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Direct serogrouping of group B streptococci from urogenital and gastric swabs with nitrous acid extraction and the Phadebact streptococcus test.

The micro-nitrous acid extraction-Phadebact coagglutination method was demonstrated to be a highly sensitive means to identify group B streptococci directly from cervical, uterine, and placental swab specimens, as well as from gastric and amniotic fluids. The swabs used for the standard culture method were used successfully for the direct extraction method. Accordingly, the direct swab extraction method yielded an identification of 92.0% of the group B streptococci identified by a standard culture procedure. Identification of the group B streptococci by the direct method was obtainable within 25 minutes of receipt of the clinical specimen in the laboratory. At this time, this highly rapid and sensitive method appears to be most applicable for the identification of group B streptococci from infants and colonized parturient women with associated risk factors before delivery, and therefore could provide a better opportunity for infant survival.

Antigens, Bacterial↗

[Mutagenic action of nitrous acid on prophage lambda].

The lethal and mutagenic effects of nitrous acid (0,1 M NaNO2 in 0,1 M acetate buffer, pH 4.6) on prophage lambda cI857 ind- were studied in the wild-type cells of Escherichia coli and in 9 repair-deficient mutants: uvrA6, uvrA6 umuC36, uvrD3, uvrE502, polA1, recA13, lexA102, recF143 and xthA9. After treatment with HNO2, the prophage was heat-induced either immediately or after 90 min incubation in broth at 32 degrees C. The prophage survival after delayed induction was considerably higher than after immediate induction. The lethal action of HNO2 was highly expressed in uvrA- and uvrE- lysogens after delayed induction. The frequency of temperature-independent c mutants forming clear plaques at 32 degrees C reached 4% in the wild-type host after immediate induction, this value being 10-15% in uvrA, uvrA umuC, uvrD, uvrE, polA and xthA mutants, 0,8% in recF- lysogen and only 0,2-0,3% in recA and lexA mutants. Under these conditions, about 90% of c mutants are generated by recA+, lexA+-dependent repair mechanism (most probably, due to W-mutagenesis). After delayed induction, mutation frequency in the wild-type host declines considerably (down to 0,1%). Analogous phenomenon of mutation frequency decline was registered in uvrA, xthA, recF, polA, uvrE and uvrD lysogens. Under conditions of delayed induction, the frequency of HNO2-induced c mutations only slightly depends on the recA+ and lexA+ gene products and mutations are, apparently, fixed by replication.

Bacteriophage lambda↗

The reaction of flavanols with nitrous acid protects against N-nitrosamine formation and leads to the formation of nitroso derivatives which inhibit cancer cell growth.

Studies have suggested that diets rich in polyphenols such as flavonoids may lead to a reduced risk of gastrointestinal cancers. We demonstrate the ability of monomeric and dimeric flavanols to scavenge reactive nitrogen species derived from nitrous acid. Both epicatechin and dimer B2 (epicatechin dimer) inhibited nitrous acid-induced formation of 3-nitrotyrosine and the formation of the carcinogenic N-nitrosamine, N-nitrosodimethylamine. The reaction of monomeric and dimeric epicatechin with nitrous acid led to the formation of mono- and di-nitroso flavanols, whereas the reaction with hesperetin resulted primarily in the formation of nitrated products. Although, epicatechin was transferred across the jejunum of the small intestine yielding metabolites, its nitroso form was not absorbed. Dimer B2 but not epicatechin monomer inhibited the proliferation of, and triggered apoptosis in, Caco-2 cells. The latter was accompanied by caspase-3 activation and reductions in Akt phosphorylation, suggesting activation of apoptosis via inhibition of prosurvival signaling. Furthermore, the dinitroso derivative of dimer B2, and to a lesser extent the dinitroso-epicatechin, also induced significant toxic effects in Caco-2 cells. The inhibitory effects on cellular proliferation were paralleled by early inhibition of ERK 1/2 phosphorylation and later reductions in cyclin D1 levels, indicating modulation of cell cycle regulation in Caco-2 cells. These effects highlight multiple routes in which dietary derived flavanols may exert beneficial effects in the gastrointestinal tract.

Absorption↗

[Transforming activity of phage T4r+ DNA, treated with ultraviolet light, nitrous acid, hydroxylamine and visible light in the presence of methylene blue].

