Urinary Proteomic Biomarkers of Trabecular Bone Volume Change during Army Basic Combat Training.

PURPOSE: Optimize a dMS-based urinary proteomic technique and evaluate the relationship between urinary proteome content and adaptive changes in bone microarchitecture during BCT. METHODS: Urinary proteomes were analyzed with an optimized dMS technique in two groups of 13 recruits (n = 26) at the beginning (Pre) and end (Post) of BCT. Matched by age (21 ± 4 yr), sex (16 W), and baseline tibial trabecular bone volume fractions (Tb.BV/TV), these groups were distinguished by the most substantial (High) and minimal (Low) improvements in Tb.BV/TV. Differential protein expression was analyzed with mixed permutation ANOVA and false discovery proportion-based adjustment for multiple comparisons. RESULTS: Tibial Tb.BV/TV increased from pre- to post-BCT in High (3.30 ± 1.64%, p < 0.0001) but not Low (-0.35 ± 1.25%, p = 0.4707). The optimized dMS technique identified 10,431 peptides from 1,368 protein groups that represented 165 integrative biological processes. 74 urinary proteins changed from pre- to post-BCT (p = 0.0019) and neutrophil mediated immunity was the most prominent ontology. Two proteins (Immunoglobulin heavy constant gamma 4 and C-type lectin domain family 4 member G) differed from pre- to post-BCT in High and Low (p = 0.0006). CONCLUSIONS: The dMS technique can identify more than 1000 urinary proteins. At least 74 proteins are responsive to BCT, and other principally immune system-related proteins show differential expression patterns that coincide with adaptive bone formation.

Acute Resistance Exercise Modifies Extracellular Vesicle miRNAs Targeting Anabolic Gene Pathways: A Prospective Cohort Study.

BACKGROUND: Resistance training confers numerous health benefits that are mediated in part by circulating factors. Towards an enhanced molecular understanding, there is growing interest in a class of signaling biomarkers called extracellular vesicles (EVs). Extracellular vesicles support physiological adaptations to exercise by transporting their cargo (e.g., microRNA [miRNA]) to target cells. Previous studies of changes in EV cargo have focused on aerobic exercise, with limited data examining the effects of resistance exercise. We examined the effect of acute resistance exercise on circulating EV miRNAs and their predicted target pathways. METHODS: Ten participants (5 men; age: 26.9 ± 5.5 y, height: 1.7 ± 0.1 m, body mass: 74.0 ± 11.1 kg, body fat: 25.7 ± 11.6 %) completed an acute heavy resistance exercise test (AHRET) consisting of six sets of 10 repetitions of back squats using 75% one-repetition maximum. Pre-/post-AHRET, EVs were isolated from plasma using size exclusion chromatography, and RNA sequencing was performed. Differentially expressed (DE) miRNAs between pre- and post-AHRET EVs were analyzed using Ingenuity Pathway Analysis to predict target messenger RNAs and their target biological pathways. RESULTS: Overall, 34 miRNAs were altered by AHRET (p < 0.05), targeting 4,895 mRNAs, with enrichment of 175 canonical pathways (p < 0.01), including 12 related to growth/metabolism (p53, IGF-I, STAT3, PPAR, JAK/STAT, growth hormone, WNT/ß-catenin, ERK/MAPK, AMPK, mTOR, and PI3K/AKT) and eight to inflammation signaling (TGF-ß, IL-8, IL-7, IL-3, IL-6, IL-2, IL-17, IL-10). CONCLUSIONS: Acute resistance exercise alters EV miRNAs targeting pathways involved in growth, metabolism, and immune function. Circulating EVs may serve as significant adaptive signaling molecules influenced by exercise training.

Assessing the physiological basis for resilience in military personnel.

Military personnel experience training and operational demands that are different from civilian populations, including frequent deployment, exposure to austere environments, and dislocation from family. These unique occupational demands may result in negative impacts on health, performance, and career success. As such, resilience, defined as a system's capacity to resist, recover, recover better, or adapt, to perturbation from a challenge or stressor, is a critical factor in assuring the health and safety of military personnel. In recent years, the Department of Defense (DoD) has funded research programs assessing the physiological basis of resilience. This review will overview research programs, review salient findings from recent studies, and highlight potential future areas of research. Physiological factors influencing or predicting resilience in US military populations, including physical performance, anthropometrics and body composition, nutrition and dietary supplements, and other biomarkers will be highlighted. Finally, this manuscript will detail potential future studies, including interventions, aimed at optimising physiological resilience in military personnel.

