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PubMed2004

Correlation of nitrites in breath condensates and lung function in asthmatic children.

We aimed to evaluate the value of exhaled breath condensates in monitoring airway inflammation in childhood asthma before and after high altitude climate therapy. Forty-eight asthmatic children on regular anti-asthma treatment with a normal FEV1 and positive skin prick test for house dust mites were recruited. All children had been referred to an alpine clinic for high altitude climate therapy, because of persistent asthmatic symptoms despite use of daily anti-inflammatory treatment. Subjects were assessed on their arrival and before departure from the alpine clinic. Spirometry, bronchial provocation tests and measurements of nitrites in breath condensates were performed. Median levels of nitrites were significantly higher before than after high altitude climate therapy (1.27 vs. 0.93 microm; p = 0.008). In addition, MEF50 improved significantly (p < 0.0005). There was a significant correlation between nitrites in breath condensates and MEF50 (r = -0.63, p < 0.0001), symptoms (r = 0.47, p = 0.0007) and airway hyper-reactivity (AHR) (r = -0.41, p = 0.004). In summary, we found a reduction in nitrites in breath condensates after a high altitude climate therapy. Significant correlations were found between nitrites and MEF50, AHR and symptoms. We conclude that the measurement of nitrites may be feasible to objectively assess airway inflammation in asthmatic children in order to detect ongoing inflammation in children with normal FEV1 but persistent symptoms.

Adolescent↗

PubMed1997

'Locked-on' and 'locked-off' signal transduction mutations in the periplasmic domain of the Escherichia coli NarQ and NarX sensors affect nitrate- and nitrite-dependent regulation by NarL and NarP.

The Escherichia coli NarX, NarQ, NarL and NarP proteins comprise a two-component regulatory system that controls the expression of many anaerobic electron-transport and fermentation-related genes in response to nitrate and nitrite. Either of the two sensor-transmitter proteins, NarX and NarQ, can activate the response-regulator proteins, NarL and NarP, which in turn are able to bind at their respective DNA regulatory sites to modulate gene expression. NarX contains a conserved 17 amino acid sequence, designated the 'P-box' element, that is essential for nitrate sensing. In this study we characterize narQ mutants that also confer altered nitrate control of NarL-dependent nitrate reductase (narGHJI) and fumarate reductase (frdABCD) gene expression. While some narQ mutations cause the constitutive activation or repression of reporter-gene expression even when the cells are grown in the absence of the nitrate signal (i.e. a 'locked-on' phenotype), other mutations abolish nitrate-dependent control (i.e. a 'locked-off' phenotype). Interestingly the narQ (A42-->T) and narQ (R50-->Q) mutations along with the analogous narX18 (A46-->T) and narX902 (R54-->E) mutations also confer a 'locked-on' or a 'locked-off' phenotype in response to nitrite, the second environmental signal detected by NarQ and NarX. Furthermore, these narQ and narX mutations also affect NarP-dependent gene regulation of nitrite reductase (nrfABCDEFG) and aeg-46.5 gene expression in response to nitrite. We therefore propose that the NarQ sensor-transmitter protein also detects nitrate and nitrite in the periplasmic space via its periplasmic domain. A signal transduction model, which we previously proposed for NarX, is now extended to NarQ, in which a nitrate- or nitrite-detection event in the periplasmic region of the cell is followed by a signal transduction event through the inner membrane to the cytoplasmic domain of NarQ and NarX proteins to modulate their protein kinase/phosphatase activities.

Alanine↗

PubMed1997

Discrimination between structurally related ligands nitrate and nitrite controls autokinase activity of the NarX transmembrane signal transducer of Escherichia coli K-12.

