreatment of the cells for 12 hr with DAHP decreased intracellular BH4 concentrations by > 95% yet inhibited the cytokine-stimulated production of nitric oxide by only 50%. However, pretreatment with DAHP plus N-acetylserotonin, an inhibitor of sepiapterin reductase, the terminal enzyme of the BH4 biosynthetic pathway, decreased biopterin levels by > 99% and inhibited nitric oxide synthesis by 90%. This inhibition could be reversed by loading the cells with dihydrobiopterin, a precursor of BH4 via the dihydrofolate reductase salvage pathway. In addition, these studies revealed that N-acetylserotonin has a direct inhibitory effect on nitric oxide synthesis, acting in a BH4-independent manner. The results presented here support previous suggestions, based on experiments with isolated enzymes, that BH4 is absolutely required for cytokine-stimulated nitric oxide production in macrophages and they suggest that only a small fraction of the total intracellular BH4 pool in macrophages is utilized in the production of fully active nitric oxide synthase.

Nitric Oxide Release in Response to Gut Injury

Abstract: We evaluated nitric oxide release in several models of intestinal inflammation through luminal nitrite concentrations In anesthetized rabbits, piglets, and guinea pigs, luminal lavages were collected from loops of normal or injured small intestine Lavages were analyzed spectrophotometrically for nitrite (Griess reagent) and protein Myeloperoxidase (MPO) content of intestinal segments was used as an index of granulocyte infiltration and intestinal inflammation Acute ileal inflammation was induced by luminal acetic acid + casein in rabbits and luminal deoxycholate in neonatal piglets and adult rabbits Chronic ileitis was induced in guinea pigs by intraluminal trinitrobenzenesulfonic acid In each model nitrite levels in ileal lavages were significantly greater than control loops/animals Increased luminal protein and intestinal MPO activity paralleled the changes in nitrite levels To determine whether nitric oxide production influenced mucosal repair, segments of ileum were perfused with the L-arginine antagonist NG-nitro-L-arginine methyl ester (L-NAME, 10 mg/ml) after acute acetic acid + casein exposure L-NAME administration reversed the decline in epithelial permeability characteristic of epithelial restitution, causing an increase in epithelial permeability which was readily reversible These results suggest that nitrite production is a useful index of gut injury and that nitric oxide may contribute to the functional repair of the epithelial barrier under acute conditions

Vascular smooth muscle contains a depletable store of a vasodilator which is light-activated and restored by donors of nitric oxide.

Abstract: Endothelium-denuded strips of rabbit thoracic aorta relax on exposure to light. This response is similar to endothelium-dependent relaxation as it is inhibited by hemoglobin and methylene blue, and is mediated by an increase in cyclic GMP. We now demonstrate that photorelaxation decreases on repeated exposure to light. The response can be restored by treating the depleted smooth muscle strips with acidified nitrite, but not nitrite alone, and with the nitric oxide donors, S-nitrosopenicillamine and glyceryl trinitrate, but not with hydralazine. These data indicate that photorelaxation is mediated in part by a "pool" of light-activated vasodilator(s) and suggest that this may act as a store of nitric oxide which could play a role in the regulation of vascular tone.

Induction of a macrophage-like nitric oxide synthase in cultured rat aortic endothelial cells. IL-1 beta-mediated induction regulated by tumor necrosis factor-alpha and IFN-gamma.

Abstract: We investigated the effects of murine rTNF-alpha, human rIL-1 beta, and rat rIFN-gamma in various concentrations and/or combinations on inducible nitric oxide (NO) production in primary cultures of rat aortic endothelial cells. Northern blot analysis of total RNA from induced and control cultures using the cloned mouse macrophage gene of inducible NO synthase as probe as well as polymerase chain reaction using a specific primer sequence gave a positive signal for activated cells only. A RNA approximately 4.4 kb of length similar to the inducible form of NO synthase in macrophages was labeled. The concentration of nitrite as a stable reaction product of NO in culture supernatants was determined 24 h after incubation with the various cytokines. IL-1 beta alone (40 to 1000 U/ml) induced formation of increasing amounts of nitrite with increasing concentrations of IL-1 beta present. Neither TNF-alpha alone (10 to 2000 U/ml) nor IFN-gamma alone 25 to 500 U/ml) showed significant effects on nitrite production. Simultaneous incubation with low concentrations of TNF-alpha ( or = 500 U/ml) led to near maximal levels of nitrite formation even at lowest IL-1 beta concentrations (40 U/ml). In addition, simultaneous incubation of endothelial cells with IFN-gamma plus IL-1 beta and/or TNF-alpha led to near maximal NO production of endothelial cells, even at lowest IFN-gamma concentrations (25 U/ml). We hypothesize that the regulating effect of TNF-alpha may in vivo help to prevent local inflammatory responses from spreading to intact sites.