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Model experiments on nitrite and nitrate in simulated primeval conditions.

In experiments on the prebiotic formation of nitric oxides, anoxic mixtures of N2 and water vapour were sparked in contact with phosphate buffer solutions at various pH values. Nitrite was found in the aqueous phase, and nitrate grew from it, presumably by reaction with H2O2. In acid solutions, these anions were reduced and destroyed by Fe2+, and the same was true of nitrite in solutions kept at a pH value similar to that of the contemporary ocean (8.2) with HEPES buffer. Nitrate was not destroyed in short-term experiments, but as in sparking nitrate is formed only vianitrite, neither anion could accumulate. In further sparking experiments with alkaline sulphide, both nitrite and nitrate were reduced entirely. It is concluded that it is unlikely that the primeval ocean contained appreciable concentrations of nitrite or nitrate either at the reducing or at the redox-neutral stage.

Anaerobiosis

Nitrite reductase system involved in the terminal oxidation of the Streptomyces griseus respiratory particle.

A nitrite reductase system which was associated with the electron transfer system of the respiratory particle in Streptomyces griseus was studied. The electron transfer pathway consisted of the cytochrome oxidase and the nitrite reductase systems under aerobic and anaerobic conditions respectively, and these systems showed the exact opposite response to 2-n-heptyl-4-hydroxyquinoline-N-oxide and azide. Azide inhibited specifically the nitrite reductase system. It seems that cytochrome d works as the nitrite reductase and the reduced cytochrome b works as an intermediate electron donor for cytochrome d respectively. The respiratory particle also had a hydroxylamine reductase activity and ammonia was identified as the product of hydroxylamine reduction by the respiratory particle. A terminal electron transfer pathway in Streptomyces griseus was proposed.

Anaerobiosis

Stoichiometry of the reaction of oxyhemoglobin with nitrite.

During the reaction of oxyhemoglobin (HbO2) with nitrite, the concentration of residual nitrite, nitrate, oxygen, and methemoglobin (Hb+) was determined successively. The results obtained at various pH values indicate the following stoichiometry for the overall reaction: 4HbO2 + 4NO2- 4H+ leads to 4Hb+ + 4NO3- + O2 + 2H2 O (Hb denotes hemoglobin monomer). NO2- binds with methemoglobin noncooperatively with a binding constant of 340 M-1 at pH 7.4 and 25 degrees C. Thus, the major part of Hb+ produced is aquomethemoglobin, not methemoglobin nitrite, when less than 2 equivalents of nitrite is used for the oxidation.

Hemoglobin A

Deciphering the effects of sulfonamide antibiotics on denitrification from a metagenomic perspective: Inhibition of nitrite reduction and succession patterns of functional microorganisms.

Limited research has thoroughly elucidated the impact mechanisms of antibiotics on the denitrification process at the genomic and gene levels, which has hindered the optimization and development of nitrogen removal technology for antibiotic-containing swine wastewater. Lab-scale sequencing batch reactors were constructed in this study to treat synthetic wastewater containing different sulfonamides and nitrate. Investigations were carried out on denitrification performance, microbial community diversity, denitrifier succession patterns, and functional gene distribution. The stress of sulfonamides inhibited the nitrite reduction process, transforming complete denitrification into partial denitrification and causing significant nitrite accumulation. The average nitrogen removal efficiency in the treatment groups decreased from 81.0% ± 2.2-40.1% ± 6.1%. Alicycliphilus and Thauera were identified as the key taxa, accounting for 32.2% and 16.9% of all potential denitrifying bacteria, respectively. Although metagenome-assembled genomes (MAGs) from Thauera were enriched with genes encoding nitrate reductases (nap, nar) and nitrite reductases (nir), this genus preferentially utilized nitrate as an electron acceptor, resulting in the preferential nitrate reduction and subsequent nitrite accumulation. In contrast, Alicycliphilus MAGs developed tolerance to the sulfonamides stress during later stages, with concomitant enrichment of associated functional genes. They replaced Thauera to reemerge as the dominant group, thereby restoring complete denitrification. This study provides new insights into the regulatory mechanisms governing complete versus partial denitrification in nitrogen removal from antibiotic-containing wastewater.

Denitrifier succession

Effects of amyl nitrite on phasic aortocoronary bypass graft blood velocity in man.