Anaerobic respiratory gene expression in Escherichia coli is differentially controlled by nitrate and nitrite through dual interacting two-component regulatory systems. The NarX sensor is one of two membrane-spanning sensor kinases that control the phosphorylation state of two DNA-binding response regulators. We have studied NarX autophosphorylation in crude membrane preparations from cells that overexpress NarX protein. The low basal autophosphorylation rate was stimulated about sixfold and threefold by nitrate and nitrite respectively. This demonstrates that nitrate and nitrite differentially activate NarX autokinase activity. We also isolated single-residue substitutions in NarX that affect its ability to respond to or discriminate between nitrate and nitrite. Most of these substitutions affect residues within the conserved P-box sequence in the periplasmic domain. We characterized several of the mutants in vivo, by monitoring ligand-regulated gene expression, and in vitro, by monitoring ligand-responsive autophosphorylation. At least one change, K491 (Lys at position 49 changed to Ile), resulted in a protein with greatly impaired ability to discriminate between nitrate and nitrite. Other changes (H45E and R59K) resulted in proteins that responded normally to nitrate but were unable to respond to nitrite. These results implicate the P-box region in discrimination between subtly different small molecules.

Bacterial Proteins↗

PubMed2001

Blood nitrite levels in patients with migraine during headache-free period.

OBJECTIVE: To investigate blood nitrite levels after migraine attacks and to assess whether or not the change in nitric oxide levels observed during acute migraine persist after the attacks. BACKGROUND: Involvement of nitric oxide has been suggested in the initiation of acute migraine. Recent studies have shown alteration in the platelet response and platelet nitrite levels during migraine attacks. METHODS: Patients with migraine with aura and patients without aura were included in the study. The study was conducted on 50 patients with migraine and 90 healthy controls. Blood from the patients was collected at least 7 +/- 0.8 days after the last attack of migraine. Nitrite levels in the polymorphonuclear leukocytes, platelets, and plasma were estimated. Platelet aggregation response in some of these patients was also studied. RESULTS: No significant change in the polymorphonuclear leukocyte, platelet, and plasma nitrite levels in patients with migraine compared to controls was observed. Patients with migraine with aura had significantly lower polymorphonuclear leukocyte nitrite levels compared to those without aura (P<.05). In addition, no significant difference in the adenosine diphosphate-induced platelet aggregation was observed in the migraineurs compared to the healthy controls. CONCLUSIONS: Results obtained indicate that the platelet aggregation response and the blood nitrite levels were not altered significantly after an attack in the patients with migraine.

Adult↗

PubMed2000

Nitrite as a substrate and inhibitor of myeloperoxidase. Implications for nitration and hypochlorous acid production at sites of inflammation.

Myeloperoxidase is a heme enzyme of neutrophils that uses hydrogen peroxide to oxidize chloride to hypochlorous acid. Recently, it has been shown to catalyze nitration of tyrosine. In this study we have investigated the mechanism by which it oxidizes nitrite and promotes nitration of tyrosyl residues. Nitrite was found to be a poor substrate for myeloperoxidase but an excellent inhibitor of its chlorination activity. Nitrite slowed chlorination by univalently reducing the enzyme to an inactive form and as a consequence was oxidized to nitrogen dioxide. In the presence of physiological concentrations of nitrite and chloride, myeloperoxidase catalyzed little nitration of tyrosyl residues in a heptapeptide. However, the efficiency of nitration was enhanced at least 4-fold by free tyrosine. Our data are consistent with a mechanism in which myeloperoxidase oxidizes free tyrosine to tyrosyl radicals that exchange with tyrosyl residues in peptides. These peptide radicals then couple with nitrogen dioxide to form 3-nitrotyrosyl residues. With neutrophils, myeloperoxidase-dependent nitration required a high concentration of nitrite (1 mM), was doubled by tyrosine, and increased 4-fold by superoxide dismutase. Superoxide is likely to inhibit nitration by reacting with nitrogen dioxide and/or tyrosyl radicals. We propose that at sites of inflammation myeloperoxidase will nitrate proteins, even though nitrite is a poor substrate, because the co-substrate tyrosine will be available to facilitate the reaction. Also, production of 3-nitrotyrosine will be most favorable when the concentration of superoxide is low.

Cells, Cultured↗

PubMed1990

The effect of dietary nitrate on nitrate and nitrite excretion in man.

