Dietary Nitrate Supplementation Research Articles

Preliminary Research on the Effects of Dietary Nitrate Supplementation from Beetroot Juice on Sprint Performance in Army ROTC Cadets

Preliminary Research on the Effects of Dietary Nitrate Supplementation from Beetroot Juice on Sprint Performance in Army ROTC Cadets

Dietary Nitrate and Physical Performance.

Nitric oxide (NO) plays a plethora of important roles in the human body. Insufficient production of NO (for example, during older age and in various disease conditions) can adversely impact health and physical performance. In addition to its endogenous production through the oxidation of l-arginine, NO can be formed nonenzymatically via the reduction of nitrate and nitrite, and the storage of these anions can be augmented by the consumption of nitrate-rich foodstuffs such as green leafy vegetables. Recent studies indicate that dietary nitrate supplementation, administered most commonly in the form of beetroot juice, can ( a) improve muscle efficiency by reducing the O2 cost of submaximal exercise and thereby improve endurance exercise performance and ( b) enhance skeletal muscle contractile function and thereby improve muscle power and sprint exercise performance. This review describes the physiological mechanisms potentially responsible for these effects, outlines the circumstances in which ergogenic effects are most likely to be evident, and discusses the effects of dietary nitrate supplementation on physical performance in a range of human populations.

Effect of dietary nitrate on enteric methane emissions, production performance and rumen fermentation of dairy cows grazing kikuyu-dominant pasture during summer

Dietary nitrate supplementation is an effective methane (CH4) mitigation strategy in total mixed ration based diets fed to ruminants. To date, limited information is available on the effect of dietary nitrate on CH4 production from grazing dairy cows. Fifty-four multiparous Jersey cows were subjected to a randomised complete block design (blocked according to milk yield, days in milk and parity) to evaluate the effect of three dietary nitrate levels on enteric CH4 emissions and cow production performance. Additionally, six rumen-cannulated cows in a replicated 3 × 3 Latin square design were used in a rumen study. Dietary treatments consisted of concentrate fed at 5.4 kg of DM/cow per day containing one of three levels of dietary nitrate: 0 g (control), 11 g (low nitrate), and 23 g of nitrate/kg of dry matter (DM; high nitrate). Cows grazed late-summer pasture containing approximately 3 g of nitrate/kg of DM. Concentrates were formulated to be isonitrogenous, by substituting urea, and isoenergetic. Cows were gradually adapted to concentrates over a 3-wk period before the onset of a 57-d experimental period. Enteric CH4 emissions and total dry matter intake (DMI) from 11 cows per treatment were measured during one 6-d measurement period using the sulphur hexafluoride tracer gas technique. Individual pasture DMI was determined using TiO2 and indigestible neutral detergent fibre (NDF). Milk yield decreased by approximately 12% when feeding the high nitrate diet compared with the control and low nitrate diets. Although total DMI was unaffected by treatment, concentrate DMI decreased linearly (5.5–3.7 kg/d) while pasture DMI increased linearly (9.1–11.4 kg/d) with increasing dietary nitrate addition. Methane production (313–280 g/d), CH4 yield (21.8–18.7 g/kg of DMI) and CH4 energy per gross energy intake (6.9–5.9%) tended to decrease linearly with increasing dietary nitrate addition. Diurnal ruminal pH of the high nitrate group was greater, for selective periods after concentrate feeding, than the control and low nitrate groups. Spot sample ruminal pH (6.2–6.3) tended to increase while total volatile fatty acid (VFA) concentration (99.9–104 mM/L) increased quadratically with increasing dietary nitrate addition. Individual VFA concentrations were unaffected by treatment. Rate of NDF disappearance (2.4–2.8%/h) after 18 h of ruminal incubation tended to increase quadratically with increasing dietary nitrate addition. Dietary nitrate fed to grazing dairy cows tended to decrease CH4 emissions while improving the fibrolytic environment of the rumen. However, when feeding high levels of dietary nitrate a decrease in milk yield could be expected due to a decrease in concentrate DMI.

Effects of dietary nitrate supplementation, from beetroot juice, on blood pressure in hypertensive pregnant women: A randomised, double-blind, placebo-controlled feasibility trial

