Nitrate Nitrite Nitric Oxide Research Articles

Risk of circulatory diseases associated with proton-pump inhibitors: a retrospective cohort study using electronic medical records in Thailand.

Proton-pump inhibitors (PPIs) are prescribed to treat gastric acid-related diseases, while they may also have potential risks to population health. Recent studies suggested that a potential mechanism explaining the association between PPIs and cardiovascular diseases (CVD) includes the inhibition of the nitrate-nitrite-nitric oxide (NO) pathway. However, previous observational studies showed controversial results of the association. In addition, the inhibition of the NO pathway due to PPIs use may lead to peripheral vascular diseases (PVD); however, none of the studies explore the PPI-PVD association. Therefore, this study aimed to evaluate the association of PPIs with circulatory diseases (CVD, ischemic strokes or IS, and PVD). We conducted a retrospective hospital-based cohort study from Oct 2010 to Sep 2017 in Songkhla province, Thailand. PPIs and histamine 2-receptor antagonists (H2RAs) prescriptions were collected from electronic pharmacy records, while diagnostic outcomes were retrieved from electronic medical records at Songklanagarind hospital. Patients were followed up with an on-treatment approach. Cox proportional hazard models were applied to measure the association comparing PPIs vs H2RAs after 1:1 propensity-score-matching. Sub-group analysis, multi-bias E-values, and array-based sensitivity analysis for some covariates were used to assess the robustness of associations. A total of 3,928 new PPIs and 3,928 H2RAs users were included in the 1:1 propensity score-matched cohort. As compared with H2RAs, the association of PPIs with CVD, IS, and PVD, the hazard ratios were 1.76 95% CI = [1.40-2.20] for CVD, 3.53 95% CI = [2.21-5.64] for ischemic strokes, and 17.07 95% CI = [13.82-76.25] for PVD. The association between PPIs and each outcome was significant with medication persistent ratio of over 50%. In addition, the association between PPIs and circulatory diseases was robust to unmeasured confounders (i.e., smoking and alcohol). PPIs were associated with circulatory diseases, particularly ischemic strokes in this hospital-based cohort study, whereas, the strength of associations was robust to unmeasured confounders.

Renal replacement therapy removes a large number of nitric oxide donors responsible for the nitrate-nitrite-nitric oxide pathway.

Nitric oxide has a broad-spectrum antibacterial property promising as a new therapeutic agent for severe acute respiratory syndrome coronavirus-2 because nitric oxide donor (such as S-nitroso-N-acetylpenicillamine) reduces the replication of coronavirus-2. Patients with coronavirus disease 2019 undergoing dialysis generally have a higher mortality rate than the general population. Although the higher mortality rate in these patients may be related to their advanced age, it has been suggested that plasma nitrite and nitrate levels (products of nitric oxide metabolism) are significantly decreased after hemodialysis which may compromise the nitrate-nitrite-nitric oxide pathway and impair nitric oxide homeostasis. It results in increased cardiovascular mortality in patients undergoing dialysis. However, the profile of nitric oxide-producing substances is poorly understood during renal replacement therapy. We simulated continuous hemodialysis and hemodiafiltration to measure the amount of nitric oxide (nitric oxide-producing substance) clearance in vitro. The results demonstrated increased nitric oxide clearance and higher clearance than creatinine (molecular weight: 113) and vitamin B12 (molecular weight: 1355) using highly efficient renal replacement therapy modes. The high nitric oxide clearance may have partly contributed to the high cardiovascular and coronavirus-2 mortality risk in patients on dialysis.

Nitrate-Nitrite-Nitric Oxide Pathway: A Mechanism of Hypoxia and Anoxia Tolerance in Plants.

