Doses Of Nitrite Research Articles

P224 EFFECT OF SODIUM NITRITE ON RENAL DAMAGE FROM 2-KIDNEY, 1-CLIP (2K-1C) HYPERTENSION IN RATS

Renovascular hypertension is characterized by oxidative stress and decreased availability of nitric oxide (NO). Nitrite is a physiologically recycled metabolite that generates NO. Treatment with sodium nitrite has antihypertensive effects due to its antioxidant capacity. However, the renal effects of this treatment are not known in 2K1C hypertension. This study aims to evaluate the effect of treatment with sodium nitrite on kidney damage caused by 2K1C hypertension. Hypertension was induced by cleavage of the left renal artery in Wistar rats. Two weeks after surgery, the animals were treated with two doses (1 mg/kg/day or 15 mg/kg/day) of sodium nitrite or vehicle for 4 weeks, by gavage. Blood pressure (BP) was checked weekly by tail plethysmography. To assess renal function, plasma urea and creatinine were determined. Kidney oxidative stress was evaluated through the activity of catalase (CAT), superoxide dismutase (SOD), levels of reduced glutathione (GSH), lipid peroxidation, and superoxide (O2-) levels. Treatment with 15 mg/kg/day of sodium nitrite reversed the increase in BP in 2K1C rats (p<0.05, 2K1C:170.5±6.542; 2K1C+Nitrite15:162.4±6.712 mmHg), and improved renal function previously impaired by hypertension (urea, p<0.05; 2K1C:76.37±24.53; 2K1C+Nitrite15:47.82±13.86; creatinine, p<0.05; 2K1C:0.540±0.105; 2K1C+Nitrite15:0.3550±0.117). SOD activity decreased in the kidneys of hypertensive animals (p<0.05, Control:0.7176±0.06426; 2K1C:0.636±0.0888; 2K1C+Nitrite1:0.703±0.0866; 2K1C+Nitrite15:0.648±0.173). As for CAT and GSH, there was a decrease in activity in hypertensive animals, with improvement in the group treated with nitrite 15 mg/kg/day (CAT p<0.05, Control:0.651±0.255; 2K1C:0.2772±0.1837; 2K1C+Nitrite15: 0.6654±0.1440 and GSH: p<0.05, Control:44.80±10.59; 2K1C: 29,62±7,253 2K1C+Nitrite15:44.25±10.07). Lipid peroxidation increased by hypertension was reduced with treatment at the highest dosage (p<0.05, Control:0.314±0.158; 2K1C:0.497±0.113; 2K1C+Nitrite15:0.278±0.084). O2- levels were decreased in all groups treated with nitrite (p<0.05, 2K1C:3.844±1.598; 2K1C+Nitrite1:0.9643±0.2063; 2K1C+Nitrite15:0.4431±0.1838). Our results suggest that sodium nitrite treatment improves 2K1C renal damage through its antioxidant activity. Acknowledgments: FAPEMIG# APQ-00946-23, CNPq# 302076/2022-0

Nitrite-dependent microbial utilization for simultaneous removal of sulfide and methane in sewers

Chemicals are commonly dosed in sewer systems to reduce the emission of hydrogen sulfide (H2S) and methane (CH4), incurring high costs and environmental concerns. Nitrite dosing is a promising approach as nitrite can be produced from urine wastewater, which is a feasible integrated water management strategy. However, nitrite dosing usually requires strict conditions, e.g., relatively high nitrite concentration (e.g., ∼200 mg N/L) and acidic environment, to inhibit microorganisms. In contrast to “microbial inhibition”, this study proposes “microbial utilization” concept, i.e., utilizing nitrite as a substrate for H2S and CH4 consumption in sewer. In a laboratory-scale sewer reactor, nitrite at a relatively low concentrations of 25–48 mg N/L was continuously dosed. Two nitrite-dependent microbial utilization processes, i.e., nitrite-dependent anaerobic methane oxidation (n-DAMO) and microbial sulfide oxidation, successfully occurred in conjunction with nitrite reduction. The occurrence of both processes achieved a 58 % reduction in dissolved methane and over 90 % sulfide removal in the sewer reactor, with microbial activities measured as 15.6 mg CH4/(L·h) and 29.4 mg S/(L·h), respectively. High copy numbers of n-DAMO bacteria and sulfide-oxidizing bacteria (SOB) were detected in both sewer biofilms and sediments. Mechanism analysis confirmed that the dosed nitrite at a relatively low level did not cause the inhibition of sulfidogenic process due to the downward migration of activity zones in sewer sediments. Therefore, the proposed “microbial utilization” concept offers a new alternative for simultaneous removal of sulfide and methane in sewers.