Chronic hypertension in pregnancy is associated with significant adverse pregnancy outcomes, increasing the risk of pre-eclampsia, fetal growth restriction and preterm birth. Dietary nitrate, abundant in green leafy vegetables and beetroot, is reduced in vivo to nitrite and subsequently nitric oxide, and has been demonstrated to lower blood pressure, improve vascular compliance and enhance blood flow in non-pregnant humans and animals.The primary aims of this study were to determine the acceptability and efficacy of dietary nitrate supplementation, in the form of beetroot juice, to lower blood pressure in hypertensive pregnant women. In this double-blind, placebo-controlled feasibility trial, 40 pregnant women received either daily nitrate supplementation (70 mL beetroot juice, n = 20) or placebo (70 mL nitrate-depleted beetroot juice, n = 20) for 8 days. Blood pressure, cardiovascular function and uteroplacental blood flow was assessed at baseline and following acute (3 h) and prolonged (8 days) supplementation. Plasma and salivary samples were collected for analysis of nitrate and nitrite concentrations and acceptability of this dietary intervention was assessed based on questionnaire feedback. Dietary nitrate significantly increased plasma and salivary nitrate/nitrite concentrations compared with placebo juice (p < 0.001), with marked variation between women. Compared with placebo, there was no overall reduction in blood pressure in the nitrate-treated group; however there was a highly significant correlation between changes in plasma nitrite concentrations and changes in diastolic blood pressure in the nitrate-treated arm only (r = −0.6481; p = 0.0042). Beetroot juice supplementation was an acceptable dietary intervention to 97% of women. This trial confirms acceptability and potential efficacy of dietary nitrate supplementation in pregnant women. Conversion of nitrate to nitrite critically involves oral bacterial nitrate reductase activities. We speculate that differences in efficacy of nitrate supplementation relate to differences in the oral microbiome, which will be investigated in future studies.

Long-Term Dietary Nitrate Supplementation Does Not Prevent Development of the Metabolic Syndrome in Mice Fed a High-Fat Diet.

Background Nitric oxide (NO) is an important vascular signaling molecule that plays a role in vascular homeostasis. A reduction in NO bioavailability is thought to contribute to endothelial dysfunction, an early risk factor for both cardiovascular disease and type 2 diabetes. Dietary nitrate, through the nitrate-nitrite-NO pathway, may provide an alternate source of NO when the endogenous eNOS system is compromised. In addition to a role in the vascular system, NO may also play a role in the metabolic syndrome including obesity and glucose tolerance. Aim To investigate the effect of long-term dietary nitrate supplementation on development of the metabolic syndrome in mice fed a high-fat diet. Methods Following 1 week of acclimatisation, male (6–8 weeks) C57BL6 mice were randomly assigned to the following groups (10/group) for 12 weeks: (i) normal chow + NaCl (1 mmol/kg/day), (ii) normal chow + NaNO3 (1 mmol/kg/day), (iii) high-fat diet + NaCl (1 mmol/kg/day), and (iv) high-fat diet + NaNO3 (1 mmol/kg/day). Body weight and food consumption were monitored weekly. A subset of mice (5/group) underwent running wheel assessment. At the end of the treatment period, all mice underwent fasting glucose tolerance testing. Caecum contents, serum, and tissues (liver, skeletal muscle, white and brown adipose, and kidney) were collected, frozen, and stored at −80°C until analysis. Results Consumption of the high-fat diet resulted in significantly greater weight gain that was not affected by dietary nitrate. Mice on the high-fat diet also had impaired glucose tolerance that was not affected by dietary nitrate. There was no difference in adipose tissue expression of thermogenic proteins or energy expenditure as assessed by the running wheel activity. Mice on the high-fat diet and those receiving dietary nitrate had reduced caecum concentrations of both butyrate and propionate. Conclusions Dietary nitrate does not prevent development of the metabolic syndrome in mice fed a high-fat diet. This may be due, in part due, to reductions in the concentration of important short-chain fatty acids.

The effect of beetroot juice supplementation on repeat-sprint performance in hypoxia

ABSTRACTThis investigation assessed the effect of dietary nitrate (NO3−) supplementation, in the form of beetroot juice (BR), on repeat-sprint performance in normoxia and normobaric hypoxia. 12 male team-sport athletes (age 22.3 ± 2.6 y, VO2peak 53.1 ± 8.7 mL.kg−1.min−1) completed three exercise trials involving a 10 min submaximal warm-up and 4 sets of cycling repeat-sprint efforts (RSE; 9 × 4 s) at sea level (CON), or at 3000 m simulated altitude following acute supplementation (140 mL) with BR (HYPBR; 13 mmol NO3−) or NO3–depleted BR placebo (HYPPLA). Peak (PPO) and mean (MPO) power output, plus work decrement were recorded during the RSE task, while oxygen consumption (VO2) was measured during the warm-up. There were no significant differences observed between HYPBR and HYPPLA for PPO or MPO; however, work decrement was reduced in the first RSE set in HYPBR compared with HYPPLA. There was a moderate effect for VO2 to be lower following BR at the end of the 10 min warm-up (ES = 0.50 ± 0.51). Dietary NO3− may not improve repeat-sprint performance in hypoxia but may reduce VO2 during submaximal exercise. Therefore, BR supplementation may be more effective for performance improvement during predominantly aerobic exercise.

