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Nitric oxide (NO) is a gaseous signaling molecule that plays an important role in myriad physiological processes, including the regulation of vascular tone, neurotransmission, mitochondrial respiration, and skeletal muscle contractile function. NO may be produced via the canonical NO synthase-catalyzed oxidation of l-arginine and also by the sequential reduction of nitrate to nitrite and then NO. The body's nitrate stores can be augmented by the ingestion of nitrate-rich foods (primarily green leafy vegetables). NO bioavailability is greatly enhanced by the activity of bacteria residing in the mouth, which reduce nitrate to nitrite, thereby increasing the concentration of circulating nitrite, which can be reduced further to NO in regions of low oxygen availability. Recent investigations have focused on promoting this nitrate-nitrite-NO pathway to positively affect indices of cardiovascular health and exercise tolerance. It has been reported that dietary nitrate supplementation with beetroot juice lowers blood pressure in hypertensive patients, and sodium nitrite supplementation improves vascular endothelial function and reduces the stiffening of large elastic arteries in older humans. Nitrate supplementation has also been shown to enhance skeletal muscle function and to improve exercise performance in some circumstances. Recently, it has been established that nitrate concentration in skeletal muscle is much higher than that in blood and that muscle nitrate stores are exquisitely sensitive to dietary nitrate supplementation and deprivation. In this review, we consider the possibility that nitrate represents an essential storage form of NO and discuss the integrated function of the oral microbiome, circulation, and skeletal muscle in nitrate-nitrite-NO metabolism, as well as the practical relevance for health and performance.

Introduction Dietary inorganic nitrate is a popular nutritional supplement, which increases nitric oxide bioavailability and may improve exercise performance. Despite over a decade of research into the effects of dietary nitrate supplementation during exercise there is currently no expert consensus on how, when and for whom this compound could be recommended as an ergogenic aid. Moreover, there is no consensus on the safe administration of dietary nitrate as an ergogenic aid. This study aimed to address these research gaps. Methods The modified Delphi technique was used to establish the views of 12 expert panel members on the use of dietary nitrate as an ergogenic aid. Over three iterative rounds (two via questionnaire and one via videoconferencing), the expert panel members voted on 222 statements relating to dietary nitrate as an ergogenic aid. Consensus was reached when > 80% of the panel provided the same answer (i.e. yes or no). Statements for which > 80% of the panel cast a vote of insufficient evidence were categorised as such and removed from further voting. These statements were subsequently used to identify directions for future research. Results The 12 panel members contributed to voting in all three rounds. A total of 39 statements (17.6%) reached consensus across the three rounds (20 yes, 19 no). In round one, 21 statements reached consensus (11 yes, 10 no). In round two, seven further statements reached consensus (4 yes, 3 no). In round three, an additional 11 statements reached consensus (5 yes, 6 no). The panel agreed that there was insufficient evidence for 134 (60.4%) of the statements, and were unable to agree on the outcome of the remaining statements. Conclusions This study provides information on the current expert consensus on dietary nitrate, which may be of value to athletes, coaches, practitioners and researchers. The effects of dietary nitrate appear to be diminished in individuals with a higher aerobic fitness (peak oxygen consumption [V̇O2peak] > 60 ml/kg/min), and therefore, aerobic fitness should be taken into account when considering use of dietary nitrate as an ergogenic aid. It is recommended that athletes looking to benefit from dietary nitrate supplementation should consume 8–16 mmol nitrate acutely or 4–16 mmol/day nitrate chronically (with the final dose ingested 2–4 h pre-exercise) to maximise ergogenic effects, taking into consideration that, from a safety perspective, athletes may be best advised to increase their intake of nitrate via vegetables and vegetable juices. Acute nitrate supplementation up to ~ 16 mmol is believed to be safe, although the safety of chronic nitrate supplementation requires further investigation. The expert panel agreed that there was insufficient evidence for most of the appraised statements, highlighting the need for future research in this area. Graphical Abstract

