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Evaluation of a new sensitive nitrite test as a reliable screening tool for bacteriuria.

A new sensitive nitrite test, the SRN test, was evaluated for its suitability as a reliable screening technique for the detection of bacteriuria. The SRN test was compared to a currently available nitrite test, the Microstix nitrite test, and the results obtained with both nitrite tests were assessed in comparison with the results of the quantitative culture method. Of 158 cases of significant bacteriuria found among 1060 randomly collected specimens, the SRN test detected 90% and the Microstix nitrite test, 30%. The higher reliability of the SRN test reflects its high capability of nitrite detection (greater than or equal to 0.1 ppm), and its ability to overcome interference by various factors, such as dark colour of the urine, presence of phenazopyridine, urobilinogen, blood, high concentration of ascorbate, and high urinary pH, all of which do interfere with the Microstix nitrite test. The high sensitivity of the SRN test allows detection of bacteriuria in urine specimens collected at random throughout the day; the test is therefore not restricted to the use of first-morning samples as are other nitrite tests. Since the SRN test was found to give a quantitative indication of the size of the bacterial population, the possibility of its use as an exact quantitative test under controlled conditions its discussed.

Bacteriuria↗

Serum concentrations of nitrite in patients with HIV-1 infection.

AIMS: To measure circulating concentrations of nitrite in patients with HIV-1 infection. METHODS: Nitrite concentrations were measured using the Griess reaction adapted to microtitre plates in the serum of 10 asymptomatic HIV-1 positive patients, 33 patients with AIDS with cerebral disorders, 17 patients with AIDS with pulmonary involvement, and in eight patients with AIDS with other disorders. Nitrite concentrations were also measured in bronchoalveolar lavage (BAL) fluid and cerebrospinal fluid (CSF) of patients with AIDS with pulmonary involvement and cerebral disorders, respectively. RESULTS: Increased serum concentrations of nitrite were observed in patients with pulmonary involvement, and in particular in serum and in BAL samples of patients with interstitial pneumonia (36.2 (26.2) mumol/l and 0.3 (0.4) mumol/l, respectively). Increased serum concentrations of nitrite were also noted in patients with retinitis caused by infection with cytomegalovirus. Serum nitrite concentrations were also raised in patients with cerebral toxoplasmosis, whereas normal serum concentrations were found in patients with HIV-1 encephalopathy and cryptococcal meningitis. Nitrite concentrations in CSF were not raised in patients with cerebral disorders. CONCLUSIONS: These results suggest that production of nitrite in patients with AIDS with concomitant opportunistic infections may be part of the host defense against opportunistic organisms.

Adult↗

Nitrite levels in breath condensate of patients with cystic fibrosis is elevated in contrast to exhaled nitric oxide.

BACKGROUND: Nitric oxide (NO) is released by activated macrophages, neutrophils, and stimulated bronchial epithelial cells. Exhaled NO has been shown to be increased in patients with asthma and has been put forward as a marker of airways inflammation. However, we have found that exhaled NO is not raised in patients with cystic fibrosis, even during infective pulmonary exacerbation. One reason for this may be that excess airway secretions may prevent diffusion of gaseous NO into the airway lumen. We hypothesised that exhaled NO may not reflect total NO production in chronically suppurative airways and investigated nitrite as another marker of NO production. METHODS: Breath condensate nitrite concentration and exhaled NO levels were measured in 21 clinically stable patients with cystic fibrosis of mean age 26 years and mean FEV1 57% and 12 healthy normal volunteers of mean age 31 years. Breath condensate was collected with a validated method which excluded saliva and nasal air contamination and nitrite levels were measured using the Griess reaction. Exhaled NO was measured using a sensitive chemiluminescence analyser (LR2000) at an exhalation rate of 250 ml/s. Fourteen patients with cystic fibrosis had circulating plasma leucocyte levels and differential analysis performed on the day of breath collection. RESULTS: Nitrite levels were significantly higher in patients with cystic fibrosis than in normal subjects (median 1.93 microM compared with 0.33 microM). This correlated positively with circulating plasma leucocytes and neutrophils (r = 0.6). In contrast, exhaled NO values were not significantly different from the normal range (median 3.8 ppb vs 4.4 ppb). There was no correlation between breath condensate nitrite and lung function and between breath condensate nitrite and exhaled NO. CONCLUSIONS: Nitrite levels in breath condensate were raised in stable patients with cystic fibrosis in contrast to exhaled NO. This suggests that nitrite levels may be a more useful measure of NO production and possibly airways inflammation in suppurative airways and that exhaled NO may not reflect total NO production.