Physicochemical properties and sensory features of ripened, industrially prepared sausages, enriched with olive leaf extract to replace nitrite and nitrate

The use of nitrite and nitrate in meat products has been questioned due to the generation of N-nitroso compounds with carcinogenic effects. Thus, research has been focused on their replacement with several alternatives. This study is focused on evaluating the effects of a commercial olive leaf extract (OLE), marketed as dietary supplement, on protein and lipid oxidation, N-nitrosamine content and sensory properties of industrial sausages at the end of the ripening period. Five different formulations were prepared: F1 (OLE: 1000 mg/kg; NO2–NO3: 0 mg/kg), F2 (OLE: 1000 mg/kg; NO2–NO3: 75-75 mg/kg), F3 (OLE: 0 mg/kg; NO2–NO3: 0 mg/kg), F4 (OLE: 0 mg/kg; NO2–NO3: 75-75 mg/kg), F5 (OLE: 500 mg/kg; NO2–NO3: 35-35 mg/kg). The use of OLE in combination with a reduced dose of nitrite and nitrate (F5) significantly reduced the lipid and protein oxidation (p < 0.05) compared to the formulation with synthetic additives alone (F4). No differences were observed in terms of textural properties with the samples without the extract. The OLE did not affect the microbiological quality and enabled a significant reduction in the N-nitrosamines content. However, the addition of OLE decreased the intensity of red colour and determined the insurgence of a slight bitter taste in the sausages.

Nitrite decreases sickle hemoglobin polymerization in vitro independently of methemoglobin formation

The root cause of sickle cell disease (SCD) is the polymerization of sickle hemoglobin (HbS) leading to sickling of red blood cells (RBC). Earlier studies showed that in patients with SCD, high-dose nitrite inhibited sickling, an effect originally attributed to HbS oxidation to methemoglobin-S even though the anti-sickling effect did not correlate with methemoglobin-S levels. Here, we examined the effects of nitrite on HbS polymerization and on methemoglobin formation in a SCD mouse model. In vitro, at concentrations higher than physiologic (>1 μM), nitrite increased the delay time for polymerization of deoxygenated HbS independently of methemoglobin-S formation, which only occurred at much higher concentrations (>300 μM). In vitro, higher nitrite concentrations oxidized 100% of normal hemoglobin A (HbA), but only 70% of HbS. Dimethyl adipimidate, an anti-polymerization agent, increased the fraction of HbS oxidized by nitrite to 82%, suggesting that polymerized HbS partially contributed to the oxidation-resistant fraction of HbS. At low concentrations (10 μM-1 mM), nitrite did not increase the formation of reactive oxygen species but at high concentrations (10 mM) it decreased sickle RBC viability. In SCD mice, 4-week administration of nitrite yielded no significant changes in methemoglobin or nitrite levels in plasma and RBC, however, it further increased leukocytosis. Overall, these data suggest that nitrite at supra-physiologic concentrations has anti-polymerization properties in vitro and that leukocytosis is a potential nitrite toxicity in vivo. Therefore, to determine whether the anti-polymerization effect of nitrite observed in vitro underlies the decreases in sickling observed in patients with SCD, administration of higher nitrite doses is required.

Effect of application rate of sodium nitrite and hexamine on the fermentation and the chemical composition of guinea grass silage harvested at different stages of maturity

