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MicroRNAs (miRNAs) regulate molecular processes governing muscle metabolism. Physical activity and energy balance influence both muscle anabolism and substrate metabolism, but whether circulating and skeletal muscle miRNAs mediate those effects remains unknown. This study assessed the impact of sustained physical activity with participants in energy balance (BAL) or deficit (DEF) on circulating and skeletal muscle miRNAs. Using a randomized cross-over design, 10 recreational active healthy males (mean ± SD, 22 ± 5 years, 87 ± 11 kg) completed 72 h of high aerobic exercise-induced energy expenditures in BAL (689 ± 852 kcal/day) or DEF (-2047 ± 920 kcal/day). Blood and muscle samples were collected under rested/fasted conditions before (PRE) and immediately after 120 min load carriage exercise bout at the end (POST) of the 72 h. Trials were separated by 7 days. Circulating and skeletal muscle miRNAs were measured using microarray RT-qPCR. Independent of energy status, 36 circulating miRNAs decreased (P 0.05), while 10 miRNAs increased and three miRNAs decreased in skeletal muscle (P 0.05) at POST compared to PRE. Of these, miR-122-5p, miR-221-3p, miR-222-3p and miR-24-3p decreased in circulation and increased in skeletal muscle. Two circulating (miR-145-5p and miR-193a-5p) and four skeletal muscle (miR-21-5p, miR-372-3p, miR-34a-5p and miR-9-5p) miRNAs had time-by-treatment effects (P 0.05). These data suggest that changes in miRNA profiles are more sensitive to increased physical activity compared to energy status, and that changes in circulating miRNAs in response to high levels of daily aerobic exercise are not reflective of changes in skeletal muscle miRNAs. KEY POINTS: Circulating and skeletal muscle miRNA profiles are more sensitive to high levels of aerobic exercise-induced energy expenditure compared to energy status. Changes in circulating miRNA in response to high levels of daily sustained aerobic exercise are not reflective of changes in skeletal muscle miRNA.

The impact of gut microbiota–targeted interventions on the incidence, duration, and severity of respiratory tract infections (RTIs) in nonelderly adults, and factors moderating any such effects, are unclear. This systematic review and meta-analysis aimed to determine the effects of orally ingested probiotics, prebiotics, and synbiotics compared with placebo on RTI incidence, duration, and severity in nonelderly adults, and to identify potential sources of heterogeneity. Studies were identified by searching CENTRAL, PubMed, Scopus, and Web of Science up to December 2021. English-language, peer-reviewed publications of randomized, placebo-controlled studies that tested an orally ingested probiotic, prebiotic, or synbiotic intervention of any dose for ≥1 wk in adults aged 18–65 y were included. Results were synthesized using intention-to-treat and per-protocol random-effects meta-analysis. Heterogeneity was explored by subgroup meta-analysis and meta-regression. Risk of bias was assessed using the Cochrane risk-of-bias assessment tool for randomized trials version 2 (RoB2). Forty-two manuscripts reporting effects of probiotics (n = 38), prebiotics (n = 2), synbiotics (n = 1) or multiple -biotic types (n = 1) were identified (n = 9179 subjects). Probiotics reduced the risk of experiencing ≥1 RTI (relative risk = 0.91; 95% CI: 0.84, 0.98; P = 0.01), and total days (rate ratio = 0.77; 95% CI: 0.71, 0.83; P

Supplemental digital content is available in the text., Introduction/Purpose Evidence suggests that carbohydrate and protein (CHO-PRO) ingestion after exercise enhances muscle glycogen repletion to a greater extent than carbohydrate (CHO) alone. However, there is no consensus at this point, and results across studies are mixed, which may be attributable to differences in energy content and carbohydrate intake relative to body mass consumed after exercise. The purpose of this study was determine the overall effects of CHO-PRO and the independent effects of energy and relative carbohydrate content of CHO-PRO supplementation on postexercise muscle glycogen synthesis compared with CHO alone. Methods Meta-analysis was conducted on crossover studies assessing the influence of CHO-PRO compared with CHO alone on postexercise muscle glycogen synthesis. Studies were identified in a systematic review from PubMed and Cochrane Library databases. Data are presented as effect size (95% confidence interval [CI]) using Hedges’ g. Subgroup analyses were conducted to evaluate effects of isocaloric and nonisocaloric energy content and dichotomized by median relative carbohydrate (high, ≥0.8 g·kg−1⋅h−1; low

