The biphasic activity of autophagy and heat shock protein response in peripheral blood mononuclear cells following acute resistance exercise in resistance-trained males.

PURPOSE: Autophagy and heat shock protein (HSP) response are proteostatic systems involved in the acute and adaptive responses to exercise. These systems may upregulate sequentially following cellular stress including acute exercise, however, currently few data exist in humans. This study investigated the autophagic and HSP responses to acute intense lower body resistance exercise in peripheral blood mononuclear cells (PBMCs) with and without branched-chain amino acids (BCAA) supplementation. METHODS: Twenty resistance-trained males (22.3 ± 1.5 yr; 175.4 ± .7 cm; 86.4 ± 15.6 kg) performed a bout of intense lower body resistance exercise and markers of autophagy and HSP70 were measured immediately post- (IPE) and 2, 4, 24, 48, and 72 h post-exercise. Prior to resistance exercise, 10 subjects were randomly assigned to BCAA supplementation of 0.22 g/kg/d for 5 days pre-exercise and up to 72 h following exercise while the other 10 subjects consumed a placebo (PLCB). RESULTS: There were no difference in autophagy markers or HSP70 expression between BCAA and PLCB groups. LC3II protein expression was significantly lower 2 and 4 h post-exercise compared to pre-exercise. LC3II: I ratio was not different at any time point compared to pre-exercise. Protein expression of p62 was lower IPE, 2, and 4 h post-exercise and elevated 24 h post-exercise. HSP70 expression was elevated 48 and 72 h post-exercise. CONCLUSIONS: Autophagy and HSP70 are upregulated in PBMCs following intense resistance exercise with autophagy increasing initially post-exercise and HSP response in the latter period. Moreover, BCAA supplementation did not affect this response.

Effect of heat stress on heat shock protein expression and hypertrophy-related signaling in the skeletal muscle of trained individuals.

Muscle mass is balanced between hypertrophy and atrophy by cellular processes, including activation of the protein kinase B-mechanistic target of rapamycin (Akt-mTOR) signaling cascade. Stressors apart from exercise and nutrition, such as heat stress, can stimulate the heat shock protein A (HSPA) and C (HSPC) families alongside hypertrophic signaling factors and muscle growth. The effects of heat stress on HSP expression and Akt-mTOR activation in human skeletal muscle and their magnitude of activation compared with known hypertrophic stimuli are unclear. Here, we show a single session of whole body heat stress following resistance exercise increases the expression of HSPA and activation of the Akt-mTOR cascade in skeletal muscle compared with resistance exercise in a healthy, resistance-trained population. Heat stress alone may also exert similar effects, though the responses are notably variable and require further investigation. In addition, acute heat stress in C2C12 muscle cells enhanced myotube growth and myogenic fusion, albeit to a lesser degree than growth factor-mediated hypertrophy. Though the mechanisms by which heat stress stimulates hypertrophy-related signaling and the potential mechanistic role of HSPs remain unclear, these findings provide additional evidence implicating heat stress as a novel growth stimulus when combined with resistance exercise in human skeletal muscle and alone in isolated murine muscle cells. We believe these findings will help drive further applied and mechanistic investigation into how heat stress influences muscular hypertrophy and atrophy.NEW & NOTEWORTHY We show that acute resistance exercise followed by whole body heat stress increases the expression of HSPA and increases activation of the Akt-mTOR cascade in a physically active and resistance-trained population.

Carbohydrate intake and resistance-based exercise: are current recommendations reflective of actual need?

Substantial research has been completed examining the impact of carbohydrate (CHO) intake on endurance exercise, whereas its role in resistance-based exercise performance, adaptation and cell signalling has yet to be fully characterised. This empirical shortcoming has precluded the ability to establish specific CHO recommendations for resistance exercise. This results in recommendations largely stemming from findings based on endurance exercise and/or anecdotal evidence despite the distinct energetic demands and molecular responses mediating adaptation from endurance- and resistance-based exercise. Moreover, the topic of CHO and exercise has become one of polarising nature with divergent views - some substantiated, others lacking evidence. Current literature suggests a moderately high daily CHO intake (3-7 g/kg per d) for resistance training, which is thought to prevent glycogen depletion and facilitate performance and adaptation. However, contemporary investigation, along with an emerging understanding of the molecular underpinnings of resistance exercise adaptation, may suggest that such an intake may not be necessary. In addition to the low likelihood of true glycogen depletion occurring in response to resistance exercise, a diet restrictive in CHO may not be detrimental to acute resistance exercise performance or the cellular signalling activity responsible for adaptation, even when muscle glycogen stores are reduced. Current evidence suggests that signalling of the mammalian target of rapamycin complex 1, the key regulatory kinase for gene translation (protein synthesis), is unaffected by CHO restriction or low muscular glycogen concentrations. Such findings may call into question the current view and subsequent recommendations of CHO intake with regard to resistance-based exercise.

