Electrochemistry and spectroelectrochemistry of iron porphyrins in the presence of nitrite

The reaction of nitrite with ferric and ferrous porphyrins was examined using visible, infrared and NMR spectroscopy. Solutions of either ferric or ferrous porphyrin were stable in the presence of nitrite, with only complexation reactions being observed. Under voltammetric conditions, though, a rapid reaction between nitrite and iron porphyrins was observed to form the nitrosyl complex, Fe(p)(NO), where Pporphyrins. The products of the reduction of ferric porphyrins in the presence of nitrite were confirmed by visible spectroelectrochemistry to be Fe(P)(NO) and [Fe(P)]2O. Visible, NMR and infrared spectroscopy were used to rule out the formation of Fe(P)(NO) by the iron-catalyzed disproportionation of nitrite. A reaction between iron porphyrins and nitrite only occurred by the presence of both oxidation states (ferric:ferrous). The kinetics of the reaction were monitored by visible spectroscopy, and the reaction was found to be first-order with respect to Fe(OEP)(Cl) and Fe(OEP). The products were the same as those observed in the spectroelectrochemical experiment. The rate was not strongly dependent upon the concentration of nitrite, indicating that the coordinated, not the free nitrite, was the reaction species. The kinetics observed were consistent with a mixed oxidation state nitrite-bridged intermediate, which carried out the oxygen transfer reaction from nitrite to the iron porphyrin. The effect of nitrite coordination on the reaction rate was examined. © 2001 Elsevier Science B.V. All rights reserved

An investigation into the unusual linkage isomerization and nitrite reduction activity of a novel tris(2-pyridyl) copper complex

The copper-containing nitrite reductases (CuNIRs) are a class of enzymes that mediate the reduction of nitrite to nitric oxide in biological systems. Metal–ligand complexes that reproduce the salient features of the active site of CuNIRs are therefore of fundamental interest, both for elucidating the possible mode of action of the enzymes and for developing biomimetic catalysts for nitrite reduction. Herein, we describe the synthesis and characterization of a new tris(2-pyridyl) copper complex ([Cu1(NO2)2]) that binds two molecules of nitrite, and displays all three of the common binding modes for NO2−, with one nitrite bound in an asymmetric quasi-bidentate κ2-ONO manner and the other bound in a monodentate fashion with a linkage isomerism between the κ1-ONO and κ1-NO2 binding modes. We use density functional theory to help rationalize the presence of all three of these linkage isomers in one compound, before assessing the redox activity of [Cu1(NO2)2]. These latter studies show that the complex is not a competent nitrite reduction electrocatalyst in non-aqueous solvent, even in the presence of additional proton donors, a finding which may have implications for the design of biomimetic catalysts for nitrite reduction

Simultaneous Determination of Nitrite and Nitrate in Milk Samples by Ion Chromatography Method and Estimation of Dietary Intake

The presence of nitrate and nitrite in foods may be considered hazardous after ingestion in the gastrointestinal tract due to their reaction with naturally occurred secondary amines to form potentially carcinogenic nitrosamines. Due to this fact, a new method was developed in this study for the simultaneous determination of nitrite and nitrate in milk samples using by ion chromatography. Proposed mobile phase composed of sodium hydrogen carbonate and sodium carbonate (1.0 and 3.2 mmol/L) with a flow rate of 0.7 ml/min. The average recoveries for nitrate and nitrite were higher than 86 and 88, respectively. The limit of detection for nitrate and nitrite were 0.24 and 0.09 mg/L, respectively. The results of 102 real milk samples showed nitrate was found in all of the samples (100) with a mean of 34 ± 11 mg/L, while nitrite was found in none of the samples. The mean intake of nitrate in all age groups was lower than World Health Organization guideline. The present assessment concludes that the maximum contaminant level was equal to 82.8 mg/L nitrate. This method was fast, sensitive and accurate and is capable of being an alternative method in food control laboratories for investigation of nitrite and nitrate content. This is the first study of the determination and survey of nitrite and nitrate and exposure assessment of the Iranian population to nitrite and nitrate level in milk, which was widely used in infants and adolescents as one of the basic food components. Copyright © Taylor & Francis Group, LLC

An observational prospective study of topical acidified nitrite for killing methicillin-resistant Staphylococcus aureus (MRSA) in contaminated wounds