With the use of a Doppler flowmeter catheter, phasic instantaneous aortocoronary saphenous vein bypass graft blood velocity was continuously measured during the inhalation of amyl nitrite in 20 closed-chest conscious subjects. Administration of amyl nitrite augmented peak diastolic and systolic graft blood velocity within 10 seconds and maximal blood velocities were recorded between 8 and 60 seconds after inhalation. Control mean (+/- 1 S.D.) bypass graft blood velocity was 25 +/- 10 cm. per second and after amyl nitrite 46 +/- 14 cm. per second, resulting in an average 84 per cent rise of blood velocity. It is concluded that amyl nitrite increases aortocoronary bypass graft blood velocity, suggesting a possible enhancement of blood flow to the distal native circulation in patients so operated upon.

Adult

Gastric-juice nitrite. A risk factor for cancer in the hypochlorhydric stomach?

Nitrite and hydrogen ion concentrations have been measured in the fasting gastric juice of 69 patients undergoing routine gastrointestinal investigations. There was an inverse relationship between nitrite concentration and hydrogen ion concentration, with a highly significant increase in gastric-juice nitrite in hypochlorhydric subjects. Thiocyanate was also found in all specimens in concentrations likely to increase nitrosamine formation, if nitrosation of amines occurs in the fasting stomach. Neutral gastric juice contains metabolically active bacteria capable both of generating nitrite from nitrate and of catalysing nitrosation. In this way an intragastric environment suitable for the formation of carcinogenic nitrosamines exists in the hypochlorhydric and achlorhydric stomach, providing a possible mechanism for the high incidence of gastric cancer in these subjects.

Achlorhydria

Electron-paramagnetic-resonance studies of the mechanism of leaf nitrite reductase. Signals from the iron-sulphur centre and haem under turnover conditions.

Low-temperature e.p.r. spectra are presented of nitrite reductase purified from leaves of vegetable marrow (Cucurbita pepo). The oxidized enzyme showed a spectrum at g=6.86, 4.98 and 1.95 corresponding to high-spin Fe(3+) in sirohaem, which disappeared slowly on treatment with nitrite. The midpoint potential of the sirohaem was estimated to be -120mV. On reduction with Na(2)S(2)O(4) or Na(2)S(2)O(4)+Methyl Viologen a spectrum at g=2.038, 1.944 and 1.922 was observed, due to a reduced iron-sulphur centre. The midpoint potential of this centre was very low, about -570mV at pH8.1, decreasing with increasing pH. On addition of cyanide, which binds to haem, and Na(2)S(2)O(4), the iron-sulphur centre became further reduced. We think that this is due to an increased midpoint potential of the iron-sulphur centre. Other ligands to haem, such as CO and the reaction product NH(3), had similar but less pronounced effects, and also changed the lineshape of the iron-sulphur signal. Samples were prepared of the enzyme frozen during the reaction with nitrite, Methyl Viologen and Na(2)S(2)O(4) in various proportions. Signals were interpreted as due to the reduced iron-sulphur centre (with slightly different g values), a haem-NO complex and reduced Methyl Viologen. In the presence of an excess of nitrite, the haem-NO spectrum was more intense, whereas in the presence of an excess of Na(2)S(2)O(4) it was weaker, and disappeared at the end of the reaction. A reaction sequence is proposed for the enzyme, in which the haem-NO complex is an intermediate, followed by other e.p.r.-silent states, leading to the production of NH(4) (+).

Electron Spin Resonance Spectroscopy

Quantification of the inhibitory effect of eriochrome black and sodium nitrite on non-specific immunofluorescent staining.

To evaluate non-specific staining (NSS) of animal tissues by FITC-labelled immunoglobulins a model system was developed. HeLa cells were treated with labelled antisalmonella globulins and the fluorescence intensity of the cells was determined quantitatively by means of a fibre optic probe system. This system was used to determine the optimal treatment conditions (adequate concentrations, duration of treatment) using the two NSS-reducing agents eriochrome black and sodium nitrite. Approximately the same inhibitory effects (40-50 per cent using nitrite; about 85 per cent using eriochrome black) were obtained by conjugates of different F/P ratio values. The fibre optic probe system was also used to determine the effects of the above-mentioned agents on the NSS of liver sections. In this system, the NSS was strongly reduced by eriochrome black whereas nitrite treatment did not induce any inhibitory effect. The applicability of nitrite and eriochrome black as NSS-reducing agents was further demonstrated by the fact that they had no influence on the specific fluorescence intensity of salmonella bacteria. The effect of eriochrome black was also studied in clinical specimens infected with salmonella or tularemia bacteria.

Animals

Effect of sodium nitrite inhibition on intracellular thiol groups and on the activity of certain glycolytic enzymes in Clostridium perfringens.