The Potential Interplay Between HIF-1α, Angiogenic, and Autophagic Signaling During Intermittent Hypoxic Exposure and Exercise.

Berkemeier QN, Deyhle MR, McCormick JJ, Escobar KA, Mermier CM. The Potential Interplay between HIF-1α, Angiogenic, and Autophagic Signaling during Intermittent Hypoxic Exposure and Exercise High Alt Med Biol. 00:000-000, 2024.-Berkemeier QN, Deyhle MR, McCormick JJ, Escobar KA, Mermier CM. The Potential Interplay between HIF-1α, Angiogenic, and Autophagic Signaling During Intermittent Hypoxic Exposure and Exercise High Alt Med Biol. 00:000-000, 2024.-Environmental hypoxia as a result of decreased barometric pressure upon ascent to high altitudes (>2,500 m) presents increased physiological demands compared with low altitudes, or normoxic environments. Competitive athletes, mountaineers, wildland firefighters, military personnel, miners, and outdoor enthusiasts commonly participate in, or are exposed to, forms of exercise or physical labor at moderate to high altitudes. However, the majority of research on intermittent hypoxic exposure is centered around hematological markers, and the skeletal muscle cellular responses to exercise in hypoxic environments remain largely unknown. Two processes that may be integral for the maintenance of cellular health in skeletal muscle include angiogenesis, or the formation of new blood vessels from preexisting vasculature and autophagy, a process that removes and recycles damaged and dysfunctional cellular material in the lysosome. The purpose of this review is to is to examine the current body of literature and highlight the potential interplay between low-oxygen-sensing pathways, angiogenesis, and autophagy during acute and prolonged intermittent hypoxic exposure in conjunction with exercise. The views expressed in this paper are those of the authors and do not reflect the official policy of the Department of Army, DOD, DOE, ORAU/ORISE or U.S. Government.

Enhancing cardiometabolic health: unveiling the synergistic effects of high-intensity interval training with spirulina supplementation on selected adipokines, insulin resistance, and anthropometric indices in obese males.

This study investigated the combined effects of 12 weeks of high-intensity interval training (HIIT) and spirulina supplementation on adipokine levels, insulin resistance, anthropometric indices, and cardiorespiratory fitness in 44 obese males (aged 25-40 years). The participants were randomly assigned to one of four groups: control (CG), supplement (SG), training (TG), or training plus supplement (TSG). The intervention involved daily administration of either spirulina or a placebo and HIIT three times a week for the training groups. Anthropometric indices, HOMA-IR, VO2peak, and circulating adipokines (asprosin and lipocalin2, omentin-1, irisin, and spexin) were measured before and after the 12-week intervention. Post-intervention analysis indicated differences between the CG and the three interventional groups for body weight, fat-free mass (FFM), percent body fat (%BF), HOMA-IR, and adipokine levels (p < 0.05). TG and SG participants had increased VO2peak (p < 0.05). Spirulina supplementation with HIIT increased VO2peak, omentin-1, irisin, and spexin, while causing decreases in lipocalin-2 and asprosin levels and improvements in body composition (weight, %fat), BMI, and HOMA-IR. Notably, the combination of spirulina and HIIT produced more significant changes in circulating adipokines and cardiometabolic health in obese males compared to either supplementation or HIIT alone (p < 0.05). These findings highlight the synergistic benefits of combining spirulina supplementation with HIIT, showcasing their potential in improving various health parameters and addressing obesity-related concerns in a comprehensive manner.

Autophagy in peripheral blood mononuclear cells is associated with body fat percentage.