Background Endogenous nitric oxide (NO) kills bacteria and other organisms as part of the innate immune response. When nitrite is exposed to low pH, NO is generated and has been used as an NO delivery system to treat skin infections. We demonstrated eradication of MRSA carriage from wounds using a topical formulation of citric acid (4.5%) and sodium nitrite (3%) creams co-applied for 5 days to 15 wounds in an observational prospective pilot study of 8 patients. Findings Following treatment with topical citric acid and sodium nitrite, 9 of 15 wounds (60%) and 3 of 8 patients (37%) were cleared of infection. MRSA isolates from these patients were all sensitive to acidified nitrite in vitro compared to methicillin-sensitive S. aureus and a reference strain of MRSA. Conclusions Nitric oxide and acidified nitrite offer a novel therapy for control of MRSA in wounds. Wounds that were not cleared of infection may have been re-contaminated or the bioavailability of acidified nitrite impaired by local factors in the tissue

Ameliorating Effect of Chloride on Nitrite Toxicity to Freshwater Invertebrates with Different Physiology: a Comparative Study Between Amphipods and Planarians

High nitrite concentrations in freshwater ecosystems may cause toxicity to aquatic animals. These living organisms can take nitrite up from water through their chloride cells, subsequently suffering oxidation of their respiratory pigments (hemoglobin, hemocyanin). Because NO2¿ and Cl¿ ions compete for the same active transport site, elevated chloride concentrations in the aquatic environment have the potential of reducing nitrite toxicity. Although this ameliorating effect is well documented in fish, it has been largely ignored in wild freshwater invertebrates. The aim of this study was to compare the ameliorating effect of chloride on nitrite toxicity to two species of freshwater invertebrates differing in physiology: Eulimnogammarus toletanus (amphipods) and Polycelis felina (planarians). The former species presents gills (with chloride cells) and respiratory pigments, whereas in the latter species these are absent. Test animals were exposed in triplicate for 168 h to a single nitrite concentration (5 ppm NO2-N for E. toletanus and 100 ppm NO2-N for P. felina) at four different environmental chloride concentrations (27.8, 58.3, 85.3, and 108.0 ppm Cl¿). The number of dead animals and the number of affected individuals (i.e., number of dead plus inactive invertebrates) were monitored every day. LT50 (lethal time) and ET50 (effective time) were estimated for each species and each chloride concentration. LT50 and ET50 values increased with increases in the environmental chloride concentration, mainly in amphipods. Results clearly show that the ameliorating effect of chloride on nitrite toxicity was more significant in amphipods than in planarians, likely because of the absence of gills (with chloride cells) and respiratory pigments in P. felina. Additionally, this comparative study indicates that the ecological risk assessment of nitrite in freshwater ecosystems should take into account not only the most sensitive and key species in the communities, but also chloride levels in the aquatic environmen

Resolution of Key Roles for the Distal Pocket Histidine in Cytochrome c Nitrite Reductases

Cytochrome c nitrite reductases perform a key step in the biogeochemical N-cycle by catalyzing the six-electron reduction of nitrite to ammonium. These multi-heme cytochromes contain a number of His/His ligated c-hemes for electron transfer and a structurally differentiated heme that provides the catalytic center. The catalytic heme has proximal ligation from lysine, or histidine, and an exchangeable distal ligand bound within a pocket that includes a conserved histidine. Here we describe properties of a penta-heme cytochrome c nitrite reductase in which the distal His has been substituted by Asn. The variant is unable to catalyze nitrite reduction despite retaining the ability to reduce a proposed intermediate in that process, namely, hydroxylamine. A combination of electrochemical, structural and spectroscopic studies reveals that the variant enzyme simultaneously binds nitrite and electrons at the catalytic heme. As a consequence the distal His is proposed to play a key role in orienting the nitrite for N-O bond cleavage. The electrochemical experiments also reveal that the distal His facilitates rapid nitrite binding to the catalytic heme of the native enzyme. Finally it is noted that the thermodynamic descriptions of nitrite- and electron-binding to the active site of the variant enzyme are modulated by the prevailing oxidation states of the His/His ligated hemes. This is behavior that is likely to be displayed by other multi-centered redox enzymes such that there are wide implications for considering the determinants of catalytic activity in this important and varied group of oxidoreductases

Role of aldehyde dehydrogenase in hypoxic vasodilator effects of nitrite in rats and humans