Oxygen (O2) is the most crucial substrate for numerous biochemical processes in plants. Its deprivation is a critical factor that affects plant growth and may lead to death if it lasts for a long time. However, various biotic and abiotic factors cause O2 deprivation, leading to hypoxia and anoxia in plant tissues. To survive under hypoxia and/or anoxia, plants deploy various mechanisms such as fermentation paths, reactive oxygen species (ROS), reactive nitrogen species (RNS), antioxidant enzymes, aerenchyma, and adventitious root formation, while nitrate (NO3−), nitrite (NO2−), and nitric oxide (NO) have shown numerous beneficial roles through modulating these mechanisms. Therefore, in this review, we highlight the role of reductive pathways of NO formation which lessen the deleterious effects of oxidative damages and increase the adaptation capacity of plants during hypoxia and anoxia. Meanwhile, the overproduction of NO through reductive pathways during hypoxia and anoxia leads to cellular dysfunction and cell death. Thus, its scavenging or inhibition is equally important for plant survival. As plants are also reported to produce a potent greenhouse gas nitrous oxide (N2O) when supplied with NO3− and NO2−, resembling bacterial denitrification, its role during hypoxia and anoxia tolerance is discussed here. We point out that NO reduction to N2O along with the phytoglobin-NO cycle could be the most important NO-scavenging mechanism that would reduce nitro-oxidative stress, thus enhancing plants’ survival during O2-limited conditions. Hence, understanding the molecular mechanisms involved in reducing NO toxicity would not only provide insight into its role in plant physiology, but also address the uncertainties seen in the global N2O budget.

Dietary nitrate and brain health. Too much ado about nothing or a solution for dementia prevention?

Dementia is a significant public health priority with approximately 55 million cases worldwide, and this number is predicted to quadruple by 2050. Adherence to a healthy diet and achieving optimal nutritional status are vital strategies to improve brain health. The importance of this area of research has been consolidated into the new term ‘nutritional psychiatry’. Dietary nitrate, closely associated with the intake of fruits and vegetables, is a compound that is increased in dietary patterns such as the Mediterranean and MIND diets and has protective effects on cognition and brain health. Nitrate is characterised by a complex metabolism and is the precursor of the nitrate–nitrite–nitric oxide (NO) pathway contributing to systemic NO generation. A higher intake of dietary nitrate has been linked to protective effects on vascular outcomes including blood pressure and endothelial function. However, the current evidence supporting the protective effects of dietary nitrate on brain health is less convincing. This article aims to provide a critical appraisal of the current evidence for dietary nitrate supplementation for improving brain health and provide suggestions for future research.

Plasma Nitrate and Nitrite Kinetics after Single Intake of Beetroot Juice in Adult Patients on Chronic Hemodialysis and in Healthy Volunteers: A Randomized, Single-Blind, Placebo-Controlled, Crossover Study

Nitric oxide (NO) contributes to maintaining normal cardiovascular and renal function. This bioactive signalling molecule is generally formed enzymatically by NO synthase in the vascular endothelium. NO bioactivity can also be attributed to dietary intake of inorganic nitrate, which is abundant in our diet, especially in green leafy vegetables and beets. Ingested nitrate is reduced to nitrite by oral commensal bacteria and further to NO systemically. Previous studies have shown that dialysis, by means of removing nitrate and nitrite from the body, can reduce NO bioactivity. Hence, dietary intervention approaches aimed to boost the nitrate–nitrite–NO pathway may be of benefit in dialysis patients. The purpose of this study was to examine the kinetics of plasma nitrate and nitrite after a single intake of nitrate-rich concentrated beetroot juice (BJ) in adult hemodialysis (HD) patients and in age-matched healthy volunteers (HV). Eight HD patients and seven HV participated in this single center, randomized, single-blind, placebo-controlled, crossover study. Each participant received a sequential single administration of active BJ (70 mL, 400 mg nitrate) and placebo BJ (70 mL, 0 mg nitrate) in a random order separated by a washout period of seven days. For the kinetic analysis, blood samples were collected at different time-points before and up to 44 h after BJ intake. Compared with placebo, active BJ significantly increased plasma nitrate and nitrite levels both in HD patients and HV. The area under the curve and the maximal concentration of plasma nitrate, but not of nitrite, were significantly higher in HD patients as compared with HV. In both groups, active BJ ingestion did not affect blood pressure or plasma potassium levels. Both BJs were well tolerated in all participants with no adverse events reported. Our data provide useful information in planning dietary nitrate supplementation efficacy studies in patients with reduced NO bioactivity.

Nitrite Generating and Depleting Capacity of the Oral Microbiome and Cardiometabolic Risk: Results from ORIGINS.

