Comparative analysis of circulating metabolomic profiles identifies shared metabolic alterations across distinct multistressor military training exercises.

Military training provides insight into metabolic responses under unique physiological demands that can be comprehensively characterized by global metabolomic profiling to identify potential strategies for improving performance. This study identified shared changes in metabolomic profiles across three distinct military training exercises, varying in magnitude and type of stress. Blood samples collected before and after three real or simulated military training exercises were analyzed using the same untargeted metabolomic profiling platform. Exercises included a 2-wk survival training course (ST, n = 36), a 4-day cross-country ski march arctic training (AT, n = 24), and a 28-day controlled diet- and exercise-induced energy deficit (CED, n = 26). Log2-fold changes of greater than ±1 in 191, 121, and 64 metabolites were identified in the ST, AT, and CED datasets, respectively. Most metabolite changes were within the lipid (57-63%) and amino acid metabolism (18-19%) pathways and changes in 87 were shared across studies. The largest and most consistent increases in shared metabolites were found in the acylcarnitine, fatty acid, ketone, and glutathione metabolism pathways, whereas the largest decreases were in the diacylglycerol and urea cycle metabolism pathways. Multiple shared metabolites were consistently correlated with biomarkers of inflammation, tissue damage, and anabolic hormones across studies. These three studies of real and simulated military training revealed overlapping alterations in metabolomic profiles despite differences in environment and the stressors involved. Consistent changes in metabolites related to lipid metabolism, ketogenesis, and oxidative stress suggest a potential common metabolomic signature associated with inflammation, tissue damage, and suppression of anabolic signaling that may characterize the unique physiological demands of military training.NEW & NOTEWORTHY The extent to which metabolomic responses are shared across diverse military training environments is unknown. Global metabolomic profiling across three distinct military training exercises identified shared metabolic responses with the largest changes observed for metabolites related to fatty acids, acylcarnitines, ketone metabolism, and oxidative stress. These changes also correlated with alterations in markers of tissue damage, inflammation, and anabolic signaling and comprise a potential common metabolomic signature underlying the unique physiological demands of military training.

Independent effects of volume and energy density manipulation on energy intake and appetite in healthy adults: A randomized, controlled, crossover study.

Consuming enough energy to meet high energy demands can be challenging for military personnel wherein logistical constraints limit food availability. Increasing dietary energy density (ED) and/or volume density (VD) of rations may be countermeasures, but whether positive linear associations between ED and energy intake (EI) hold at moderate-to-high ED and VD is unclear. This study examined the effects of covertly increasing the ED and VD of moderate ED (≥1.6 kcal/g) foods on appetite and energy intake. Twenty healthy men completed four 2-day treatments in random order by consuming a standardized diet containing three experimental food items (EXP) engineered using leavening, physical compression and fat manipulation to be isovolumetric but lower (L) or higher (H) in ED and VD creating four treatments: LED/LVD, LED/HVD, HED/LVD, HED/HVD. Consumption of EXP was compulsory during two meals and a snack, but remaining intake was self-selected (SSF). Results failed to show any ED-by-VD interactions. During LVD, EI was lower for EXP (-417 kcal [95%CI: 432, -402], p < 0.01) and TOTAL (SSF + EXP) (-276 kcal [95%CI: 470, -83], p = 0.01) compared to HVD, while SSF EI did not differ (140 kcal [-51, 332], p = 0.15). During LED, EI for EXP (-291 kcal [95%CI: 306, -276], p < 0.01) was lower than HED, while SSF EI was higher than HED (203 kcal 95%CI: [12, 394], p = 0.04) and TOTAL EI did not differ (-88 kcal [-282, 105], p = 0.36). Thus, when a small isovolumetric portion of the diet was manipulated, increasing the VD of moderate ED foods failed to elicit compensatory reductions in ad libitum EI while increasing the ED of moderate ED foods did. Findings may support VD manipulation of moderate ED foods as a strategy to promote increased short-term EI in environments wherein logistical burden may limit food volume.

Sex differences in body composition and serum metabolome responses to sustained, physical training suggest enhanced fat oxidation in women compared with men.