Activities of glyceraldehyde-3-phosphate dehydrogenase (EC 1.2.1.12) (GAP-DH) and aldolase (EC 4.1.2.13) in cells of Clostridium perfringens that had been inhibited with sodium nitrite were investigated. A complete loss in GAP-DH activity and a 67% decrease in aldolase activity were observed when growth of C. perfringens was inhibited. There was also a 91% decrease in the concentration of free sulfhydryl groups of soluble cellular components. Dithiothreitol restored some activity to inactive GAP-DH from sodium nitrite-inhibited cells, indicating that a loss of reduced sulfhydryl groups was involved in the inactivation of the enzyme. The evidence presented suggests that sodium nitrite inhibition of C. perfringens may involve an interaction of sodium nitrite as nitrous acid with sulfhydryl-containing constituents of the bacterial cell.

Cell Wall

Comparison of nitrite and dipslide methods for bacteriuria screening.

The results of screening for bacteriuria by the nitrite reaction and the dipslide method were compared. The nitrite reaction was specific in 99.8%, its sensitivity was 93.1% and its reproducibility, 100%. Specificity of the dipslide method was 94.5%. The nitrite reactions is considered most suited for screening, owing to its specificity, satisfactory sensitivity, short time requirement and low cost. The dipslide method is not suited for screening, as apart from being expensive it yielded 5.5% false positive reactions, but proved excellent for estimating the bacterial count in nitrite positive samples.

Adolescent

[Effect of the triazine herbicide herbazine-50 on the nitrate-nitrite concentration of corn].

Studied were the changes in the nitrate-nitrite content of maize grown on areas treated with herbazine-50 and artificial nitrogen fertilizers. The chemical analysis of various parts of the plant (stem. leaves, flowers and grains) revealed that the content of nitrates and nitrites was highest in the ears, and lowest in the leaves. Highest was also the content of nitrates and nitrites in all parts of the plant in maize from areas treated with herbazine-50 at the rate of 4 kg of a. i. per hectare as well as with artificial nitrogen fertilizers (ammonium nitrate and superphosphate at 450 kg/ha), followed by maize obtained from areas where ammonium nitrate and superphosphate at 450 kg/ha were introduced only. Slightly lower amounts of nitrates and nitrites were found in maize from areas treated with herbazine-50 at 4 kg/ha only, and lowest was their content in maize from areas that were neither fertilized nor treated with herbazine-50.

Herbicides

Comparison of two extraction procedures for recovering nitrite from cured meat products.

A study was undertaken to determine the extent of loss of nitrite during the analytical workup of cured meats for nitrite analysis. Two extraction techniques were compared: (A) a relatively long (2 hr) and hot (80 degrees C) procedure carried out at the natural pH of the sample, and (B) the method of Sen and Donaldson that specifies an alkaline solution and much milder conditions (20 min at 50 degrees C at pH 8) for extraction. The data obtained suggest that considerable losses of nitrite can occur with the former procedure, especially in acidic samples. The overall percentage recoveries of added nitrite by the 2 methods ranged between 20 and 112, and 80 and 114, respectively. Average percentage recoveries for the acidic (pH less than 5) samples only were 47.2 for Method A as compared with 100.2 for Method B.

Meat

[Absorption kinetics of nitrate and nitrite in the intestine of rats (author's transl)].

In situ perfusions into the small intestine of rats showed the absorption kinetics of nitrate and nitrite. This absorption is important since the rate rises to 50% for nitrates and 80% for nitrites in our experimental conditions. These compounds are excreted in the biliary secretion and, with the nitrate perfusions, nitrite appearance in the perfusion medium indicates some reduction potentiality either in the liver or in the entero-cells. These results are all the more interesting as men daily ingest nitrates and nitrites with many foods.

Animals

Acquisition deficits induced by sodium nitrite in rats and mice.

Graded doses of sodium nitrite were administered shortly before or immediately after a single training trial in an inhibitory avoidance task. A retention test was given at 24 h (rats) or 72 h (mice) after training. The results indicate that sodium nitrite impairs acquisition of an inhibitory avoidance response in rats and mice. Retention was impaired with doses lower than those required for producing hypoxic effects, as measured by the EEG, methemoglobin levels in the blood, blood pressure, and heart rate. Since hypoxia was not involved, it is likely that the effects were mediated by some direct action of sodium nitrite on the CNS.

Amnesia

Comparative transcriptomic analysis of the gills and hepatopancreas of freshwater-cultured Litopenaeus vannamei under chronic nitrite stress.