BackgroundThe enterosalivary nitrate–nitrite–nitric oxide (NO3–NO2–NO) pathway generates NO following oral microbiota‐mediated production of salivary nitrite, potentially linking the oral microbiota to reduced cardiometabolic risk. Nitrite depletion by oral bacteria may also be important for determining the net nitrite available systemically. We examine if higher abundance of oral microbial genes favoring increased oral nitrite generation and decreased nitrite depletion is associated with a better cardiometabolic profile cross‐sectionally.Methods and ResultsThis study includes 764 adults (mean [SD] age 32 [9] years, 71% women) enrolled in ORIGINS (Oral Infections, Glucose Intolerance, and Insulin Resistance Study). Microbial DNA from subgingival dental plaques underwent 16S rRNA gene sequencing; PICRUSt2 was used to estimate functional gene profiles. To represent the different components and pathways of nitrogen metabolism in bacteria, predicted gene abundances were operationalized to create summary scores by (1) bacterial nitrogen metabolic pathway or (2) biochemical product (NO2, NO, or ammonia [NH3]) formed by the action of the bacterial reductases encoded. Finally, nitrite generation‐to‐depletion ratios of gene abundances were created from the above summary scores. A composite cardiometabolic Z score was created from cardiometabolic risk variables, with higher scores associated with worse cardiometabolic health. We performed multivariable linear regression analysis with cardiometabolic Z score as the outcome and the gene abundance summary scores and ratios as predictor variables, adjusting for sex, age, race, and ethnicity in the simple adjusted model. A 1 SD higher NO versus NH3 summary ratio was inversely associated with a −0.10 (false discovery rate q=0.003) lower composite cardiometabolic Z score in simple adjusted models. Higher NH3 summary score (suggestive of nitrite depletion) was associated with higher cardiometabolic risk, with a 0.06 (false discovery rate q=0.04) higher composite cardiometabolic Z score.ConclusionsIncreased net capacity for nitrite generation versus depletion by oral bacteria, assessed through a metagenome estimation approach, is associated with lower levels of cardiometabolic risk.

Effect of Vitamin C and Protein Supplementation on Plasma Nitrate and Nitrite Response following Consumption of Beetroot Juice.

Beetroot juice is a food high in nitrate and is associated with cardiometabolic health benefits and enhanced exercise performance through the production of nitric oxide in the nitrate–nitrite–nitric oxide pathway. Since various food components influence this pathway, the aim of this trial was to study the effect of beetroot juice alone and in conjunction with vitamin C or protein on the acute response to plasma nitrate and nitrite levels in healthy middle- to older-aged adults. In this cross-over trial, each participant received, in a randomized order, a single dose of Beet It Sport® alone; Beet It Sport®, plus a 200 mg vitamin C supplement; and Beet It Sport® plus 15 g of whey protein. Plasma levels of nitrate and nitrite were determined prior to and at 1 and 3 h after intervention. Log plasma nitrate and nitrite was calculated to obtain data that were normally distributed, and these data were analyzed using two-way within-factors ANOVA, with time and treatment as the independent factors. There were no statistically significant differences for log plasma nitrate (p = 0.308) or log plasma nitrite (p = 0.391) values across treatments. Log plasma nitrate increased significantly from pre-consumption levels after 1 h (p < 0.001) and 3 h (p < 0.001), but plasma nitrate was lower at 3 h than 1 h (p < 0.001). Log plasma nitrite increased from pre to 1 h (p < 0.001) and 3 h (p < 0.001) with log values at 3 h higher than at 1 h (p = 0.003). In this cohort, we observed no differences in log plasma nitrate and nitrite at 1 h and 3 h after co-ingesting beetroot juice with vitamin C or a whey protein supplement compared to beetroot juice alone. Further research needs to be undertaken to expand the blood-sampling time-frame and to examine factors that may influence the kinetics of the plasma nitrate to nitrite efficacy, such as differences in fluid volume and osmolarity between treatments employed.

