The metabolic recovery of marathon runners: an untargeted 1H-NMR metabolomics perspective.

Introduction: Extreme endurance events may result in numerous adverse metabolic, immunologic, and physiological perturbations that may diminish athletic performance and adversely affect the overall health status of an athlete, especially in the absence of sufficient recovery. A comprehensive understanding of the post-marathon recovering metabolome, may aid in the identification of new biomarkers associated with marathon-induced stress, recovery, and adaptation, which can facilitate the development of improved training and recovery programs and personalized monitoring of athletic health/recovery/performance. Nevertheless, an untargeted, multi-disciplinary elucidation of the complex underlying biochemical mechanisms involved in recovery after such an endurance event is yet to be demonstrated. Methods: This investigation employed an untargeted proton nuclear magnetic resonance metabolomics approach to characterize the post-marathon recovering metabolome by systematically comparing the pre-, immediately post, 24, and 48 h post-marathon serum metabolite profiles of 15 athletes. Results and Discussion: A total of 26 metabolites were identified to fluctuate significantly among post-marathon and recovery time points and were mainly attributed to the recovery of adenosine triphosphate, redox balance and glycogen stores, amino acid oxidation, changes to gut microbiota, and energy drink consumption during the post-marathon recovery phase. Additionally, metabolites associated with delayed-onset muscle soreness were observed; however, the mechanisms underlying this commonly reported phenomenon remain to be elucidated. Although complete metabolic recovery of the energy-producing pathways and fuel substrate stores was attained within the 48 h recovery period, several metabolites remained perturbed throughout the 48 h recovery period and/or fluctuated again following their initial recovery to pre-marathon-related levels.

Polyphenol-rich tart cherries (Prunus Cerasus, cv Montmorency) improve sustained attention, feelings of alertness and mental fatigue and influence the plasma metabolome in middle-aged adults: a randomised, placebo-controlled trial.

Tart Montmorency cherries (MC) are a particularly rich source of anthocyanins and other polyphenols that have been shown to elicit antioxidant, anti-inflammatory and vasomodulatory actions. The current study aimed to determine the influence of chronic MC supplementation on cognitive function and mood. In a 3-month double-blinded, placebo-controlled parallel study, middle-aged adults (mean ± sd: 48 ± 6 years) were randomly assigned to either 30 ml twice daily of MC (n 25) or the same amount of an isoenergetic placebo (n 25). Cognitive function and mood were assessed before and after supplementation using a computerised cognitive task battery and visual analogue scales. Cerebral blood flow was also monitored by near-infrared spectroscopy during the task battery, and questionnaires were administered to determine subjective sleep and health status and plasma metabolomics were analysed before and after supplementation. After 3 months, the MC resulted in higher accuracy in digit vigilance (mean difference: 3·3, 95 % CI: 0·2, 6·4 %) with lower number of false alarms (mean difference: -1·2, 95 % CI: -2·0, -0·4) compared with the placebo. There was also a treatment effect for higher alertness (mean difference: 5·9, 95 % CI: 1·3, 10·5 %) and lower mental fatigue ratings (mean difference -9·5, 95 % CI: -16·5, -2·5 %) with MC. Plasma metabolomics revealed an increase in a number of amino acids in response to MC intake, but not placebo. These data suggest an anti-fatiguing effect of MC supplementation as well as the ability to improve sustained attention during times of high cognitive demand, this could be related to changes in amino acid metabolism.

Plasma uptake of selected phenolic acids following New Zealand blackcurrant extract supplementation in humans.

New Zealand blackcurrant (NZBC) extract is a rich source of anthocyanins and in order to exert physiological effects, the anthocyanin-derived metabolites need to be bioavailable in vivo. We examined the plasma uptake of selected phenolic acids following NZBC extract supplementation alongside maintaining a habitual diet (i.e. not restricting habitual polyphenol intake). Twenty healthy volunteers (nine females, age: 28 ± 7 years, height 1.73 ± 0.09 m, body mass 73 ± 11 kg) consumed a 300 mg NZBC extract capsule (CurraNZ®; anthocyanin content 105 mg) following an overnight fast. Venous blood samples were taken pre and 1, 1.5, 2, 3, 4, 5, and 6 h post-ingestion of the capsule. Reversed-phase high-performance liquid chromatography (HPLC) was used for analysis of two dihydroxybenzoic acids [i.e. vanillic acid (VA) and protocatechuic acid (PCA)] and one trihydroxybenzoic acid [i.e. gallic acid (GA)] in plasma following NZBC extract supplementation. Habitual anthocyanin intake was 168 (95%CI:68-404) mg⋅day-1 and no associations were observed between this and VA, PCA, and GA plasma uptake by the NZBC extract intake. Plasma time-concentration curves revealed that GA, and PCA were most abundant at 4, and 1.5 h post-ingestion, representing a 261% and 320% increase above baseline, respectively, with VA remaining unchanged. This is the first study to demonstrate that an NZBC extract supplement increases the plasma uptake of phenolic acids GA, and PCA even when a habitual diet is followed in the days preceding the experimental trial, although inter-individual variability is apparent.