Grass ensilability varies with maturity stage, mainly due to changing concentrations of dry matter and soluble carbohydrates with progressing forage maturity. Consequently, the required dose of silage additive to prevent the development of undesirable microorganisms may change with maturity stage. The objective of this study was to verify whether the application rate of an additive containing sodium nitrite and hexamine interacts with guinea grass maturity to alter silage fermentation and chemical composition. Four fields of guinea grass (0.5–0.7 ha each field) were mowed and divided into two plots per field. After 5 wk, one plot of each field was mowed again to establish differences in stage of maturity. Ten weeks after the first mowing, the grass plots with 5- and 10-wk regrowth were manually harvested and used for the trial. The grass from each plot (approx. 30 kg) was chopped and divided into 3 piles, totaling to 24 piles, as result of four fields, two maturities, and three additive treatments: control (without additive), low dose of sodium nitrite (0.5 g/kg) + hexamine (0.325 g/kg) (NHL), and high dose of sodium nitrite (1 g/kg) + hexamine (0.65 g/kg) (NHH). After 90 d of storage, the silos were opened and silages sampled to determine dry matter (DM) loss, microbial counts, fermentation end-products, aerobic stability, chemical composition, and in vitro DM digestibility. Guinea grass harvested at 10-wk regrowth had a lower content of crude protein (P < 0.001) and a greater content of cell wall components (P < 0.001), resulting in a more lignified (P < 0.001) and less digestible (P < 0.001) forage than that harvested at 5 wk. There were interactions between plant maturity and additive dose for several silage traits (P < 0.05), likely due to the slightly greater fermentability coefficient (+5.1 points) for the more mature grass (P < 0.001). Within each maturity stage, silage pH and fermentation end-products associated with clostridia metabolism (i.e., n-butyric acid, propionic acid, i-butyric acid, i-valeric acid, n-valeric acid, ammonia, and 2,3-butanediol) linearly decreased (P < 0.001) with additive application rate, but the magnitude of improvement was slightly greater for 5-wk than 10-wk regrowth. Application of additive linearly decreased silage DM loss at both 5-wk (95.2, 46.7, and 20.6 g/kg DM, P < 0.001) and 10-wk (66.5, 31.7, and 13.6 g/kg DM, P < 0.001) regrowth stages, but only silages treated with NHH had n-butyric acid concentration < 3 g/kg DM. The proportion of rumen undegradable protein (P < 0.001), soluble carbohydrates concentration (P < 0.001), and in vitro DM digestibility (P < 0.001) were linearly increased with additive dose within each maturity stage. As treated silages were better conserved, silage aerobic stability was linearly reduced (P < 0.001) with additive dose, although all silages were aerobically stable for ≥ 4.7 d. In conclusion, the additive based on sodium nitrite and hexamine, applied at a regular dose, was able to largely restrict Clostridium development and DM losses during fermentation of guinea grass silage at both maturity stages. However, harvesting more mature grass markedly impaired its chemical composition and digestibility, rendering it no feasible strategy to reduce the additive application rate by half.

Nitrite Toxicity: Chemical Analysis, Metabolism, and Health Effects

Nitrites can be formed from and away by the nitrate-nitrite-nitric oxide pathway. The properties of nitrite oxidation and metHb formation are carefully studied in mechanism, forming either N-binding or O-binding structures. Apart from nitrate and nitric oxide, nitrites can also form carcinogenic nitrosamines in acidic environments. MetHb can cause hypoxia and vasodilation, while symptoms are revealed in different degrees under recalled or present hypoxic conditions. The study thoroughly studied nitrite’s metabolic properties, chemical pathways, and dosage effects on health. The cancer risks of consuming dietary nitrite need more statistical support, while its metabolite N-nitrosodimethylamine and NDMA concentration are well considered with increasing cancer risks. ED50 of human vasodilation is identified, and lethal doses on juvenile pike-perch can be further utilized to predict related doses for humans. More studies should be done to investigate relative nitrite doses to boost utilization and studies about this chemical.

Effects of low-dose sodium nitrite on the structure of yak meat myoglobin during wet curing.

The effects of low doses of sodium nitrite on yak meat colouring, myoglobin oxygenation status, myoglobin aggregation and myoglobin structure were evaluated using Fourier transform infrared spectroscopy, laser micro-Raman spectroscopy and liquid chromatography-electrospray ionization tandem mass spectrometry. The results showed that the yak meat redness value increased steadily relative to that of the control after the addition of low dose sodium nitrite. The nitrosomyoglobin level gradually increased and was significantly higher in the sodium nitrite-treated group than in the control group. The secondary structures were also transformed. The Cα-N bond extended and then contracted, the area of the haem core decreased and then increased and the frequency of contraction increased. A total of 34 nitrosylated peptides were identified, of which 15 were stable and 19 were unstable. These findings show that low doses of sodium nitrite facilitated the dynamic transformation of the myoglobin nitrosylated peptide fragment, which in turn preserved the colour of the meat.

Effects of nitrite concentrations on the quality and protein oxidation of salted meat.