흰쥐에서 고용량 아질산이온 함유 마늘 발효농축액에 의한 뇌혈류 변화

Nitrate–nitrite–nitric oxide (NO) pathway is a major alternative source of NO and is essential for NO - dependent physiological functions in body. Food supplements having nitrate/nitrite can improve metabolic syndromes including hypertension through antioxidant activity or vasodilation. The purpose of this study was to observe the effects of fermented garlic (F. garlic) having high concentration of NO₂ SUP-/SUP on changes in blood flow and nitric oxide synthesis in the cerebral cortex of rodents. The generation of nitric oxide detected by a chemi-luminescence detector was higher in F. Garlic compared with NaNO₂ solution under artificial gastric juice with pH 2.0. Ether F. garlic or NaNO₂ diluted with artificial cerebrospinal fluid was directly applied into around the needle probe of laser Doppler flow meter that was located on epidural surface of the cortex. Direct application of F. garlic resulted in increase of cerebral blood flow detected by a laser Doppler flow meter with a dose-dependent manner. Compared with NaNO₂ solution, F. garlic produced changes in cerebral blood flow at lower concentration of NO₂SUP-/SUP. Pretreatment of methylene blue, a guanylyl cyclase inhibitor prevented upregulation of cerebral blood flow by the treatment of F. garlic. In addition, the application of F. garlic with 250, 500ppm of NO₂SUP-/SUP caused significantly the production of NO in the cortical tissue but NaNO₂ solution with 500ppm of NO₂SUP-/SUP did not. In summary, these results suggested that F. garlic with high content of NO₂SUP-/SUP induce increase in cerebral blood flow through nitric oxide-dependent signal pathway.

Antiseptic mouthwash, the nitrate-nitrite-nitric oxide pathway, and hospital mortality: a hypothesis generating review.

Meta-analyses and several large cohort studies have demonstrated that antiseptic mouthwashes are associated with mortality in hospitalized patients. A clear pathogenic mechanism is lacking, leading to controversy and a reluctance to abandon or limit the use of antiseptic mouthwashes. Here, we generate the hypothesis that a disturbance in nitric oxide homeostasis by antiseptic mouthwashes may be responsible for the observed increase in mortality risk. Nitric oxide is essential in multiple physiological processes, and a reduction in nitric oxide bioavailability is associated with the occurrence or worsening of pathologies, such as atherosclerosis, diabetes, and sepsis. Oral facultative anaerobic bacteria are essential for the enterosalivary nitrate–nitrite–nitric oxide pathway due to their capacity to reduce nitrate to nitrite. Nitrate originates from dietary sources or from the active uptake by salivary glands of circulating nitrate, which is then excreted in the saliva. Because antiseptic mouthwashes eradicate the oral bacterial flora, this nitric oxide-generating pathway is abolished, which may result in nitric oxide-deficient conditions potentially leading to life-threatening complications such as ischaemic heart events or sepsis.

Effects of Dietary Nitrates on Time Trial Performance in Athletes with Different Training Status: Systematic Review.

Much research has been done in sports nutrition in recent years as the demand for performance-enhancing substances increases. Higher intake of nitrates from the diet can increase the bioavailability of nitric oxide (NO) via the nitrate–nitrite–NO pathway. Nevertheless, the increased availability of NO does not always lead to improved performance in some individuals. This review aims to evaluate the relationship between the athlete’s training status and the change in time trial performance after increased dietary nitrate intake. Articles indexed by Scopus and PubMed published from 2015 to 2019 were reviewed. Thirteen articles met the eligibility criteria: clinical trial studies on healthy participants with different training status (according to VO2max), conducting time trial tests after dietary nitrate supplementation. The PRISMA guidelines were followed to process the review. We found a statistically significant relationship between VO2max and ergogenicity in time trial performance using one-way ANOVA (p = 0.001) in less-trained athletes (VO2 < 55 mL/kg/min). A strong positive correlation was observed in experimental situations using a chronic supplementation protocol but not in acute protocol situations. In the context of our results and recent histological observations of muscle fibres, there might be a fibre-type specific role in nitric oxide production and, therefore, supplement of ergogenicity.

Grapefruit juice enhances the systolic blood pressure-lowering effects of dietary nitrate-containing beetroot juice.