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.

Military tactical adaptive decision making during simulated military operational stress is influenced by personality, resilience, aerobic fitness, and neurocognitive function.

Laboratory-based studies designed to mimic combat or military field training have consistently demonstrated deleterious effects on warfighter's physical, cognitive, and emotional performance during simulated military operational stress (SMOS). Purpose: The present investigation sought to determine the impact of a 48-h simulated military operational stress (SMOS) on military tactical adaptive decision making, and the influence of select psychological, physical performance, cognitive, and physiological outcome measures on decision making performance. Methods: Male (n = 48, 26.2 ± 5.5 years, 177.7 ± 6.6 cm, 84.7 ± 14.1 kg.) subjects currently serving in the U.S. military were eligible to participate in this study. Eligible subjects completed a 96-h protocol that occurred over five consecutive days and four nights. Day 2 (D2) and day 3 (D3) consisted of 48-h of SMOS wherein sleep opportunity and caloric needs were reduced to 50%. Differences in SPEAR total block score from baseline to peak stress (D3 minus D1) were calculated to assess change in military tactical adaptive decision making and groups were stratified based on increase (high adaptors) or decrease (low adaptors) of the SPEAR change score. Results: Overall, military tactical decision-making declined 1.7% from D1 to D3 (p < 0.001). High adaptors reported significantly higher scores of aerobic capacity (p < 0.001), self-report resilience (p = 0.020), extroversion (p < 0.001), and conscientiousness (p < 0.001). at baseline compared to low adaptors, while low adaptors reported greater scores in Neuroticism (p < 0.001). Conclusion: The present findings suggest that service members whose adaptive decision making abilities improved throughout SMOS (i.e., high adaptors) demonstrated better baseline psychological/self-reported resilience and aerobic capacity. Further, changes in adaptive decision-making were distinct from those of lower order cognitive functions throughout SMOS exposure. With the transition of future military conflicts placing higher priority on enhancing and sustaining cognitive readiness and resiliency, data presented here demonstrates the importance of measuring and categorizing baseline measures inherent to military personnel, in order to change and train one's ability to suffer less of a decline during high stress conditions.

Reliability and Validity of a Flume-Based Maximal Oxygen Uptake Swimming Test.

A mode-specific swimming protocol to assess maximal aerobic uptake (VO2maxsw) is vital to accurately evaluate swimming performance. A need exists for reliable and valid swimming protocols that assess VO2maxsw in a flume environment. The purpose was to assess: (a) reliability and (b) "performance" validity of a VO2maxsw flume protocol using the 457-m freestyle pool performance swim (PS) test as the criterion. Nineteen males (n = 9) and females (n = 10) (age, 28.5 ± 8.3 years.; height, 174.7 ± 8.2 cm; mass, 72.9 ± 12.5 kg; %body fat, 21.4 ± 5.9) performed two flume VO2maxsw tests (VO2maxswA and VO2maxswB) and one PS test [457 m (469.4 ± 94.7 s)]. For test-retest reliability (Trials A vs. B), moderately strong relationships were established for VO2maxsw (mL·kg-1·min-1)(r= 0.628, p = 0.002), O2pulse (mL O2·beat-1)(r = 0.502, p = 0.014), VEmax (L·min-1) (r = 0.671, p = 0.001), final test time (sec) (0.608, p = 0.004), and immediate post-test blood lactate (IPE (BLa)) (0.716, p = 0.001). For performance validity, moderately strong relationships (p < 0.05) were found between VO2maxswA (r =-0.648, p = 0.005), O2pulse (r= -0.623, p = 0.008), VEmax (r = -0.509 p = 0.037), and 457-m swim times. The swimming flume protocol examined is a reliable and valid assessment of VO2maxsw., and offers an alternative for military, open water, or those seeking complementary forms of training to improve swimming performance.

Efforts toward the continuous monitoring of molecular markers of performance.

