Dry blood spot samples to monitor immune-associated mRNA expression in intervention studies: Impact of Baker's yeast beta glucan.

Monitoring immunological response to physical stressors in a field setting is challenging because existing methods require a laboratory visit and traditional blood collection via venipuncture. The purpose of this study was to determine if our optimized dry blood spot (DBS) methodology yields sufficient total RNA to quantify the effect of Baker's Yeast Beta Glucan supplementation (BYBG; Wellmune; 250 mg/d) on post-exercise mRNA expression. Participants had venous DBS samples collected prior to (PRE), and immediately (POST), 2 (2H), and 4 (4H) hrs after completion of a 90 min run/walk trial in a hot, humid environment. Total RNA extracted from DBS was analyzed using a 574-plex Human Immunology mRNA panel (Nanostring). BYBG supplementation was associated with the increased expression of 12 mRNAs (LTB4R, PML, PRFM1, TNFRSF14, LCK, MYD88, STAT3, CCR1, TNFSF10, LILRB3, MME, and STAT6) and decreased expression of 4 mRNAs (MAP4K1, IKBKG, CD5, and IL4R) across all post-exercise time points. In addition to individually changed mRNA targets, we found eleven immune-response pathways that were significantly enriched by BYBG following exercise (TNF Family signaling, immunometabolism, oxidative stress, toll-like receptor (TLR) signaling, Treg differentiation, autophagy, chemokine signaling, complement system, Th2 differentiation, cytokine signaling, and innate immune). The present approach showed that DBS samples can be used to yield useful information about mRNA biomarkers in an intervention study. We have found that BYBG supplementation induces changes at the mRNA level that support the immune system and reduce susceptibility to opportunistic infection (i.e., upper respiratory tract infection) and facilitate improved physical recovery from exercise. Future studies may look to use DBS sampling for testing other nutritional, health, or medical interventions.

Inhaled diesel exhaust particles result in microbiome-related systemic inflammation and altered cardiovascular disease biomarkers in C57Bl/6 male mice.

BACKGROUND: The gut microbiota plays a vital role in host homeostasis and is associated with inflammation and cardiovascular disease (CVD) risk. Exposure to particulate matter (PM) is a known mediator of inflammation and CVD and is reported to promote dysbiosis and decreased intestinal integrity. However, the role of inhaled traffic-generated PM on the gut microbiome and its corresponding systemic effects are not well-characterized. Thus, we investigated the hypothesis that exposure to inhaled diesel exhaust particles (DEP) alters the gut microbiome and promotes microbial-related inflammation and CVD biomarkers. 4-6-week-old male C57Bl/6 mice on either a low-fat (LF, 10% fat) or high-fat (HF, 45% fat) diet were exposed via oropharyngeal aspiration to 35 µg DEP suspended in 35 µl saline or saline only (CON) 2x/week for 30 days. To determine whether probiotics could prevent diet or DEP exposure mediated alterations in the gut microbiome or systemic outcomes, a subset of animals on the HF diet were treated orally with 0.3 g/day (~ 7.5 × 108 CFU/day) of Winclove Ecologic® Barrier probiotics throughout the study. RESULTS: Our results show that inhaled DEP exposure alters gut microbial profiles, including reducing Actinobacteria and expanding Verrucomicrobia and Proteobacteria. We observed increased circulating LPS, altered circulating cytokines (IL-1α, IL-3, IL-13, IL-15, G-CSF, LIF, MIP-2, and TNF-α), and CVD biomarkers (siCAM, PAI-1, sP-Selectin, thrombomodulin, and PECAM) in DEP-exposed and/or HF diet mice. Furthermore, probiotics attenuated the observed reduction of Actinobacteria and expansion of Proteobacteria in DEP-exposed and HF-diet mice. Probiotics mitigated circulating cytokines (IL-3, IL-13, G-CSF, RANTES, and TNF- α) and CVD biomarkers (siCAM, PAI-1, sP-Selectin, thrombomodulin, and PECAM) in respect to DEP-exposure and/or HF diet. CONCLUSION: Key findings of this study are that inhaled DEP exposure alters small intestinal microbial profiles that play a role in systemic inflammation and early CVD biomarkers. Probiotic treatment in this study was fundamental in understanding the role of inhaled DEP on the microbiome and related systemic inflammatory and CVD biomarkers.

Optimized Curcumin, Pomegranate Extract, and Methylsulfonylmethane Reduce Acute, Systemic Inflammatory Response to a Half-marathon Race.