Dietary nitrate and nitrite may affect colonic pathophysiology. These anions influence fermentation, and nitrite has been shown to augment sodium absorption by the colon and participate in the formation of N-nitroso compounds. There is, however, no general agreement as to how much dietary nitrate and nitrite reaches the colon. To help resolve this question, balance studies were performed on six healthy ileostomy subjects who were given diets that varied in nitrate content from 0.83 to 5.20 mmol/d. Nitrate and nitrite excretion in ileal effluent and urine were measured by anion-exchange chromatography with conductivity detection. There was no significant nitrite in the diets, urine or ileal effluent. Dietary nitrate was largely excreted in urine (1.31-4.25 mmol/d). The urinary excretion findings indicated net synthesis of nitrate at low dietary intakes and net catabolism of nitrate at high intakes. Nitrate losses in ileal effluent were very low (0.03-0.05 mmol/d, 0.03-0.06 mmol/kg) and unrelated to intake for all the diets. It is concluded that dietary nitrate and nitrite do not enter the colon from the small intestine in amounts that would affect fermentation and mucosal metabolism in man. The possibility of significant amounts of nitrate reaching the colon via blood in normal subjects has not been excluded.

Aged↗

PubMed2003

Cerebrospinal fluid nitrite and malondialdehyde levels in patients with motor neuron disease.

Nitric oxide (NO) mediated oxidative damage may be involved in the pathogenesis of neuronal degeneration in motor neuron disease (MND). The present study was undertaken to evaluate the role of NO and oxidative stress in MND by estimating nitrite and malondialdehyde (MDA) levels in the cerebrospinal fluid (CSF) in 22 patients of MND and 20 control subjects suffering from neurological disorders not known to affect NO metabolism. There was no significant change in the CSF nitrite and MDA levels in MND. The nitrite and MDA levels did not have any significant correlation with age, duration of illness, or severity of disease. Univariate analysis of the clinical features in patients with MND and the nitrite levels revealed that two patients with a positive family history had significantly higher CSF nitrite levels as compared to those with a negative family history. There was no correlation between the CSF nitrite and MDA levels. Results of the present study did not indicate significant alterations in the MDA and NO levels in the CSF of MND patients. However, involvement of NO in MND with positive family history is suggested by the results obtained.

Adult↗

PubMed2005

Nitrites, nitrates and N-nitrosoamines in Estonian cured meat products: intake by Estonian children and adolescents.

The contents of nitrate, nitrite and N-nitrosoamines in commercial cured meat products on the Estonian market were determined for 2000-01 and 2003-04 as part of the Estonian food safety monitoring programme and the Estonian Science Foundation grant research activities. The maximum permitted levels of residual nitrites and nitrates were not exceeded in the samples analysed. However, a great variation in the content of nitrate, nitrite and N-nitrosoamines was found for all the products. The concentrations of these compounds in domestic cured meat products showed a decrease from year to year. The mean intake of nitrate, nitrite and N-nitrosoamines by Estonian children (n=346) from cured meat products was calculated on the basis of individual intake data. The mean daily intake of nitrates was 1.7 mg, that of nitrites was 0.83 mg and that of N-nitrosoamines was 0.073 microg. In the 2000-01 study, the calculated nitrite intake exceeded the acceptable daily intake by up to 140% for 1-6-year-old children and up to 105% in 2003-04.

Adolescent↗

PubMedDate not supplied

Calculated dietary intakes of nitrate and nitrite by young Finns.

Dietary intakes of nitrate and nitrite of 1212 Finns aged 9, 12, 15, 18, 21, and 24 years were calculated using food consumption data obtained by the 48-hour recall method in 1986, in connection with the Study on Cardiovascular Risk in Young Finns. Files on nitrate and nitrite content of foods and water were compiled for this study. The mean daily intakes of nitrate (NO3-) and nitrite (NO2-) from food were 54.0 mg and 1.4 mg, respectively. Vegetables including potatoes contributed 86% of nitrate intake and meat products 69% of the nitrite intake. If two litres of water within the mode concentration class of nitrate and nitrite were used, the intake of nitrate from water would have been between 1.0 and 2.0 mg and the intake of nitrite between 0.0 and 0.2 mg.

Body Weight↗

PubMed1999

Nitrate and nitrite in vegetables on the Danish market: content and intake.