The objective of this study was to investigate the effects of different concentrations of sodium nitrite on the quality and protein oxidation of salted meat during 21 days of curing. The salted meat was treated with sodium nitrite at 50, 100, and 150 mg/kg for curing, and salted meat without sodium nitrite was used as a control. The results showed that in salted meat added with sodium nitrite, the carbonyl group, disulfide bond, dityrosine, surface hydrophobicity, and the transformation rate from α-helix to β-sheet were all significantly reduced, whereas the sulfhydryl group content of myofibrillar proteins was significantly increased compared to the control. Meanwhile, the total volatile basic nitrogen and aerobic plate content were significantly decreased, while both the pH and a* value were significantly increased with an increase in nitrite concentration compared to the control group. Importantly, this phenomenon was also observed in salted meat treated with low doses of sodium nitrite (50 mg/kg). In conclusion, the quality of salted beef can be improved by adding low-dose sodium nitrite to inhibit protein oxidation during the curing process. PRACTICAL APPLICATION: A low dose of sodium nitrite inhibited the rate of α-helix to β-sheet transformation of myofibrillar proteins in salted meat, reducing the exposure of hydrophobic groups and decreasing the production of protein oxidation products and TVB-N to improve the quality of salted meat. These results provided a theoretical basis and technical guidance for the application of low-dose sodium nitrite in meat processing enterprises.

Effect of subchronic use of sodium nitrite on the functional parameters of an isolated rat heart

Introduction. The active development of agriculture with the widespread use of nitrogen-containing fertilizers, increased consumption of processed meat products containing sodium nitrite as a preservative, inevitably leads to an increase in the nitrogen load on the human body and can cause the development of cardiovascular diseases. Materials and methods. The cardiotoxic effect of sodium nitrite was evaluated on a model of an isolated rat heart with subchronic intragastric administration for 28 days at doses of 0.03; 0.3; 3.0 mg/kg. Results. The use of sodium nitrite leads to dilation of the coronary vessels and myocardium, inhibition of the contractile activity of the heart, disruption of its electrical activity in the form of a shortening of the QT interval associated with an increased risk of ventricular fibrillation, and a tendency to increase the amplitude of the T wave, which indicates myocardial ischemia. In a stress test with total ischemia, it was revealed that the magnitude of the change in the parameters characterizing the restoration of myocardial function under reperfusion conditions is inversely proportional to the dose of sodium nitrite used. Limitations. The study was performed only on male white rats more prone to the development of cardiovascular pathology. Conclusion. With subchronic administration, sodium nitrite has cardiotoxicity at a dose of 0.028 mg/kg, which is twice lower than the WHO-established permissible daily intake for humans (0.06 mg/kg).

Acute Nitrite Exposure Induces Dysfunction and Oxidative Damage in Grass Carp Isolated Hemocytes.

To evaluate the effects of nitrite on the oxidative damage of blood cells of Grass Carp Ctenopharyngodon idella, the isolated hemocytes were exposed to nitrite (0, 1, 10, or 100 mg/L) for up to 24 h. Hemoglobin (Hb) and methemoglobin (MetHb) concentrations, reactive oxygen species (ROS) and malondialdehyde (MDA) levels, mitochondrial membrane potential (∆Ψm), and antioxidant enzyme activity were assayed to assess hematological parameters and the antioxidant defense mechanism. Results showed a remarkable decrease in Hb concentration with increasing nitrite concentration after a 24-h exposure, while the MetHb concentration increased significantly in nitrite exposure groups. The levels of ROS, ∆Ψm, and MDA increased to varying degrees with increases in nitrite exposure concentration and time. The total antioxidant capacity, catalase (CAT) activity, glutathione peroxidase (GPx) activity, and glutathione content showed a trend of rising initially and then decreasing with prolonged exposure time. Superoxide dismutase (SOD) activity was higher in the 1-mg/L nitrite exposure group and lower in the 100-mg/L group than in the control. The relative messenger RNA expression ratios of cat, sod1, and gpx were up-regulated significantly in the 1- and 10-mg/L groups and then declined in the 100-mg/L group. Therefore, it can be concluded that nitrite exposure activates the antioxidant defense mechanism of Grass Carp hemocytes and that the balance of oxidant-antioxidant homeostasis will be undermined by higher nitrite doses or longer exposure periods.

Supplementation with dietary omega-3 PUFA mitigates fetal brain inflammation and mitochondrial damage caused by high doses of sodium nitrite in maternal rats.