Context: Endurance running places substantial physiological strain on the body, which can develop into chronic inflammation and overuse injuries, negatively affecting subsequent training and performance. A recent study found that dietary polyphenols and methlysulfonylmethane (MSM) can reduce systemic inflammation and oxidative stress without adverse side effects. Objective: The purpose was to identify a set of candidate protein and RNA biomarkers that are associated with improved outcomes related to inflammation and muscle injury, when athletes used 3 proprietary supplements both prior to and during early recovery from a half-marathon race. Design: The study was an open-label pilot study. Setting: The study was field based, with sample analysis conducted in the Applied Physiology Laboratory in the Department of Kinesiology, Health Promotion and Recreation at the University of North Texas in Denton, Texas. Participants: Participants were 15 young, exercise-trained men and women. Intervention: The intervention group consumed 1000 mg/d of a proprietary 50-50 mix of optimized curcumin and pomegranate extract for 26 days. The group also consumed 500 mg/d of a proprietary MSM for the same period. Three days prior to and one day after a race, the daily dosage was doubled. The control group received no supplements. Outcome Measures: Venous blood samples were collected at pre-race and at 4h and 24h after running a half-marathon race. The research team evaluated results for target proteins that have been associated with inflammation and muscle injury in the scientific literature. The team also performed an analysis of RNA biomarkers. Results: At the 4h and 24h time points, a significant treatment-response was observed that included increases in proteins: (1) osteonectin/SPARC-osteonectin/secreted protein acidic and rich in cysteine and (2) BDNF-brain-derived neurotrophic factor. At the same points, the study also found increased RNA: (1) PACER-P50-associated COX-2 extragenic RNA, (2) PTGES-prostaglandin E synthase, (3) MYD88-innate immune signal transduction adaptor MYD88, (4) TNFS14-tumor necrosis factor (TNF) superfamily member 14, (5) THRIL-TNF and heterogeneous nuclear ribonucleoprotein L (HNRNPL)-related immunoregulatory long noncoding RNA, (6) TRAF6-TNF receptor associated factor 6, (7) CX3CL1-C-X3-C motif chemokine ligand 1, (8) MALAT1-metastasis-associated lung adenocarcinoma transcript 1, and (9) LINC00305-long intergenic nonprotein coding RNA 305. Conclusions: The combination of polyphenol and MSM supplementation resulted in a systemic response that may translate to an accelerated rate of muscle recovery, allowing participants return to exercise and normal activities more quickly. This pilot study is the foundation for a larger investigation in the research team's laboratory.

Combining single molecule counting with bead-based multiplexing to quantify biological inflammation time course following skeletal muscle injury.

Bead-based analysis methods allow for the exploration of a variety of complex biological processes. In particular, these techniques can be applied to better understand how peripheral muscle injury contributes to systemic inflammation. Understanding how these two processes affect one another can give additional insight concerning how changes in inflammation effect readiness to perform in exercise and work environments. The present method sought to combine the strengths of bead-based multiplexing with the precision and low-end detection of single molecule counting (SMC) methods. We used performance of an extreme aerobic exercise session (i.e. half-marathon race) to cause a defined quantity of lower body muscle injury and a systemic inflammatory response lasting up to 24 h. Using a high-sensitivity, multiplex assay (Milliplex; Millipore-Sigma) we were able to identify 9 of 21 cytokines that were significantly elevated at either 4 or 24 h post half-marathon performance. Despite the known role of IL-1ß, IL-6, and TNF-α in the pro-inflammatory response, they did not appear to change based on the multiplex analysis. We thus, conducted further analysis using an SMC assay and found increases in IL-1ß, IL-6, and TNF-α at 4 h compared to 24 h post exercise. This method approach demonstrates how combining two common, bead-based protein assays can increase the amount of meaningful biological information that can be collected. We anticipate that this approach will be useful in a variety of inflammation-associated disease states.

Combined bead-based multiplex detection of RNA and protein biomarkers: Implications for understanding the time course of skeletal muscle injury and repair.

Biological response to skeletal muscle injury time course is generally classified as initial (elevated within first 4-h), delayed (elevated at 24-h), and/or prolonged (elevated at 4-h and sustained to 24-h). Accurate description of this process requires the ability to measure a robust set of RNA and protein biomarkers, yet such an approach is not common and not always feasible. This method proposes a novel experimental approach that focuses on the use of bead-based multiplex detection to measure mRNA, lncRNA, cytokines, soluble cytokine receptors, and myokines at 4-h and 24-h post muscle injury. We used an extreme aerobic exercise session (half-marathon race) to create a consistent muscle injury stimulus via oxidative stress and eccentric contractions. Venous blood samples were analyzed to determine the change in 90 targets. Specifically, we identified 14 mRNA, 2 lncRNA, 4 cytokines, and 5 myokines that had only an initial response (change at 4-h). We identified 2 mRNA, 2 cytokines, 13 soluble cytokine receptors, and 1 myokine that had only a delayed response (change at 24-h). Finally, we identified 18 mRNA, 4 lncRNA, 6 myokines and 15 cytokines that had a prolonged response (change at 4-h and sustained at 24-h). We found 4 targets to be undetectable or having no response relative to muscle injury recovery. These findings demonstrate the interplay between RNA and protein biomarkers in response to skeletal muscle injury. This novel experimental application of bead-based multiplexing is applicable to a variety of clinical models that involve muscle injury and/or wasting.

Resistance exercise-induced hormonal response promotes satellite cell proliferation in untrained men but not in women.