PPARα-independent effects of nitrate supplementation on skeletal muscle metabolism in hypoxia

Hypoxia is a feature of many disease states where convective oxygen delivery is impaired, and is known to suppress oxidative metabolism. Acclimation to hypoxia thus requires metabolic remodelling, however hypoxia tolerance may be aided by dietary nitrate supplementation. Nitrate improves tissue oxygenation and has been shown to modulate skeletal muscle tissue metabolism via transcriptional changes, including through the activation of peroxisome proliferator-activated receptor alpha (PPARα), a master regulator of fat metabolism. Here we investigated whether nitrate supplementation protects skeletal muscle mitochondrial function in hypoxia and whether PPARα is required for this effect. Wild-type and PPARα knockout (PPARα−/−) mice were supplemented with sodium nitrate via the drinking water or sodium chloride as control, and exposed to environmental hypoxia (10% O2) or normoxia for 4 weeks. Hypoxia suppressed mitochondrial respiratory function in mouse soleus, an effect partially alleviated through nitrate supplementation, but occurring independently of PPARα. Specifically, hypoxia resulted in 26% lower mass specific fatty acid-supported LEAK respiration and 23% lower pyruvate-supported oxidative phosphorylation capacity. Hypoxia also resulted in 24% lower citrate synthase activity in mouse soleus, possibly indicating a loss of mitochondrial content. These changes were not seen, however, in hypoxic mice when supplemented with dietary nitrate, indicating a nitrate dependent preservation of mitochondrial function. Moreover, this was observed in both wild-type and PPARα−/− mice. Our results support the notion that nitrate supplementation can aid hypoxia tolerance and indicate that nitrate can exert effects independently of PPARα.

Oral Nitrate Supplementation Differentially Modulates Cerebral Artery Blood Velocity and Prefrontal Tissue Oxygenation During 15 km Time-Trial Cycling in Normoxia but Not in Hypoxia.

Background: Nitrate is a precursor of nitric oxide (NO), an important regulator of cerebral perfusion in normoxic and hypoxic conditions. Nitrate supplementation could be used to improve cerebral perfusion and oxygenation during exercise in hypoxia. The effects of dietary nitrate supplementation on cerebral haemodynamics during exercise in severe hypoxia (arterial O2 saturation < 70%) have not been explored.Methods: In twelve trained male cyclists, we measured blood pressure (BP), middle cerebral artery blood velocity (MCAv), cerebrovascular resistance (CVR) and prefrontal oxyhaemoglobin and deoxyhaemoglobin concentration (O2Hb and HHb, respectively) during 15 km cycling time trials (TT) in normoxia and severe hypoxia (11% inspired O2, peripheral O2 saturation ∼66%) following 3-day oral supplementation with placebo or sodium nitrate (0.1 mmol/kg/day) in a randomised, double-blinded manner. We tested the hypothesis that dietary nitrate supplementation increases MCAv and cerebral O2Hb during TT in severe hypoxia.Results: During TT in normoxia, nitrate supplementation lowered MCAv by ∼2.3 cm/s and increased cerebral O2Hb by ∼6.8 μM and HHb by ∼2.1 μM compared to normoxia placebo (p ≤ 0.01 for all), while BP tended to be lowered (p = 0.06). During TT in severe hypoxia, nitrate supplementation elevated MCAv (by ∼2.5 cm/s) and BP (by ∼5 mmHg) compared to hypoxia placebo (p < 0.01 for both), while it had no effect on cerebral O2Hb (p = 0.98), HHb (p = 0.07) or PETCO2 (p = 0.12). Dietary nitrate had no effect of CVR during TT in normoxia or hypoxia (p = 0.19).Conclusion: Our findings indicate that during normoxic TT, the modulatory effect of dietary nitrate on regional and global cerebral perfusion is heterogeneous. Meanwhile, the lack of major changes in cerebral perfusion with dietary nitrate during hypoxic TT alludes to an exhausted cerebrovascular reserve.

Dietary nitrate supplementation reduces low frequency blood pressure fluctuations in rats following distal middle cerebral artery occlusion.

It is known that high blood pressure variability (BPV) in acute ischemic stroke is associated with adverse outcomes, yet there are no therapeutic treatments to reduce BPV. Studies have found increasing nitric oxide (NO) bioavailability improves neurological function following stroke, but whether dietary nitrate supplementation could reduce BPV remains unknown. We investigated the effects of dietary nitrate supplementation on heart rate (HR), blood pressure (BP), and beat-to-beat BPV using wireless telemetry in a rat model of distal middle cerebral artery occlusion. Blood pressure variability was characterized by spectral power analysis in the low frequency (LF; 0.2-0.6 Hz) range prestroke and during the 7 days poststroke in a control group ( n = 8) and a treatment group ( n = 8, 183 mg/l sodium nitrate in drinking water). Dietary nitrate supplementation moderately reduced systolic BPV in the LF range by ~11% compared with the control group ( P = 0.03), while resting BP and HR were not different between the two groups ( P = 0.28 and 0.33, respectively). Despite systolic BPV being reduced with dietary nitrate, we found no difference in infarct volumes between the treatment and the control groups (1.59 vs. 1.62 mm3, P = 0.86). These findings indicate that dietary nitrate supplementation is effective in reducing systolic BPV following stroke without affecting absolute BP. In light of mounting evidence linking increased BPV with poor stroke patient outcome, our data support the role of dietary nitrate as an adjunct treatment following ischemic stroke. NEW & NOTEWORTHY Using a rat model of stroke, we found that dietary nitrate supplementation reduced low frequency blood pressure fluctuations following stroke without affecting absolute blood pressure values. Since blood pressure fluctuations are associated with poor clinical outcome in stroke patients, our findings indicate that dietary nitrate could be an effective strategy for reducing blood pressure fluctuations, which could help reduce stroke severity and improve patient recovery.