Purpose: Dietary nitrate has been shown to enhance muscle contractile function and has, therefore, been linked to increased muscle power and sprint exercise performance. However, the impact of dietary nitrate supplementation on maximal strength, performance and muscular endurance remains to be established.Methods: Fifteen recreationally active males (25 +/- 4 y, BMI 24 +/- 3 kg/m(2)) participated in a randomized double-blinded cross-over study comprising two 6-d supplementation periods; 140 mL/d nitrate-rich (BR; 985 mg/d) and nitrate-depleted (PLA; 0.37 mg/d) beetroot juice. Three hours following the last supplement, we assessed countermovement jump (CMJ) performance, maximal strength and power of the upper leg by voluntary isometric (30 degrees and 60 degrees angle) and isokinetic contractions (60, 120, 180 and 300 degrees center dot s(-1)), and muscular endurance (total workload) by 30 reciprocal isokinetic voluntary contractions at 180 degrees center dot s(-1).Results: Despite differences in plasma nitrate (BR: 879 +/- 239 vs. PLA: 33 +/- 13 mu mol/L,P 0.50 for both angles) and isokinetic knee extension power (P > 0.33 for all velocities) did not differ between treatments. Isokinetic knee flexion power was significantly higher following BR compared with PLA ingestion at 60 degrees center dot s(-1)(P = 0.001), but not at 120 degrees center dot s(-1)(P = 0.24), 180 degrees center dot s(-1)(P = 0.066), and 300 degrees center dot s(-1)(P = 0.36).Conclusion: Nitrate supplementation does not improve maximal strength, countermovement jump performance and muscular endurance in healthy, active males.

Purpose Strenuous exercise induces intestinal injury, which is likely related to splanchnic hypoperfusion and may be associated with gastrointestinal complaints commonly reported during certain exercise modalities. Increasing circulating nitric oxide (NO) levels or inducing postprandial hyperemia may improve splanchnic perfusion, thereby attenuating intestinal injury during exercise. Therefore, we investigated the effects of both dietary nitrate ingestion and sucrose ingestion on splanchnic perfusion and intestinal injury induced by prolonged strenuous cycling.Methods In a randomized crossover manner, 16 well-trained male athletes (age, 28 7 yr; W-max, 5.0 0.3 Wkg(-1)) cycled 60 min at 70% W-max after acute ingestion of sodium nitrate (NIT; 800 mg NO3), sucrose (SUC; 40 g), or a water placebo (PLA). Splanchnic perfusion was assessed by determining the gap between gastric and arterial pCO(2) (gap(g-a)pCO(2)) using gastric air tonometry. Plasma intestinal fatty acid-binding protein (I-FABP) concentrations, reflecting enterocyte damage, were assessed every 20 min during and up to 60 min of postexercise recovery.Results The exercise protocol resulted in splanchnic hypoperfusion, as gap(g-a)pCO(2) levels increased during exercise (P Conclusion Sucrose but not nitrate ingestion lowers intestinal injury evoked during prolonged strenuous cycling. These results suggest that sucrose ingestion, but not nitrate, prevents hypoperfusion-induced gastrointestinal damage during exercise and, as such, may help to lower exercise-related gastrointestinal complaints.

Background Emerging evidence suggests that increasing dietary nitrate intake may be an effective approach to reduce blood pressure. Beetroot juice is often used to supplement dietary nitrate, whereas nitrate intake levels from habitual diet are low. An increase in the habitual intake of nitrate-rich vegetables may represent an alternative to nitrate supplementation. However, the effectiveness and acceptability of a nitrate–rich-vegetables diet remain to be established. Objective The aim was to investigate the effect and feasibility of two different intervention strategies to increase dietary nitrate intake, on plasma nitrate/nitrite concentrations and blood pressure. Design A randomized, crossover trial was used. Participants Participants were healthy men and women (both n=15; age: 24±6 years) from the Netherlands. Intervention Participants were instructed to consume ∼400 mg nitrate at lunch, provided through nitrate-rich vegetables and dietary counseling, or beetroot juice supplementation. Both interventions lasted 1 week, with 1-week washout (January to April 2017). Main outcome Plasma nitrate and nitrite concentrations and resting systolic and diastolic blood pressure were measured in an overnight fasted state (before and after intervention) and ∼2.5 hours after lunch (before and throughout intervention on day 1, 4, and 7). Statistical analysis Two-factor (time × treatment) repeated-measures analyses of variance were performed. Results Mean plasma nitrate concentrations increased with both interventions, with a larger increase in beetroot juice vs nitrate-rich vegetables, both in a fasted state and ∼2.5 hours after lunch (day 1, beetroot juice: 2.31±0.56 mg/dL [373±90 μmol/L] vs nitrate-rich vegetables: 1.71±0.83 mg/dL [277±134 μmol/L]; P