The efficiency of phages T4rIIB-638v+ and T4rIIB-638v- transformation by native and denatured DNA treated with UV, nitrous acid, hydroxylamine and visible light in the presence of methylene blue is studied. A greater transformation efficiency of UV-irradiated T4r+ phage native and denatured DNA was observed in the v+ recipient as compared with v- recipients. Denatured donor DNA treated with nitrous acid has higher transformation activity in spheroplasts infected with T4v+ phage than in those infected with T4v- phage. Native donor DNA, treated with methylene blue and visible light-irradiated, developed a decrease of the transformation activity in T4v- phage-infected spheroplasts as compared with T4v+ phage-infected spheroplasts. Hydroxylamine treatment of native and denatured donor DNA did not reveal any differences in the transforming activity for v+ and v- recipients. Denatured donor DNA was more resistant to the effect of hydroxylamine than native DNA.

Coliphages↗

Improvement of bioinsecticides production through mutagenesis of Bacillus thuringiensis by u.v. and nitrous acid affecting metabolic pathways and/or delta-endotoxin synthesis.

AIMS: The present work aimed to obtain bioinsecticide over-producing mutants through classical mutagenesis of vegetative cells of Bacillus thuringiensis (Bt) by using u.v. and nitrous acid, and to evidence the involvement of cell metabolism in delta-endotoxin synthesis. METHODS AND RESULTS: Vegetative cells of Bt were treated by nitrous acid (0.17 mg ml(-1)) or exposed to u.v. rays (emitted at a wave length of 240 nm). The isolated survivors were screened on the basis of the production of delta-endotoxins and biomass in glucose and/or in gruel-based media at two aeration conditions. Bioinsecticide over-producing mutants were obtained with high frequencies because random mutations were shown to affect cell metabolism at different pathways related to the regulation of delta-endotoxin synthesis. CONCLUSIONS: Classical mutagenesis of Bt cells lead to the isolation of a large variety of delta-endotoxin over-producing mutants that could be classified into six groups based on the location of the mutations, particularly in metabolism pathways and delta-endotoxin synthesis. SIGNIFICANCE AND IMPACT OF THE STUDY: High frequencies of delta-endotoxin over-producing mutants of Bt could be obtained through classical mutagenesis of vegetative cells. This should contribute to a significant reduction of production and utilization costs of Bt bioinsecticides.

Bacillus thuringiensis↗

Mutagenicity arising from boiled rice on treatment with nitrous acid.

When an aqueous homogenate of boiled rice was treated with nitrous acid at pH 3, direct-acting mutagens were formed. The presence of the mutagens was demonstrated by isolating the mutagenic fractions through blue-rayon adsorption, a method used to extract polycyclic compounds, and subsequent high-performance liquid chromatography (HPLC). The mutagens were active in Salmonella typhimurium TA100 and TA98 without metabolic activation. Several different mutagens were formed, as judged from the HPLC profile.

Chromatography, High Pressure Liquid↗

Reaction of deoxyguanosine with nitrous acid.

We reinvestigated the reaction of dGuo in the presence of nitrous acid by reversed phase HPLC (RPLC) analysis. When 10 mM deoxyguanosine (dGuo) was treated with 100 mM NaNO2 in 3.0 M acetate buffer (pH 3.7) at 37 degrees C, unknown peak appeared in large amount (yield 19.7% at 2 h, 28.7% at 6 h). The yield of this newly found compound, denoted as 1, arose with increasing H+ concentration and was maximal at 50 degrees C. 1 has small depurination rate constant. If no enzymatic repair system for 1 exists, 1 would have strong mutagenic ability.

Chromatography, High Pressure Liquid↗

Prophage Activation as an Overlooked Mechanism Underlying the Biocidal Effect of Free Nitrous Acid in Sewers.

Biogenic hydrogen sulfide produced in sewer systems causes odor nuisance and concrete corrosion, necessitating effective biocidal control. Free nitrous acid (FNA) has emerged as a promising biocide, but its unclear mechanisms complicate dosage optimization and risk assessment. Here, using Desulfovibrio vulgaris as a model lysogenic bacterium, we demonstrate that low-dose FNA (0.2-4.0 mg N/L) induces bacterial inactivation via prophage activation-associated lysis in addition to chemical oxidation. Reactive nitrogen species (RNS) scavenging tests revealed that RNS-mediated oxidative stress was closely associated with prophage activation. Activated phages further infected new hosts, reducing the viability of freshly cultured D. vulgaris cells by 25.7% and increasing total phage production 15.2-fold. The phenomenon was further validated in real sewage biofilms, where increased phage production and decreased bacterial viability were observed at a low FNA dose of 0.2 mg N/L, below the dose required for cell destruction by chemical oxidation. Furthermore, metagenomic analysis of 896 sewage samples worldwide revealed that 71.9% of recovered sewage-derived microbial genomes harbor prophages, indicating a widespread genomic basis for prophage activation-mediated bacterial inactivation. Overall, this study expands current understanding of the biocidal mechanisms of FNA and contributes to the development of environmentally sustainable biocidal strategies.