Sex differences in energy metabolism during acute, submaximal exercise are well documented. Whether these sex differences influence metabolic and physiological responses to sustained, physically demanding activities is not well characterized. This study aimed to identify sex differences within changes in the serum metabolome in relation to changes in body composition, physical performance, and circulating markers of endocrine and metabolic status during a 17-day military training exercise. Blood was collected, and body composition and lower body power were measured before and after the training on 72 cadets (18 women). Total daily energy expenditure (TDEE) was assessed using doubly labeled water in a subset throughout. TDEE was greater in men (4,085 ± 482 kcal/d) than in women (2,982 ± 472 kcal/d, P < 0.001), but not after adjustment for dry lean mass (DLM). Men tended to lose more DLM than women (mean change [95% CI]: -0.2[-0.3, -0.1] vs. -0.0[-0.0, 0.0] kg, P = 0.063, Cohen's d = 0.50) and have greater reductions in lower body power (-244[-314, -174] vs. -130[-209, -51] W, P = 0.085, d = 0.49). Reductions in DLM and lower body power were correlated (r = 0.325, P = 0.006). Women demonstrated greater fat oxidation than men (Δfat mass/DLM: -0.20[-0.24, -0.17] vs. -0.15[-0.17, -0.13] kg, P = 0.012, d = 0.64). Metabolites within pathways of fatty acid, endocannabinoid, lysophospholipid, phosphatidylcholine, phosphatidylethanolamine, and plasmalogen metabolism increased in women relative to men. Independent of sex, changes in metabolites related to lipid metabolism were inversely associated with changes in body mass and positively associated with changes in endocrine and metabolic status. These data suggest that during sustained military training, women preferentially mobilize fat stores compared with men, which may be beneficial for mitigating loss of lean mass and lower body power.NEW & NOTEWORTHY Women preferentially mobilize fat stores compared with men in response to sustained, physically demanding military training, as evidenced by increased lipid metabolites and enhanced fat oxidation, which may be beneficial for mitigating loss of lean mass and lower body power.

In vitro gut microbiome response to carbohydrate supplementation is acutely affected by a sudden change in diet.

BACKGROUND: Interactions between diet, stress and the gut microbiome are of interest as a means to modulate health and performance. Here, in vitro fermentation was used to explore the effects of a sudden change in diet, 21 days sole sustenance on the Meal, Ready-to-Eat (MRE) U.S. military combat ration, on inter-species competition and functional potential of the human gut microbiota. Human fecal samples collected before and after MRE intervention or consuming a habitual diet (HAB) were introduced to nutrient-rich media supplemented with starch for in vitro fermentation under ascending colon conditions. 16S rRNA amplicon and Whole-metagenome sequencing (WMS) were used to measure community composition and functional potential. Specific statistical analyses were implemented to detect changes in relative abundance from taxa, genes and pathways. RESULTS: Differential changes in relative abundance of 11 taxa, Dorea, Lachnospira, Bacteroides fragilis, Akkermansia muciniphila, Bifidobacterium adolescentis, Betaproteobacteria, Enterobacteriaceae, Bacteroides egerthii, Ruminococcus bromii, Prevotella, and Slackia, and nine Carbohydrate-Active Enzymes, specifically GH13_14, over the 24 h fermentation were observed as a function of the diet intervention and correlated to specific taxa of interest. CONCLUSIONS: These findings suggest that consuming MRE for 21 days acutely effects changes in gut microbiota structure in response to carbohydrate but may induce alterations in metabolic capacity. Additionally, these findings demonstrate the potential of starch as a candidate supplemental strategy to functionally modulate specific gut commensals during stress-induced states.

Biomarkers of oxidative stress, diet and exercise distinguish soldiers selected and non-selected for special forces training.

INTRODUCTION: The metabolomic profiles of Soldiers entering the U.S. Special Forces Assessment and Selection course (SFAS) have not been evaluated. OBJECTIVES: To compare pre-SFAS blood metabolomes of Soldiers selected during SFAS versus those not selected, and explore the relationships between the metabolome, physical performance, and diet quality. METHODS: Fasted blood samples and food frequency questionnaires were collected from 761 Soldiers prior to entering SFAS to assess metabolomic profiles and diet quality, respectively. Physical performance was assessed throughout SFAS. RESULTS: Between-group differences (False Discovery Rate < 0.05) in 108 metabolites were detected. Selected candidates had higher levels of compounds within xenobiotic, pentose phosphate, and corticosteroid metabolic pathways, while non-selected candidates had higher levels of compounds potentially indicative of oxidative stress (i.e., sphingomyelins, acylcarnitines, glutathione, amino acids). Multiple compounds higher in non-selected versus selected candidates included: 1-carboxyethylphenylalanine; 4-hydroxy-nonenal-glutathione; α-hydroxyisocaproate; hexanoylcarnitine; sphingomyelin and were associated with lower diet quality and worse physical performance.  CONCLUSION: Candidates selected during SFAS had higher pre-SFAS levels of circulating metabolites that were associated with resistance to oxidative stress, higher physical performance and higher diet quality. In contrast, non-selected candidates had higher levels of metabolites potentially indicating elevated oxidative stress. These findings indicate that Soldiers who were selected for continued Special Forces training enter the SFAS course with metabolites associated with healthier diets and better physical performance. Additionally, the non-selected candidates had higher levels of metabolites that may indicate elevated oxidative stress, which could result from poor nutrition, non-functional overreaching/overtraining, or incomplete recovery from previous physical activity.