Hypoxic conditions favour the reduction of nitrite to nitric oxide (NO) to elicit vasodilatation, but the mechanism(s) responsible for bioconversion remains ill defined. In the present study, we assess the role of aldehyde dehydrogenase 2 (ALDH2) in nitrite bioactivation under normoxia and hypoxia in the rat and human vasculature

Physico-chemical factors and bacteria in fish ponds

Analyses of pond water and mud samples show that nitrifying bacteria (including ammonifying bacteria, nitrite bacteria, nitrobacteria and denitrifying bacteria) are in general closely correlated with various physico-chemical factors, ammonifying bacteria are mainly correlated with dissolved oxygen; denitrifying bacteria are inversely correlated with phosphorus; nitrite bacteria are closely correlated with nitrites, nitrobacteria are inversely correlated with ammoniac nitrogen. The nitrifying bacteria are more closely correlated with heterotrophic bacteria. Nitrobacteria are inversely correlated with anaerobic heterotrophic bacteria. The correlation is quite weak between all the nitrite bacteria which indicates that the nitrite bacteria have a controlling and regulating function in water quality and there is no interdependence as each plays a role of its own. The paper also discusses how the superficial soil (pond mud down to 3.5 cm deep) and different layers of the mud affect the biomass of bacteria. The study shows that the top superficial layer (down to 1.5 cm deep) is the major area for decomposing and converting organic matter

Characteristics of Se'i (Rotenesse Smoked Meat) Treated with Coconut Shell Liquid Smoked and Citrus Aurantifolia Extract

The objective of this study was to investigate the effect of Citrus aurantifolia extract (CAE), coconut shell liquid smoke (CSLS) and the combination of CAE and CSLS (CACS) on se'i characteristics. A completely randomized design was assigned in this experiment. Treatments used were: se'i treated with 5% (v/v) CAE, CSLS 5% (v/v), (CAE : CSL 1:1) )/ (CACS) and untreated se'i as a control (C). Parameters measured were: aroma, color, taste, pH, residual nitrite, total bacterial count, Coliform, Staphylococcus aureus, Escherichia coli, and Salmonella. The data of aroma, color, and taste were analyzed by using Kruskal-Wallis test followed by Mann-Whitney test. The pH, residual nitrite, and bacterial data were analyzed with analysis of variance (ANOVA) followed by least significant differences test. Results showed that CAE caused the highest score at both aroma and taste (P<0.05). CSLS caused the lowest residual nitrite (27 ppm) (P<0.05). Application of CAE and CACS could reduce total bacteria (P<0.05) at least 1 log. Color, pH, and Coliform number were not significantly different. S. aureus, E. coli, and Salmonella were negative in all se'i samples. CAE gives the best organoleptics and bacteriological characteristics while CSLS is more effective in reducing nitrite

Nitric oxide and nitrous oxide production and cycling during dissimulatory nitrite reduction by Pseudomonas perfectomarina

The denitrifier Pseudomonas perfectomarina reduced nitrite under conditions of kinetic competition between cells and gas sparging for extracellular dissolved nitric and nitrous oxides, NOaq and N2Oaq, in a chemically defined marine medium. Time courses of nitrite reduction and NOg and N2Og removal were integrated to give NOg , and N2Og yields. At high sparging rates, the NOg yield was >50% of nitrite-N reduced, and the yield of NOg + N2Og was ~75%. Hence interrupted denitrification yields NOaq and N2Oaq as major products. The yields varied with sparging rates in agreement with a quantitative model of denitrification (Betlach, M. P., and Tiedje, J. M. (1981) Appl. Environ. Microbiol. 42, 1074-1084) that applies simplified Michaelis-Menten kinetics to NO2 > NOaq > N2Oaq > N2. The fit gave an estimate of the maximum scavengeable NOaq yield of 73 ± 8% of nitrite-N. Thus a minor path independent of NOaq is also required. The fit of the model to data at lower sparging rates, where normal denitrification products predominate, implies that the extracellular NOaq pool yield is independent of gas sparging rate. Thus in P. perfectomarina NOaq and N2Oaq are intermediates, or facilely equilibrate with true intermediates, during complete denitrification. The recovery of most nitrite-N as NO and/or N20 under perturbed conditions is not an artifact of irreversible product removal, but an attribute of denitrification in this species, and most probably it is characteristic of denitrification in other species as well