Methodological Considerations for a Vascular Function Test Battery.

There is a dearth of information regarding the reliability of non-invasive measures of vascular function taken in a single testing session. This study aimed to determine the test-retest reliability of a test battery of vascular function measures: automated blood pressure (BP), laser Doppler imaging with iontophoresis (LDI), digital volume pulse (DVP), pulse wave velocity (PWV), augmentation index (AIx) measured by pulse wave analysis (PWA) and flow-mediated dilation (FMD) taken within and between sessions. Measures were taken in 21 non-smoking males intra-session and again inter-session (one week apart) to determine repeatability and reproducibility, respectively. There was moderate to excellent repeatability (ICC: 0.53-0.93; CV=2.2-18.1%) and reproducibility (ICC: 0.71-0.96; CV 1.9-14.2%) for BP, DVP stiffness index, PWV, AIx, AIx normalised to heart rate (75 bpm), absolute and percentage FMD. Repeatability of the DVP reflection index was moderate (ICC: 0.64; CV=9.5%) but there was poor reproducibility (ICC: 0.17; CV=15.1%). Moreover, the repeatability and reproducibility of the LDI measures ranged from poor to good (ICC: 0.31-0.84; CV=28.4-36.7%). These data indicated that there was considerable variability in the repeatability and reproducibility of measurements of endothelial function and arterial stiffness taken in a battery of measurements, which needs careful consideration in future research designs.

Minimal muscle damage after a marathon and no influence of beetroot juice on inflammation and recovery.

This study examined whether beetroot juice (BTJ) would attenuate inflammation and muscle damage following a marathon. Using a double blind, independent group design, 34 runners (each having completed ca. ∼16 previous marathons) consumed either BTJ or an isocaloric placebo (PLA) for 3 days following a marathon. Maximal isometric voluntary contractions (MIVC), countermovement jumps (CMJ), muscle soreness, serum cytokines, leucocytosis, creatine kinase (CK), high sensitivity C-reactive protein (hs-CRP), and aspartate aminotransferase (AST) were measured pre, post, and 2 days after the marathon. CMJ and MIVC were reduced after the marathon (P < 0.05), but no group differences were observed (P > 0.05). Muscle soreness was increased in the day after the marathon (BTJ; 45 ± 48 vs. PLA; 46 ± 39 mm) and had returned to baseline by day 2, irrespective of supplementation (P = 0.694). Cytokines (interleukin-6; IL-6, interleukin-8, tumour necrosis factor-α) were increased immediately post-marathon but apart from IL-6 had returned to baseline values by day 1 post. No interaction effects were evident for IL-6 (P = 0.213). Leucocytes increased 1.7-fold after the race and remained elevated 2 days post, irrespective of supplement (P < 0.0001). CK peaked at 1 day post marathon (BTJ: 965 ± 967, and PLA: 1141 ± 979 IU·L-1) and like AST and hs-CRP, was still elevated 2 days after the marathon (P < 0.05); however, no group differences were present for these variables. Beetroot juice did not attenuate inflammation or reduce muscle damage following a marathon, possibly because most of these indices were not markedly different from baseline values in the days after the marathon.

The plasma bioavailability of nitrate and betanin from Beta vulgaris rubra in humans.

PURPOSE: To evaluate the plasma bioavailability of betanin and nitric oxide (NOx) after consuming beetroot juice (BTJ) and whole beetroot (BF). BTJ and BF were also analysed for antioxidant capacity, polyphenol content (TPC) and betalain content. METHODS: Ten healthy males consumed either 250 ml of BTJ, 300 g of BF or a placebo drink, in a randomised, crossover design. Venous plasma samples were collected pre (baseline), 1, 2, 3, 5 and 8 h post-ingestion. Betanin content in BTJ, BF and plasma was analysed with reverse-phase high-performance liquid chromatography (HPLC) and mass spectrometry detection (LCMS). Antioxidant capacity was estimated using the Trolox equivalent antioxidant capacity (TEAC) and polyphenol content using Folin-Ciocalteu colorimetric methods [gallic acid equivalents (GAE)] and betalain content spectrophotometrically. RESULTS: TEAC was 11.4 ± 0.2 mmol/L for BTJ and 3.4 ± 0.4 µmol/g for BF. Both BTJ and BF contained a number of polyphenols (1606.9 ± 151 mg/GAE/L and 1.67 ± 0.1 mg/GAE/g, respectively), betacyanins (68.2 ± 0.4 mg/betanin equivalents/L and 19.6 ± 0.6 mg/betanin equivalents/100 g, respectively) and betaxanthins (41.7 ± 0.7 mg/indicaxanthin equivalents/L and 7.5 ± 0.2 mg/indicaxanthin equivalents/100 g, respectively). Despite high betanin contents in both BTJ (~194 mg) and BF (~66 mg), betanin could not be detected in the plasma at any time point post-ingestion. Plasma NOx was elevated above baseline for 8 h after consuming BTJ and 5 h after BF (P < 0.05). CONCLUSIONS: These data reveal that BTJ and BF are rich in phytonutrients and may provide a useful means of increasing plasma NOx bioavailability. However, betanin, the major betalain in beetroot, showed poor bioavailability in plasma.