Ketone Monoester Plus Carbohydrate Supplementation Does Not Alter Exogenous and Plasma Glucose Oxidation or Metabolic Clearance Rate During Exercise in Men Compared with Carbohydrate Alone.

BACKGROUND: Increasing ß-hydroxybutyrate (ßHB) availability through ketone monoester (KE) plus carbohydrate supplementation is suggested to enhance physical performance by sparing glucose use during exercise. However, no studies have examined the effect of ketone supplementation on glucose kinetics during exercise. OBJECTIVES: This exploratory study primarily aimed to determine the effect of KE plus carbohydrate supplementation on glucose oxidation during steady-state exercise and physical performance compared with carbohydrate alone. METHODS: Using a randomly assigned, crossover design, 12 men consumed 573 mg KE/kg body mass plus 110 g glucose (KE+CHO) or 110 g glucose (CHO) before and during 90 min of steady-state treadmill exercise [54 ± 3% peak oxygen uptake (V˙O2peak)] wearing a weighted vest (30% body mass; 25 ± 3 kg). Glucose oxidation and turnover were determined using indirect calorimetry and stable isotopes. Participants performed an unweighted time to exhaustion (TTE; 85% V˙O2peak) after steady-state exercise and a weighted (25 ± 3 kg) 6.4 km time trial (TT) the next day after consuming a bolus of KE+CHO or CHO. Data were analyzed by paired t-tests and mixed model ANOVA. RESULTS: ßHB concentrations were higher (P < 0.05) after exercise [2.1 mM (95% CI: 1.6, .6)] and the TT [2.6 mM (2.1, 3.1)] in KE+CHO compared with CHO. TTE was lower [-104 s (-201, -8)], and TT performance was slower [141 s (19,262)] in KE+CHO than in CHO (P < 0.05). Exogenous [-0.01 g/min (-0.07, 0.04)] and plasma [-0.02 g/min (-0.08, 0.04)] glucose oxidation and metabolic clearance rate {MCR [0.38 mg·kg-1·min-1 (-0.79, 1.54)]} were not different, and glucose rate of appearance [-0.51 mg·kg-1·min-1 (-0.97, -0.04)], and disappearance [-0.50 mg·kg-1·min-1 (-0.96, -0.04)] were lower (P < 0.05) in KE+CHO compared with CHO during steady-state exercise. CONCLUSIONS: In the current study, rates of exogenous and plasma glucose oxidation and MCR were not different between treatments during steady-state exercise, suggesting blood glucose utilization is similar between KE+CHO and CHO. KE+CHO supplementation also results in lower physical performance compared with CHO alone. This trial was registered at www. CLINICALTRIALS: gov as NCT04737694.

Circulating and skeletal muscle microRNA profiles are more sensitive to sustained aerobic exercise than energy balance in males.

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.

Metabolomics of testosterone enanthate administration during severe-energy deficit.