OBJECTIVES: Technologies supporting the continuous, real-time measurement of blood oxygen saturation and plasma glucose levels have improved our ability to monitor performance status. Our ability to monitor other molecular markers of performance, however, including the hormones known to indicate overtraining and general health, has lagged. That is, although a number of other molecular markers of performance status have been identified, we have struggled to develop viable technologies supporting their real-time monitoring in the body. Here we review biosensor approaches that may support such measurements, as well as the molecules potentially of greatest interest to monitor. DESIGN: Narrative literature review. METHOD: Literature review. RESULTS: Significant effort has been made to harness the specificity, affinity, and generalizability of biomolecular recognition in a platform technology supporting continuous in vivo molecular measurements. Most biosensor approaches, however, are either not generalizable to most targets, or fail when challenged in the complex environments found in vivo. Electrochemical aptamer-based sensors, in contrast, are the first technology to simultaneously achieve both of these critical attributes. In an effort to illustrate the potential of this platform technology, we both critically review the literature describing it and briefly survey some of the molecular performance markers we believe will prove advantageous to monitor using it. CONCLUSIONS: Electrochemical aptamer-based sensors may be the first truly generalizable technology for monitoring specific molecules in situ in the body and how adaptation of the platform to subcutaneous microneedles will enable the real-time monitoring of performance markers via a wearable, minimally invasive device.

Less daytime sleepiness and slow wave activity during sleep predict better physical readiness in military personnel.

BACKGROUND: Military personnel must maintain physical performance despite exposure to operational stressors such as sleep loss, caloric restriction and high cognitive load. Habitual sleep and specific sleep features are positively associated with fitness and may contribute to physical performance in operational settings. Further, by affecting muscle recovery, sleep may contribute to the ability to maintain performance across multiple days of exposure to operational stressors. OBJECTIVES: We examined the role of individual differences in baseline sleep on baseline physical performance and on change in physical performance throughout exposure to simulated military operational stress (SMOS). METHODS: Military personnel (36 male, 9 female, 26.3 ± 5.3 years) completed a 5-day SMOS protocol during which they completed a tactical mobility test daily. Sleep questionnaires were administered at intake and sleep was monitored each night with polysomnography. Lasso regressions were used to identify meaningful predictors of physical performance at baseline and of change in physical performance across SMOS. RESULTS: Better aerobic fitness, lower daytime sleepiness (Epworth Sleepiness Scale), and lower absolute slow wave activity (0.5-4 Hz) predicted better physical performance at baseline (66.1% of variance explained), but did not relate to changes in performance. CONCLUSIONS: Collectively, higher daytime sleepiness and slow wave activity may reflect more chronic exposure to insufficient sleep and higher baseline sleep drive, which in turn led to compromised physical performance. The findings suggest that low self-report sleepiness and low objective slow wave activity may reflect two quantifiable markers of healthy sleep behaviors that have implications for operational performance.

Effects of energy balance on cognitive performance, risk-taking, ambulatory vigilance and mood during simulated military sustained operations (SUSOPS).

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. METHODS: 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. RESULTS: 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. CONCLUSION: 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.

Prevention of Lower Extremity Musculoskeletal Injuries in Tactical and First Responder Populations: A Systematic Review and Meta-Analysis of Randomized Trials From 1955 to 2020.

ABSTRACT: Sinnott, AM, Krajewski, KT, LaGoy, AD, Beckner, ME, Proessl, F, Canino, MC, Nindl, BC, Turner, RL, Lovalekar, MT, Connaboy, C, and Flanagan, SD. Prevention of lower extremity musculoskeletal injuries in tactical and first responder populations: A systematic review and meta-analysis of randomized trials from 1955 to 2020. J Strength Cond Res 37(1): 239-252, 2023-Lower extremity musculoskeletal injuries (LEMSIs) impose a significant burden on tactical and first responder populations. To determine the effectiveness of LEMSI prevention strategies, we performed a systematic review and meta-analysis of randomized controlled trials published in English from 1955 to 2020 (PROSPERO: CRD42018081799). MEDLINE, EMBASE, Cochrane, CINAHL, ProQuest, and DTIC databases were searched for trials that assigned military service members, police, firefighters, or paramedics to LEMSI prevention interventions with a minimum surveillance period of 12 weeks. Evidence was synthesized as odds ratios (OR) for LEMSI occurrence between individuals assigned to interventions and those assigned to standard activities. Risk of bias was assessed with the Cochrane Risk of Bias tool 2.0. Random-effects meta-analyses were conducted for (a) physical training and (b) footwear modifications to reduce LEMSI and (c) footwear modifications to reduce stress fractures specifically. Certainty in the body of evidence was determined with the GRADE approach. Of 28,499 records, 18 trials comprised of more than 11,000 subjects were synthesized. Interventions included physical training (8, N = 6,838), footwear modifications (8, N = 3,792), nutritional supplementation (1, N = 324), and training modifications (1, N = 350). Overall risk of bias was generally moderate ( N = 7 of 18) or high ( N = 9 of 18). Physical training (OR = 0.87, 95% CI [0.71, 1.08], p = 0.22, I 2 = 58.4%) and footwear modification (OR = 1.13, 95% CI [0.85, 1.49], p = 0.42, I 2 = 0.0%) did not reduce LEMSI or stress fractures (OR = 0.76, 95% CI [0.45, 1.28], p = 0.30, I 2 = 70.7%). Our results indicate that there is weak evidence to support current LEMSI prevention strategies. Future efforts will benefit from longer surveillance periods, assessment of women and nonmilitary populations, improved methodological rigor, and a greater breadth of approaches.