CONTEXT: Numerous chronic conditions including obesity exhibit autophagic dysfunction. Association of immune cell autophagic marker regulation by body fat percentage (%BF) is unknown. OBJECTIVE: Investigate autophagy activity in peripheral blood mononuclear cells (PBMCs) of adults with distinct %BFs and obesity-related circulating inflammatory markers. MATERIALS AND METHODS: Sixteen individuals (eight males) with %BF above (n = 8, 36.9 ± 3.6 years, 27.1 ± 8.1%BF) and below (n = 8, 37.1 ± 3.7 years, 13.3 ± 3.7%BF) their age- and sex-specific 50th percentile value based on the American College Sports Medicine guidelines participated. Body fat percentage was calculated from hydrostatic weighing. PBMCs were isolated from venous blood, and PBMC autophagic flux markers (LC3-I, LC3-II, and p62) were measured via Western blot. CRP, resistin, leptin, and adiponectin were measured via ELISA. RESULTS: LC3-II/LC3-I ratio correlated with %BF (r=-0.56, p=.023). Insulin (p=.05) and CRP (p=.018) were higher in high %BF participants. DISCUSSION AND CONCLUSIONS: Autophagic activity markers in PBMCs correlate with %BF, but are not different between %BF groups.

Spirulina Supplementation with High-Intensity Interval Training Decreases Adipokines Levels and Cardiovascular Risk Factors in Men with Obesity.

Adiposity, a state characterized by excessive accumulation of body fat, is closely linked to metabolic complications and the secretion of specific adipokines. This study explores the potential of exercise and Spirulina supplementation to mitigate these complications and modulate adipokine release associated with obesity. The primary objective of this investigation was to examine the impact of a 12-week regimen of high-intensity training combined with Spirulina supplementation on adipokine concentrations and lipid profiles in male individuals with obesity (N = 44). The participants were randomly distributed into four groups, each consisting of 11 participants: a control group (CG), a supplement group (SG), a training group (TG), and a training plus supplement group (TSG). The intervention comprised a 12-week treatment involving Spirulina supplementation (6 g capsule daily), a 12-week high-intensity interval training (HIIT) protocol with three sessions per week, or a combined approach. Following the interventions, metabolic parameters, anthropometric measurements, cardiorespiratory indices, and circulating adipokines [CRP, Sema3C, TNF-α, IL-6, MCP1, IL-8] were assessed within 48 h of the before and final training session. Statistical analyses revealed significant differences across all measures among the groups (p < 0.05). Notably, post hoc analyses indicated substantial disparities between the CG and the three interventional groups regarding body weight (p < 0.05). The combined training and supplementation approach led to noteworthy reductions in low-density lipoprotein (LDL), total cholesterol (TC), and triglyceride (TGL) levels (all p < 0.0001), coupled with an elevation in high-density lipoprotein-cholesterol (HDL-C) levels (p = 0.0001). Furthermore, adipokine levels significantly declined in the three intervention groups relative to the CG (p < 0.05). The findings from this 12-week study demonstrate that Spirulina supplementation in conjunction with high-intensity interval training reduced adipokine levels, improved body weight and BMI, and enhanced lipid profiles. This investigation underscores the potential of Spirulina supplementation and high-intensity interval training as a synergistic strategy to ameliorate obesity-related complications and enhance overall cardiometabolic well-being in obese males.

Astaxanthin Supplementation Augments the Benefits of CrossFit Workouts on Semaphorin 3C and Other Adipokines in Males with Obesity.

Regular physical activity and the use of nutritional supplements, including antioxidants, are recognized as efficacious approaches for the prevention and mitigation of obesity-related complications. This study investigated the effects of 12 weeks of CrossFit training combined with astaxanthin (ASX) supplementation on some plasma adipokines in males with obesity. Sixty-eight males with obesity (BMI: 33.6 ± 1.4 kg·m-2) were randomly assigned into four groups: the control group (CG; n = 11), ASX supplementation group (SG; n = 11), CrossFit group (TG; n = 11), and training plus supplement group (TSG; n = 11). Participants underwent 12 weeks of supplementation with ASX or placebo (20 mg/day capsule daily), CrossFit training, or a combination of both interventions. Plasma levels of semaphorin 3C (SEMA3C), apelin, chemerin, omentin1, visfatin, resistin, adiponectin, leptin, vaspin, and RBP4 were measured 72 h before the first training session and after the last training session. The plasma levels of all measured adipokines were significantly altered in SG, TG, and TSG groups (p < 0.05). The reduction of resistin was significantly higher in TSG than in SG (p < 0.05). The plasma levels of omentin1 were significantly higher in both training groups of TG and TSG than SG (p < 0.05), although such a meaningful difference was not observed between both training groups (p > 0.05). Significant differences were found in the reductions of plasma levels of vaspin, visfatin, apelin, RBP4, chemerin, and SEMA3C between the SG and TSG groups (p < 0.05). The study found that a 12-week intervention using ASX supplementation and CrossFit exercises resulted in significant improvements in several adipokines among male individuals with obesity. Notably, the combined approach of supplementation and training had the most pronounced results. The findings presented in this study indicate that the supplementation of ASX and participation in CrossFit exercise have the potential to be effective therapies in mitigating complications associated with obesity and enhancing metabolic health.