INTRODUCTION: Testosterone administration attenuates reductions in total body mass and lean mass during severe energy deficit (SED). OBJECTIVES: This study examined the effects of testosterone administration on the serum metabolome during SED. METHODS: In a double-blind, placebo-controlled clinical trial, non-obese men were randomized to receive 200-mg testosterone enanthate/wk (TEST) (n = 24) or placebo (PLA) (n = 26) during a 28-d inpatient, severe exercise- and diet-induced energy deficit. This study consisted of three consecutive phases. Participants were free-living and provided a eucaloric diet for 14-d during Phase 1. During Phase 2, participants were admitted to an inpatient unit, randomized to receive testosterone or placebo, and underwent SED for 28-d. During Phase 3, participants returned to their pre-study diet and physical activity habits. Untargeted metabolite profiling was conducted on serum samples collected during each phase. Body composition was measured using dual-energy X-ray absorptiometry after 11-d of Phase 1 and after 25-d of Phase 2 to determine changes in fat and lean mass. RESULTS: TEST had higher (Benjamini-Hochberg adjusted, q < 0.05) androgenic steroid and acylcarnitine, and lower (q < 0.05) amino acid metabolites after SED compared to PLA. Metabolomic differences were reversed by Phase 3. Changes in lean mass were associated (Bonferroni-adjusted, p < 0.05) with changes in androgenic steroid metabolites (r = 0.42-0.70), acylcarnitines (r = 0.37-0.44), and amino acid metabolites (r = - 0.36-- 0.37). Changes in fat mass were associated (p < 0.05) with changes in acylcarnitines (r = - 0.46-- 0.49) and changes in urea cycle metabolites (r = 0.60-0.62). CONCLUSION: Testosterone administration altered androgenic steroid, acylcarnitine, and amino acid metabolites, which were associated with changes in body composition during SED.

Multiple Dietary Vitamin K Forms Are Converted to Tissue Menaquinone-4 in Mice.

BACKGROUND: Vitamin K is a term that comprises a family of structurally related quinones, phylloquinone (PK) and the menaquinones (MKn), that share a common naphthoquinone ring but vary in sidechain length (n) and saturation. Dietary PK is a biosynthetic precursor to tissue menaquinone-4 (MK4), but little is known about the absorption and metabolism of dietary MKn. OBJECTIVE: To characterize the absorption and metabolism of dietary MKn relative to PK. METHODS: In the 4-week diet study, 10-week-old male and female C57BL/6 mice were pair-fed a vitamin K deficient diet (control) or a diet supplemented with 5.0 µmol/kg total PK, MK4, and/or MK9 (separately and in combination). In the 1-week stable isotope study, 12-week-old mice were pair-fed diets containing 2.2 µmol/kg PK (unlabeled control), 2H7PK, 13C11MK4, 2H7MK7, or 2H7MK9. Vitamin K tissue content was quantified by HPLC and/or LC-MS, and concentrations were compared by sex and diet group using 2-factor ANOVA. RESULTS: Regardless of the form(s) of vitamin K provided in the diet, tissue MK4 concentrations did not differ across equimolar supplemented groups in the kidney, adipose, reproductive organ, bone, or pancreas in either males or females in the diet study (all P values > 0.05). Isotopic labeling confirmed the naphthoquinone ring of MK4 in tissues originated from the administered dietary PK or MKn. Despite equimolar supplementation, accumulation of the administered dietary form differed across diet groups in small intestinal segments (all P values < 0.002) and the liver (P < 0.001). Female mice had greater total vitamin K than males in every tissue examined (P < 0.05). CONCLUSIONS: Dietary PK, MK4, MK7, and MK9 all served as precursors to tissue MK4 in mice. This study expands our understanding of vitamin K metabolism and supports a common conversion mechanism of all dietary vitamin K forms to MK4. Further investigation of the metabolism and physiological roles of MK4 that may be independent of classical vitamin K function is warranted.

The Fecal Metabolome Links Diet Composition, Foacidic positive ion conditions, chromatographicallyod Processing, and the Gut Microbiota to Gastrointestinal Health in a Randomized Trial of Adults Consuming a Processed Diet.