The contents of nitrate and nitrite in lettuce, leek, potato, beetroot, Chinese cabbage and white cabbage on the Danish market were determined for 3 years in the period 1993-1997 as part of the Danish food monitoring programme. These vegetables are supposed to provide the major contribution to the intake of nitrate from the diet. Results for nitrate and nitrite in fresh and frozen spinach are also shown. The highest content of nitrate was found in lettuce followed by beetroot, Chinese cabbage, fresh spinach, leek, frozen spinach, white cabbage and potatoes. For all the products a great variation in the content of nitrate was found. For lettuce a characteristic variation throughout the year is clearly seen with the highest content in the winter period the lowest content in the summer period. Generally, the content of nitrite was low but in spinach high contents were found, probably due to improper storage conditions during transportation. The intake of nitrate and nitrite from these vegetables is calculated on the basis of two different consumption surveys. For both surveys the average intake of nitrate from the vegetables included in the monitoring programme is estimated to be approximately 40 mg day-1, whereas for nitrite the average intake is approximately 0.09 mg day-1. The total intake of nitrate and nitrite is estimated to be respectively 61 mg day-1 and 0.5 mg day-1.

Denmark↗

PubMed2004

Adaptation of the Griess reaction for detection of nitrite in human plasma.

The determination of nitrite in human plasma or serum has been most frequently used as a marker of nitric oxide (NO) production. In addition, it has recently been suggested that nitrite could act as a vasodilating agent at physiological concentrations by NO delivery. Therefore, nitrite determination in biological fluids is becoming increasingly important. The most frequently used method to measure nitrite is based on the spectrophotometric analysis of the azo dye obtained after reaction with the Griess reagent. This method has some limitations regarding detection limit and sensitivity, thus resulting unsuitable for nitrite detection in plasma. We have identified some drawbacks and modified the original procedure to overcome these problems. By the use of the newly developed method, we measured 221+/-72 nM nitrite in human plasma from healthy donors.

Health↗

PubMed1989

Urinary excretion of nitrate, nitrite and N-nitroso compounds in Schistosomiasis and bilharzia bladder cancer patients.

Saliva and 24-h urine samples were collected from male Schistosomiasis (bilharzia) patients with S. haematobium infection and possible concurrent S. mansoni infection without diagnosed bladder cancer (n = 27), bilharzia patients with diagnosed bladder cancer (n = 23) as well as a comparative control group (n = 27) of healthy Egyptian volunteers with no current bilharzia infection and/or bacterial urinary tract infections from the Nile Delta area of Egypt. Saliva samples were analysed for the presence of nitrate and nitrite; urine samples were analysed for the presence of nitrate, nitrite, volatile and non-volatile N-nitroso compounds. Bilharzia patients prior to, and after, diagnosed bladder cancer regularly excreted free nitrite as well as volatile nitrosamines (N-nitrosodimethylamine, N-nitrosodiethylamine, N-nitrosopiperidine and N-nitrosopyrrolidine) in addition to which elevated concentrations of non-volatile N-nitrosamino acids (N-nitrosoproline, N-nitrososarcosine, N-nitrosothiazolidine-4-carboxylic acid and its 2-methyl derivative) were also present. Total urinary excretion of volatile N-nitroso compounds (0.32 +/- 0.64 micrograms/day; mean +/- SD) and non-volatile N-nitroso compounds (31.20 +/- 22.07 micrograms/day) was observed in the Egyptian control group. Significantly higher concentrations were found in bilharzia patients: 3.47 +/- 6.42 (P less than 0.05) and 62.91 +/- 21.96 (P less than 0.05); as well as in bilharzia patients with diagnosed bladder cancer: 1.71 +/- 1.96 (P less than 0.02) and 44.94 +/- 7.31 respectively. Free nitrite was found in the urine of two volunteers in the Egyptian control group (1.7 and 3.0 micrograms/day), urinary nitrite was significantly increased in bilharzia patients (5.18 +/- 9.11 micrograms/day, P less than 0.02) and in bladder cancer patients (1.75 +/- 2.81 micrograms/day, P less than 0.05). Nitrate concentrations were elevated from 139.3 +/- 82.2 in the control group to 143.6 +/- 136.3 and 175 +/- 190 in the bilharzia and bladder cancer groups respectively. These results indicate that significant in vivo formation of nitrite and volatile N-nitroso compounds occurs in the urinary bladder of bilharzia patients and this may be an oetiological factor in the induction of bilharzial bladder cancer associated with S. haematobium infection.