Acute dietary nitrate supplementation does not attenuate oxidative stress or the hemodynamic response during submaximal exercise in hypobaric hypoxia

The purpose of this study was to investigate changes in oxidative stress, arterial oxygen saturation (SaO2), blood pressure (BP), and heart rate (HR) during exercise in hypobaric hypoxia following acute dietary nitrate supplementation. Nine well-trained (maximal oxygen consumption, 60.8 ± 7.8 mL·kg-1·min-1) males (age, 29 ± 7 years) visited the laboratory on 3 occasions, each separated by 1 week. Visit 1 included a maximal aerobic cycling test and five 5-min increasing-intensity exercise bouts in a normobaric environment (1600 m). A single dose of either a nitrate-depleted placebo (PL) or a nitrate-rich beverage (NR; 12.8 mmol nitrate) was consumed 2.5 h prior to exercise during visits 2 and 3 (3500 m) in a double-blind, placebo-controlled, crossover study consisting of a 5-min cycling warm-up and 4 bouts, each 5 min in duration, separated by 4-min periods of passive rest. Exercise wattages were determined during visit 1 and corresponded to 25%, 40%, 50%, 60%, and 70% of normobaric maximal oxygen consumption. Catalase and 8-isoprostane were measured before and after exercise (immediately before and 1 h postexercise, respectively). NR increased plasma nitrite (1.53 ± 0.83 μmol·L-1) compared with PL (0.88 ± 0.56 μmol·L-1) (p < 0.05). In both conditions, postexercise (3500 m) 8-isoprostane (PL, 23.49 ± 3.38 to 60.90 ± 14.95 pg·mL-1; NR, 23.23 ± 4.12 to 52.11 ± 19.76 pg·mL-1) and catalase (PL, 63.89 ± 25.69 to 128.15 ± 41.80 mmol·min-1·mL-1; NR, 78.89 ± 30.95 to 109.96 ± 35.05 mmol·min-1·mL-1) were elevated compared with baseline resting values (p < 0.05). However, both 8-isoprostane and catalase were similar in the 2 groups (PL and NR) (p = 0.217 and p = 0.080, respectively). We concluded that an acute, pre-exercise dose of dietary nitrate yielded no beneficial changes in oxidative stress, SaO2, BP, or HR in healthy, aerobically fit men exercising at 3500 m.

Effect of dietary nitrate supplementation on conduit artery blood flow, muscle oxygenation, and metabolic rate during handgrip exercise.

Dietary nitrate supplementation has positive effects on mitochondrial and muscle contractile efficiency during large muscle mass exercise in humans and on skeletal muscle blood flow (Q̇) in rats. However, concurrent measurement of these effects has not been performed in humans. Therefore, we assessed the influence of nitrate supplementation on Q̇ and muscle oxygenation characteristics during moderate- (40 %peak) and severe-intensity(85% peak) handgrip exercise in a randomized, double-blind, crossover design. Nine healthy men (age: 25 ± 2 yr) completed four constant-power exercise tests (2/intensity) randomly assigned to condition [nitrate-rich (nitrate) or nitrate-poor (placebo) beetroot supplementation] and intensity (40 or 85% peak). Resting mean arterial pressure was lower after nitrate compared with placebo (84 ± 4 vs. 89 ± 4 mmHg, P < 0.01). All subjects were able to sustain 10 min of exercise at 40% peak in both conditions. Nitrate had no effect on exercise tolerance during 85% peak (nitrate: 358 ± 29; placebo: 341 ± 34 s; P = 0.3). Brachial artery Q̇ was not different after nitrate at rest or any time during exercise. Deoxygenated [hemoglobin + myoglobin] was not different for 40% peak ( P > 0.05) but was elevated throughout 85% peak ( P < 0.05) after nitrate. The metabolic cost (V̇o2) was not different at the end of exercise; however, the V̇o2 primary amplitude at the onset of exercise was elevated after nitrate for the 85% peak work rate (96 ± 20 vs. 72 ± 12 ml/min, P < 0.05) and had a faster response. These findings suggest that an acute dose of nitrate reduces resting blood pressure and speeds V̇o2 kinetics in young adults but does not augment Q̇ or reduce steady-state V̇o2 during small muscle mass handgrip exercise. NEW & NOTEWORTHY We show that acute dietary nitrate supplementation via beetroot juice increases the amplitude and speed of local muscle V̇o2 on kinetics parameters during severe- but not moderate-intensity handgrip exercise. These changes were found in the absence of an increased blood flow response, suggesting that the increased V̇o2 was attained via improvements in fractional O2 extraction and/or spatial distribution of blood flow within the exercising muscle.