BACKGROUND: Food processing alters diet digestibility and composition, thereby influencing interactions between host biology, diet, and the gut microbiota. The fecal metabolome offers insight into those relations by providing a readout of diet-microbiota interactions impacting host health. OBJECTIVES: The aims were to determine the effects of consuming a processed diet on the fecal metabolome and to explore relations between changes in the fecal metabolome with fecal microbiota composition and gastrointestinal health markers. METHODS: This was a secondary analysis of a randomized controlled trial wherein healthy adults [94% male; 18-61 y; BMI (kg/m2): 26 ± 3] consumed their usual diet [control (CON), n = 27] or a Meal, Ready-to-EatTM (Ameriqual Packaging) military ration diet composed of processed, shelf-stable, ready-to-eat items for 21 d (MRE; n = 27). Fecal metabolite profiles, fecal microbiota composition, biomarkers of intestinal barrier function, and gastrointestinal symptoms were measured before and after the intervention. Between-group differences and associations were assessed using nonparametric t tests, partial least-squares discriminant analysis, correlation, and redundancy analysis. RESULTS: Fecal concentrations of multiple dipeptides [Mann-Whitney effect size (ES) = 0.27-0.50] and long-chain SFAs (ES = 0.35-0.58) increased, whereas plant-derived compounds (ES = 0.31-0.60) decreased in MRE versus CON (P < 0.05; q < 0.20). Changes in dipeptides correlated positively with changes in fecal concentrations of Maillard-reaction products (ρ = 0.29-0.70; P < 0.05) and inversely with changes in serum prealbumin (ρ = -0.30 to -0.48; P ≤ 0.03). Multiple bile acids, coffee and caffeine metabolites, and plant-derived compounds were associated with both fecal microbiota composition and gastrointestinal health markers, with changes in fecal microbiota composition explaining 26% of the variability within changes in gastrointestinal health-associated fecal metabolites (P = 0.001). CONCLUSIONS: Changes in the fecal metabolomes of adults consuming a Meal, Ready-to-EatTM diet implicate interactions between diet composition, diet digestibility, and the gut microbiota as contributing to variability within gastrointestinal responses to the diet. Findings underscore the need to consider both food processing and nutrient composition when investigating the impact of diet-gut microbiota interactions on health outcomes. This trial was registered at www. CLINICALTRIALS: gov as NCT02423551.

Supplemental Protein and a Multinutrient Beverage Speed Wound Healing after Acute Sleep Restriction in Healthy Adults.

BACKGROUND: Physiologic and psychologic stress slow healing from experimental wounds by impairing immune function. OBJECTIVES: We aimed to determine whether supplemental protein and multinutrient supplementation improved wound healing markers after acute stress induced by acute sleep restriction. METHODS: In this single-blind, crossover study in generally healthy young adults (18 males/2 females; mean ± SD age: 19.7 ± 2.30 y), experimental wounds were created by removing the top layer of forearm blisters induced via suction after 48 h of 72-h sleep restriction (2-h nightly sleep), a protocol previously shown to delay wound healing. Skin barrier restoration (measured by transepidermal water loss) assessed wound healing ≤10 d postblistering, and local immune responses were evaluated by serial measurement of cytokine concentrations in fluid collected at wound sites for 48 h postblistering. Participants consumed controlled, isocaloric diets with either 0.900 g · kg-1 · d-1 protein plus placebo (PLA) or 1.50 g · kg-1 · d-1 protein plus multinutrient beverage [l-arginine: 20.0 g/d; l-glutamine: 30.0 g/d; omega-3 (n-3) fatty acids: 1.00 g/d; zinc sulfate: 24.0 mg/d; cholecalciferol: 800 IU/d; and vitamin C: 400 mg/d] (NUT) during sleep restriction and for 4 d afterwards. RESULTS: Skin barrier restoration (primary outcome) was shorter for NUT (median: 3.98 d; IQR: 1.17 d) than for PLA (median: 5.25 d; IQR: 1.05 d) (P = 0.001). Cytokines from wound fluid (secondary outcome) increased over time (main effect of time P ≤ 0.001), except IL-13 (P = 0.07); however, no effects of treatment were observed. CONCLUSIONS: Supplemental nutrition may promote wound healing after sleep restriction in healthy adults including military personnel, the latter of which also have a high incidence of wounds and infection.This trial was registered at clinicaltrials.gov as NCT03525184.

Weight management behaviours mediate the relationship between weight cycling, BMI and diet quality among US Army Soldiers.

Weight cycling is prevalent in sports/professions with body composition standards, and has been associated with weight management behaviours that may contribute to suboptimal diet quality and weight gain. US Army Soldiers may be at increased risk of weight cycling relative to civilians due to mandated body composition standards. However, the relationship between weight cycling, weight management behaviours, BMI and diet quality among Soldiers is unknown. In this cross-sectional study, 575 Soldiers (89 % enlisted, 90 % male, 23 ± 4 years) at Army installations at Joint Base Elmendorf-Richardson, AK, Joint-Base Lewis McChord, WA, and Fort Campbell, KY completed questionnaires on food frequency, health-related behaviours and history of weight cycling (≥ 3 weight fluctuations ≥ 5 % body weight). Weight cycling was reported by 33 % of Soldiers. Those who reported weight cycling reported higher BMI (27 ± 4 v. 25 ± 3 kg/m2, P < 0·001) and higher prevalence of engaging in weight management behaviours prior to body weight screening but did not report lower dietary quality (Healthy Eating Index-2015 (HEI) scores 59 ± 10 v 59 ± 11, P = 0·46) relative to those who did not report weight cycling. Results of mediation analyses suggested that weight cycling may affect BMI both directly (c' = 1·19, 95 % CI: 0·62, 1·75) and indirectly (ab = 0·45, 95 % CI: 0·19, 0·75), and HEI scores indirectly (ab = 0·69, 95 % CI: 0·20, 1·35) through the adoption of weight management behaviours. Weight cycling is common in Soldiers and is associated with higher BMI and higher prevalence of engaging in weight management behaviours that mediate associations between weight cycling, BMI and diet quality.

