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We aimed to assess the association between gastrointestinal (GI) injury, complaints, and food intake in 60-km ultramarathon runners. Thirty-three ultramarathon runners provided pre- and post-race blood samples for assessment of GI injury by intestinal fatty-acid binding protein (I-FABP), and inflammatory response by interleukin (IL)-6, IL-8, tumour necrosis factor alpha (TNF-α), and C-reactive protein (CRP). GI complaints and nutritional intake were reported by a post-race questionnaire. GI complaints were reported by 73% of the runners, of which 20% reported 1 or 2 severe complaints. IL-6, IL8, TNF-α, and CRP increased significantly from pre- to post-race (P < 0.001 for all biomarkers), while I-FABP did not (1375 [IQR: 1264–2073] to 1726 [IQR: 985–3287] pg/mL; P = 0.330). The ‘GI complaints score’, as the integral of the number and severity of GI complaints, did not correlate with ΔI-FABP (rs: –0.050, P = 0.790) or energy intake (rs: 0.211, P = 0.260). However, there was a significant negative correlation between energy intake and ΔI-FABP (rs: –0.388, P = 0.031). In conclusion, GI complaints were neither associated with food intake nor GI injury as assessed by plasma I-FABP response. Energy intake, however, was inversely related to the I-FABP response to exercise. This finding suggests that substantial energy intakes during exercise may prevent exercise-induced GI injury as assessed by the I-FABP response. Novelty: No association between gastrointestinal complaints and gastrointestinal injury (I-FABP response) or food intake was present. There was an inverse correlation between energy intake and plasma I-FABP response, suggesting that higher energy intakes may prevent gastrointestinal injury as assessed by the I-FABP response.

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.

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.

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

While the majority of studies reporting ergogenic effects of dietary nitrate have used a multiday supplementation protocol, some studies suggest that a single dose of dietary nitrate before exercise can also improve subsequent performance. We aimed to compare the impact of acute and 6-day sodium nitrate supplementation on oxygen uptake (V̇O2) and time-trial performance in trained cyclists. Using a randomized, double-blind, cross-over design, 17 male cyclists (25 ± 4 y, V̇O2peak 65 ± 4 ml·kg-1·min-1, Wmax 411 ± 35 W) were subjected to 3 different trials; 5 days placebo and 1 day sodium nitrate supplementation (1-DAY); 6 days sodium nitrate supplementation (6-DAY); 6 days placebo supplementation (PLA). Nitrate was administered as 1097 mg sodium nitrate providing 800 mg (~12.9 mmol) nitrate per day. Three hours after ingestion of the last supplemental bolus, indirect calorimetry was performed while subjects performed 30 min of exercise at 45% Wmax and 30 min at 65% Wmax on a cycle ergometer, followed by a 10 km time-trial. Immediately before exercise, plasma [nitrate] and [nitrite] increased to a similar extent during the 6-DAY and 1-DAY trial, but not with PLA (plasma nitrite: 501 ± 205, 553 ± 278, and 239 ± 74 nM, respectively; p < .001). No differences were observed between interventions in V̇O2 during submaximal exercise, or in time to complete the time-trial (6-DAY: 1004 ± 61, 1-DAY: 1022 ± 72, PLA: 1017 ± 71 s; p = .28). We conclude that both acute and 6-days of sodium nitrate supplementation do not alter V̇O2 during submaximal exercise or improve time-trial performance in highly trained cyclists, despite increasing plasma [nitrate] and [nitrite].

Dietary protein intake should be optimized in all athletes to ensure proper recovery and enhance the skeletal muscle adaptive response to exercise training. In addition to total protein intake, the use of specific proteincontaining food sources and the distribution of protein throughout the day are relevant for optimizing protein intake in athletes. In the present study, we examined the daily intake and distribution of various proteincontaining food sources in a large cohort of strength, endurance and team-sport athletes. Well-trained male (n=327) and female (n=226) athletes completed multiple web-based 24-hr dietary recalls over a 2-4 wk period. Total energy intake, the contribution of animal- and plant-based proteins to daily protein intake, and protein intake at six eating moments were determined. Daily protein intake averaged 108±33 and 90±24 g in men and women, respectively, which corresponded to relative intakes of 1.5±0.4 and 1.4±0.4 g/kg. Dietary protein intake was correlated with total energy intake in strength (r=0.71, p r=0.79, p r=0.77, p 1.2 g protein/kg/d, but the distribution throughout the day may be suboptimal to maximize the skeletal muscle adaptive response to training.

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