Adult↗

The effect of nitrite on aerobic phosphate uptake and denitrifying activity of phosphate-accumulating organisms.

An anaerobic/aerobic/anoxic/aerobic sequencing batch reactor (SBR) was operated with municipal wastewater to investigate the effect of nitrite on biological phosphorus removal (BPR). When nitrite accumulated, aerobic phosphate uptake activity significantly decreased and, in case of hard exposure to nitrite, BPR severely deteriorated. The interesting observation was that the relative anoxic activity of phosphate accumulating organisms (PAOs) increased after nitrite exposure. Moreover batch tests of aerobic phosphate uptake in the presence/absence of nitrite indicated that PAOs with the higher relative anoxic activity are less sensitive to nitrite exposure. From these results, we concluded that BPR is sensitive to nitrite exposure, but BPR containing PAOs with the higher relative anoxic activity is possibly more stable against nitrite than BPR containing PAOs with the lower relative anoxic activity.

Bacteria, Anaerobic↗

Effect of inorganic carbon on nitrite accumulation in an aerobic granule reactor.

Pilot scale experiments were performed to evaluate the potential of nitrite type nitrification process with an airlift reactor and granular biomass. Initially, oxygen limitation was used as the main control parameter for accumulating nitrite in the effluent. After 30 d operation, the maximum nitrite conversion rate reached 2.5 kgNO2-N m(-3) d(-1), average diameter of the granule was 0.7 mm. Nitrite type reaction continued over 100 d, but nitrate formation increased after 150 d of operation. Once nitrate formation increased, oxygen limitation could not eliminate nitrite oxidising bacteria from granule. To overcome nitrate formation, laboratory scale batch experiments were conducted and it revealed a high concentration of inorganic carbon which had a significant effect on nitrite accumulation. Following this new concept, inorganic carbon was fed to the pilot scale reactor by changing pH adjustment reagent from NaOH to Na2CO3 and nitrite accumulation was recovered successfully without changing DO concentration. These results show that a high concentration of inorganic carbon is one of the control parameters for accumulating nitrite in biofilm nitrification system.

Aerobiosis↗

Mitochondria recycle nitrite back to the bioregulator nitric monoxide.

Nitric monoxide (NO) exerts a great variety of physiological functions. L-Arginine supplies amino groups which are transformed to NO in various NO-synthase-active isoenzyme complexes. NO-synthesis is stimulated under various conditions increasing the tissue of stable NO-metabolites. The major oxidation product found is nitrite. Elevated nitrite levels were reported to exist in a variety of diseases including HIV, reperfusion injury and hypovolemic shock. Denitrifying bacteria such as Paracoccus denitrificans have a membrane bound set of cytochromes (cyt cd1, cyt bc) which were shown to be involved in nitrite reduction activities. Mammalian mitochondria have similar cytochromes which form part of the respiratory chain. Like in bacteria quinols are used as reductants of these types of cytochromes. The observation of one-e- divergence from this redox-couple to external dioxygen made us to study whether this site of the respiratory chain may also recycle nitrite back to its bioactive form NO. Thus, the aim of the present study was therefore to confirm the existence of a reductive pathway which reestablishes the existence of the bioregulator NO from its main metabolite NO2-. Our results show that respiring mitochondria readily reduce added nitrite to NO which was made visible by nitrosylation of deoxyhemoglobin. The adduct gives characteristic triplet-ESR-signals. Using inhibitors of the respiratory chain for chemical sequestration of respiratory segments we were able to identify the site where nitrite is reduced. The results confirm the ubiquinone/cyt be1 couple as the reductant site where nitrite is recycled. The high affinity of NO to the heme-iron of cytochrome oxidase will result in an impairment of mitochondrial energy-production. "Nitrite tolerance" of angina pectoris patients using NO-donors may be explained in that way.

Animals↗

Nitrite accumulation in the treatment of wastewaters with high ammonia concentration.