Metabolomic profiles are reflective of hypoxia-induced insulin resistance during exercise in healthy young adult males.

Hypoxia-induced insulin resistance appears to suppress exogenous glucose oxidation during metabolically matched aerobic exercise during acute (<8 h) high-altitude (HA) exposure. However, a better understanding of this metabolic dysregulation is needed to identify interventions to mitigate these effects. The objective of this study was to determine if differences in metabolomic profiles during exercise at sea level (SL) and HA are reflective of hypoxia-induced insulin resistance. Native lowlanders (n = 8 males) consumed 145 g (1.8 g/min) of glucose while performing 80-min of metabolically matched treadmill exercise at SL (757 mmHg) and HA (460 mmHg) after 5-h exposure. Exogenous glucose oxidation and glucose turnover were determined using indirect calorimetry and dual tracer technique ([13C]glucose and [6,6-2H2]glucose). Metabolite profiles were analyzed in serum as change (Δ), calculated by subtracting postprandial/exercised state SL (ΔSL) and HA (ΔHA) from fasted, rested conditions at SL. Compared with SL, exogenous glucose oxidation, glucose rate of disappearance, and glucose metabolic clearance rate (MCR) were lower (P < 0.05) during exercise at HA. One hundred and eighteen metabolites differed between ΔSL and ΔHA (P < 0.05, Q < 0.10). Differences in metabolites indicated increased glycolysis, tricarboxylic acid cycle, amino acid catabolism, oxidative stress, and fatty acid storage, and decreased fatty acid mobilization for ΔHA. Branched-chain amino acids and oxidative stress metabolites, Δ3-methyl-2-oxobutyrate (r = -0.738) and Δγ-glutamylalanine (r = -0.810), were inversely associated (P < 0.05) with Δexogenous glucose oxidation. Δ3-Hydroxyisobutyrate (r = -0.762) and Δ2-hydroxybutyrate/2-hydroxyisobutyrate (r = -0.738) were inversely associated (P < 0.05) with glucose MCR. Coupling global metabolomics and glucose kinetic data suggest that the underlying cause for diminished exogenous glucose oxidative capacity during aerobic exercise is acute hypoxia-mediated peripheral insulin resistance.

Challenging traditional carbohydrate intake recommendations for optimizing performance at high altitude.

PURPOSE OF REVIEW: To highlight emerging evidence challenging traditional recommendations to increase carbohydrate intake to optimize performance at high altitude. RECENT FINDINGS: Several studies have now clearly demonstrated that, compared with sea level, exogenous carbohydrate oxidation during aerobic exercise is blunted in lowlanders during initial exposure to high altitude. There is also no apparent ergogenic effect of ingesting carbohydrate during aerobic exercise on subsequent performance at high altitude, either initially after arriving or even after up to 22 days of acclimatization. The inability to oxidize and functionally benefit from exogenous carbohydrate intake during exercise after arriving at high altitude coincides with hyperinsulinemia, accelerated glycogenolysis, and reduced peripheral glucose uptake. Collectively, these responses are consistent with a hypoxia-mediated metabolic dysregulation reflective of insulin resistance. Parallel lines of evidence have also recently demonstrated roles for the gut microbiome in host metabolism, bioenergetics, and physiologic responses to high altitude, implicating the gut microbiome as one potential mediator of hypoxia-mediated metabolic dysregulation. SUMMARY: Identification of novel and well tolerated nutrition and/or pharmacological approaches for alleviating hypoxia-mediated metabolic dysregulation and enhancing exogenous carbohydrate oxidation may be more effective for optimizing performance of lowlanders newly arrived at high altitude than traditional carbohydrate recommendations.

Effects of energy balance on appetite and physiological mediators of appetite during strenuous physical activity: secondary analysis of a randomised crossover trial.

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.