Changes in Oxidative Stress and Resting Metabolic Rate after Acute Dietary Nitrate Supplementation

BACKGROUND: Cardiovascular disease is the leading cause of preventable death in the United States and the Western world. A major component of cardiovascular disease is the deterioration of the cardiovascular system by increased oxidative stress. Dietary nitrate supplementation could increase nitric oxide bioavailability, decreasing oxidative stress. PURPOSE: The purpose of the present study is twofold: 1) to determine the impact of an acute dietary nitrate supplementation on oxidative stress and 2) to assess the oxygen dependent nature of a dietary nitrate supplementation. METHODS: Eighteen (7 females and 11 males) apparently healthy subjects, aged 18-30 years of age, participated in a randomized, double blind, placebo-controlled crossover study. Following a 12-hour fast and adherence to an NIH-approved low-nitrate diet for 48 hours, subjects visited the lab on 4 occasions (identical procedures) within 2 weeks. A resting blood draw preceded 15 minutes of supine rest followed by a 20-minute period of metabolic gas analysis for determination of resting metabolic rate; RMR (True One, ParvoMedics, Sandy, Utah, USA). Visits 2 and 4 served as baseline controls for the placebo or dietary nitrate treatments (negligible and 800mg nitrate, respectively) which were consumed 2.5 hours prior to visits 3 and 5. The resting concentration of the oxidative stress marker 8-isoprostane (8-ISO) was determined by ELISA testing using a commercial available kit (Cayman Chemical, USA). A 2-way repeated measures ANOVA was used to determine differences between protocols and over time, with an alpha of 0.05. RESULTS: There was no significant difference between placebo and dietary nitrate supplementation in oxidative stress (Placebo: 158±59 vs. 181±62 pg/ml; Active: 176±74 vs. 171±59 pg/ml, p=0.84) or in RMR (Placebo: 1839±308 vs. 1790±307 kcal/day; Active: 1828±318 vs. 1859±324 kcal/day, p=0.20). CONCLUSION: These data showed that acute dietary nitrate supplementation does not decrease resting oxidative stress. The conversion of nitrate to nitrite to nitric oxide has been previously described to be oxygen-independent. Our data supported this statement, as RMR did not change following the dietary nitrate supplementation.

Acute Dietary Nitrate Supplementation has no Significant Effect on Wasted Left Ventricular Energy in Young Healthy Individuals

Acute Dietary Nitrate Supplementation has no Significant Effect on Wasted Left Ventricular Energy in Young Healthy Individuals

Ergogenic effects of beetroot juice supplementation during severe-intensity exercise in obese adolescents.

Previous studies showed a higher O2 cost of exercise, and therefore, a reduced exercise tolerance in patients with obesity during constant work rate (CWR) exercise compared with healthy subjects. Among the ergogenic effects of dietary nitrate ([Formula: see text]) supplementation in sedentary healthy subjects, a reduced O2 cost and enhanced exercise tolerance have often been demonstrated. The aim of this study was to evaluate the effects of beetroot juice (BR) supplementation, rich in [Formula: see text], on physiological variables associated with exercise tolerance in adolescents with obesity. In a double-blind, randomized crossover study, 10 adolescents with obesity (8 girls, 2 boys; age = 16 ± 1 yr; body mass index = 35.2 ± 5.0 kg/m2) were tested after 6 days of supplementation with BR (5 mmol [Formula: see text] per day) or placebo (PLA). Following each supplementation period, patients carried out two repetitions of 6-min moderate-intensity CWR exercise and one severe-intensity CWR exercise until exhaustion. Plasma [Formula: see text] concentration was significantly higher in BR versus PLA (108 ± 37 vs. 15 ± 5 μM, P < 0.0001). The O2 cost of moderate-intensity exercise was not different in BR versus PLA (13.3 ± 1.7 vs. 12.9 ± 1.1 ml·min-1·W-1, P = 0.517). During severe-intensity exercise, signs of a reduced amplitude of the O2 uptake slow component were observed in BR, in association with a significantly longer time to exhaustion (561 ± 198 s in BR vs. 457 ± 101 s in PLA, P = 0.0143). In obese adolescents, short-term dietary [Formula: see text] supplementation is effective in improving exercise tolerance during severe-intensity exercise. This may prove to be useful in counteracting early fatigue and reduced physical activity in this at-risk population.

Effects of Dietary Inorganic Nitrate Supplementation on Exercise Performance in Patients With Heart Failure: Protocol for a Randomized, Placebo-Controlled, Cross-Over Trial.