Effects of Montmorency tart cherry (Prunus Cerasus L.) consumption on vascular function in men with early hypertension.

BACKGROUND: Tart cherries contain numerous polyphenolic compounds that could potentially improve endothelial function and reduce cardiovascular disease risk. OBJECTIVE: We sought to examine the acute effects of Montmorency tart cherry (MC) juice on vascular function in subjects with early hypertension. DESIGN: A placebo-controlled, blinded, crossover, randomized Latin square design study with a washout period of ≥14 d was conducted. Fifteen men with early hypertension [systolic blood pressure (SBP) ≥130 mm Hg, diastolic blood pressure ≥80 mm Hg, or both] received either a 60-mL dose of MC concentrate or placebo. Microvascular reactivity (laser Doppler imaging with iontophoresis), arterial stiffness (pulse wave velocity and analysis), blood pressure, and phenolic acid absorption were assessed at baseline and at 1, 2, 3, 5, and 8 h postconsumption. RESULTS: MC consumption significantly lowered SBP (P < 0.05) over a period of 3 h, with peak reductions of mean ± SEM 7 ± 3 mm Hg 2 h after MC consumption relative to the placebo. Improvements in cardiovascular disease risk factors were closely linked to increases in circulating protocatechuic and vanillic acid at 1-2 h. CONCLUSIONS: MC intake acutely reduces SBP in men with early hypertension. These benefits may be mechanistically linked to the actions of circulating phenolic acids. This study provides information on a new application of MCs in health maintenance, particularly in positively modulating SBP. This trial was registered at clinicaltrials.gov as NCT02234648.

Phytochemical uptake following human consumption of Montmorency tart cherry (L. Prunus cerasus) and influence of phenolic acids on vascular smooth muscle cells in vitro.

PURPOSE: To investigate the phytochemical uptake following human consumption of Montmorency tart cherry (L. Prunus cerasus) and influence of selected phenolic acids on vascular smooth muscle cells in vitro. METHODS: In a randomised, double-blinded, crossover design, 12 healthy males consumed either 30 or 60 mL of Montmorency tart cherry concentrate. Following analysis of the juice composition, venous blood samples were taken before and 1, 2, 3, 5 and 8 h post-consumption of the beverage. In addition to examining some aspects of the concentrate contents, plasma concentrations of protocatechuic acid (PCA), vanillic acid (VA) and chlorogenic (CHL) acid were analysed by reversed-phase high-performance liquid chromatography (HPLC) with diode array for quantitation and mass spectrometry detection (LCMS) for qualitative purposes. Vascular smooth muscle cell migration and proliferation were also assessed in vitro. RESULTS: Both the 30 and 60 mL doses of Montmorency cherry concentrate contained high amounts of total phenolics (71.37 ± 0.11; 142.73 ± 0.22 mg/L) and total anthocyanins (62.47 ± 0.31; 31.24 ± 0.16 mg/L), as well as large quantities of CHL (0.205 ± 0.24; 0.410 ± 0.48 mg/L) and VA (0.253 ± 0.84; 0.506 ± 1.68 mg/L). HPLC/LCMS identified two dihydroxybenzoic acids (PCA and VA) in plasma following MC concentrate consumption. Both compounds were most abundant 1-2 h post-initial ingestion with traces detectable at 8 h post-ingestion. Cell migration was significantly influenced by the combination of PCA and VA, but not in isolation. There was no effect of the compounds on cell proliferation. CONCLUSIONS: These data show new information that phenolic compounds thought to exert vasoactive properties are bioavailable in vivo following MC consumption and subsequently can influence cell behaviour. These data may be useful for the design and interpretation of intervention studies investigating the health effects of Montmorency cherries.