Sensitivity and reliability of zinc transporter and metallothionein gene expression in peripheral blood mononuclear cells as indicators of zinc status: responses to ex vivo zinc exposure and habitual zinc intake in humans.

Zn is an essential nutrient for humans; however, a sensitive biomarker to assess Zn status has not been identified. The objective of this study was to determine the reliability and sensitivity of Zn transporter and metallothionein (MT) genes in peripheral blood mononuclear cells (PBMCs) to Zn exposure ex vivo and to habitual Zn intake in human subjects. In study 1, human PBMCs were cultured for 24 h with 0-50 µm ZnSO4 with or without 5 µm N,N,N',N'-tetrakis(2-pyridylmethyl)ethylenediamine (TPEN), and mRNA expression of SLC30A1-10, SLC39A1-14, MT1 subtypes (A, B, E, F, G, H, L, M and X), MT2A, MT3 and MT4 mRNA was determined. In study 2, fifty-four healthy male and female volunteers (31·9 (sd 13·8) years, BMI 25·7 (sd 2·9) kg/m2) completed a FFQ, blood was collected, PBMCs were isolated and mRNA expression of selected Zn transporters and MT isoforms was determined. Study 1: MT1E, MT1F, MT1G, MT1H, MT1L, MT1M, MT1X, MT2A and SLC30A1 increased with increasing concentrations of Zn and declined with the addition of TPEN. Study 2: Average daily Zn intake was 16·0 (sd 5·3) mg/d (range: 9-31 mg/d), and plasma Zn concentrations were 15·5 (SD 2·8) µmol/l (range 11-23 µmol/l). PBMC MT2A was positively correlated with dietary Zn intake (r 0·306, P = 0·03) and total Zn intake (r 0·382, P < 0·01), whereas plasma Zn was not (P > 0·05 for both). Findings suggest that MT2A mRNA in PBMCs reflects dietary Zn intake in healthy adults and may be a component in determining Zn status.

Testosterone supplementation upregulates androgen receptor expression and translational capacity during severe energy deficit.

Testosterone supplementation during energy deficit promotes whole body lean mass accretion, but the mechanisms underlying that effect remain unclear. To elucidate those mechanisms, skeletal muscle molecular adaptations were assessed from muscle biopsies collected before, 1 h, and 6 h after exercise and a mixed meal (40 g protein, 1 h postexercise) following 14 days of weight maintenance (WM) and 28 days of an exercise- and diet-induced 55% energy deficit (ED) in 50 physically active nonobese men treated with 200 mg testosterone enanthate/wk (TEST) or placebo (PLA) during the ED. Participants (n = 10/group) exhibiting substantial increases in leg lean mass and total testosterone (TEST) were compared with those exhibiting decreases in both of these measures (PLA). Resting androgen receptor (AR) protein content was higher and fibroblast growth factor-inducible 14 (Fn14), IL-6 receptor (IL-6R), and muscle ring-finger protein-1 gene expression was lower in TEST vs. PLA during ED relative to WM (P < 0.05). Changes in inflammatory, myogenic, and proteolytic gene expression did not differ between groups after exercise and recovery feeding. Mechanistic target of rapamycin signaling (i.e., translational efficiency) was also similar between groups at rest and after exercise and the mixed meal. Muscle total RNA content (i.e., translational capacity) increased more during ED in TEST than PLA (P < 0.05). These findings indicate that attenuated proteolysis at rest, possibly downstream of AR, Fn14, and IL-6R signaling, and increased translational capacity, not efficiency, may drive lean mass accretion with testosterone administration during energy deficit.

Severe negative energy balance during 21 d at high altitude decreases fat-free mass regardless of dietary protein intake: a randomized controlled trial.