Supplemental digital content is available in the text., Introduction Recently, it has been speculated that protein supplementation may further augment the adaptations to chronic endurance exercise training. We assessed the effect of protein supplementation during chronic endurance exercise training on whole-body oxidative capacity (V˙O2max) and endurance exercise performance. Methods In this double-blind, randomized, parallel placebo-controlled trial, 60 recreationally active males (age, 27 ± 6 yr; body mass index, 23.8 ± 2.6 kg·m−2; V˙O2max, 47 ± 6 mL·min−1·kg−1) were subjected to 12 wk of triweekly endurance exercise training. After each session and each night before sleep, participants ingested either a protein supplement (PRO; 28.7 g casein protein) or an isoenergetic carbohydrate placebo (PLA). Before and after the 12 wk of training, V˙O2max and endurance exercise performance (~10-km time trial) were assessed on a cycle ergometer. Muscular endurance (total workload achieved during 30 reciprocal isokinetic contractions) was assessed by isokinetic dynamometry and body composition by dual-energy x-ray absorptiometry. Mixed-model ANOVA was applied to assess whether training adaptations differed between groups. Results Endurance exercise training induced an 11% ± 6% increase in V˙O2max (time effect, P < 0.0001), with no differences between groups (PRO, 48 ± 6 to 53 ± 7 mL·min−1·kg−1; PLA, 46 ± 5 to 51 ± 6 mL·min−1·kg−1; time–treatment interaction, P = 0.50). Time to complete the time trial was reduced by 14% ± 7% (time effect, P < 0.0001), with no differences between groups (time–treatment interaction, P = 0.15). Muscular endurance increased by 6% ± 7% (time effect, P < 0.0001), with no differences between groups (time–treatment interaction, P = 0.84). Leg lean mass showed an increase after training (P < 0.0001), which tended to be greater in PRO compared with PLA (0.5 ± 0.7 vs 0.2 ± 0.6 kg, respectively; time–treatment interaction, P = 0.073). Conclusion Protein supplementation after exercise and before sleep does not further augment the gains in whole-body oxidative capacity and endurance exercise performance after chronic endurance exercise training in recreationally active, healthy young males.

Background: Dietary nitrate is receiving increased attention due to its reported ergogenic and cardioprotective properties. The extent to which ingestion of various nitrate-rich vegetables increases postprandial plasma nitrate and nitrite concentrations and lowers blood pressure is currently unknown. Objective: We aimed to assess the impact of ingesting different nitrate-rich vegetables on subsequent plasma nitrate and nitrite concentrations and resting blood pressure in healthy normotensive individuals. Methods: With the use of a semirandomized crossover design, 11 men and 7 women [mean +/- SEM age: 28 +/- 1 y; mean +/- SEM body mass index (BMI, in kg/m(2)): 23 +/- 1; exercise: 1-10 h/wk] ingested 4 different beverages, each containing 800 mg (similar to 12.9 mmol) nitrate: sodium nitrate (NaNO3), concentrated beetroot juice, a rocket salad beverage, and a spinach beverage. Plasma nitrate and nitrite concentrations and blood pressure were determined before and up to 300 min after beverage ingestion. Data were analyzed using repeated-measures ANOVA. Results: Plasma nitrate and nitrite concentrations increased after ingestion of all 4 beverages (P