Combined effects of time-of-day and simulated military operational stress on perception-action coupling performance.

Perception-action coupling, the ability to 'read and react' to the environment, is essential for military personnel to operate within complex and unpredictable environments. Exposure to military operational stressors (e.g., caloric restriction, sleep loss, physical exertion), including around-the-clock operations, may compromise perception-action coupling, thereby impacting performance and safety. We examined the combined effects of simulated military operational stress (SMOS) and time-of-day on perception-action coupling. Fifty-seven active duty and reservist military personnel (45 M; 26.4 ± 5.6 years) completed a 5-day SMOS protocol that included two consecutive days of caloric restriction, and sleep restriction, and disruption. Participants completed a tablet-based perception-action coupling task (PACT) that involves perceiving whether virtual balls fit through virtual apertures. Familiarization occurred on day 0. Eight trials across day 1 (18:00, 22:00), 2 (04:00, 18:00, 22:00) and 3 (04:00, 18:00, 22:00) were analyzed. Mixed models were run to examine the interactive and main effects of day, and time-of-day on PACT response speed and accuracy outcomes. PACT response speed and accuracy outcomes improved at 18:00 and 22:00, whereas performance at 04:00 deteriorated across days. Perception-action coupling performance was resilient to SMOS, except in the early morning when the circadian drive for sleep is high, and the effects of sleep loss are more prominent.

Psychological and physiological changes during basic, underwater, demolition/SEAL training.

This longitudinal study examines the growth of psychological characteristics and adaptation of physiological markers of stress during a six-month assessment and selection course for U.S. Navy SEALs. Resilience, hardiness, and grit instruments were used to evaluate the psychological characteristics. Blood samples were taken to determine physiological markers related to stress adaptation; specifically, evaluating DHEA, DHEA-to-cortisol ratio, BDNF, NPY, and cortisol. Data was collected at four timepoints throughout the assessment and selection course from 353 students over three classes. Results indicated that resilience and hardiness grow after an initial decline, DHEA and DHEA-to-cortisol increased suggesting physiological adaptation. However, psychological and physiological markers do not exhibit the same growth patterns for participants in the course. This study enhances the understanding of psychological growth and physiological adaptation in a high-stress environment over an extended duration.

Resistance exercise differentially alters extracellular vesicle size and subpopulation characteristics in healthy men and women: an observational cohort study.

Extracellular vesicles (EVs) are established mediators of adaptation to exercise. Currently, there are no published data comparing changes in EVs between men and women after resistance exercise. We tested the hypothesis that EV profiles would demonstrate a sex-specific signature following resistance exercise. Ten men and 10 women completed an acute heavy resistance exercise test for back squats using 75% of their one-repetition maximum. Blood was drawn before and immediately after exercise. EVs were isolated from plasma using size exclusion chromatography and stained with antibodies associated with exosomes (CD63), microvesicles (VAMP3), apoptotic bodies (THSD1), and a marker for skeletal muscle EVs (SGCA). CD63+ EV concentration and proportion of total EVs increased 23% (P = 0.006) and 113% (P = 0.005) in both sexes. EV mean size declined in men (P = 0.020), but not in women, suggesting a relative increase in small EVs in men. VAMP3+ EV concentration and proportion of total EVs increased by 93% (P = 0.025) and 61% (P = 0.030) in men and women, respectively. SGCA+ EV concentration was 69% higher in women compared with men independent of time (P = 0.007). Differences were also observed for CD63, VAMP3, and SGCA median fluorescence intensity, suggesting altered surface protein density according to sex and time. There were no significant effects of time or sex on THSD1+ EVs or fluorescence intensity. EV profiles, particularly among exosome-associated and muscle-derived EVs, exhibit sex-specific differences in response to resistance exercise which should be further studied to understand their relationship to training adaptations.