Oregano (Origanum vulgare) Consumption Reduces Oxidative Stress and Markers of Muscle Damage after Combat Readiness Tests in Soldiers.

Military activities often involve high-intensity exercise that can disrupt antioxidant capacity. We investigated the effects of oregano supplementation on muscle damage, oxidative stress, and plasma antioxidant markers of soldiers performing the army combat readiness test (ACRT). Twenty-four healthy male soldiers (age: 24 ± 3 years, height: 167 ± 14 cm, mass: 66 ± 3 kg) were randomized into a placebo group (n = 12) or an oregano supplementation group (n = 12). The participants consumed a capsule containing 500 mg Origanum vulgare immediately after completing the ACRT. Blood sampling was taken before exercise, immediately after exercise, and 60 and 120 min after oregano consumption. Plasma levels of creatine kinase (CK), lactate dehydrogenase (LDH), malondialdehyde (MDA), superoxide dismutase (SOD), total antioxidant capacity (TAC), and glutathione peroxidase (GPX) were measured at the four time points. The time × group interactions were found for CK (p < 0.0001, d = 3.64), LDH (p < 0.0001, d = 1.64), MDA (p < 0.0001, d = 9.94), SOD (p < 0.0001, d = 1.88), TAC (p < 0.0001, d = 5.68) and GPX (p < 0.0001, d = 2.38). In all variables, the difference between placebo and oregano groups were significant at 60 (p < 0.0001) and 120 (p < 0.0001) minutes after ACRT test. The main effect of time was also significant for all the variables (p < 0.0001). Our results suggest that oregano supplementation has the potential to reduce muscle damage and increase oxidative capacity following ACRT. Supplementation with oregano may serve as a dietary strategy to increase preparedness and promote recovery in military recruits.

Impact of varying doses of omega-3 supplementation on muscle damage and recovery after eccentric resistance exercise.

BACKGROUND: Exercise-induced muscle damage (EIMD) commonly occurs following intense resistance exercise and is associated with decrements in exercise performance and delayed muscle recovery. Thus, practical methods to attenuate EIMD would prove useful to both training and athletic populations. Omega-3 (n-3) supplementation has been shown to mitigate EIMD with evidence of increasing efficacy at higher doses (up to 6 g/day). However, data of its efficacy in trained individuals is limited. Therefore, this study investigated the effects of 6 and 8 g of n-3 supplementation on markers of muscle damage and muscle recovery after eccentric resistance exercise in resistance-trained males. METHODS: Using a double-blind, randomized, placebo-controlled design, 26 resistance trained males (23 ± 4 years; 173.6 ± 20.5 cm; 81.9 ± 9.7 kg; 14.2 ± 3.7% body fat) supplemented with 6 (n=10) or 8 g (n=7) of n-3 polyunsaturated fatty acids, or placebo (n=9) for 33 days. On day 30, participants performed a lower body muscle-damaging eccentric resistance exercise bout. Measures of muscle performance, soreness, and damage were taken pre-exercise on day 30 as well as on days 31-33, including vertical jump height (VJH), perceived muscle soreness (PMS), hip and knee range of motion (ROM), repetitions to fatigue (RTF) at 70% 1-RM, and serum creatine kinase (CK) while participants continued to supplement until day 33. RESULTS: There were significant differences in VJH, PMS, and serum CK following the muscle-damaging exercise bout compared to pre-exercise (p<0.05). However, there were no significant (p>0.05) differences between supplementation groups (6 g, 8 g, and placebo) at any time point post-exercise (day 31-33). There were no changes in hip and knee ROM or RTF at any time point or between groups. Vertical jump height and PMS returned to pre-exercise levels despite CK remaining elevated post-exercise. CONCLUSIONS: Thirty-three days of 6 and 8 g of n-3 supplementation did not attenuate EIMD or enhance muscle recovery following muscle-damaging eccentric resistance exercise in resistance-trained males. Further research using various n-3 supplementation durations, doses, and eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) concentrations may be needed to establish its efficacy in attenuating EIMD, which may vary between trained and untrained individuals. Furthermore, while circulating CK is commonly used to assess muscle damage, elevated CK levels may not reflect muscle recovery status following muscle-damaging exercise.