Dietary nitrate from sources such as beetroot juice lowers blood pressure (BP) via the nitrate-nitrite-nitric oxide (NO) pathway. However, NO and nitrite are inactivated via reoxidation to nitrate, potentially limiting their activity. Cytochrome P450-3A4 inhibition with troleandomycin prevents nitrite re-oxidation to nitrate in rodent liver. Grapefruit juice contains the CYP3A4 inhibitor furanocoumarin. We therefore hypothesized that grapefruit juice would enhance BP-lowering with beetroot juice by maintaining circulating [nitrite]. We performed a randomized, placebo-controlled, 7-hour crossover study in 11 healthy volunteers, attending on 3 occasions, receiving: a 70-mL shot of active beetroot juice (Beet-It) and either (i) 250 mL grapefruit juice (Active Beet+GFJ), or (ii) 250 mL water (Buxton, Active Beet+H2 O); or (iii) Placebo Beet+GFJ. The addition of grapefruit juice to active beetroot juice lowered systolic BP (SBP): Active Beet+GFJ vs Active Beet+H2 O (P = .02), and pulse pressure, PP (P = .0003). Peak mean differences in SBP and PP were seen at T = 5 hours: -3.3 mmHg (95% confidence interval [CI] -6.43 to -0.15) and at T = 2.5 hours: -4.2 mmHg (95% CI -0.3 to -8.2), respectively. Contrary to the hypothesis, plasma [nitrite] was lower with Active Beet+GFJ vs Active Beet+H2 O (P = .006), as was salivary nitrite production (P = .002) and saliva volume (-0.34 mL/min [95% CI -0.05 to -0.68]). The taste score of Beet+GFJ was 1.4/10 points higher than Beet+H2 O (P = .03). Grapefruit juice enhanced beetroot juice's effect on lowering SBP and PP despite decreasing plasma [nitrite]. Besides suggesting more complex mechanisms, there is potential for maximising the clinical benefit of dietary nitrate and targeting isolated systolic hypertension.

Nitrous Oxide Emissions from Paddies: Understanding the Role of Rice Plants.

Paddies are a potential source of anthropogenic nitrous oxide (N2O) emission. In paddies, both the soil and the rice plants emit N2O into the atmosphere. The rice plant in the paddy is considered to act as a channel between the soil and the atmosphere for N2O emission. However, recent studies suggest that plants can also produce N2O, while the mechanism of N2O formation in plants is unknown. Consequently, the rice plant is only regarded as a channel for N2O produced by soil microorganisms. The emission of N2O by aseptically grown plants and the distinct dual isotopocule fingerprint of plant-emitted N2O, as reported by various studies, support the production of N2O in plants. Herein, we propose a potential pathway of N2O formation in the rice plant. In rice plants, N2O might be formed in the mitochondria via the nitrate–nitrite–nitric oxide (NO3–NO2–NO) pathway when the cells experience hypoxic or anoxic stress. The pathway is catalyzed by various enzymes, which have been described. So, N2O emitted from paddies might have two origins, namely soil microorganisms and rice plants. So, regarding rice plants only as a medium to transport the microorganism-produced N2O might be misleading in understanding the role of rice plants in the paddy. As rice cultivation is a major agricultural activity worldwide, not understanding the pathway of N2O formation in rice plants would create more uncertainties in the N2O budget.

Nitrate partially inhibits lipopolysaccharide-induced inflammation by maintaining mitochondrial function.

ObjectiveNitrate has been reported to protect cells via the nitrate-nitrite-nitric oxide (NO) pathway. Most studies tend to use nitrite to investigate the mechanisms of this pathway. However, the latest studies have confirmed that mammals can directly degrade nitrate via xanthine oxidoreductase (XOR). The hypothesis is that nitrate could play a protective role in inflammatory responses independent of bacterial nitrate reductases.MethodsMouse RAW264.7 macrophages were pre-incubated with sodium nitrate (10, 100, and 500 µM) for 2 hours, and then treated with lipopolysaccharide (LPS) for 2 hours to induce inflammation. The Quantikine Immunoassay was used to measure interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) concentrations in the supernatant. The fluorescence intensity ratio of red/green from JC-1 was used to assay mitochondrial membrane potential. The fluorescence intensity of MitoSOX Red was used to indicate the generation of mitochondrial reactive oxygen species.ResultsNitrate partially reduced IL-6 and TNF-α secretion via reducing NF-κB signaling in LPS-induced macrophages. Nitrate also reduced the generation of mitochondrial reactive oxygen species by regulating mitochondrial function. These effects depended on XOR-derived NO but were independent of inducible nitric oxide synthase-derived NO.ConclusionNitrate regulates mitochondrial function via XOR-derived NO to partially inhibit LPS-induced inflammation.