ObjectiveFood safety and nutrition during pregnancy are important concerns related to fetal brain development. In the present study, we aimed to explore the effects of omega-3 polyunsaturated fatty acids (PUFA ω-3) on exogenous sodium nitrite intervention-induced fetal brain injury in pregnant rats.MethodsDuring pregnancy, rats were exposed to water containing sodium nitrite (0.05%, 0.15%, and 0.25%) to establish a fetal rat brain injury model. Inflammatory factors and oxidative stress levels were detected using enzyme-linked immunosorbent assay (ELISA) or flow cytometry. Subsequently, animals were divided into three groups: control, model, and 4% PUFA ω-3. Pregnancy outcomes were measured and recorded. Hematoxylin-eosin (H&E) staining and immunohistochemistry (IHC) were utilized to observe brain injury. ELISA, quantitative real-time PCR (qRT-PCR), western blot, flow cytometry, and transmission electron microscopy (TEM) were adopted to measure the levels of inflammatory factors, the NRF1/HMOX1 signaling pathway, and mitochondrial and oxidative stress damage.ResultsWith the increase of sodium nitrite concentration, the inflammatory factors and oxidative stress levels increased. Therefore, the high dose group was set as the model group for the following experiments. After PUFA ω-3 treatment, the fetal survival ratio, average body weight, and brain weight were elevated. The cells in the PUFA ω-3 group were more closely arranged and more round than the model. PUFA ω-3 treatment relieved inflammatory factors, oxidative stress levels, and mitochondria damage while increasing the indicators related to brain injury and NRF1/HMOX1 levels.ConclusionsSodium nitrite exposure during pregnancy could cause brain damage in fetal rats. PUFA ω-3 might help alleviate brain inflammation, oxidative stress, and mitochondrial damage, possibly through the NRF1/HMOX1 signaling pathway. In conclusion, appropriately reducing sodium nitrite exposure and increasing PUFA omega-3 intake during pregnancy may benefit fetal brain development. These findings could further our understanding of nutrition and health during pregnancy.

Gut microbiota mediated the toxicity of high concentration of dietary nitrite in C57BL/6 mice

Growing evidence indicates that exposure to high levels of nitrite for a prolonged time has adverse health effects. Although gut microbiota is responsible for the transformation of nitrite in the gut, the evidence concerning whether gut microbiota mediates the toxicity of nitrite is still lacking. The present study addressed the long-term effects of dietary nitrite on male C57BL/6 mice and employed fecal microbiota transplantation (FMT) to reveal whether gut microbiota mediated the effects of nitrite. Furthermore, the effect of azoxymethane (AOM) on gut microbiota was detected for mice drinking normal or nitrite-containing water. High nitrite had toxic effects on C57BL/6 mice. Meanwhile, high nitrite induced skin lesions in mice, accompanied with increased serum ALT, colon IL-6, TNF-α, and MDA levels, together with decreased serum Cr, colon sIgA, and T-AOC levels. After fecal microbiota was transplanted into the normal mice, the nitrite-regulated gut microbiota could also induce skin lesions, coupled with reduced serum Cr, and increased colon MDA. The high dose of nitrite caused the upregulations of Alistipes, Prevotella, and Ruminococcus, which could be transplanted into normal mice through FMT. Inversely, gut microbiota from normal mice reduced the effects of nitrite on serum ALT and Cr, together with colon sIgA and MDA. Gut microbiota from normal mice could also upregulate metabolic genes and downregulate stress genes in the nitrite-treated mice. It might due to the upregulation of Akkermansia and Parabacteroides caused by FMT from normal water-treated mice to nitrite-treated mice. In addition, AOM exhibited to be more toxic to the colon in the nitrite-treated mice in comparison with normal water-treated mice, and it might be due to the expression of Hspa1a and Hspa1b in the colon. Interestingly, gut microbiota was more influenced by AOM in the normal water-treated mice than the nitrite-treated mice. Overall, these data demonstrated that gut microbiota mediated the toxicity of a high concentration of dietary nitrite.