Different operational parameters of the nitritation process were investigated in both jar tests and pilot scale Sequencing Batch Reactors (SBRs). In the laboratory study, 100-1,200 mg N l(-1) of ammonia was used. The pH and temperature were varied. Batch experiments were done on municipal sludge, pectin industrial sludge and sludge from a reject water treatment unit. Ammonia oxidation was observed with relative nitrite accumulations from 2% to 100% and ammonia oxidation rates from 0.01 to 0.58 g N g VSS(-1) d(-1). The nitritation process and relative nitrite accumulation were highly affected by pH, temperature and the sludge type. pH 8.0-8.5 and temperature 30 degrees C were found favourable for the nitritation. Pilot SBR systems for treating reject water achieved 100% of nitrite accumulation under the operational conditions of pH 7.5-8.0, temperature 30 degrees C and dissolved oxygen (DO) 1.0 mg 1(-1). Six months of operation revealed that pH regulations were essential to avoid the inhibitions by either free ammonia or nitrous acid. At an unionized ammonia concentration of approximately 20 mg NH3-N l(-1), half of the normal nitritation ability still remained. Total inhibition occurred when the concentration of nitrous acid reached 3.0 mg HNO2-N l(-1). However, both types of inhibitions were reversible in the SBR with a proper operation control. Stable and controllable nitritation could be achieved in pilot scale.

Ammonia↗

Nitrite oxidation inhibition by hydroxylamine: experimental and model evaluation.

A proposed approach for biological nitrogen removal significantly reduces cost by reducing biomass production and carbon requirements via inhibition of nitrite oxidation (NO2- to NO3-). Batch experiments were conducted to examine the effect of hydroxylamine (HM) on nitrite oxidizers, ammonia oxidizers, and nitrite reducers. Hydroxylamine effect experiments were done at initial pH values of 7.4-8.4, nitrogen concentrations of 100 mg N/L, biomass concentrations of 100-400 mg VSS/L and HM dosages up to 43 mg/L. Nitrite oxidizer activity was completely inhibited by HM at dosages of 7.0 and 8.9 mg/L for pH values of 8.4 and 7.6, respectively. Relatively low HM concentrations (0.35-5.5 mg/L) can be used to completely inhibit nitrite oxidation, but do not significantly affect ammonia oxidizers and nitrite reducers. A model developed to describe the effect of pH on nitrite oxidation rate fits the data well (R2 = 0.89) with values for Vmax of 0.372 (mg N/mg VSS-hr), pH* of 7.72, and the inhibition constant Kh of 0.154. Incorporation of HM inhibition into the model provided a good fit to relative nitrite oxidation rate as a function of undissociated HM concentration (R2 = 0.80, Vmax = 0.028 mg N/mg VSS-hr, pH = 7.89, Kh = 0.302, a = 0.195, and Ki= 0.277 mg/L).

Biomass↗

Catalysis of nitrosyl transfer reactions by a dissimilatory nitrite reductase (cytochrome c,d1).

The dissimilatory nitrite reductase (cytochrome c,d1) from Pseudomonas aeruginosa was observed at pH 7.5 to catalyze nitrosyl transfer (nitrosation) between [15N]nitrite and several N-nucleophiles or H2 18O, with rate enhancement of the order of 10(8) relative to analogous chemical reactions. The reducing system (ascorbate, N,N,N',N'-tetramethylphenylenediamine) could reduce nitrite (but not NO) enzymatically and had essentially no direct chemical reactivity toward nitrite or NO. The N-nitrosations showed saturation kinetics with respect to the nucleophile and, while exhibiting Vmax values which varied by about 40-fold, nevertheless showed little or no dependence of Vmax on nucleophile pKa. The N-nitrosations and NO-2/H2O-18O exchange required the reducing system, whereas NO/H2O-18O exchange was inhibited by the reducing system. NO was not detected to serve as a nitrosyl donor to N-nucleophiles. These and other kinetic observations suggest that the enzymatic nitrosyl donor is an enzyme-bound species derived from reduced enzyme and one molecule of nitrite, possibly a heme-nitrosyl compound (E-FeII X NO+) for which there is precedence. Nitrosyl transfer to N-nucleophiles may occur within a ternary complex of enzyme, nitrite, and nucleophile. Catalysis of nitrosyl transfer by nitrite reductase represents a new class of enzymatic reactions and may present another example of electrophilic catalysis by a metal center. The nitrosyl donor trapped by these reactions is believed to represent an intermediate in the reduction of nitrite by cytochrome c,d1.

Cytochromes↗

The sequential determination of nitrite, N-nitroso compounds and nitrate and its application.