BackgroundChronic heart failure is characterized by an inability of the heart to pump enough blood to meet the demands of the body, resulting in the hallmark symptom of exercise intolerance. Chronic underperfusion of the peripheral tissues and impaired nitric oxide bioavailability have been implicated as contributors to the decrease in exercise capacity in these patients. nitric oxide bioavailability has been identified as an important mediator of exercise tolerance in healthy individuals, but there are limited studies examining the effects in patients with chronic heart failure.ObjectiveThe proposed trial is designed to determine the effects of chronic inorganic nitrate supplementation on exercise tolerance in both patients with heart failure preserved ejection fraction (HFpEF) and heart failure reduced ejection fraction (HFrEF) and to determine whether there are any differential responses between the 2 cohorts. A secondary objective is to provide mechanistic insights into the 2 heart failure groups’ exercise responses to the nitrate supplementation.MethodsPatients with chronic heart failure (15=HFpEF and 15=HFrEF) aged 40 to 85 years will be recruited. Following an initial screen cardiopulmonary exercise test, participants will be randomly allocated in a double-blind fashion to consume either a nitrate-rich beetroot juice (16 mmol nitrate/day) or a nitrate-depleted placebo (for 5 days). Participants will continue daily dosing until the completion of the 4 testing visits (maximal cardiopulmonary exercise test, submaximal exercise test with echocardiography, vascular function assessment, and vastus lateralis muscle biopsy). There will then be a 2-week washout period after which the participants will cross over to the other treatment and complete the same 4 testing visits.ResultsThis study is funded by National Heart Foundation of Australia and Victoria University. Enrolment has commenced and the data collection is expected to be completed in mid 2018. The initial results are expected to be submitted for publication by the end of 2018.ConclusionsIf inorganic nitrate supplementation can improve exercise tolerance in patients with chronic heart failure, it has the potential to aid in further refining the treatment of patients in this population.Trial RegistrationAustralian New Zealand Clinical Trials Registry ACTRN12615000906550; https://www.anzctr.org.au/Trial/Registration/TrialReview.aspx?id=368912 (Archived by WebCite at http://www.webcitation.org/6xymLMiFK)

Effect of Dietary Nitrate on Blood Pressure and Vascular Control in Post‐Menopausal Hypertensive Women

Individuals with hypertension exhibit abnormal pressor responses during rest‐to‐exercise transitions, significantly increasing the likelihood of sudden cardiac events. Dietary nitrate supplementation can lower resting blood pressure in patients with hypertension and improve bulk muscle blood flow in older healthy adults. However, investigations using dietary nitrate as a non‐pharmacological intervention in post‐menopausal women diagnosed with hypertension are lacking. Therefore, the purpose of the current investigation was to determine if acute dietary nitrate supplementation would improve the pressor response to exercise and increase forearm blood flow during steady‐state hand‐grip exercise in post‐menopausal hypertensive women. It was hypothesized that the pressor response to exercise would be attenuated and muscle blood flow would be improved in the nitrate‐rich (NR) versus nitrate‐poor (NP) treatment.METHODSTen hypertensive post‐menopausal women [age 56 ± 3 years (mean±SD); height 165 ± 2 cm; mass 84 ± 13 kg; body mass index 31 ± 5 kg/m2] agreed to participate in the current study. Patients underwent a randomized double‐blind placebo‐controlled trial with a NR or NP beet root juice supplement. Blood samples were taken at the beginning of each testing session (~2 h post ingestion) to determine plasma nitrite levels. Forearm blood flow (FBF) was measured via ultrasonography at the brachial artery of the exercising limb. Beat‐by‐beat blood pressure and heart rate were recorded throughout the trial on the non‐exercising limb. After resting measurements were completed, patients performed hand‐grip exercise at 20% maximal voluntary contraction (MVC) for five minutes to obtain steady‐state blood flow and blood pressure response to exercise.RESULTSPlasma nitrite levels were significantly higher during the NR (809 ± 146 nM) than the NP (79 ± 19 nM) condition. No differences were found between conditions for resting blood flow (NR: 64±2; NP: 62 ± 4 ml min−1; P &gt; 0.05), systolic (NR: 144 ± 2; NP: 145 ± 2 mmHg−1; P &gt; 0.05) or diastolic blood pressure (NR: 78 ± 1; NP: 80 ± 1 mmHg−1; P &gt; 0.05). In addition, there were no significant differences across 10‐second bins (e.g., 0–10, 11–20 seconds) during the first minute pressor response to hand‐grip exercise in either condition (systolic: P &gt; 0.05; diastolic: P &gt; 0.05). As such, the rate pressure product was not significantly different during steady‐state between conditions (NR: 11,562 ± 437; NP: 11,728 ± 437 bpm · mmHg−1; P &gt; 0.05). During steady‐state exercise, FBF (NR: 189 ± 8; NP: 218 ± 8 ml min−1; P = 0.03) and forearm vascular conductance (FVC) (NR: 157 ± 4; NP: 183 ± 4 ml min−1; P = 0.04) were significantly lower during the steady‐state NR condition.CONCLUSIONAcute dietary nitrate supplementation did not improve the pressor response to hand‐grip exercise in hypertensive post‐menopausal women. However, it is important to note that steady‐state FBF (~30 ml min−1) and FVC (~27 ml min mmHg−1) were significantly reduced during the nitrate‐rich condition, which suggests a reduced O2 cost for a given workload because blood flow typically matches metabolic rate.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Abstract P242: Dietary Nitrate Does not Reduce Resting Metabolic Rate or Oxidative Stress in Healthy Males