Neuroendocrine, inflammatory, and extracellular vesicle responses during the Navy Special Warfare Screener Selection Course.

Military operational stress is known to increase adrenal hormones and inflammatory cytokines, while decreasing hormones associated with the anabolic milieu and neuroendocrine system. Less is known about the role of extracellular vesicles (EVs), a form of cell-to-cell communication, in military operational stress and their relationship to circulating hormones. The purpose of this study was to characterize the neuroendocrine, cytokine, and EV response to an intense. 24-h selection course known as the Naval Special Warfare (NSW) Screener and identify associations between EVs and cytokines. Blood samples were collected the morning of and following the NSW Screener in 29 men (18-26 yr). Samples were analyzed for concentrations of cortisol, insulin-like growth factor I (IGF-I), neuropeptide-Y (NPY), brain-derived neurotrophic factor (BDNF), α-klotho, tumor necrosis factor-α (TNFα), and interleukins (IL) -1ß, -6, and -10. EVs stained with markers associated with exosomes (CD63), microvesicles (VAMP3), and apoptotic bodies (THSD1) were characterized using imaging flow cytometry and vesicle flow cytometry. The selection event induced significant changes in circulating BDNF (-43.2%), IGF-I (-24.6%), TNFα (+17.7%), and IL-6 (+13.6%) accompanied by increases in intensities of THSD1+ and VAMP3+ EVs (all P < 0.05). Higher concentrations of IL-1ß and IL-10 were positively associated with THSD1+ EVs (P < 0.05). Military operational stress altered the EV profile. Surface markers associated with apoptotic bodies were positively correlated with an inflammatory response. Future studies should consider a multiomics assessment of EV cargo to discern canonical pathways that may be mediated by EVs during military stress.

Utility of extracellular vesicles as a potential biological indicator of physiological resilience during military operational stress.

Extracellular vesicles (EVs) transport biological content between cells to mediate physiological processes. The association between EVs and resilience, the ability to cope with stress, is unknown. Using unbiased machine learning approaches, we aimed to identify a biological profile of resilience. Twenty servicemen (27.8 ± 5.9 years) completed the Connor Davidson Resilience (CD-RISC) questionnaire and were exposed to daily physical and cognitive exertion with 48-hr sleep and caloric restriction. Blood samples from baseline and the second day of stress were analyzed for neuroendocrine biomarkers impacted by military stress. EVs were isolated from plasma and stained with antibodies associated with exosomes (CD63), microvesicles (VAMP3), and apoptotic bodies (THSD1). Individuals were separated into high (n = 10, CD-RISC > 90) and low (n = 10, CD-RISC < 79) resilience. EV features were stratified by size, then down-selected using regression trees and compared between groups. Diagnostic accuracy was assessed using receiver operating characteristic curves. Compared to low resilience, high resilience demonstrated a greater increase in variability of THSD1 local bright spot intensities among large-sized EVs in response to stress (p = 0.002, Hedges' g = 1.59). Among medium-sized EVs, high resilience exhibited a greater decrease in side scatter intensity (p = 0.014, Hedges' g = 1.17). Both features demonstrated high to moderate diagnostic accuracy for high resilience (AUC = 0.90 and 0.79). In contrast, neuroendocrine biomarker concentrations were similar between groups. The increase in variability among THSD1 + EVs in high, but not low, resilient individuals following stress may suggest high resilience is accompanied by stress-triggered apoptotic adaptations to the environment that are not detected in neuroendocrine biomarkers.

Men and women display distinct extracellular vesicle biomarker signatures in response to military operational stress.