The selective determination of nitrite, N-nitroso compounds as a group and nitrate can be accomplished directly on most food or other matrices by quantifying the nitric oxide released following the additions in sequence of acetic acid, hydrogen bromide and titanous chloride under an atmosphere of nitrogen. Most of the many other compounds potentially formed from nitrite in a biological system are either decomposed to nitric oxide in the refluxing solvent system alone or else do not yield nitric oxide at any stage throughout the determinations of nitrite and N-nitroso compounds. Only nitrolic acids and S-nitrothiols, like nitrosamines and nitrosamides, require the use of hydrogen bromide for breakdown to nitric oxide. The application of the present procedure to the determination of nitrite and N-nitroso compounds (as a group) in a series of animal diets has shown the presence of levels of nitrite below the limit of detection of more conventional methods, even in those diets not deliberately treated with nitrite. Evidence was also obtained for the presence of non-volatile, non-extractable N-nitroso compounds in diets both treated and untreated with high levels of nitrite. Confirmation was provided in the case of one diet not treated with nitrite, in that it was found to convert morpholine into its N-nitroso derivative, which was detected by gas chromatography with a Thermal Energy Analyzer detector.

Animals↗

Rat, mouse and human neutrophils stimulated by a variety of activating agents produce much less nitrite than rodent macrophages.

The role of reactive nitrogen intermediates (RNI) in the antimicrobial activities of neutrophils from various mammalian species is unclear. However, it has been reported that rodent neutrophils possess the inducible form of nitric oxide synthase and that inflammatory neutrophils from rats produce potentially antimicrobial levels of RNI. In the present study, neutrophils from humans, rats and mice were evaluated for production of nitrite, a stable end-product of RNI. Human neutrophil preparations (> 95% neutrophils) isolated from peripheral blood were stimulated for 2-24 hr with agents known to trigger the Ca(2+)-dependent constitutive nitric oxide synthase, or to stimulate synthesis of the inducible nitric oxide synthase. Superoxide dismutase was added to some cultures to decrease the levels of superoxide, a compound reported to react with RNI and yield products other than nitrite. Even though the cells were viable and responsive to stimuli, they did not produce nitrite concentrations indicative of antimicrobial potential. Preparations of inflammatory (casein-elicited) mouse neutrophils also failed to produce high concentrations of nitrite. Inflammatory rat neutrophils (2.5 x 10(6)/ml) produced nitrite concentrations of approximately 40 microM in 24-hr cultures, but plots of nitrite production versus cell number for neutrophil and macrophage preparations indicated that contaminating macrophages could account for all the nitrite production in the neutrophil preparations. Thus, neutrophils from rats, mice and humans seem comparable in their inability to produce high levels of nitrite in response to a variety of stimuli. This suggests that in most circumstances the constitutive nitric oxide synthase known to be present in these cells is limited to the production of low levels of nitric oxide for intercellular signalling. In addition, this raises questions about the presence or functional status of inducible nitric oxide synthase in rodent neutrophils.

Adult↗

Modulation of nitrate-nitrite conversion in the oral cavity.

The formation of nitrite from ingested nitrate can give rise to the induction of methemoglobinemia and endogenous nitrosation resulting in the formation of carcinogenic N-nitroso compounds. We investigated the possibility of modulation of the conversion of nitrate into nitrite in the oral cavity in order to seek ways of reducing the formation of the deleterious nitrite. We investigated the effectiveness of several mouthwash solutions with antibacterial constituents on the reduction of nitrate into nitrite in the oral cavity. In 15 studied subjects, the mean percentage of salivary nitrate reduced to nitrite after ingestion of 235 mg (3.8 mmol) nitrate was found to be 16.1 +/- 6.2%. The use of an antiseptic mouthwash with active antibacterial constituent chlorhexidine resulted in an almost complete decrease of the mean percentage of reduced nitrate, to 0.9 +/- 0.8%. Mouthwash solutions with antibacterial component triclosan or antimicrobial enzymes amyloglucosidase and glucose oxidase did not affect the reduction of nitrate into nitrite. A toothpaste with active components triclosan and zinc citrate with synergistic antiplaque activity was also without effect. Use of a pH-regulating chewing gum resulted in a rise in the pH in the oral cavity from 6.8 to 7.3. At 30 min after nitrate ingestion, this rise was accompanied by a significant increase in the salivary nitrite concentration, which might be explained by the pH being close to the optimal pH for nitrate reductase of 8. In conclusion, a limited number of possibilities of modulation of the conversion of nitrate into nitrite in the oral cavity are available.