Effects of chronic dietary nitrate supplementation on longevity, vascular function and cancer incidence in rats

RationaleDietary nitrate and nitrite have a notoriously bad reputation because of their proposed association with disease, in particular cancer. However, more recent lines of research have challenged this dogma suggesting that intake of these anions also possess beneficial effects after in vivo conversion to the vital signaling molecule nitric oxide. Such effects include improvement in cardiovascular, renal and metabolic function, which is partly mediated via reduction of oxidative stress. A recent study even indicates that low dose of dietary nitrite extends life span in fruit flies. MethodsIn this study, 200 middle-aged Wistar rats of both sexes were supplemented with nitrate or placebo in the drinking water throughout their remaining life and we studied longevity, biochemical markers of disease, vascular reactivity along with careful determination of the cause of death. ResultsDietary nitrate did not affect life span or the age-dependent changes in markers of oxidative stress, kidney and liver function, or lipid profile. Ex vivo examination of vascular function, however, showed improvements in endothelial function in rats treated with nitrate. Neoplasms were not more common in the nitrate group. ConclusionWe conclude that chronic treatment with dietary nitrate does not affect life span in rats nor does it increase the incidence of cancer. In contrast, vascular function was improved by nitrate, possibly suggesting an increase in health span.

Swine behavioral and physiological response to increasing sodium nitrite oral drench administration and resulting tissue residues

Objectives: This study aimed to evaluate the physiological and behavioral responses of pigs administered sodium nitrite, determine an ideal dosing rate by oral drenching of sodium nitrite for depopulation events, and evaluate the nitrite residue present in the ocular fluid and skeletal muscle after sodium nitrite administration. Materials and methods: Four groups of 10 market weight pigs (40 market weight pigs total) and 1 group of 10 sows were used. Each group of market weight animals received a different oral drench dose of sodium nitrite solution (1× [400-441 mg/kg], 2× [800-882 mg/kg], 2.5× [1000-1102 mg/kg], and 3× [1200-1323 mg/kg]) and was observed for distress behaviors. Two market weight animals in each treatment group were implanted with a monitor to measure body temperature, heart rate, and activity levels. The dosing rate with apparent best behavioral and physiological response was applied to the 10 sows and the same behaviors monitored. After death was confirmed, ocular fluid and skeletal muscle samples were collected from the sows. Results: An increased dosage of sodium nitrite greatly reduced the time to distress with a significant linear relationship. A higher frequency of vocalizations and the most frequent spikes in activity levels were observed in the lowest dosing group. No correlation was found between ocular fluid nitrite and skeletal muscle sodium nitrite concentrations. Implications: Oral drenching of sodium nitrite is a viable method for swine depopulation events. Higher doses of sodium nitrite have better welfare associations. Ocular fluid nitrite anion concentrations do not correlate with sodium nitrite skeletal muscle concentrations.

Antioxidant tempol modulates the increases in tissue nitric oxide metabolites concentrations after oral nitrite administration

Nitric oxide (NO) metabolites have physiological and pharmacological importance and increasing their tissue concentrations may result in beneficial effects. Tempol (4-hydroxy-2,2,6,6-tetramethylpiperidine-N-oxyl) has antioxidant properties that may improve NO bioavailability. Moreover, tempol increases oral nitrite-derived gastric formation of S-nitrosothiols (RSNO). We hypothesized that pretreatment with tempol may further increase tissue concentrations of NO-related species after oral nitrite administration and therefore we carried out a time-dependent analysis of how tempol affects the concentrations of NO metabolites in different tissues after oral nitrite administration to rats. NO metabolites (nitrate, nitrite and RSNO) were assessed by ozone-based reductive chemiluminescence assays in plasma, stomach, aorta, heart and liver samples obtained from anesthetized rats at baseline conditions and 15 min, 30 min, 2 h or 24 h after oral nitrite (15 mg/kg) was administered to rats pretreated with tempol (18 mg/kg) or vehicle 15 min prior to nitrite administration. Aortic protein nitrosation was assessed by resin-assited capture (SNO-RAC) method. We found that pretreatment with tempol transiently enhanced nitrite-induced increases in nitrite, RSNO and nitrate concentrations in the stomach and in the plasma (all P < 0.05), particularly for 15–30 min, without affecting aortic protein nitrosation. Pretreatment with tempol enhanced nitrite-induced increases in nitrite (but not RSNO or nitrate) concentrations in the heart (P < 0.05). In contrast, tempol attenuated nitrite-induced increases in nitrite, RSNO or nitrate concentrations in the liver. These findings show that pretreatment with tempol affects oral nitrite-induced changes in tissue concentrations of NO metabolites depending on tissue type and does not increase nitrite-induced vascular nitrosation. These results may indicate that oral nitrite therapy aiming at achieving increased nitrosation of cardiovascular targets requires appropriate doses of nitrite and is not optimized by tempol.