To investigate the differences in molecular responses between the gills and hepatopancreas of freshwater-cultured Litopenaeus vannamei under chronic nitrite stress, a 30-day chronic stress experiment was conducted with a control group and a stress group. Transcriptomic analysis of the gills and hepatopancreas was performed using Illumina sequencing; differentially expressed genes (DEGs) were identified, and GO, KEGG, GSEA, PPI, and RT-qPCR validation were carried out. The results showed that 196 DEGs (161 up-regulated and 35 down-regulated) were identified in the gills, and 287 DEGs (199 up-regulated and 88 down-regulated) in the hepatopancreas, with only 18 DEGs shared between the two tissues. DEGs in the gills were enriched in oxidoreductase activity, glycerophospholipid metabolism, and tyrosine metabolism; DEGs in the hepatopancreas were enriched in lipid transporter activity, phagosome, ECM-receptor interaction, and riboflavin metabolism. GSEA revealed significant suppression of the mTOR pathway in the gills and the Polycomb complex pathway in the hepatopancreas. PPI network analysis identified hub genes P5CS and eEF2 in the gills, and PER, TUBB1, SHMT, and TUBB4B in the hepatopancreas. RT-qPCR validation was consistent with the RNA-seq results (R2 = 0.764). This study indicates that, under chronic nitrite stress, the gill response is centered on redox regulation and inhibition of growth metabolism, whereas the hepatopancreas response primarily involves lipid transport, cytoskeletal remodeling, and phagosome activation. The two tissues synergistically adapt through fundamental biosynthetic and motor protein pathways. This research provides molecular evidence for deciphering the nitrite tolerance mechanisms in freshwater-cultured shrimp.

Animals

[Mutagenic activity of basfungin in its combined action with sodium nitrite].

The mutagenic activity of basfungin, when acting in combination with sodium nitrite, was studied in a subacute experiment, using the method of bone marrow cytogenetic analysis. The results were compared with those in groups in which basfungin and sodium nitrite were applied separately, as well as with the results of the parallel control. The mutagenic effect in the combined action group did not essentially differ from the effect of application of basfungin alone, both qualitatively and diaulitatively. Interesting results were obtained in the group receiving sodium nitrite alone - the percentage of cells with chromosomal aberrations was significantly than the control values.

Animals

Comparison of studies on saccharin and sodium nitrite.

A review of long term animal studies of saccharin and sodium nitrite was undertaken to assess the effect of variability of selected protocol elements on the results obtained. These elements were divided into 4 general categories: design, including selection of test animals, basal diet, dosage form and doses of test substance, route of administration, and duration of exposure; observations, including gross observations during life and at necropsy, clinical tests, and histopathology; performance, including conduct of the test and animal husbandry; and analytical procedures, including chemical and statistical analyses. Because many of the protocol elements are not fully discussed in study reports, it was often impossible to determine what actually had been done. The review of various saccharin studies suggests that bladder tumors resulted following in utero exposure. In utero exposure with sodium nitrite did not appear to cause reticuloendothelial changes. The numerous variations in protocol elements in the nitrite studies precluded identification of a prime element responsible for the variation in reticuloendothelial changes observed. It can be concluded from this review that achievement of reproducibility in long term studies requires minimal variation of protocol elements for the new study.

Animals

Analysis of nitrosamines in aqueous and biological fluids based on measurement of photochemically liberated nitrite.

A method is described for the analysis of nitrosamines in aqueous solution and in biological fluids (blood, plasma, and rat liver microsomal suspensions). The method is based on photochemical degradation of the nitrosamine in a controlled environment to yield the corresponding amine and nitrite ion, and the latter is subsequently used to form a chromophoric or fluorescent product. The analysis scheme is a modular three-component system consisting of a column to remove contaminating nitrite prior to photolysis, a photochemical reactor, and a chemical reactor. Additional modules are used to accommodate biological samples or large-volume (5--50 ml) aqueous samples. In this study, N-nitrosopyrrolidine, N-nitrosodimethylamine, and N,N-diethanolnitrosamine were utilized as substrates. Because of intersubstrate variability in the photochemical decomposition rate and overall nitrite yield, the structure (i.e., photochemical behavior) of the particular nitrosamine in the sample must be known prior to analysis. With a colorimetric readout, the sensitivity for analysis of N-nitrosopyrrolidine was 800 ng/ml for a 5-ml sample and the measurement precision was +/- 6% in the biological fluids. Fluorometric analysis improved sensitivity to 4 ng/ml with a precision of +/- 10% in biological media.

Animals