Sustained operations (SUSOPS) require military personnel to conduct combat and training operations while experiencing physical and cognitive stress and limited sleep. These operations are often conducted in a state of negative energy balance and are associated with degraded cognitive performance and mood. Whether maintaining energy balance can mitigate these declines is unclear. This randomized crossover study assessed the effects of energy balance on cognitive performance, risk-taking propensity, ambulatory vigilance, and mood during a simulated 72-h SUSOPS.Ten male Soldiers (mean ± SE; 22.4 ± 1.7 y; body weight 87.3 ± 1.1 kg) completed two, 72-h simulated SUSOPS in random order, separated by 7 days of recovery. Each SUSOPS elicited ∼4500 kcal/d total energy expenditure and restricted sleep to 4 h/night. During SUSOPS, participants consumed either an energy-balanced or restricted diet that induced a 43 ± 3% energy deficit. A cognitive test battery was administered each morning and evening to assess: vigilance, working memory, grammatical reasoning, risk-taking propensity, and mood. Real-time ambulatory vigilance was assessed each morning, evening, and night via a wrist-worn monitoring device.Participants exhibited heightened risk-taking propensity (p = 0.047) with lower self-reported self-control (p = 0.021) and fatigue (p = 0.013) during energy deficit compared to during energy balance. Vigilance accuracy (p 0.001) and working memory (p = 0.040) performance decreased, and vigilance lapses increased (p 0.001) during SUSOPS, but did not differ by diet. Percentage of correct responses to ambulatory vigilance stimuli varied during SUSOPS (p = 0.019) independent of diet, with generally poorer performance during the morning and night. Total mood disturbance (p = 0.001), fatigue (p 0.001), tension (p = 0.003), and confusion (p = 0.036) increased whereas vigor decreased (p 0.001) during SUSOPS, independent of diet.Prolonged physical activity combined with sleep restriction is associated with impaired vigilance, memory, and mood state. Under such conditions, maintaining energy balance prevents increased risk-taking and improves self-control, but does not improve other aspects of cognitive function or mood. Given the small sample in the present study, replication in a larger cohort is warranted.

OBJECTIVES: Secretory IgA (SIgA) is a critical component of mucosal immunity and a first line of defense against pathogens. Intense physical exercise, lack of sleep, and inadequate energy intake are frequently observed during military training and operations. These factors are associated with a decline in SIgA and may increase the risk of infection; however, to what degree each of these factors contributes to immune dysfunction is unclear. This study aimed to determine the effect of severe energy deficit on mucosal immunity (SIgA) during a multi-day period of intense training. METHODS: The parent study was a randomized, crossover trial in healthy males (n = 10, 22.4 ± 5.4 y, 87.3 ± 10.9 kg) to assess the effect of severe negative energy balance on inflammation, iron absorption, and other physiological and cognitive outcomes during a simulated sustained military operation (SUSOPS; high energy expenditure with repeated bouts of intense exercise). Participants completed two SUSOPS trials and were randomized to consume ± 10% of estimated total daily energy expenditure (TDEE, energy balance) or 45% of TDEE (energy deficit). At 0500 on each SUSOPS day (D1: baseline, D2:24 h, D3:48 h), participants placed polyester oral swabs under their tongue for 3-mins. A second swab was collected (i.e., placed under the tongue until saturation) to ensure adequate sample volume. SIgA secretion rate (μg/min) was calculated from SIgA concentration (μg/mL; enzyme-linked immunosorbent assay) and salivary flow rate (mL/min). Dependent variables were log10 transformed due to non-normal distribution and data were analyzed using linear mixed models. RESULTS: Independent of treatment, a main effect of time (P = 0.01) was observed where SIgA secretion rate declined by 20% from D2 [1.77 ± 0.34 μg/min] to D3 [1.41 ± 0.51 μg/min], P = 0.001, with no significant treatment by time interactions. A main effect of time (P = 0.01) was also found wherein SIgA concentration declined by 13% from D2 [2.67 ± 0.32 μg/mL] to D3 [2.33 ± 0.37 μg/mL], P = 0.001. There were no main or treatment effects with regard to SIgA flow rate. CONCLUSIONS: Mucosal immune response, as measured by SIgA, declined in response to SUSOPS. Severe energy deficit did not exacerbate the decline in SIgA secretion rate observed in response to the high intensity, multi-stressor training scenario. FUNDING SOURCES: US Army Medical Research and Development Command.