Extracellular vesicles (EVs) are mediators of physiological changes that occur during physical exertion. This study examined the effects of physical exertion with and without sleep and caloric restriction on EV size, concentration, and surface proteins in men and women. Twenty participants (10 men) completed a 5-day simulated military operational stress protocol with daily physical exertion. Blood was drawn before and immediately after exertion at baseline (D1) and following 48-h of sleep and caloric restriction (D3). EV size and concentration were assessed using nanoparticle tracking analysis. EVs were identified with markers associated with exosomes (CD63), microvesicles (VAMP3), apoptotic bodies (THSD1), and skeletal muscle-derived EVs (SGCA) and quantified using imaging flow cytometry. Interactive and main effects of sex, day, and time on EVs were assessed using three-way ANOVAs. EV concentration declined pre to postexertion in women on D1 and D3 but was stable in men. EV size increased from pre to postexertion and from D1 to D3 in men and women. Physical exertion following sleep and caloric restriction increased CD63+ EV concentration, proportion of total EVs, and CD63 surface protein expression regardless of sex. The proportion of SGCA+ EVs increased in men and women following exertion and from D1 to D3 but was higher in women than in men. No differences were observed in VAMP3+ and THSD1+ EVs. This study identified sexually dimorphic EV profiles in response to various stressors. Further investigations are necessary to determine if dimorphic EV responses affect health and performance outcomes during stress.NEW & NOTEWORTHY Sex is understudied in EV research, and most studies limit EV analysis to single stress conditions such as exercise. Multistress conditions consisting of physical exertion and sleep and caloric restriction are common in real-world settings. We demonstrate that physical exertion results in sex-specific EV signatures and that EV profiles vary according to single versus multistress conditions. Our data highlight important biological and ecological characteristics that should be considered in EV research.

Effects of Multi-ingredient Preworkout Supplements on Physical Performance, Cognitive Performance, Mood State, and Hormone Concentrations in Recreationally Active Men and Women.

ABSTRACT: Beckner, ME, Pihoker, AA, Darnell, ME, Beals, K, Lovalekar, M, Proessl, F, Flanagan, SD, Arciero, PJ, Nindl, BC, and Martin, BJ. Effects of multi-ingredient preworkout supplements on physical performance, cognitive performance, mood state, and hormone concentrations in recreationally active men and women. J Strength Cond Res 36(9): 2493-2501, 2022-Performance enhancement supplement research has primarily focused on the effectiveness of individual ingredients, rather than the combination. This study investigated the acute effects of 2 multi-ingredient preworkout supplements (MIPS), with beta-alanine and caffeine (BAC) and without (NBAC), compared with placebo (PLA) on anaerobic performance, endurance capacity, mood state, cognitive function, vascular function, and anabolic hormones. Thirty exercise-trained individuals (24.4 ± 4.9 years, 15 men and 15 women) completed a fatiguing exercise protocol on 3 separate occasions, 30 minutes after ingestion of BAC, NBAC, or PLA. Outcomes were analyzed using one-way or two-way repeated-measures analysis of variance, as appropriate (alpha = 0.05). Anaerobic power was greater when supplementing with NBAC (10.7 ± 1.2 W·kg -1 ) and BAC (10.8 ± 1.4 W·kg -1 ) compared with PLA (10.4 ± 1.2 W·kg -1 ) ( p = 0.014 and p = 0.022, respectively). BAC improved V̇ o2 peak time to exhaustion ( p = 0.006), accompanied by an increase in blood lactate accumulation ( p < 0.001), compared with PLA. Both NBAC and BAC demonstrated improved brachial artery diameter after workout ( p = 0.041 and p = 0.005, respectively), but PLA did not. L-arginine concentrations increased from baseline to postsupplement consumption of BAC ( p = 0.017). Reaction time significantly decreased after exercise for all supplements. There was no effect of supplement on mood states. Exercise-trained individuals looking to achieve modest improvements in power and endurance may benefit from consuming MIPS before exercise.

A trait of mind: stability and robustness of sleep across sleep opportunity manipulations during simulated military operational stress.