A Comparison of Muscle Activation Among the Front Squat, Overhead Squat, Back Extension and Plank.

The purpose of this study was to compare the muscle activation of the scapula, leg, and trunk among the front squat (FS), overhead squat (OHS), back extension (BE) and plank (PL). Seven recreationally trained men (age: 28 ± 3.6 years, body mass: 92 ± 26.1 kg, height: 175 ± 5.3 cm, 3-RM front squat test: 125 ± 49.8 kg, 3-RM overhead squat test: 91 ± 15.5 kg) participated in this within-subject crossover design. Two isometric exercises (plank and Biering-Sorenson back extension) were also included for trunk musculature comparisons. Neuromuscular activitation of the vastus lateralis (VL), biceps femoris (BF), thoracic region of erector spinae (ES), middle trapezius (MT), rectus abdominis (RA), external oblique (EO), serratus anterior (SA), and anterior deltoid (AD). The neuromuscular activity of the FS and OHS were analyzed using a 2 × 3 (squat variation X intensity) repeated measures analysis of variance (ANOVA). Effects were further analyzed by Bonferroni corrected paired t-tests. Results showed that AD activity was significantly greater (p < .05) during the FS compared to OHS at 65 and 95% of the 3-RM, while MT activity was significantly greater (p < .05) during the OHS than the FS at 80 and 95% of the 3-RM. ES activity was significantly greater (p< .05) during both the FS and OHS compared to the BE, but PL elicited significantly greater EO and RA activity than both the FS and OHS. These findings reveal that the FS and OHS can help facilitate the activation of muscles supporting the shoulder complex, scapula and lower back.

Physical Fitness Evaluation of Career Urban and Wildland Firefighters.

OBJECTIVE: Examine the physical fitness of career firefighters and compare their results to normative data and suggested standards for their profession. METHODS: Eighty firefighters completed a body composition analysis, maximal aerobic capacity (VO2max) test, and fitness testing battery, with results compared with normative value tables. Maximal aerobic capacity was correlated to fitness measures and differences between VO2max quartiles were examined. RESULTS: Twenty-two firefighters met the suggested standard for VO2max. Seventy percent of participants were classified as overweight or obese based on body mass index, while 25% were classified as having either "poor" or "very poor" body fat levels. Firefighters were above average for muscular strength. CONCLUSION: The firefighters had low aerobic fitness and higher than optimal body fat levels. Training programs may be necessary to assist firefighters in achieving optimal fitness levels.

Impact of Varying Dosages of Fish Oil on Recovery and Soreness Following Eccentric Exercise.

Fish oils (FOs) are rich in omega-3 long-chain polyunsaturated fatty acids, which have been purported to enhance recovery of muscular performance and reduce soreness post-exercise. However, the most effective FO dose for optimizing recovery remains unclear. The purpose of this investigation was to examine the effect of FO supplementation dosing on the recovery of measures of muscular performance, perceived soreness, and markers of muscle damage following a rigorous bout of eccentric exercise. Thirty-two college-aged resistance-trained males (~23.6 years, 71.6 kg, 172.1 cm) were supplemented with 2, 4, 6 g/day (G) FO or placebo (PL) for ~7.5 weeks. Following 7 weeks of supplementation, pre-exercise (PRE) performance assessments of vertical jump (VJ), knee extensor strength, 40-yard sprint, T-test agility, and perceived soreness were completed prior to a bout of muscle-damaging exercise and were repeated immediately post (IP), 1-, 2-, 4-, 24-, 48-, and 72-h (H) post-exercise. Repeated measures analysis of variance indicated a treatment × time interaction (p < 0.001) for VJ and perceived soreness, but no group differences were observed at any time point. VJ returned to PRE (54.8 ± 7.9 cm) by 1H (51.8 ± 6.5 cm, p = 0.112) for 6G, while no other groups returned to baseline until 48H. Lower soreness scores were observed in 6G compared to PL at 2H (mean difference [MD] = 2.74, p = 0.046), at 24H (MD: 3.45, p < 0.001), at 48H (MD = 4.45, p < 0.001), and at 72H (MD = 3.00, p = 0.003). Supplementation with 6G of FO optimized the recovery of jump performance and muscle soreness following a damaging bout of exercise.