Supplementation of dietary nitrate attenuated oxidative stress and endothelial dysfunction in diabetic vasculature through inhibition of NADPH oxidase

The metabolic disorders in diabetes, which are usually accompanied by oxidative stress and impaired nitric oxide (NO) bioavailability, increase the risk of detrimental cardiovascular complications. Herein, we investigated the therapeutic potential of dietary nitrate, which is found in high content in green leafy vegetables, on vascular oxidative stress and endothelial dysfunction in diabetic mice induced by high-fat diet and streptozotocin injection. Dietary nitrate in drinking water fuelled a nitrate–nitrite–NO pathway, which inhibited vascular oxidative stress, endothelial dysfunction and many features of metabolic syndrome in diabetic mice. These beneficial effects of nitrate on diabetic mice were abolished by PTIO (NO scavenger) treatment and significantly prevented by febuxostat (xanthine oxidoreductase inhibitor), demonstrating the central importance of NO in bioactivation of nitrate. The favorable effects of nitrate were not further influenced by apocynin (NADPH oxidase inhibitor), suggesting NADPH oxidase as a possible target. In high glucose-incubated vascular endothelial cells, NO donor attenuated oxidative stress and endothelial dysfunction via the inhibition of NADPH oxidase, where a heme oxygenase-1 (HO-1)-dependent mechanism was demonstrated for the antioxidant abilities of NO. Altogether, boosting this nitrate–nitrite–NO signaling pathway resulted in the decreases of NADPH oxidase-derived oxidative stress, endothelial dysfunction and metabolic disorders in diabetic vasculature. These findings may have novel implications for the preventive strategy against diabetes-induced vascular dysfunction and associated complications.

Effects of inorganic nitrate in a rat model of monocrotaline-induced pulmonary arterial hypertension.

The nitrate-nitrite-nitric oxide (NO) pathway represents an alternative source of NO generation, which is independent of NO synthase and potentiated by hypoxia. Augmentation of this pathway by dietary nitrate has proven favourable effects in several cardiovascular disease models. However, less is known regarding its potential value in pulmonary arterial hypertension (PAH). The aim of this study was to assess the effects of oral inorganic nitrate administration in monocrotaline (MCT)-induced PAH. Male 12-week-old Wistar rats were injected subcutaneously with monocrotaline (MCT, 60mg/kg). Nitrate treatment (0.3 or 1mmol/kg/d; drinking water) commenced on day 12 following the MCT injection and continued for 16days. Nitrate administration did not attenuate right ventricular (RV) hypertrophy, increased lung weight and up-regulated mRNA expression of brain natriuretic peptide. Plasma nitrate and nitrite levels were significantly increased as well as lung nitrate level, whereas nitrite lung level was decreased following nitrate treatment (1mmol/kg/d). MCT-induced PAH resulted in an increased MnSOD protein level, which was not observed following nitrate treatment. MCT-associated up-regulation of nNOS in the lung appeared to be dose-dependently prevented by nitrate treatment. Western blot analysis did not reveal any differences in eNOS, iNOS, XO or gp91phox expression in the lungs among the groups. In conclusion, nitrate treatment did not significantly attenuate pathological RV and lung remodelling in the rat MCT model of PAH. The suppression of MnSOD and nNOS expression by nitrate could be interpreted as reduced demand of endogenous antioxidant defence in this model.

Next Generation Sequencing Discoveries of the Nitrate-Responsive Oral Microbiome and Its Effect on Vascular Responses.

Cardiovascular disease is a worldwide human condition which has multiple underlying contributing factors: one of these is long-term increased blood pressure—hypertension. Nitric oxide (NO) is a small nitrogenous radical species that has a number of physiological functions including vasodilation. It can be produced enzymatically through host nitric oxide synthases and by an alternative nitrate–nitrite–NO pathway from ingested inorganic nitrate. It was discovered that this route relies on the ability of the oral microbiota to reduce nitrate to nitrite and NO. Next generation sequencing has been used over the past two decades to gain deeper insight into the microbes involved, their location and the effect of their removal from the oral cavity. This review article presents this research and comments briefly on future directions.

Effects of dietary nitrate on inflammation and immune function, and implications for cardiovascular health

Inorganic dietary nitrate, found abundantly in green leafy and some root vegetables, elicits several beneficial physiological effects, including a reduction in blood pressure and improvements in blood flow through nitrate-nitrite-nitric oxide signaling. Recent animal and human studies have shown that dietary nitrate and nitrite also modulate inflammatory processes and immune cell function and phenotypes. Chronic low-grade inflammation and immune dysfunction play a critical role in cardiovascular disease. This review outlines the current evidence on the efficacy of nitrate-rich plant foods and other sources of dietary nitrate and nitrite to counteract inflammation and promote homeostasis of the immune and vascular systems. The data from these studies suggest that immune cells and immune-vasculature interactions are important targets for dietary interventions aimed at improving, preserving, or restoring cardiovascular health.