The intake of nitrate from vegetables or dietary supplements that contain red beet juice can reduce blood pressure and improve sports performance. This dissertation focused on how nitrate supplementation can be optimised to improve sports performance and the extent to which nitrate supplementation can be used in professional sports. Nitrate-rich vegetables such as red beets, arugula and spinach proved to be the most effective sources of nitrate. Performance improvement following nitrate intake is primarily seen in extremely high-intensity sports, such as repeated sprints, and after multi-day nitrate intake, even among professional athletes. Together with NOC*NSF, we are currently working on sport-specific guidelines for nitrate supplementation to improve the athletic performance of professional athletes.

Purpose: There is an ongoing debate whether highly trained athletes are less responsive to the ergogenic properties of nitrate. We assessed the effects of nitrate supplementation on plasma nitrate and nitrite concentrations and repeated-sprint performance in recreational, competitive and elite sprint athletes. Methods: In a randomized double-blinded cross-over design, recreational cyclists (n=20), national talent speed-skaters (n=22) and Olympic-level track cyclists (n=10) underwent two 6-day supplementation periods; 140mL/d nitrate-rich (BR; approximate to 800mg/d) and nitrate-depleted (PLA; approximate to 0.5mg/d) beetroot juice. Blood samples were collected and three 30-s Wingate tests were performed. Results: Plasma nitrate and nitrite concentrations were higher following BR vs PLA (P.10). Peak power over the three Wingates was not different between BR and PLA (1338 +/- 30 vs 1333 +/- 30 W; P=.62), and there was no interaction between treatment (BR-PLA) and Wingate number (1-2-3; P=.48). Likewise, mean power did not differ between BR and PLA (P=.86). In contrast, time to peak power improved by approximate to 2.8% following BR vs PLA (P=.007). This improvement in BR vs PLA was not different between Wingate 1, 2 and 3.Moreover, the effects of BR vs PLA did not differ between sport levels for any Wingate parameter (all P>.30). Conclusion: The plasma and repeated-sprint performance responses to beetroot juice supplementation do not differ between recreational, competitive and elite sprint athletes. Beetroot juice supplementation reduces time to reach peak power, which may improve the capacity to accelerate during high-intensity and sprint tasks in recreational as well as elite athletes.

It has been shown that nitrate supplementation can enhance endurance exercise performance. Recent work suggests that nitrate ingestion can also increase intermittent type exercise performance in recreational athletes. We hypothesized that six days of nitrate supplementation can improve high-intensity intermittent type exercise performance in trained soccer players. Thirty-two male soccer players (age: 23 +/- 1 years, height: 181 +/- 1 m, weight: 77 +/- 1 kg, playing experience: 15.2 +/- 0.5 years, playing in the first team of a 2nd or 3rd Dutch amateur league club) participated in this randomized, double-blind cross-over study. All subjects participated in two test days in which high-intensity intermittent running performance was assessed using the Yo-Yo IR1 test. Subjects ingested nitrate-rich (140 mL; +/- 800 mg nitrate/day; BR) or a nitrate-depleted beetroot juice (PLA) for six subsequent days, with at least eight days of wash-out between trials. The distance covered during the Yo-Yo IR1 was the primary outcome measure, while heart rate (HR) was measured continuously throughout the test, and a single blood and saliva sample were collected just prior to the test. Six days of BR ingestion increased plasma and salivary nitrate and nitrite concentrations in comparison to PLA (p

nitrate rich beetroot juice has become a popular sports supplement because of its proposed ergogenic properties. The effects of dietary nitrate supplementation on exercise performance have been attributed to its capacity to increase nitric oxide (NO) bioavailability. NO modulates many processes that

to the editor: We thank the authors of the commentaries (see Ref. [1][1]) on our Viewpoint ([2][2]) for further enlightening several interesting aspects concerning nitrate supplementation in the elite athlete, a topic that evidently carries great interest. Given the large number of commentaries we