Adult↗

PubMedDate not supplied

Are true-negative nitrite results affected by a two-hour delay in dipstick/pad analysis of urine in incontinence pads?

OBJECTIVES: To determine if true-negative nitrite results of a urine dipstick pressed into an incontinence pad (dipstick/pad method) are affected by a 2-hour delay in analysis. DESIGN: A quantitative study. SETTING AND SUBJECTS: Clean-catch urine specimens from a convenience sample of clinic patients, staff, and long-term care facility residents. INSTRUMENT: Changes from negative to positive for each group of urine specimens were evaluated using percentages within the groups. METHODS: Urine specimens were collected and a portion was cultured. Urine specimens negative for a urinary tract infection were included in the study. A portion of the specimen was poured into an incontinence pad. Initial nitrite results were determined using a dipstick pressed into an incontinence pad. Pads with true-negative dipstick/pad nitrite results were tested 2 hours later in the same manner. Urine culture results determined groups: mixed colonies; 50,000 to 75,000 colony-forming units per milliliter of a single type of uropathogen; greater than or equal to 50,000 cfu/mL diphtheroids; no significant growth; and no growth. RESULTS: Of the 443 urine cultures negative for a urinary tract infection, 441 initial dipstick/pad nitrite results were negative. Two initial true-negative nitrite dipstick/pad results, or 0.5%, changed from negative to positive over the 2-hour period: 1 in the "mixed colonies" group (0.4%) and 1 in the "no growth" group (0.7%). CONCLUSIONS: Results of this study indicate that true-negative nitrite results of a dipstick pressed into urine in an incontinence pad do not appear to be affected by a 2-hour delay in analysis.

Adolescent↗

PubMed1984

Pyruvate and ethanol as electron donors for nitrite reduction by Escherichia coli K12.

Pyruvate and ethanol were both effective electron donors for nitrite reduction by Escherichia coli K12. The pyruvate-dependent rate decreased by approximately 50% when either a cysG mutation, which results in loss of NADH-dependent nitrite reductase activity (EC 1.6.6.4), or a chl mutation, which results in loss of the formate-nitrite oxidoreductase activity, was introduced into the prototrophic parental strain CGSC4315. A double mutant deficient in both of these previously described activities retained only 2% of the rate of nitrite reduction of the parental strain after growth on glucose or 5% after growth on pyruvate. We conclude that any third pathway for nitrite reduction contributes little to the in vivo rate of nitrite reduction by wild-type strains.

Escherichia coli↗

PubMed2004

Mechanistic insight into the peroxidase catalyzed nitration of tyrosine derivatives by nitrite and hydrogen peroxide.

Peroxidases perform the nitration of tyrosine and tyrosyl residues in proteins, in the presence of nitrite and hydrogen peroxide. The nitrating species is still unknown but it is usually assumed to be nitrogen dioxide. In the present investigation, the nitration of phenolic compounds derived from tyrosine by lactoperoxidase and horseradish peroxidase was studied, with the aim of elucidating the mechanism of the reaction. The results indicate that nitrogen dioxide cannot be the only nitrating species and suggest the presence of two simultaneously operative pathways, one proceeding through enzyme-generated nitrogen dioxide and another through a more reactive species, assumed to be complexed peroxynitrite, which is generated by reaction of hydrogen peroxide with the enzyme-nitrite complex. The importance of the two pathways depends on peroxide and nitrite concentrations. With lactoperoxidase, nitration through the highly reactive intermediate is preferred except at very low nitrite concentration, while with horseradish peroxidase, the nitrogen dioxide driven mechanism is preferred except at very high nitrite concentration. The preferred mechanism for the two enzymes is that operative in the physiological nitrite concentration range.

Animals↗

PubMed2006

Nitrite concentration influences the population structure of Nitrospira-like bacteria.