Reciprocal regulation of sulfite oxidation and nitrite reduction by mitochondrial sulfite oxidase

The oxygen-independent nitrate-nitrite-nitric oxide (NO) pathway is considered as a substantial source of NO in mammals. Dietary nitrate/nitrite are distributed throughout the body and reduced to NO by the action of various enzymes. The intermembrane spaced (IMS), molybdenum cofactor-dependent sulfite oxidase (SO) was shown to catalyze such a nitrite reduction. In this study we asked whether the primary function of SO – sulfite oxidation – and its novel function – nitrite reduction – impact each other. First, we utilized benzyl viologen as artificial electron donor to investigate steady state NO synthesis by SO and found fast (kcat = 14 s−1) nitrite reduction of SO full-length and its isolated molybdenum domain at pH 6.5. Next, we determined the impact of nitrite on pre-steady state kinetics in SO catalysis and identified nitrite as a pH-dependent inhibitor of SO reductive and oxidative half reaction. Finally, we report on the time-dependent formation of the paramagnetic Mo(V) species following nitrite reduction and demonstrate that sulfite inhibits nitrite reduction. In conclusion, we propose a pH-dependent reciprocal regulation of sulfite oxidation and nitrite reduction by each substrate, thus facilitating quick responses to hypoxia induced changes in the IMS, which may function in protecting the cell from reactive oxygen species production.

Nitrate increases in skeletal muscle peak power do not involve changes in sarcomeric protein phosphorylation and require whole‐body physiological pathways

Acute dietary nitrate supplementation in humans (within 2 hours) increases limb muscle peak power. In old mice, chronic (14 days) dietary nitrate supplementation increases diaphragm peak power. However, mechanisms of dietary nitrate effects on contractile mechanics are not fully understood. It is unclear whether nitrate ions directly or products downstream of the nitrate‐nitrite‐nitric oxide (NO) reductive pathway are responsible for changes in muscle contractile function. Increases in power likely involve post‐translational modification of myofibrillar proteins – phosphorylation being an established effect of NO signaling. We determined the phosphorylation status of myofibrillar proteins from diaphragms of 24 month old mice supplemented with nitrate (n = 8) or control (n = 6) and found no difference for myosin regulatory light chain, myosin binding protein c, tropomyosin, desmin or troponin I. Regulatory light chain phosphorylation signal normalized to total protein was 1.696 ± 0.139 and 1.699 ± 0.105 for control and nitrate‐supplemented animals, respectively. To examine a direct effect of nitrate on muscle cells, we exposed isolated diaphragm bundles from young mice in vitro to increasing concentrations of NaNO3. In vitro NaNO3 (15 min) had no effect on diaphragm peak power or isometric force (P &gt; 0.05). Specifically, power (% control) was −1.06 ± 0.20 (0.5 mM), −1.73 ± 0.79 (1 mM), −2.64 ± 2.14 (2 mM), and −2.40 ± 2.40 (4 mM). Overall, our study shows that the increase in diaphragm power of aged mice elicited by chronic dietary nitrate supplementation does not involve changes in phosphorylation of key sarcomeric proteins that regulate contractile function. Moreover, our data from in vitro experiments suggest no direct effect of nitrate ions on mammalian skeletal muscle contractile function. Instead, dietary nitrate effects on muscle function appear to require whole‐body physiological mechanisms and actions of molecules downstream of nitrate.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Chewing Well During Meals May Benefit Health Via the Enterosalivary Nitrate–Nitrite–Nitric Oxide Pathway

Saliva contains nitrates recycled from ingested foods (e.g., vegetables) via the enterosalivary pathway. During meals, thorough chewing enhances salivary flow and the reduction of salivary nitrate to nitrite by oral commensal bacteria and thus leads to a nitrite-rich gastric environment. Subsequently, nitric oxide (NO) and S-nitrosothiols, rather than N-nitroso compounds, are generated in the acidic stomach. These molecules may confer gastric and systemic health benefits by transducing NO-mediated signals, particularly in patients with vascular endothelial dysfunction and metabolic syndrome, which is associated with reduced endogenous NO generation consequent to impaired NO synthase activity. This article reviews literature suggesting that the thorough chewing of food produces a nitrite-rich gastric environment and promotes health benefits by enhancing NO bioavailability via the enterosalivary nitrate-nitrite-NO pathway.