Showing papers on "Nitrite published in 2000"

Role of circulating nitrite and S-nitrosohemoglobin in the regulation of regional blood flow in humans

Abstract: To determine the relative contributions of endothelial-derived nitric oxide (NO) vs. intravascular nitrogen oxide species in the regulation of human blood flow, we simultaneously measured forearm blood flow and arterial and venous levels of plasma nitrite, LMW-SNOs and HMW-SNOs, and red cell S-nitrosohemoglobin (SNO-Hb). Measurements were made at rest and during regional inhibition of NO synthesis, followed by forearm exercise. Surprisingly, we found significant circulating arterial-venous plasma nitrite gradients, providing a novel delivery source for intravascular NO. Further supporting the notion that circulating nitrite is bioactive, the consumption of nitrite increased significantly with exercise during the inhibition of regional endothelial synthesis of NO. The role of circulating S-nitrosothiols and SNO-Hb in the regulation of basal vascular tone is less certain. We found that low-molecular-weight S-nitrosothiols were undetectable and S-nitroso-albumin levels were two logs lower than previously reported. In fact, S-nitroso-albumin primarily formed in the venous circulation, even during NO synthase inhibition. Whereas SNO-Hb was measurable in the human circulation (brachial artery levels of 170 nM in whole blood), arterial-venous gradients were not significant, and delivery of NO from SNO-Hb was minimal. In conclusion, we present data that suggest (i) circulating nitrite is bioactive and provides a delivery gradient of intravascular NO, (ii) S-nitroso-albumin does not deliver NO from the lungs to the tissue but forms in the peripheral circulation, and (iii) SNO-Hb and S-nitrosothiols play a minimal role in the regulation of basal vascular tone, even during exercise stress.

Study on the Relationship Between Plasma Nitrite and Nitrate Level and Salt Sensitivity in Human Hypertension: Modulation of Nitric Oxide Synthesis by Salt Intake

Abstract: Background—High salt intake suppresses the effect of nitric oxide (NO) in the peripheral resistance vessels in animal models. We tested the hypothesis that the modulation of endogenous NO is relate...

Electrochemical detection of nitrate and nitrite at a copper modified electrode

Abstract: The development of a reagentless electrode system for the determination of nitrate is presented. The approach is based upon the deposition of a macroporous copper deposit which shows marked selectivity for nitrate ion under mildly acidic conditions (pH 3) with a linear range extending from 10 to 200 μM nitrate. Fabrication of the layer is both inexpensive and simple and is relatively substrate independent (Cu, Au, glassy carbon) as the response is attributed to the electrodeposited support. The influence of potential interferents such as chloride, nitrite and complexing agents such as citric acid were examined. In particular, the reduction of nitrite was found to occur at potentials significantly less negative than those required for nitrate reduction and as a result clear resolution of both voltammetric peaks is obtainable. The lack of direct interference between the two ions combined with wide peak separation (200 mV) provides a system that may offer some potential in speciation studies. The analytical applicability of the proposed system was demonstrated through the standard addition determination of nitrate in a number of authentic samples with the nitrate concentrations independently corroborated using established Griess assay spectroscopic protocols.

Nitrite-dependent nitric oxide production pathway: implications for involvement of active nitrogen species in photoinhibition in vivo.

Abstract: Air pollution studies have shown that nitric oxide (NO), a gaseous free radical, is a potent photosynthetic inhibitor that reduces CO 2 uptake activity in leaves. It is now recognized that NO is not only an air pollutant but also an endogenously produced metabolite, which may play a role in regulating plant cell functions. Although many studies have suggested the presence of mammalian–type NO synthase (NOS) in plants, the source of NO is still not clear. There has been a number of studies indicating that plant cells possess a nitrite–dependent NO production pathway which can be distinguished from the NOS–mediated reaction. Nitrate reductase (NR) has been recently found to be capable of producing NO through oneelectron reduction of nitrite using NAD(P)H as an electron donor. This review focuses on current understanding of the mechanism for the nitrite–dependent NO production in plants. Impacts of NO produced by NR on photosynthesis are discussed in association with photo–oxidative stress in leaves.

Anaerobic naphthalene degradation by microbial pure cultures under nitrate-reducing conditions

Abstract: Pure bacterial cultures were isolated from a highly enriched denitrifying consortium previously shown to anaerobically biodegrade naphthalene. The isolates were screened for the ability to grow anaerobically in liquid culture with naphthalene as the sole source of carbon and energy in the presence of nitrate. Three naphthalenedegrading pure cultures were obtained, designated NAP-3-1, NAP-3-2, and NAP-4. Isolate NAP-3-1 tested positive for denitrification using a standard denitrification assay. Neither isolate NAP-3-2 nor isolate NAP-4 produced gas in the assay, but both consumed nitrate and NAP-4 produced significant amounts of nitrite. Isolates NAP-4 and NAP-3-1 transformed 70 to 90% of added naphthalene, and the transformation was nitrate dependent. No significant removal of naphthalene occurred under nitrate-limited conditions or in cell-free controls. Both cultures exhibited partial mineralization of naphthalene, representing 7 to 20% of the initial added 14 C-labeled naphthalene. After 57 days of incubation, the largest fraction of the radiolabel in both cultures was recovered in the cell mass (30 to 50%), with minor amounts recovered as unknown soluble metabolites. Nitrate consumption, along with the results from the 14 C radiolabel study, are consistent with the oxidation of naphthalene coupled to denitrification for NAP-3-1 and nitrate reduction to nitrite for NAP-4. Phylogenetic analyses based on 16S ribosomal DNA sequences of NAP-3-1 showed that it was closely related to Pseudomonas stutzeri and that NAP-4 was closely related to Vibrio pelagius. This is the first report we know of that demonstrates nitrate-dependent anaerobic degradation and mineralization of naphthalene by pure cultures.