Introduction: Nitric oxide (NO) is a vasodilator that increases blood flow by promoting relaxation of endothelium. Dietary nitrate supplementation increases plasma nitrite, a marker of overall NO bioavailability. Previously, acute dietary nitrate supplementation has been shown to reduce oxygen consumption and improve tolerance during submaximal exercise in healthy populations. Less is known about the effect of dietary nitrate on oxygen consumption at rest. Hypothesis: We hypothesized that dietary nitrate supplementation would reduce resting metabolic rate (RMR) and oxidative stress (8-isoprostane) at rest, via enhanced NO bioavailability via the oxygen independent Nitrate-Nitrite-Nitric Oxide pathway in healthy, young males. Methods: In a randomized, double-blind, cross-over study, ten healthy, young males (21 ± 2 years) visited the laboratory on 5 separate occasions. Participants completed informed consent paperwork and underwent protocol familiarization during visit 1. Prior to visits 2 and 4, participants fasted for 12 hours and adhered to an NIH-approved low-nitrate diet for 48 hours. During visits 2 and 4, an initial blood draw was performed to analyze baseline plasma nitrite and 8-isoprostane. Participants then completed a 30-minute resting metabolic rate (RMR) test. Two hours prior to visits 3 and 5, participants consumed either a placebo or dietary nitrate supplement (negligible and 6.2 mmol nitrate, respectively). During visits 3 and 5, participants also had blood drawn for analysis of the previously stated measurements, and completed an RMR test. Visits 2 and 3 were on consecutive days, followed by a week-long washout period between visit 3 and visit 4, while visit 4 and 5 also occurred on consecutive days. Results: Plasma nitrite significantly increased following dietary nitrate consumption compared to baseline values (27.56 ± 7.58 and 1.25 ± 1.51 arbitrary units, respectively). No difference was present between nitrate and baseline measurements for 8-isoprostane (155.75 ± 57.01 and 198.42 ± 66.44 pg/mL, respectively; p=0.55) and RMR (2086.60 ± 202.23 and 2050.00 ± 209.23 kcal/day, respectively; p=0.13). Conclusion: Dietary nitrate supplementation increases plasma nitrite, but does not change resting metabolic rate following an acute dose of dietary nitrate in healthy males. Individuals consuming dietary nitrate as an ergogenic aid during exercise may not, however, experience similar changes in their resting metabolism. The lack of change in oxidative stress may have been associated with the overall health of the cohort examined. Future research should investigate the clinical implications of dietary nitrate in populations with decreased NO bioavailability and associated endothelial disfunction (and elevated oxidative stress). In such populations, dietary nitrate may provide benefit. However, in a healthy cohort, dietary nitrate exerts minimal effects.

Abstract P243: Endothelial Function, Arterial Stiffness and Central Blood Pressure are Not Affected by a Single Dose of Dietary Nitrate in Healthy Normotensive Females

Intro: Hypertension is a prominent risk factor for cardiovascular disease and arterial stiffness is a related predictor of both mortality and longitudinal onset of hypertension in normotensive individuals. Both vascular compliance and blood pressure have been shown to improve following dietary nitrate supplementation in some healthy and diseased individuals. Dietary nitrate supplementation increases plasma nitrite and may, therefore, enhance bioavailability of endothelium derived nitric oxide. Chronic increased flow mediated dilation (FMD) and reduction of pulse wave velocity (PWV) reduce a person’s long-term risk for developing endothelial dysfunction, atherosclerosis and hypertension. Hypothesis: We hypothesized that an acute dose of dietary nitrate would improve FMD, decrease PWV and reduce central blood pressure (cBP) in healthy, young females. Methods: Seven healthy, young (21 ± 2 years) females participated in a randomized, double blind, placebo controlled, crossover protocol. Participants had endothelial function measured via FMD, arterial stiffness via PWV and applanation tonometry, and cBP was determined non-invasively using pulse wave analysis following a 12-hour fasting period and 48-hour NIH approved low nitrate diet. Baseline values were measured the day before each treatment visit (placebo or nitrate-rich, separated by a 1-week washout period). Participants consumed either a placebo or nitrate-rich supplement (negligible and 6.2 mmol nitrate, respectively) 2 hours prior to their laboratory visit. Upon arrival, blood was drawn and participants assumed a supine position for 15 minutes. Then brachial artery FMD was assessed via high definition ultrasound imaging following 5 minutes of proximal forearm occlusion, followed by PWV and central blood pressure measurements. Results: Plasma nitrite was elevated following consumption of the nitrate rich supplement compared to placebo (26.33 ± 15.76 and 1.18 ± 0.96 arbitrary units, respectively; p&lt;0.05). However, there was no difference between nitrate rich and placebo treatments for endothelial function as measured by FMD (10.33 ± 1.82 and 9.11 ± 5.43%, respectively; p=0.70). No difference was present between nitrate and placebo groups for PWV (5.99 ± 0.85 and 6.16 ± 0.80 m/s, respectively; p=0.90), systolic cBP (101.57 ± 9.29 and 96.90 ± 20.95 mmHg, respectively; p=0.40) and diastolic cBP (72.57 ± 7.67 and 73.20 ± 5.01 mmHg, respectively; p=0.97). Conclusion: While a single dose of dietary nitrate does increase plasma nitrite, supplementation has minimal effects on FMD, PWV and cBP in young healthy females. Future research should examine longer duration supplementation protocols as well as the changes in vascular health of patients with known disease such as hypertension or peripheral artery disease.

