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At least 307 records · Page 17Linked to original sources
PubMed1989

Nitrate Reductase mRNA Regulation in Nicotiana plumbaginifolia Nitrate Reductase-Deficient Mutants.

Light and substrate regulation of nitrate reductase (NR) expression were compared in wild type and mutant lines of Nicotiana plumbaginifolia. Mutants affected in the NR structural gene (nia) or in the biosynthesis of the NR molybdenum cofactor (cnx) were examined. nia mutants expressing a defective apoenzyme, as well as cnx mutants, overexpressed NR mRNA, whereas nia mutants devoid of detectable NR protein had reduced or undetectable NR mRNA levels. Diurnal fluctuations of NR mRNA were specifically abolished in nia and cnx mutants, suggesting that the integrity of NR catalytic activity is required for the expression of diurnal oscillations. Unlike some fungal mutants, the nia and cnx mutants examined retained nitrate inducibility of NR expression. The possibility of autogenous control of NR expression in higher plants is discussed.

Journal Article↗
PubMed1991

A high-performance liquid chromatographic assay for phenylmercuric nitrate in the presence of zinc ions and its application to an assessment of the stability of phenylmercuric nitrate in zinc sulphate and zinc sulphate and adrenaline eye drops.

A high-performance liquid chromatographic assay for phenylmercuric nitrate in the presence of zinc ions has been developed. Application of the assay to zinc sulphate and zinc sulphate and adrenaline drops before and after autoclaving demonstrates that the phenylmercuric nitrate is chemically degraded in the zinc sulphate and adrenaline drops.

Chromatography, High Pressure Liquid↗
PubMedDate not supplied

A comparison of butoconazole nitrate cream with econazole nitrate cream for the treatment of vulvovaginal candidiasis.

In a randomized, single-blind, parallel study the safety and efficacy of 2% butoconazole nitrate cream used for 3 days were compared with those of 1% econazole nitrate cream used for 7 days at night in patients with vulvovaginal candidiasis. Patients in both treatment groups had positive potassium hydroxide smears and fungal cultures, and were similar in age, disease duration and history, obstetric history and contraceptive use. Of the 75 patients enrolled, 63 with a Candida albicans infection were included in the efficacy analyses. Evaluations were made at the start of the study (visit 1), after 10-23 days (visit 2) and after 24-45 days (visit 3). Both drugs significantly reduced all signs and symptoms, and at visits 2 and 3 the percentages of patients considered microbiologically, clinically and therapeutically cured were consistently higher for butoconazole- than for econazole-treated patients, although differences were not statistically different. Although both drugs were safe and well tolerated, it is concluded that butoconazole because of its much shorter regimen and superior clinical and microbiological performance was the drug of choice.

Administration, Intravaginal↗
PubMed2006

Mechanism of nitrate reduction by Desulfovibrio desulfuricans nitrate reductase--a theoretical investigation.

The oxidative half-reaction of oxygen atom transfer from nitrate to an Mo(IV) complex has been investigated at various levels of theory. Two models have been used to simulate the enzyme active site. In the second, more advanced model, additional amino acid residues capable of significantly affecting the catalytic efficiency of the enzyme were included. B3LYP/6-31+G*, ONIOM, and orbital-free embedding approaches have been used to construct the potential energy profile and to qualitatively compare the results of a QM/MM study with those obtained by a full quantum mechanical strategy. The study has confirmed the utility of the orbital-free embedding method in the description of enzymatic processes.

Binding Sites↗
PubMed1989

Monoclonal antibody-based immunoaffinity chromatography for purifying corn and squash NADH: nitrate reductases. Evidence for an interchain disulfide bond in nitrate reductase.