Adult↗

[The in vitro effect of ascorbic acid on sodium nitrite intoxication].

The in vitro effect of ascorbic acid on the content of sodium nitrite in relation to pH and time of incubation and also effect of sodium nitrite on the total content of vitamin C as a sum of both reduced and oxidized forms was demonstrated. Ascorbic acid in respective quantities of 60 mg, 120 mg, 180 mg and sodium nitrite in quantity of 12 mg were added to the in vitro simulated stomach content at pH 1.5, 2.5, 3.5. The control samples were tested without addition of ascorbic acid. The Griess method for determination of nitrite and the Tillmans method with Pijanowski's modification were used for the determination of vitamin C. It was found that the choice of lower pH significantly decreased the level of nitrite in the control samples produced the disappearance of nitrite increased depending on the quantity of ascorbic acid added to the sample and on pH. The total content of vitamin C did not change after adding of sodium nitrite to the samples. The performed studies suggested a positive effect of ascorbic acid decreasing of the nitrite level.

Ascorbic Acid↗

Spinach ferredoxin-nitrite reductase: characterization of catalytic activity and interaction of the enzyme with substrates.

The steady-state kinetic parameters of the enzymatic reduction of nitrite by spinach ferredoxin-nitrite reductase [EC 1.7.7.1] were measured under anaerobic conditions. The maximum velocity of ferredoxin-linked activity was essentially the same as for the methyl viologen-linked activity of the enzyme. The initial velocity patterns of the oxidation of reduced ferredoxin suggested a sequential reaction scheme by which nitrite and reduced ferredoxin bind to the free enzyme. The binding of nitrite and ferredoxin to the enzyme was also investigated by different spectra produced by the complex formed by the enzyme with the substrates. Nitrite and ferredoxin each gave a 1: 1 complex with the enzyme. The dissociation constant (Kd) of the enzyme-nitrite complex agreed well with the Km value for the ferredoxin-linked activity, whereas the Kd of the enzyme-ferredoxin complex differed from the Km value for the enzyme activity. It was concluded that our preparation of spinach ferredoxin-nitrite reductase differs from both the complex (Mr = 85,000) and the modified (Mr = 61,000) forms of the enzyme reported by Hirasawa et al. [J. Biol. Chem. 262, 12428-12433 (1987)].

Catalysis↗

Biochemical and crystallographic studies of the Met144Ala, Asp92Asn and His254Phe mutants of the nitrite reductase from Alcaligenes xylosoxidans provide insight into the enzyme mechanism.

Dissimilatory nitrite reductase catalyses the reduction of nitrite (NO(2)(-)) to nitric oxide (NO). Copper-containing nitrite reductases contain both type 1 and type 2 Cu sites. Electron transfer from redox partners is presumed to be mediated via the type 1 Cu site and used at the catalytic type 2 Cu centre along with the substrate nitrite. At the type 2 Cu site, Asp92 has been identified as a key residue in substrate utilisation, since it hydrogen bonds to the water molecule at the nitrite binding site. We have also suggested that protons enter the catalytic site via Asp92, through a water network that is mediated by His254. The role of these residues has been investigated in the blue copper nitrite reductase from Alcaligenes xylosoxidans (NCIMB 11015) by a combination of point mutation, enzymatic activity measurement and structure determination.In addition, it has been suggested that the enzyme operates via an ordered mechanism where an electron is transferred to the type 2 Cu site largely when the second substrate nitrite is bound and that this is controlled via the lowering of the redox potential of the type 2 site when it is loaded with nitrite. Thus, a small perturbation of the type 1 Cu site should result in a significant effect on the activity of the enzyme. For this reason a mutation of Met144, which is the weakest ligand of the type 1 Cu, is investigated. The structures of H254F, D92N and M144A have been determined to 1.85 A, 1.9 A and 2.2 A resolution, respectively. The D92N and H254F mutants have negligible or no activity, while the M144A mutant has 30 % activity of the native enzyme. Structural and spectroscopic data show that the loss of activity in H254F is due to the catalytic site being occupied by Zn while the loss/reduction of activity in D92N/M144A are due to structural reasons. The D92N mutation results in the loss of the Asp92 hydrogen bond to the Cu-ligated water. Therefore, the ligand is no longer able to perform proton abstraction. Even though the loss of activity in H254F is due to lack of catalytic Cu, the mutation does cause the disruption of the water network, confirming its key role in proton channel. The structure of the H254F mutant is the first case where full occupancy Zn at the type 2 Cu site is observed, but despite the previously noted similarity of this site to the carbonic anhydrase catalytic site, no carbonic anhydrase activity is observed. The H254F and D92N mutant structures provide, for the first time, observation of surface Zn sites which may act as a Zn sink and prevent binding of Zn at the catalytic Cu site in the native enzyme.