Supplemental nitrite increases choroidal neovascularization in mice

Low doses of nitrite, close to physiological levels, increase blood flow in normal and ischemic tissues through a nitric oxide (NO) dependent mechanism. Given that nitrite therapy and dietary supplementation with vegetables high in nitrate (e.g. beets) are gaining popularity we decided to determine if low doses of nitrite impact the development of choroidal neovascularization (CNV), a key feature of wet age related macular degeneration (AMD). Sodium nitrite (at 50 mg/L, 150 mg/L, and 300 mg/L), nitrate (1 g/L) or water alone were provided in the drinking water of C57BL/6 J mice aged 2 or 12 months. Mice were allowed to drink ad libitum for 1 week at which time laser-induced choroidal neovascularization (L-CNV) was induced. The mice continued to drink the supplemented water ad libitum for a further 14 days at which point optical coherence tomography (OCT) was performed to determine the volume of the CNV lesion. Blood was drawn to determine nitrite and nitrate levels and eyes taken for histology. CNV volume was 2.86 × 107 μm3 (±0.4 × 107) in young mice on water alone but CNV volume more than doubled to >6.9 × 107 μm3 (±0.8 × 107) in mice receiving 300 mg/L nitrite or 7.34 × 107 μm3 (±1.4 × 107) in 1 g/L nitrate (p < 0.01). A similar trend was observed in older mice. CNV volume was 5.3 × 107 μm3 (±0.5 × 107) in older mice on water alone but CNV volume almost doubled to approximately 9.3 × 107 μm3 (±1.1 × 107) in mice receiving 300 mg/L nitrite or 8.7 × 107 μm3 (±0.9 × 107) 1 g/L nitrate (p < 0.01). Plasma nitrite levels were highest in young mice receiving 150 mg/L in the drinking water with no changes in plasma nitrate observed. In older mice, drinking water nitrite did not significantly change plasma nitrite, but plasma nitrate was increased. Plasma nitrate was elevated in both young and old mice provided with nitrate supplemented drinking water. Our data demonstrate that the CNV lesion is larger in older mice compared to young and that therapeutic levels of oral nitrite increase the volume of CNV lesions in both young and older mice. Therapeutic nitrite or nitrate supplementation should be used with caution in the elderly population prone to CNV.

Use of physiological activities to estimate the population growth of rotifer (Brachionus calyciflorus) under the stress of toxic Microcystis and nitrite

Microcystis blooms disrupt aquatic systems and adversely affect zooplankton growth. Brachionus calyciflorus Pallas (rotifer) was introduced to different combinations of toxic Microcystis aeruginosa (0, 2 × 105, 2 × 106, and 2 × 107 cells mL−1) and nitrite (0, 2, 4, and 6 mg L−1) to evaluate their physiological activities and population growth under stress. Survival rate (S), population growth rate (r), grazing rate (G), antioxidant response, and metabolic and digestive enzyme activities were determined. Results revealed that G declined with the increasing nitrite doses and grazing time upon exposure to a certain Microcystis concentration. Toxic M. aeruginosa and nitrite inhibited the S, r, glutathione content, total antioxidant capacity level, and activities of alkaline phosphatase, xanthine oxidase, lactate dehydrogenase, and cellulase (p < 0.05) but increased the reactive oxygen species level, malondialdehyde content, and amylase activity (p < 0.05). The activities of superoxide dismutase, catalase, and pepsase were also increased in single low doses of nitrite solutions (p < 0.05). Therefore, the grazing intensity of rotifers affected B. calyciflorus physiological activities, which are useful in the estimation of its population growth in eutrophic water environments.

Improvement of the performance of simultaneous nitrification denitrification and phosphorus removal (SNDPR) system by nitrite stress

This study investigated a new way to improve the performance of simultaneous nitrification denitrification and phosphorus removal (SNDPR) system by regularly changing the anaerobic/micro-aerobic/anoxic mode to the anaerobic/anoxic mode with 30 mg/L of nitrite dosing. The results indicated that the removal efficiency of total inorganic nitrogen and PO43−-P was improved from 75.44% and 85.14% to 98.89% and 98.17%, respectively. And the good performance of the SNDPR showed a long-time sustainability when the C/N ratio was 5. The results of microbial community illustrated that the abundance of the main nitrite-oxidizing bacteria (NOB), Nitrospira sp., dropped from 5.71% to 0.85% and the abundance of denitrifying polyphosphate-accumulating organisms (DPAOs), Pseudomonas sp. and Acinetobacter sp., increased by 5 times after nitrite stress. The high level of nitric oxide (NO) and free nitrite acid produced by addition of nitrite strongly suppressed the undesired organisms NOB and ordinary heterotrophic denitrifying organisms, and promoted the enrichment of DPAOs. The NO accumulated in the nitrite denitrification process could inhibit NOB and promote AOB. This study revealed that NO plays an important role in regulating the microbial community in the SNDPR system.