NADH: nitrate reductase (EC 1.6.6.1) (NR) is present in small amounts in plant tissues and its polypeptide in inherently labile. Consequently, NR is difficult to purify. We have generated 20 monoclonal antibodies (McAb) for corn and squash NR and selected two for use in immunoaffinity chromatography. Squash McAb CM 15(11) and corn McAb ZM 2(69)9, which both bind corn and squash NR, were covalently coupled to Sepharose and used for purification of NR with elution of the purified enzyme by a pH 11 buffer. Although this procedure yielded highly purified NR, its activity was diminished by the pH 11 treatment. When corn leaf crude extract was applied to McAb CM 15(11)-Sepharose, NR bound and could be eluted in homogeneous form by its substrate, NADH. Corn leaf NR prepared by substrate elution retained a high level of NADH: NR activity. Immunoaffinity-purified corn and squash NR were shown to have an interchain disulfide bond as well as a reactive thiol group. These results are discussed in relation to the recently obtained sequences of NR clones and suggestions made for site-directed mutagenesis experiments to aid in identifying the cysteine residues of NR associated with these features of the enzyme.

Antibodies, Monoclonal↗
PubMed2002

The stability of the Chlorella nitrate reductase mRNA is determined by the secondary structure of the 5'-UTR: implications for posttranscriptional regulation of nitrate reductase.

Nitrate reductase (NR; EC 1.6.1.1-3) can be controlled at both transcriptional and posttranscriptional levels. Here we describe stability of NR mRNA as a mechanism of control. The NR gene in Chlorella vulgaris (Warburg strain) transcribes a stable mRNA and an unstable mRNA. In-vitro-synthesized transcripts representing these mRNAs show the same stability characteristics. The unstable mRNA is 30 nucleotides longer at the 5'-UTR compared to the stable mRNA. Using an RNA-folding program the 5'-UTR of the longer unstable RNA showed a more extensive stem-loop structure compared to the more linear form of the shorter stable mRNA. Transcripts representing RNAs with intermediate 5'-UTRs folded similarly to the long form and were unstable, or similarly to the short form and were more stable. Thus the secondary structure of the 5'-UTR of NR mRNA is important in the stability of NR transcripts in Chlorella and allows the cell to respond to changes in nitrogen source in an energy-efficient manner.

5' Untranslated Regions↗
PubMed2002

Effects of nitrate pulses on BnNRT1 and BnNRT2 genes: mRNA levels and nitrate influx rates in relation to the duration of N deprivation in Brassica napus L.

A de-repression mechanism based on the disappearance of 'signals' down-regulating N transporter activity has been proposed in the literature to explain the transient increase of NO(3)(-) uptake by the roots following N deprivation in higher plants. This hypothesis was investigated at the physiological and molecular levels by measuring NO(3)(-) influx into roots of Brassica napus L. grown under low or high external concentrations of KNO(3) following N deprivation. Parallel measurements were made of endogenous NO(3)(-), amino acid concentrations and abundance of mRNA for BnNRT1 and BnNRT2, genes encoding nitrate-inducible transport proteins. The effect of NO(3)(-) pulsing on NO(3)(-) transport components in N-deprived plants was also investigated by measuring influx of high- and low-affinity transport system (HATS and LATS) and assaying mRNA levels. Influx of NO(3)(-) via HATS and LATS, and transcript levels of BnNRT2 and BnNRT1 decreased with the duration of N deprivation. The results suggested that the absence of de-repression of NO(3)(-) influx and BnNRT2 gene expression following N starvation was related to a high amino acid status. Pulsing with NO(3)(-) induced a large increase in BnNRT2 mRNA level, but a comparatively small increase in NO(3)(-) influx via HATS. The level of BnNRT1 mRNA also increased, but there was no effect on LATS uptake activity. The absence of a strict correlation between the NO(3)(-) transport activity and the mRNA BnNRT1 and BnNRT2 levels is discussed in terms of possible post-transcriptional regulation by the amino acids.

Amino Acids↗
PubMed1987

Comparative induction of nitrate reductase by nitrate and nitrite in barley leaves.