Salivary nitrite production is elevated in individuals with a higher abundance of oral nitrate-reducing bacteria.

Nitric oxide (NO) can be generated endogenously via NO synthases or via the diet following the action of symbiotic nitrate-reducing bacteria in the oral cavity. Given the important role of NO in smooth muscle control there is an intriguing suggestion that cardiovascular homeostasis may be intertwined with the presence of these bacteria. Here, we measured the abundance of nitrate-reducing bacteria in the oral cavity of 25 healthy humans using 16S rRNA sequencing and observed, for 3.5 h, the physiological responses to dietary nitrate ingestion via measurement of blood pressure, and salivary and plasma NO metabolites. We identified 7 species of bacteria previously known to contribute to nitrate-reduction, the most prevalent of which were Prevotella melaninogenica and Veillonella dispar. Following dietary nitrate supplementation, blood pressure was reduced and salivary and plasma nitrate and nitrite increased substantially. We found that the abundance of nitrate-reducing bacteria was associated with the generation of salivary nitrite but not with any other measured variable. To examine the impact of bacterial abundance on pharmacokinetics we also categorised our participants into two groups; those with a higher abundance of nitrate reducing bacteria (> 50%), and those with a lower abundance (< 50%). Salivary nitrite production was lower in participants with lower abundance of bacteria and these individuals also exhibited slower salivary nitrite pharmacokinetics. We therefore show that the rate of nitrate to nitrite reduction in the oral cavity is associated with the abundance of nitrate-reducing bacteria. Nevertheless, higher abundance of these bacteria did not result in an exaggerated plasma nitrite response, the best known marker of NO bioavailability. These data from healthy young adults suggest that the abundance of oral nitrate-reducing bacteria does not influence the generation of NO through the diet, at least when the host has a functional minimum threshold of these microorganisms.

Influence of dietary nitrate supplementation on lung function and exercise gas exchange in COPD patients

BackgroundDuring exercise as pulmonary blood flow rises, pulmonary capillary blood volume increases and gas exchange surface area expands through distention and recruitment. We have previously demonstrated that pulmonary capillary recruitment is limited in COPD patients with poorer exercise tolerance. Hypoxia and endothelial dysfunction lead to pulmonary vascular dysregulation possibly in part related to nitric oxide related pathways. PurposeTo determine if increasing dietary nitrate might influence lung surface area for gas exchange and subsequently impact exercise performance. MethodsSubjects had stable, medically treated COPD (n = 25), gave informed consent, filled out the St George Respiratory Questionnaire (SGRQ), had a baseline blood draw for Hgb, performed spirometry, and had exhaled nitric oxide (exNO) measured. Then they performed the intra-breath (IB) technique for lung diffusing capacity for carbon monoxide (DLCO) as well as pulmonary blood flow (Qc). Subsequently they completed a progressive semi-recumbent cycle ergometry test to exhaustion with measures of oxygen saturation (SpO2) and expired gases along with DLCO and Qc measured during the 1st work load only. Subjects were randomized to nitrate supplement group (beetroot juice) or placebo group (black currant juice) for 8 days and returned for repeat of the above protocol. ResultsExhaled nitric oxide levels rose >200% in the nitrate group (p < 0.05) with minimal change in placebo group. The SGRQ suggested a small fall in perceived symptom limitation in the nitrate group, but no measure of resting pulmonary function differed post nitrate supplementation. With exercise, there was no influence of nitrate supplementation on peak VO2 or other measures of respiratory gas exchange. There was a tendency for the exercise DLCO to increase slightly in the nitrate group with a trend towards a rise in the DLCO/Qc relationship (p = 0.08) but not in the placebo group. The only other significant finding was a fall in the exercise blood pressure in the nitrate group, but not placebo group (p < 0.05). ConclusionDespite evidence of a rise in exhaled nitric oxide levels with nitrate supplementation, there was minimal evidence for improvement in exercise performance or pulmonary gas exchange surface area in a stable medically treated COPD population.