Energy deficit is common during prolonged periods of strenuous physical activity and limited sleep, but the extent to which appetite suppression contributes is unclear. The aim of this randomised crossover study was to determine the effects of energy balance on appetite and physiological mediators of appetite during a 72-h period of high physical activity energy expenditure (about 9·6 MJ/d (2300 kcal/d)) and limited sleep designed to simulate military operations (SUSOPS). Ten men consumed an energy-balanced diet while sedentary for 1 d (REST) followed by energy-balanced (BAL) and energy-deficient (DEF) controlled diets during SUSOPS. Appetite ratings, gastric emptying time (GET) and appetite-mediating hormone concentrations were measured. Energy balance was positive during BAL (18 (sd 20) %) and negative during DEF (–43 (sd 9) %). Relative to REST, hunger, desire to eat and prospective consumption ratings were all higher during DEF (26 (sd 40) %, 56 (sd 71) %, 28 (sd 34) %, respectively) and lower during BAL (–55 (sd 25) %, −52 (sd 27) %, −54 (sd 21) %, respectively; Pcondition < 0·05). Fullness ratings did not differ from REST during DEF, but were 65 (sd 61) % higher during BAL (Pcondition < 0·05). Regression analyses predicted hunger and prospective consumption would be reduced and fullness increased if energy balance was maintained during SUSOPS, and energy deficits of ≥25 % would be required to elicit increases in appetite. Between-condition differences in GET and appetite-mediating hormones identified slowed gastric emptying, increased anorexigenic hormone concentrations and decreased fasting acylated ghrelin concentrations as potential mechanisms of appetite suppression. Findings suggest that physiological responses that suppress appetite may deter energy balance from being achieved during prolonged periods of strenuous activity and limited sleep.

OBJECTIVES: Iron status declines with military training; however, the reason for the decline is not known. The objective of this study was to determine whether dietary iron absorption is reduced following military training and whether energy deficit during training modifies the effect. METHODS: This was a randomized, cross-over, controlled-feeding trial in healthy, active duty military males (n = 10, age 22.4 ± 5.4 y, weight 87.3 ± 10.9 kg) with normal iron status (serum ferritin 77.0 ± 36.7 ng/mL). Following a rest day (no exercise), participants completed a 72-h simulated sustained military operations (SUSOPS) followed by a 7-d recovery period. SUSOPS was comprised of military tasks designed to elicit high energy expenditures, muscle damage, and sleep deprivation. During SUSOPS, participants were randomized to consume ± 10% of estimated total daily energy expenditure (BAL) or 45% of total daily energy expenditure to induce severe negative energy balance (NEG BAL), but an equivalent amount of dietary iron. Two hours after rest, BAL, and NEG BAL participants consumed a beverage containing 3.8 mg of a stable iron isotope and plasma isotope appearance and hepcidin were determined 0, 20, 40, 60, 120, 240, and 360 min later. RESULTS: BAL maintained weight (−0.03 ± 0.8 kg) and muscle glycogen (4.1 ± 68.1% change), while NEG BAL lost weight (−2.38 ± 1.7 kg, P

BACKGROUND Strenuous physical activity promotes inflammation and depletes muscle glycogen, which may increase the iron regulatory hormone hepcidin. Hepcidin reduces dietary iron absorption and may contribute to declines in iron status frequently observed following strenuous physical activity. OBJECTIVES To determine the effects of strenuous physical activity on hepcidin and dietary iron absorption and whether energy deficit compared with energy balance modifies those effects. METHODS This was a randomized, cross-over, controlled-feeding trial in healthy male subjects (n = 10, mean ± SD age: 22.4 ± 5.4 y, weight: 87.3 ± 10.9 kg) with sufficient iron status (serum ferritin 77.0 ± 36.7 ng/mL). Rest measurements were collected before participants began a 72-h simulated sustained military operation (SUSOPS), designed to elicit high energy expenditure, glycogen depletion, and inflammation, followed by a 7-d recovery period. Two 72-h SUSOPS trials were performed where participants were randomly assigned to consume either energy matched (±10%) to their individual estimated total daily energy expenditure (BAL) or energy at 45% of total daily energy expenditure to induce energy deficit (DEF). On the rest day and at the completion of BAL and DEF, participants consumed a beverage containing 3.8 mg of a stable iron isotope, and plasma isotope appearance was measured over 6 h. RESULTS Muscle glycogen declined during DEF and was preserved during BAL (-188 ± 179 mmol/kg, P-adjusted