The comparative induction of nitrate reductase (NR) by ambient NO3- and NO2- as a function of influx, reduction (as NR was induced) and accumulation in detached leaves of 8-day-old barley (Hordeum vulgare L.) seedlings was determined. The dynamic interaction of NO3- influx, reduction and accumulation on NR induction was shown. The activity of NR, as it was induced, influenced its further induction by affecting the internal concentration of NO3-. As the ambient concentration of NO3- increased, the relative influences imposed by influx and reduction on NO3- accumulation changed with influx becoming a more predominant regulant. Significant levels of NO3- accumulated in NO2(-)-fed leaves. When the leaves were supplied cycloheximide or tungstate along with NO2-, about 60% more NO3- accumulated in the leaves than in the absence of the inhibitors. In NO3(-)-supplied leaves NR induction was observed at an ambient concentration of as low as 0.02 mM. No NR induction occurred in leaves supplied with NO2- until the ambient NO2- concentration was 0.5 mM. In fact, NR induction from NO2- solutions was not seen until NO3- was detected in the leaves. The amount of NO3- accumulating in NO2(-)-fed leaves induced similar levels of NR as did equivalent amounts of NO3- accumulating from NO3(-)-fed leaves. In all cases the internal concentration of NO3-, but not NO2-, was highly correlated with the amount of NR induced. The evidence indicated that NO3- was a more likely inducer of NR than was NO2-.

Cycloheximide↗
PubMed1992

Nitrates in different vascular beds, nitrate tolerance, and interactions with endothelial function.

The favorable anti-ischemic effect of nitrates is based on the unique distribution pattern of vascular relaxation that they evoke in different vascular sections. Nitrovasodilators reduce cardiac preload and wall tension, and thus myocardial oxygen consumption. They increase precollateral coronary perfusion pressure, thereby augmenting oxygen delivery to ischemic sections, especially to the subendocardial layers. These vasodilator actions are caused by the nitric oxide (NO)-induced activation of soluble guanylyl cyclase, which augments vascular cyclic guanosine monophosphate (cGMP) levels to suppress intracellular Ca2+ concentrations. After some metabolic steps NO is finally cleaved from all nitrovasodilators and is probably identical with, or very closely related to, endothelium-derived relaxing factor (EDRF). A dinitrosyl-iron complex may serve under biologic conditions to stabilize the NO- radical, which has an extremely short half-life. NO derived from nitrovasodilators is used therapeutically to substitute for a deficient endothelium-mediated vascular control and autacoid production.

Animals↗
PubMed2005

Spatial variability of shallow groundwater level, electrical conductivity and nitrate concentration, and risk assessment of nitrate contamination in North China Plain.

In recent years, nitrate (NO3) contamination of groundwater has become a growing concern for people in rural areas in North China Plain (NCP) where groundwater is used as drinking water. The objective of this study was to evaluate groundwater resource level, to determine groundwater quality and to assess the risk of NO3 pollution in groundwater in Quzhou County in the NCP. Ordinary Kriging (OK) method was used to analyze the spatial variability of shallow groundwater level, groundwater electrical conductivity (EC) and NO3-N concentrations, and Indictor Kriging (IK) method was used to analyze the data with NO3-N concentrations equal or greater than the groundwater NO3 pollution threshold (20 mg L(-1)). The results indicated that groundwater level averaged 9.81 m, a level 6 m lower than in 1990. The spatial correlation distances for groundwater level, EC and NO3-N concentration were 21.93, 2.19 and 3.55 km, respectively. The contour map showed that shallow groundwater level areas extended from north to south across the County. Groundwater EC was above 3 dS m(-1) in the most part of the northern county. Groundwater NO3 pollution (NO3-N> or =20 mg L(-1)) mainly occurred in the County Seat areas due to wastewater irrigation and excessive fertilizer leaching from agricultural fields. At Henantuang town, besides suburban of the County Seat, groundwater was also contaminated by NO3 shown by the map generated using the IK method, which was not reflected in the map generated using the OK method. The map generated using the OK method could not reflect correctly the groundwater NO3 pollution status. The IK method is useful to assess the risk of NO3 pollution by giving the conditional probability of NO3 concentration exceeding the threshold value. It is suggested that risk assessment of NO3 pollution is useful for better managing groundwater resource, preventing soil salinization and minimizing NO3 pollution in groundwater.