Nitrous Acid↗

Intermediate in reaction of 2'-deoxycytidine with nitrous acid.

Using reversed phase (RP) HPLC analysis, we reinvestigated the reaction of dCyd with nitrous acid. When 10 mM 2'-deoxycytidine (dCyd) was incubated with 100 mM NaNO2 in 1.0 M acetate buffer (pH 3.7) at 37 degrees C, an unknown product (referred to compound 1) was detected. The time course of the yield of compound 1 showed profile characteristic of an intermediate. Under the physiological conditions, compound 1 was converted exclusively to dUrd with a life time of ca. 300 h. Elucidation of genotoxic effects of compound 1 may be an important subject of the future study.

Chromatography, High Pressure Liquid↗

Kinetics of nitrous acid formation in nitric acid solutions under the effect of power ultrasound.

Sonochemical nitrous acid formation was investigated in 0.1-4.0 mol dm(-3) aqueous nitric acid solutions under the effect of power ultrasound with 20 kHz frequency. HNO2 steady-state concentration was obtained under long-time sonication; the excess HNO2 formed is decomposed and evoluted from the solution as NO and NO2 gases. The HNO2 steady-state concentration and the HNO2 initial formation rate depend linearly on the HNO3 concentration and acoustic intensity (1.8-3.5 W cm(-2)) and decrease with rising temperature in the range 21-50 degrees C. The HNO2 formation rate depends on the type of saturating gas as follows: Ar > N2 > He > air. NO and O2 are the major gaseous products of HNO3 sonication. The NO2 accumulation of in the gas phase is observed only when the decomposition of HNO2 formed becomes noticeable. The gaseous products formation rates depend on the HNO3 concentration, acoustic intensity and the type of saturating gas. The mechanism of HNO2 sonochemical formation is assumed to be the thermal decomposition of HNO3 in the gaseous vicinity of collapsing bubbles or in the overheated liquid reaction zone surrounding the cavitational bubbles.

Journal Article↗

Polarographic determination of metyrosine through treatment with nitrous acid.

A simple and sensitive polarographic method is described for the determination of metyrosine through treatment with nitrous acid. The different experimental parameters affecting the derivatization process, as well as the polarographic analysis were studied. The derivatization product was found to be reducible at the dropping mercury electrode over the whole pH range in Britton Robinson buffers. At pH 5, a well-defined diffusion-controlled cathodic wave was produced. The limiting current versus the concentration plot was linear over the range 8-80 mumol/l in the direct current mode with a detection limit of 0.2 mumol/l. The method was then applied to the determination of metyrosine capsules, and the results obtained were in good agreement with those given by the USP method.

Methyltyrosines↗

Compounds formed by treatment of corn (Zea mays) with nitrous acid.

Nitrohexane has been identified as a major product formed following treatment of corn (Zea mays) with nitrous acid. Preliminary evidence suggests that another compound isolated from the nitrosated corn is an unsaturated nitrolic acid. As an aid to the analysis of N-nitro compounds, we have characterized the response of a chemiluminescence detector (Thermal Energy Analyzer) as a function of pyrolysis chamber temperature for several nitrosamines and for an aliphatic C-nitroso compound, an aromatic C-nitro compound, a nitramine and an alkyl nitrite. The response-temperature profiles are valuable in distinguishing among the various compounds and in optimizing the sensitivity of the detector for use in chromatography. Other tests, including photolysis and stability toward nitrite-scavenging reagents, further aid in distinguishing among the various compounds.

Chromatography, Gas↗

Polarographic determination of prenalterol hydrochloride through treatment with nitrous acid.

A simple and sensitive polarographic method is described for the determination of prenalterol hydrochloride through treatment with nitrous acid. The different experimental parameters affecting the derivatization process, as well as the polarographic analysis were studied. The derivatization product was found to be reducible at the dropping mercury electrode over the whole pH range in Britton-Robinson buffers. At pH 5, a well defined diffusion-controlled cathodic wave was produced. The limiting current vs. the concentration plot was linear over the range 0.015-0.15 mM in the direct current mode, with a minimum detectability of 0.8 microM. The proposed method was applied successfully to the determination of prenalterol hydrochloride either in the pure form or in its dosage forms.