In this 2-phase randomized controlled study, we examined whether consuming a higher-protein (HP) diet would attenuate fat-free mass (FFM) loss during energy deficit (ED) at high altitude (HA) in 17 healthy males (mean ± sd: 23 ± 6 yr; 82 ± 14 kg). During phase 1 at sea level (SL, 55 m), participants consumed a eucaloric diet providing standard protein (SP; 1.0 g protein/kg,) for 21 d. During phase 2, participants resided at HA (4300 m) for 22 d and were randomly assigned to either an SP or HP (2.0 g protein/kg) diet designed to elicit a 40% ED. Body composition, substrate oxidation, and postabsorptive whole-body protein kinetics were measured. Participants were weight stable during SL and lost 7.9 ± 1.9 kg ( P < 0.01) during HA, regardless of dietary protein intake. Decrements in whole-body FFM (3.6 ± 2.4 kg) and fat mass (3.6 ± 1.3 kg) were not different between SP and HP. HP oxidized 0.95 ± 0.32 g protein/kg per day more than SP and whole-body net protein balance was more negative for HP than for SP ( P < 0.01). Based on changes in body energy stores, the overall ED was 70% (-1849 ± 511 kcal/d, no group differences). Consuming an HP diet did not protect FFM during severe ED at HA.-Berryman, C. E., Young, A. J., Karl, J. P., Kenefick, R. W., Margolis, L. M., Cole, R. E., Carbone, J. W., Lieberman, H. R., Kim, I.-Y., Ferrando, A. A., Pasiakos, S. M. Severe negative energy balance during 21 d at high altitude decreases fat-free mass regardless of dietary protein intake: a randomized controlled trial.

Serum and Erythrocyte Biomarkers of Nutrient Status Correlate with Short-Term Α-Carotene, Β-Carotene, Folate, and Vegetable Intakes Estimated by Food Frequency Questionnaire in Military Recruits.

OBJECTIVE: Food frequency questionnaires (FFQs) estimate habitual dietary intake and require evaluation in populations of interest in order to determine accuracy. Thus, the purpose of this study was to determine agreement between circulating biomarkers and FFQ estimated dietary intake in a military population consuming all meals in a dining facility over 12 weeks. METHODS: 2014 Block FFQs were administered and fasted blood samples were drawn to assess nutritional biomarkers at the end of a 12-week training period in male (n = 141) and female (n = 125) Marine recruits undergoing initial military training. FFQ estimates of alpha- and beta-carotene, folate, and fruit and vegetable intake and circulating concentrations of serum alpha- and beta-carotene and serum and erythrocyte folate were measured. Partial correlations were used in the full model, and weighted kappa coefficients were used to determine agreement between ranking quartiles of dietary intake estimates with corresponding biomarker status quartiles. RESULTS: Serum and dietary intake of alpha-carotene were positively associated in males (p = 0.009) and females (p < 0.001), as was serum and intake of beta-carotene (males, p = 0.002; females, p < 0.001). Alpha-carotene was positively associated with vegetable intake in males (p = 0.02) and beta-carotene with vegetable intake in females (p = 0.003). Serum folate in males (p = 0.002) and erythrocyte folate in females (p = 0.02) were associated with dietary folate intake. In females, the relationships between biomarker and dietary estimates yielded significant kappa coefficients. In males, a significant kappa coefficient was observed for erythrocyte folate and dietary intake of folate only. The kappa coefficient for serum and estimated intake of beta-carotene was not significant in males. CONCLUSION: Twelve-week habitual intake of alpha-and beta-carotene and folate were correlated with circulating biomarkers in a military training population. The 2014 Block FFQ was able to accurately rank females into quartiles of nutrient status based on intake, while males were ranked less accurately than females.

Self-reported eating behaviors of military recruits are associated with body mass index at military accession and change during initial military training.

Eating behaviors such as eating fast and ignoring internal satiety cues are associated with overweight/obesity, and may be influenced by environmental factors. This study examined changes in those behaviors, and associations between those behaviors and BMI, cardiometabolic biomarkers, and diet quality in military recruits before and during initial military training (IMT), an environment wherein access to food is restricted. Eating rate and reliance on internal satiety cues were self-reported, and BMI, body fat, cardiometabolic biomarkers, and diet quality were measured in 1389 Army, Air Force and Marine recruits (45% female, mean ±â€¯SEM BMI = 24.1 ±â€¯0.1 kg/m2) before and after IMT. Pre-IMT, habitually eating fast relative to slowly was associated with a 1.1 ±â€¯0.3 kg/m2 higher BMI (P < 0.001), but not with other outcomes; whereas, habitually eating until no food is left (i.e., ignoring internal satiety cues) was associated with lower diet quality (P < 0.001) and, in men, 1.6 ±â€¯0.6% lower body fat (P = 0.03) relative to those that habitually stopped eating before feeling full. More recruits reported eating fast (82% vs 39%) and a reduced reliance on internal satiety cues (55% vs 16%) during IMT relative to pre-IMT (P < 0.001). Findings suggest that eating behaviors correlate with body composition and/or diet quality in young, predominantly normal-weight recruits entering the military, and that IMT is associated with potentially unfavorable changes in these eating behaviors.