Chemolithoautotrophic nitrite oxidizers of the genus Nitrospira are a monophyletic but diverse group of organisms, are widely distributed in many natural habitats, and play a key role in nitrogen elimination during biological wastewater treatment. Phylogenetic analyses of cloned 16S rRNA genes and fluorescence in situ hybridization with newly developed rRNA-targeted oligonucleotide probes revealed coexistence of uncultured members of sublineages I and II of the genus Nitrospira in biofilm and activated sludge samples taken from nitrifying wastewater treatment plants. Quantitative microscopic analyses of their spatial arrangement relative to ammonia oxidizers in the biofilm and activated sludge flocs showed that members of the Nitrospira sublineage I occurred significantly more often in immediate vicinity to ammonia oxidizers than would be expected from random community assembly while such a relationship was not observed for Nitrospira sublineage II. This spatial distribution suggested a niche differentiation of these coexisting Nitrospira populations with respect to their preferred concentrations of nitrite. This hypothesis was tested by mathematical modelling of nitrite consumption and resulting nitrite gradients in nitrifying biofilms and by quantifying the abundance of sublineage I and II Nitrospira in activated sludge during incubations with nitrite in different concentrations. Consistent with the observed localization patterns, a higher nitrite concentration selected for sublineage I but suppressed sublineage II Nitrospira.

Ammonia↗

PubMedDate not supplied

Nitrate and nitrite intake and the risk for type 1 diabetes in Finnish children. Childhood Diabetes in Finland Study Group.

The intakes of nitrate and nitrite of children and their parents from food and drinking water were estimated in a Finnish nation-wide case-control study on the epidemiology of Type 1 diabetes. The study population consisted of 684 case and 595 control children; 548 case-control pairs of fathers; and 620 case-control pairs of mothers. The consumption frequencies of foods which are important sources of nitrate and nitrite were assessed by structured questionnaire. Nitrate and nitrite concentration data were collected from Finnish water works. Diabetic children's and their mothers' daily dietary intake of nitrite was greater compared with that of control children and mothers (for case and control children 0.9 mg vs 0.8 mg, for case and control mothers 0.9 mg vs 0.8 mg, p < 0.001). Case mothers compared with control mothers received less (p < 0.05) nitrate from their diet. No differences were observed in the intake of nitrate or nitrite from drinking water. Dietary nitrite intake of children (odds ratios and 95% confidence intervals for the second, third, and fourth quartile 1.16, 0.82-1.65; 1.49, 1.06-2.10; 2.32, 1.67-3.24, respectively) and mothers (odds ratios and 95% confidence intervals for the second, third, and fourth quartile 1.15, 0.76-1.74; 1.29, 0.87-1.91; 1.98, 1.35-2.90, respectively) was positively associated with the risk for Type 1 diabetes independently from length of mother's education, child's or mother's age, place of residence or mother's smoking status.(ABSTRACT TRUNCATED AT 250 WORDS)

Adolescent↗

PubMedDate not supplied

Influence of Pneumocystis carinii on nitrite production by rat alveolar macrophages.

Nitrite production by rat alveolar macrophages was studied to determine the role of L-arginine oxidation in the interaction between these cells and Pneumocystis carinii. Alveolar macrophages from rats obtained from two different breeders were used: rats from Janvier breeder had latent P. carinii infection, while those from Charles River breeder were bred in a germ-free environment. Pneumocystis carinii increased in vitro nitrite generation by unstimulated alveolar macrophages from Janvier rats only, and this was blocked by NG-monomethyl-L-arginine. Incubation of cells from Janvier and Charles River rats with lipopolysaccharide and/or interferon-gamma increased nitrite production to a similar extent. Pneumocystis carinii partially decreased nitrite release by activated alveolar macrophages, and this was still inhibited by NG-monomethyl-L-arginine. In the presence of P. carinii, superoxide dismutase used as a superoxide anion scavenger had no effect on nitrite production by activated cells. These results show that prior exposure to P. carinii leads to nitric oxide production by rat alveolar macrophages. Although the magnitude of this production seems to be moderate, it is of biological significance since cells of P. carinii-naive rats do not generate nitrite whereas those of latently infected rats do.

Animals↗