Adrenergic beta-Agonists↗

A new instrument to measure gaseous nitrous acid (HONO) in the atmosphere.

A new in situ instrument (LOPAP: long path absorption photometer) to measure gaseous nitrous acid (HONO) using wet chemical sampling and photometric detection has been developed. This instrument is aimed to overcome the known problems with current HONO measurement techniques and was designed to be a cheap, sensitive, compact, and continuouslyworking HONO monitorfor ambient air measurements in the troposphere or for measurements of higher concentrations e.g. in smog chambers, in exhaust gases, and in indoor environments. Laboratory investigations were carried outto characterize the instrument components with respect to collection efficiency, optimum dye formation, optimum detection, and interfering species. Detection limits ranging from approximately 3 to 50 pptV have been obtained with response times from 4 to 1.5 min, respectively, using different instrument parameters. The accuracy of the measurements is in the range between +/-(10-15)%. The validation of the instrument was performed in the laboratory for HONO concentrations of 3 and 30 ppbV using ion chromatography and with a DOAS (differential optical absorption spectrometer) instrument in a large outdoor smog chamber in the range from 0.1 to 20 ppbV. The deviations were well within the errors of the measurements; however, when comparing the data with the DOAS instrument systematically higher values were found with the LOPAP instrument.

Air Pollutants↗

Nitrous acid interference with passive NO2 measurement methods and the impact on indoor NO2 data.

Recent research has demonstrated that nitrous acid (HONO) is produced in indoor environments by NO2 reacting with interior surfaces, and is also emitted directly by some combustion sources. We have recently characterized the interference by HONO to NO2 measurements made by several commonly used continuous NO2 monitors. This paper reports on the effect of HONO on NO2 measurements made by passive sampling devices. The objective of this study was to evaluate this interference, and the accuracy and precision of passive samplers used for indoor NO2 measurements. We report that HONO interferes quantitatively with three of the four NO2 passive samplers tested.

Air Pollution, Indoor↗

Formation of 2'-deoxyoxanosine from 2'-deoxyguanosine and nitrous acid: mechanism and intermediates.

The reaction mechanism for the formation of 2'-deoxy-oxanosine from 2'-deoxyguanosine by nitrous acid was explored using methyl derivatives of guanosine and an isolated intermediate of the reaction. When 1-methylguanosine was incubated with NaNO(2)under acidic conditions, N (5) -methyloxanosine and 1-methylxanthosine were generated, whereas the same treatment of N (2), N (2)-dimethylguanosine generated no product. In a similar experiment without NO(2)(-), participation of a Dimroth rearrangement was ruled out. In the guanosine-HNO(2)reaction system, an intermediate with a half-life of 5.6 min (pH 7.0, 20 degrees C) was isolated and tentatively identified as a diazoate derivative of guanosine. The diazoate intermediate was converted into oxanosine and xanthosine at a molar ratio (oxanosine:xanthosine) of 0.26 at pH 7.0 and 20 degrees C. The ratio was not affected by the incubation pH between 2 and 10, but increased linearly with temperature from 0.22 (0 degrees C) to 0.32 (50 degrees C). The addition of acetone also increased the ratio up to 0.85 (98% acetone). Based on these results, a con-ceivable pathway for the formation of 2'-deoxyoxanosine from 2'-deoxyguanosine by HNO(2)is proposed.

Cations↗

Nitrous acid mutagenesis of duplex DNA as a three-component system.

Purified native Hemophilus influenzae DNA is relatively insusceptible to nitrous acid (NA) mutagenesis in vitro, but is readily mutated following denaturation. NA mutagenicity for duplex DNA is significantly increased in the presence of various alcohols, glycols, phenols or primary amines. Phenol-extracted DNA contains dissociable contaminants of low molecular weight that enhance NA mutagenesis. Enhancement of NA mutagenesis by phenol and by spermine is due to the formation of unstable molecular species. We propose that reactive organic nitroso compounds are formed which then serve as delivery vehicles to promote mutagenicity of native DNA, perhaps via transnitrosation reactions. Similar reactions probably occur in vivo to promote NA-induced base substitution (but not frameshift) mutations in Salmonella typhimurium and in Escherichia coli. The possible significance of these observations to carcinogenesis is discussed.

DNA, Bacterial↗