Nitrate and nitrite absorption, recycling and retention in tissues of sheep

Dietary nitrate is of increasing interest both for the pharmacological effects of its metabolites as well as its capacity to inhibit methanogenesis in the gut. A sequence of three experiments was conducted to investigate the absorption, metabolism and excretion of nitrate and nitrite through the gastrointestinal tract (GIT) of sheep, and to determine the fate of nitrate and nitrite in body fluids, tissues and faeces after intravenous dosing with 15N-labeled potassium nitrate (K15NO3) and with 15N-labeled sodium nitrite (Na15NO2). In Experiment 1, twelve female Merino sheep were assigned to one of two dietary treatments and adapted to the experimental diet over two weeks. Six sheep were fed a control diet of wheaten chaff mixture (600 g wheaten chaff plus 200 g wheat grain, CON) and six sheep were fed the CON diet with the inclusion of 18 g nitrate/kg DM (Nitrate). After acclimation to the diets, all sheep received a single intravenous dose of K15NO3 and were placed in metabolic cages for daily collection of total faeces and urine over 6 days. Experiment 2 studied movement of an intravenous dose of 15N in body fluids and tissues. Two sheep not adapted to dietary nitrate were dosed intravenously with K15NO3 or Na15NO2 and body fluids and tissue samples were collected 60 min after dosing. Finally, Experiment 3 was conducted to identify and quantify the major sites of nitrate and nitrite transfer within the body, focusing on absorption, partitioning and secretion into the GIT of anaesthetised sheep. A single dose of sodium nitrate (NaNO3) or sodium nitrite (NaNO2) was introduced into the rumen or abomasum or small intestine, and changes in nitrate and nitrite concentrations in other pools, including plasma, urine and saliva, were determined. Results from Experiment 1 showed that urinary recovery of 15N dose in urea after 46 h and total urinary recovery of 15N 141 h after dosing were greater in sheep fed the Nitrate diet relative to CON (P < 0.05). Recoveries of 15N in tissues indicated that nitrate and nitrite principally accumulated in the skin and muscle of sheep (Experiment 2). Finally, Experiment 3 indicated that nitrate and nitrite were rapidly absorbed from the rumen, abomasum and small intestine into the bloodstream. Nitrite was oxidized in plasma and the resultant nitrate was concentrated and recycled via saliva. Appearance of 15N in urinary urea confirmed the passage of plasma nitrate to the digestive tract, via saliva or transruminal flow to be reduced by gut biota to ammonia.

Protective Effects of Topical Application of Nitrite on Testicular Ischemia-Reperfusion Injury in Rats.

Testicular torsion is a urologic emergency induced by torsion of the spermatic cord, interrupting blood circulation to the testis. Therapeutic options for testicular torsion, except surgical restoration of testis, are rarely applied in clinical practice. This study, therefore, investigated whether topical application of nitrite (NO2−) is beneficial in tissue damage due to testicular ischemia-reperfusion (I/R) injury in rats. Pubertal Sprague-Dawley rats were assigned to seven groups: group A, sham-operated control group; group B, I/R with no treatment; groups C, D, and E, I/R followed by treatment with three different doses of nitrite; group F, I/R followed by administration of nitrite and a NO scavenger, C-PTIO (2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide potassium salt); and group G, I/R followed by administration of nitrate (NO3−). Unilateral testicular ischemia was maintained for 5 h, followed by reperfusion for 24 h. Nitrite and nitrate were topically administered before reperfusion. Compared to group A, germ cell apoptosis, oxidative stress, antioxidant enzymatic function, and lipid peroxidation were significantly increased, along with abnormal morphology and impaired spermatogenesis in group B (P < 0.05). In contrast, testicular damage was generally attenuated in the nitrite treatment groups due to a reduction in superoxide and peroxynitrite levels and the inhibition of caspase-3-dependent apoptosis (P < 0.05 vs. group B). These therapeutic effects of nitrite-derived NO were suppressed after injection of C-PTIO, which showed in group F. Taken together, our results demonstrate that topical application of nitrite may be one of the therapeutic strategies for testicular ischemia-reperfusion injury.