Simultaneous derivatization and quantification of the nitric oxide metabolites nitrite and nitrate in biological fluids by gas chromatography/mass spectrometry.

Abstract: Simultaneous quantification of nitrite and nitrate, the major oxidative metabolites of L-arginine-derived nitric oxide (NO), in biological fluids by GC or GC/MS methods is currently impossible. The separate analysis of these anions is associated with severe methodological problems. Therefore, a GC/MS method was developed which allows, for the first time, simultaneous quantification of nitrite and nitrate in various biological fluids. The method involves a single derivatization procedure, by which endogenous nitrite and nitrate and their externally added 15N-labeled analogues are simultaneously converted in aqueous acetone by pentafluorobenzyl bromide to the nitro and nitric acid ester pentafluorobenzyl derivatives, respectively, and a single GC/MS analysis. Nitrite and nitrate concentrations measured in plasma and urine of humans by this method correlated excellently with those from quantification of nitrite and nitrate in these matrixes using a previously reported GC/MS method that, however, requires reduction of nitrate to nitrite. Also, the present method enables discrimination between S-nitro- and S-nitroso-glutathione, which have identical chromatographic and spectrophotometric properties. The method is very useful to routinely study metabolism and reactions of NO and its metabolites in vitro and in vivo. It is accurate, interference-free, sensitive-50 fmol of [15N]-nitrite and [15N]nitrate were detected at signal-to-noise ratios of 870:1 and 95:1, respectively-and should be a reference method for nitrite and nitrate measurements.

Catalytic reduction of nitrates and nitrites in water solution on pumice-supported Pd–Cu catalysts

Abstract: Two series of pumice-supported palladium and palladium–copper catalysts, prepared by impregnation with different palladium and copper precursors, were tested for the hydrogenation of aqueous nitrate and nitrite solutions. Measurements were performed in a stirred tank reactor, operating in batch conditions, in buffered water solution at atmospheric pressure and at 293 K. The activities of the catalysts were calculated in terms of nitrate and/or nitrite removal. With the monometallic Pd/pumice, the reduction of nitrite is highly selective; only 0.2% of the initial nitrite content is converted to ammonium ions. The activity in terms of turn over frequency (TOF) is higher as compared to a catalyst of Pd on silica. Addition of copper to the palladium catalyst is essential for the reduction of nitrates, although it decreases the nitrite reduction activity and increases the production of ammonium ions. Nitrate reduction appears to be structure-insensitive and a volcano-type dependence of the activity versus the overall Cu atomic weight percentage is observed for the two series of catalysts.

The nrfA and nirB Nitrite Reductase Operons in Escherichia coli Are Expressed Differently in Response to Nitrate than to Nitrite

Abstract: Escherichia coli possesses two distinct nitrite reductase enzymes encoded by the nrfA and nirB operons. The expression of each operon is induced during anaerobic cell growth conditions and is further modulated by the presence of either nitrite or nitrate in the cells' environment. To examine how each operon is expressed at low, intermediate, and high levels of either nitrate or nitrite, anaerobic chemostat culture techniques were employed using nrfA-lacZ and nirB-lacZ reporter fusions. Steady-state gene expression studies revealed a differential pattern of nitrite reductase gene expression where optimal nrfA-lacZ expression occurred only at low to intermediate levels of nitrate and where nirB-lacZ expression was induced only by high nitrate conditions. Under these conditions, the presence of high levels of nitrate suppressed nrfA gene expression. While either NarL or NarP was able to induce nrfA-lacZ expression in response to low levels of nitrate, only NarL could repress at high nitrate levels. The different expression profile for the alternative nitrite reductase operon encoded by nirBDC under high-nitrate conditions was due to transcriptional activation by either NarL or NarP. Neither response regulator could repress nirB expression. Nitrite was also an inducer of nirB and nrfA gene expression, but nitrate was always the more potent inducer by >100-fold. Lastly, since nrfA operon expression is only induced under low-nitrate concentrations, the NrfA enzyme is predicted to have a physiological role only where nitrate (or nitrite) is limiting in the cell environment. In contrast, the nirB nitrite reductase is optimally synthesized only when nitrate or nitrite is in excess of the cell's capacity to consume it. Revised regulatory schemes are presented for NarL and NarP in control of the two operons.