Alcaligenes↗

Anaerobic nitrite production by plant cells and tissues: evidence for two nitrate pools.

Tobacco (Nicotiana tabacum L. cv. Xanthi) XD cells containing nitrate and nitrate reductase stopped producing nitrite after approximately 1 hour when incubated under anaerobic conditions. The cessation of nitrite production was not due to an inactivation of the nitrate reducing system. This was shown by the ability of the cells to resume anaerobic nitrite production at a rate similar to the initial rate of nitrite production upon exposure to nitrate, monohydroxy alcohols or pyrazole. Cessation of nitrite production also could not be attributed to leakage of nitrate from the cells. Although some nitrate did leak from the cells, most of the nitrate was still in the cells by the time anaerobic nitrite production ceased. We infer the existence of a small metabolic pool and a large storage pool of nitrate, such that nitrite production ceases when the metabolic pool is depleted of nitrate. The metabolic pool of nitrate in tobacco cells decreased 170-fold as the culture aged from 3 to 5 days. However, total cellular nitrate during this period remained relatively constant.Anaerobic nitrite production by barley (Hordeum vulgare) aleurone layers and corn (Zea mays) leaf sections also ceased after only a small fraction of endogenous nitrate was reduced and resumed again upon addition of exogenous nitrate. In contrast to that found with tobacco cells, the metabolic pool of nitrate in corn leaf sections remained constant with age, while total endogenous nitrate increased. These results were interpreted to mean that higher plants in general contain metabolic and storage pools of nitrate, the properties of which vary with species and physiological variables.

Journal Article↗

Dependence of nitrite reduction on electron transport chloroplasts.

Methyl viologen and phenazine methosulfate (photosystem I electron acceptors), 3-(3,4-dichlorophenyl)-1, 1-dimethylurea (DCMU, electron-transport inhibitor), and methylamine (photophosphorylation uncoupler) were used to study the dependence of nitrite reduction on electron transport in chloroplasts.DCMU, methyl viologen, and phenazine methosulfate markedly inhibited, whereas methylamine stimulated NO(2) (-) reduction in isolated, intact spinach (Spinacia oleracea L.) chloroplasts. The addition of DCMU to leaf sections of spinach and corn, (Zea mays L. var. XL81), incubated with No(3) (-), caused no inhibition of nitrate reduction but inhibited nitrite reduction leading to the accumulation of NO(2) (-) in the light. The addition of methylamine to comparable leaf sections did not affect either nitrate or nitrite reduction.WE CONCLUDED THAT: (a) nitrite reduction is functionally associated with the electron transport arising from the light reactions of the chloroplast and this provides additional support for the localization of nitrite reductase in the chloroplast; (b) nitrite reduction is associated with photosystem I and ferredoxin is the most likely donor in leaf tissue; and (c) ATP is not involved directly in nitrite reduction. However, ATP synthesis, by regulating electron flow to photosystem I, can affect nitrite reduction in the light.

Journal Article↗

Control of Nitrite Reductase Activity in Excised Embryos of Agrostemma githago.

When excised embryos of Agrostemma githago were incubated with nitrate, the activities of both nitrate reductase and nitrite reductase were enhanced. By contrast, benzyladenine induced nitrate reductase only. Our data suggest that nitrate affected nitrite reductase activity directly, without first being reduced to nitrite. When the endogenous nitrite production was increased by raising the level of nitrate reductase through simultaneous treatment with nitrate and benzyladenine, the activity of nitrite reductase was not higher than in embryos treated with nitrate alone. On the other hand, tungstate given together with nitrate drastically inhibited the development of nitrate reductase activity without reducing the enhancement of nitrite reductase activity. Nitrite enhanced nitrite reductase activity, though less efficiently than nitrate.

Journal Article↗