Autophagy and aging: Maintaining the proteome through exercise and caloric restriction.

Accumulation of dysfunctional and damaged cellular proteins and organelles occurs during aging, resulting in a disruption of cellular homeostasis and progressive degeneration and increases the risk of cell death. Moderating the accrual of these defunct components is likely a key in the promotion of longevity. While exercise is known to promote healthy aging and mitigate age-related pathologies, the molecular underpinnings of this phenomenon remain largely unclear. However, recent evidences suggest that exercise modulates the proteome. Similarly, caloric restriction (CR), a known promoter of lifespan, is understood to augment intracellular protein quality. Autophagy is an evolutionary conserved recycling pathway responsible for the degradation, then turnover of cellular proteins and organelles. This housekeeping system has been reliably linked to the aging process. Moreover, autophagic activity declines during aging. The target of rapamycin complex 1 (TORC1), a central kinase involved in protein translation, is a negative regulator of autophagy, and inhibition of TORC1 enhances lifespan. Inhibition of TORC1 may reduce the production of cellular proteins which may otherwise contribute to the deleterious accumulation observed in aging. TORC1 may also exert its effects in an autophagy-dependent manner. Exercise and CR result in a concomitant downregulation of TORC1 activity and upregulation of autophagy in a number of tissues. Moreover, exercise-induced TORC1 and autophagy signaling share common pathways with that of CR. Therefore, the longevity effects of exercise and CR may stem from the maintenance of the proteome by balancing the synthesis and recycling of intracellular proteins and thus may represent practical means to promote longevity.

Effect of Branched-Chain Amino Acid Supplementation on Recovery Following Acute Eccentric Exercise.

This study investigated the effect of branched-chain amino acid (BCAA) supplementation on recovery from eccentric exercise. Twenty males ingested either a BCAA supplement or placebo (PLCB) prior to and following eccentric exercise. Creatine kinase (CK), vertical jump (VJ), maximal voluntary isometric contraction (MVIC), jump squat (JS) and perceived soreness were assessed. No significant (p > 0.05) group by time interaction effects were observed for CK, soreness, MVIC, VJ, or JS. CK concentrations were elevated above baseline (p < 0.001) in both groups at 4, 24, 48 and 72 hr, while CK was lower (p = 0.02) in the BCAA group at 48 hr compared to PLCB. Soreness increased significantly from baseline (p < 0.01) in both groups at all time-points; however, BCAA supplemented individuals reported less soreness (p < 0.01) at the 48 and 72 hr time-points. MVIC force output returned to baseline levels (p > 0.05) at 24, 48 and 72 hr for BCAA individuals. No significant difference between groups (p > 0.05) was detected for VJ or JS. BCAA supplementation may mitigate muscle soreness following muscle-damaging exercise. However, when consumed with a diet consisting of ~1.2 g/kg/day protein, the attenuation of muscular performance decrements or corresponding plasma CK levels are likely negligible.

The Effect of a Moderately Low and High Carbohydrate Intake on Crossfit Performance.

CrossFit is a metabolically demanding strength and conditioning method which performance may benefit from a carbohydrate (CHO)-rich diet. This study investigated the effect of three consecutive days of high CHO intake on CrossFit performance and corresponding metabolically -related variables in strength trained individuals. Eighteen subjects with a CHO intake of <6 g/kg/day were randomly assigned into a CHO (n = 9) or control (C) group (n =9) and underwent a 9-day training protocol. During days 1, 5, and 9, performance was measured as repetitions completed during a 12 minute CrossFit workout. Oxygen consumption (VO2), respiratory exchange ratio (RER), and blood lactate (BL) were also measured. Days 6-8, the CHO group increased CHO intake from <6 g/kg/day to 6-8 g/kg/day; the C group maintained their current intake of <6 g/kg/day. On days 6 and 7 both groups performed CrossFit workouts followed by a day of rest prior to day 9. There was a significant increase in repetitions completed in both groups in day 9 (vs. means score of day 1 + 5) (p = 0.002), but no differences between C and CHO groups (p = 0.111). However, the CHO group displayed a 15.2 repetition increase (+10.9%) in day 9, compared to 5.7 (+4.2%) by the C group. VO2, RER, and BL were not influenced by the experimental intervention. Our results suggest that the CrossFit-embraced practice of moderately-low CHO diets may be adequate in CHO during short periods of training, however, given the noted trend, extended training periods may be effected.