Mitochondria recycle nitrite back to the bioregulator nitric monoxide.

Abstract: Nitric monoxide (NO) exerts a great variety of physiological functions. L-Arginine supplies amino groups which are transformed to NO in various NO-synthase-active isoenzyme complexes. NO-synthesis is stimulated under various conditions increasing the tissue of stable NO-metabolites. The major oxidation product found is nitrite. Elevated nitrite levels were reported to exist in a variety of diseases including HIV, reperfusion injury and hypovolemic shock. Denitrifying bacteria such as Paracoccus denitrificans have a membrane bound set of cytochromes (cyt cd1, cyt bc) which were shown to be involved in nitrite reduction activities. Mammalian mitochondria have similar cytochromes which form part of the respiratory chain. Like in bacteria quinols are used as reductants of these types of cytochromes. The observation of one-e- divergence from this redox-couple to external dioxygen made us to study whether this site of the respiratory chain may also recycle nitrite back to its bioactive form NO. Thus, the aim of the present study was therefore to confirm the existence of a reductive pathway which reestablishes the existence of the bioregulator NO from its main metabolite NO2-. Our results show that respiring mitochondria readily reduce added nitrite to NO which was made visible by nitrosylation of deoxyhemoglobin. The adduct gives characteristic triplet-ESR-signals. Using inhibitors of the respiratory chain for chemical sequestration of respiratory segments we were able to identify the site where nitrite is reduced. The results confirm the ubiquinone/cyt be1 couple as the reductant site where nitrite is recycled. The high affinity of NO to the heme-iron of cytochrome oxidase will result in an impairment of mitochondrial energy-production. "Nitrite tolerance" of angina pectoris patients using NO-donors may be explained in that way.

Degradation of n-Hexadecane and Its Metabolites by Pseudomonas aeruginosa under Microaerobic and Anaerobic Denitrifying Conditions

Abstract: A strategy for sequential hydrocarbon bioremediation is proposed. The initial O2-requiring transformation is effected by aerobic resting cells, thus avoiding a high oxygen demand. The oxygenated metabolites can then be degraded even under anaerobic conditions when supplemented with a highly water-soluble alternative electron acceptor, such as nitrate. To develop the new strategy, some phenomena were studied by examining Pseudomonas aeruginosa fermentation. The effects of dissolved oxygen (DO) concentration on n-hexadecane biodegradation were investigated first. Under microaerobic conditions, the denitrification rate decreased as the DO concentration decreased, implying that the O2-requiring reactions were rate limiting. The effects of different nitrate and nitrite concentrations were examined next. When cultivated aerobically in tryptic soy broth supplemented with 0 to 0.35 g of NO2−-N per liter, cells grew in all systems, but the lag phase was longer in the presence of higher nitrite concentrations. However, under anaerobic denitrifying conditions, even 0.1 g of NO2−-N per liter totally inhibited cell growth. Growth was also inhibited by high nitrate concentrations (>1 g of NO3−-N per liter). Cells were found to be more sensitive to nitrate or nitrite inhibition under denitrifying conditions than under aerobic conditions. Sequential hexadecane biodegradation by P. aeruginosa was then investigated. The initial fermentation was aerobic for cell growth and hydrocarbon oxidation to oxygenated metabolites, as confirmed by increasing dissolved total organic carbon (TOC) concentrations. The culture was then supplemented with nitrate and purged with nitrogen (N2). Nitrate was consumed rapidly initially. The live cell concentration, however, also decreased. The aqueous-phase TOC level decreased by about 40% during the initial active period but remained high after this period. Additional experiments confirmed that only about one-half of the derived TOC was readily consumable under anaerobic denitrifying conditions.

Influence of Denitrification on the Corrosion of Iron and Stainless Steel Powder

Abstract: The purpose of this research was to address the anoxic oxidation of metallic iron and stainless steel powder by nitrate, nitrite, and anaerobic mixed cultures. In sterile batch reactors, both nitrate and nitrite(10 mg N/L) could chemically oxidize metallic iron, with a concomitant reduction to ammonium. Nitrate or nitrite reduction coupled to metal corrosion was not observed in the case of stainless steel powder. Combination of an anaerobic mixed culture and metallic iron led to (cathodically produced) Hg consumption and a complete nitrate or nitrite reduction (mainly to NH4+). This caused a slightly enhanced metal oxidation. In the case of stainless steel, corrosion caused by denitrifying microorganisms was evidenced by data on nitrate/nitrite ions and solubilized iron. Experiments with increasing nitrite concentrations indicated that nitrite in the range of 50 mg of NO2- -N/L inhibited the corrosion processes. Moreover, at concentrations above 140 mg NO2- -N/L, a significant production of nitric oxide (NO) was detected. Differences between iron and stainless steel powder at low concentrations of nitrate or nitrite are most probably due to differences in kinetics: metallic iron exhibited faster chemical than biological reactions as opposed to stainless steel. It is postulated that the inhibitory effect of higher nitrite concentrations could partly be due to the chemical formation of NO and its toxic effect on the microorganisms acting at the steel surface.