STUDY OBJECTIVES: Within-subject stability of certain sleep features across multiple nights is thought to reflect the trait-like behavior of sleep. However, to be considered a trait, a parameter must be both stable and robust. Here, we examined the stability (i.e. across the same sleep opportunity periods) and robustness (i.e. across sleep opportunity periods that varied in duration and timing) of different sleep parameters. METHODS: Sixty-eight military personnel (14 W) spent 5 nights in the sleep laboratory during a simulated military operational stress protocol. After an adaptation night, participants had an 8-hour sleep opportunity (23:00-07:00) followed by 2 consecutive nights of sleep restriction and disruption which included two 2-hour sleep opportunities (01:00-03:00; 05:00-07:00) and, lastly, another 8-hour sleep opportunity (23:00-07:00). Intra-class correlation coefficients were calculated to examine differences in stability and robustness across different sleep parameters. RESULTS: Sleep architecture parameters were less stable and robust than absolute and relative spectral activity parameters. Further, relative spectral activity parameters were less robust than absolute spectral activity. Absolute alpha and sigma activity demonstrated the highest levels of stability that were also robust across sleep opportunities of varying duration and timing. CONCLUSIONS: Stability and robustness varied across different sleep parameters, but absolute NREM alpha and sigma activity demonstrated robust trait-like behavior across variable sleep opportunities. Reduced stability of other sleep architecture and spectral parameters during shorter sleep episodes as well as across different sleep opportunities has important implications for study design and interpretation.

Circulating biomarkers associated with performance and resilience during military operational stress.

Adaptation to military operational stress is a complex physiological response that calls upon the sympathetic nervous system (SNS), hypothalamic pituitary adrenal (HPA) axis and immune system, to create a delicate balance between anabolism and catabolism and meet the demands of an ever-changing environment. As such, resilience, the ability to withstand and overcome the negative impact of stress on military performance, is likely grounded in an appropriate biological adaptation to encountered stressors. Neuroendocrine [i.e. cortisol, epinephrine (EPI), norepinephrine (NE), neuropeptide-Y (NPY), and brain derived neurotropic factor (BDNF)], inflammatory [i.e. interleukin 6 (IL-6), IL-1ß, IL-4, IL-10 and tumour necrosis factor (TNF)-α], as well as growth and anabolic [i.e. insulin-like growth factor-I (IGF-I), testosterone, and dehydroepiandrosterone (DHEA)] biomarkers independently and interactively function in stress adaptations that are associated with a soldier's physical and psychological performance. In this narrative review, we detail biomarkers across neuroendocrine, inflammatory, and growth stimulating domains to better elucidate the biological basis of a resilient soldier. The findings from the reviewed studies indicate that military readiness and resiliency may be enhanced through better homeostatic control, better regulated inflammatory responses, and balanced anabolic/catabolic processes. It is unlikely that one class of biomarkers is better for assessing physiological resilience. Therefore, a biomarker panel that can account for appropriate balance across these domains may be superior in developing monitoring frameworks. Real-time physiological monitoring to assess biomarkers associated with resilience will be possible pending more sophisticated technologies and provide a field-expedient application for early identification and intervention of at-risk soldiers to improve military resiliency.

Sex differences in the physical performance, physiological, and psycho-cognitive responses to military operational stress.

Combat roles are physically demanding and expose service personnel to operational stressors such as high levels of physical activity, restricted nutrient intake, sleep loss, psychological stress, and environmental extremes. Women have recently integrated into combat roles, but our knowledge of the physical, physiological, and psycho-cognitive responses to these operational stressors in women is limited. The aim of this narrative review was to evaluate the evidence for sex-specific physical, physiological, and psycho-cognitive responses to real, and simulated, military operational stress. Studies examining physical and cognitive performance, body composition, metabolism, hypothalamic-pituitary-gonadal axis, and psychological health outcomes were evaluated. These studies report that women expend less energy and lose less body mass and fat-free mass, but not fat mass, than men. Despite having similar physical performance decrements as men during operational stress, women experience greater physiological strain than men completing the same physical tasks, but this may be attributed to differences in fitness. From limited data, military operational stress suppresses hypothalamic-pituitary-gonadal, but not hypothalamic-pituitary-adrenal, axis function in both sexes. Men and women demonstrate different psychological and cognitive responses to operational stress, including disturbances in mood, with women having a higher risk of post-traumatic stress symptoms compared with men. Based on current evidence, separate strategies to maximize selection and combat training are not warranted until further data directly comparing men and women are available. However, targeted exercise training programmes may be advisable to offset the physical performance gap between sexes and optimize performance prior to inevitable declines caused by intense military operations.