China↗
PubMed2002

Lack of tyrosine nitration by hypochlorous acid in the presence of physiological concentrations of nitrite. Implications for the role of nitryl chloride in tyrosine nitration in vivo.

Elevated levels of reactive nitrogen species (RNS) such as peroxynitrite have been implicated in over 50 diverse human diseases as measured by the formation of the RNS biomarker 3-nitrotyrosine. Recently, an additional RNS was postulated to contribute to 3-nitrotyrosine formation in vivo; nitryl chloride formed from the reaction of nitrite and neutrophil myeloperoxidase-derived hypochlorous acid (HOCl). Whether nitryl chloride nitrates intracellular protein is unknown. Therefore, we exposed intact human HepG2 and SW1353 cells or cell lysates to HOCl and nitrite and examined each for 3-nitrotyrosine formation by: 1) Western blotting, 2) using a commercial 3-nitrotyrosine enzyme-linked immunosorbent assay kit, 3) flow cytometric analysis, and 4) confocal microscopic analysis. With each approach, no significant 3-nitrotyrosine formation was observed in either whole cells or cell lysates. However, substantial 3-nitrotyrosine was observed when peroxynitrite (100 microm) was added to cells or cell lysates. These data suggest that nitryl chloride formed from the reaction of nitrite with HOCl does not contribute to the elevated levels of 3-nitrotyrosine observed in human diseases.

Enzyme-Linked Immunosorbent Assay↗
PubMed2000

Regulation of high-affinity nitrate transporter genes and high-affinity nitrate influx by nitrogen pools in roots of barley.

To investigate the regulation of HvNRT2, genes that encode high-affinity NO(3)(-) transporters in barley (Hordeum vulgare) roots, seedlings were treated with 10 mM NO(3)(-) in the presence or absence of amino acids (aspartate, asparagine, glutamate [Glu], and glutamine [Gln]), NH(4)(+), and/or inhibitors of N assimilation. Although all amino acids decreased high-affinity (13)NO(3)(-) influx and HvNRT2 transcript abundance, there was substantial interconversion of administered amino acids, making it impossible to determine which amino acid(s) were responsible for the observed effects. To clarify the role of individual amino acids, plants were separately treated with tungstate, methionine sulfoximine, or azaserine (inhibitors of nitrate reductase, Gln synthetase, and Glu synthase, respectively). Tungstate increased the HvNRT2 transcript by 20% to 30% and decreased NO(3)(-) influx by 50%, indicating that NO(3)(-) itself does not regulate transcript abundance, but may exert post-transcriptional effects. Experiments with methionine sulfoximine suggested that NH(4)(+) may down-regulate HvNRT2 gene expression and high-affinity NO(3)(-) influx by effects operating at the transcriptional and post-transcriptional levels. Azaserine decreased HvNRT2 transcript levels and NO(3)(-) influx by 97% and 95%, respectively, while decreasing Glu and increasing Gln levels. This suggests that Gln (and not Glu) is responsible for down-regulating HvNRT2 expression, although it does not preclude a contributory effect of other amino acids.

Anion Transport Proteins↗
PubMed1980

[Evolution of the nitrate content of water collected in the Department of Aisne, and search for side basins permitting correlations between surface events and nitrate contents].

The aim of this study is to search for small underground side basins in order to propose them to agricultural businessmen. Those basins should be apt to a successful experiment of cultural techniques to reduce nitrogen leakages toward aquifers. They should allow to identify and try to quantify the reasons of nitrogen enrichment of the underground waters by a critical examination of the past of some reappearances and the use of their recharge basin. This work consisted, after drawing up evolution graphs of nitrate rates, in selecting springs at first according to their interannual variations amplitude with increasing nitrogen rates, secondly according to their moderate flow-having unimportant side basins and thin storage aquifer and therefore reacting rapidly to the surface attraction. Finally, the springs were selected according to their specified pollution so that the collected water should be a representative sample of the aquifer. Eleven spots were thus selected. We hope they will allow to draw an experiment over the 3 or 5 next years.

France↗