The purpose of this work was to determine the effect of resistance exercise (RE)-induced hormonal changes on the satellite cell (SC) myogenic state in response to muscle damage. Untrained men (n = 10, 22 ± 3 yr) and women (n = 9, 21 ± 4 yr) completed 2 sessions of 80 unilateral maximal eccentric knee extensions followed by either an upper body RE protocol (EX) or a 20-min rest (CON). Muscle samples were collected and analyzed for protein content of Pax7, MyoD, myogenin, cyclin D1, and p21 before (PRE), 12 h, and 24 h after the session was completed. Serum testosterone, growth hormone, cortisol, and myoglobin concentrations were analyzed at PRE, post-damage, immediately after (IP), and 15, 30, and 60 min after the session was completed. Testosterone was significantly (P < 0.05) higher immediately after the session in EX vs. CON for men. A significant time × sex × condition interaction was found for MyoD with an increase in EX (men) and CON (women) at 12 h. A significant time × condition interaction was found for Pax7, with a decrease in EX and increase in CON at 24 h. A significant time effect was found for myogenin, p21, and cyclin D1. Myogenin and p21 were increased at 12 and 24 h, and cyclin D1 was increased at 12 h. These results suggest that the acute RE-induced hormonal response can be important for men to promote SC proliferation after muscle damage but had no effect in women. Markers of SC differentiation appeared unaffected by the hormonal response but increased in response to muscle damage.

Flow cytometry what you see matters: Enhanced clinical detection using image-based flow cytometry.

Image-based flow cytometry combines the throughput of traditional flow cytometry with the ability to visually confirm findings and collect novel data that would not be possible otherwise. Since image-based flow cytometry borrows measurement parameters and analysis techniques from microscopy, it is possible to collect unique measures (i.e. nuclear translocation, co-localization, cellular synapse, cellular endocytosis, etc.) that would not be possible with traditional flow cytometry. The ability to collect unique outcomes has led many researchers to develop novel assays for the monitoring and detection of a variety of clinical conditions and diseases. In many cases, investigators have innovated and expanded classical assays to provide new insight regarding clinical conditions and chronic disease. Beyond human clinical applications, image-based flow cytometry has been used to monitor marine biology changes, nano-particles for solar cell production, and particle quality in pharmaceuticals. This review article summarizes work from the major scientists working in the field of image-based flow cytometry.

Consumption of a high-fat, high-calorie meal is associated with an increase in intracellular co-localization of PPAR-γ mRNA and protein in monocytes.

Acute and habitual dietary habits contribute to the onset and progression of many forms of cardiovascular disease. Circulating peripheral blood monocytes have been a target of pre-clinical research related to the risk of atherosclerosis. Specifically, when monocytes migrate into the subendothelial space and endocytosize modified LDL (i.e. acLDL or oxLDL) they phenotypically transform into foam cells. The endocytosis of modified LDL is mediated by the scavenger receptor CD36, whose expression is in tern regulated by the transcription factor PPAR-γ. In this report, we describe a novel technique for the simultaneous measurement of intracellular PPAR-γ mRNA and protein in peripheral blood monocytes collected from human subjects in fasted state or 3 and 5-h after consuming a high-calorie (65% of daily calorie needs), high-fat meal. Intracellular detection and co-localization of PPAR-γ was made possible using a combination of image-based flow cytometry (MilliporeSigma FlowSight) and an amplified mRNA FISH staining technique (Affymetrix/eBioscience PrimeFlow). Consumption of a high-calorie, high-fat meal increased the percentage of co-localization at both 3 and 5-h post prandial compared to pre-meal. No obvious difference in co-localization was observed when cells were treated by acLDL in vitro. More research is needed to determine how to best use this method to study pre-clinical risk of atherosclerosis.

Using image-based flow cytometry to monitor satellite cells proliferation and differentiation in vitro.

Muscle satellite cells can proliferate and differentiate into myocytes; this biological function has important implications for muscle development, aging, repair, and wasting diseases. Established analytical methods, including western blotting, PCR, and immunohistochemistry, has been used to characterize the stages of satellite cells growth and development but there is currently a lack of methods to simultaneously monitor cell proliferation and identify changes in cell properties (i.e., expression of myogenic regulatory factors) for each individual cell during the progression to become myocytes. Image-based flow cytometry has the capability to simultaneously obtain morphometric data, monitor cell division, and detect expression of multiple proteins of interest. In this article we demonstrate the use of image-based flow cytometry and cell-trace dye to monitor satellite cells proliferation and to identify different stages of satellite cell differentiation.

Resistance Training Is Associated With Higher Bone Mineral Density Among Young Adult Male Distance Runners Independent of Physiological Factors.

Duplanty, AA, Levitt, DE, Hill, DW, McFarlin, BK, DiMarco, NM, and Vingren, JL. Resistance training is associated with higher bone mineral density among young adult male distance runners independent of physiological factors. J Strength Cond Res 32(6): 1594-1600, 2018-Low bone mineral density (BMD) in male distance runners is common and could be modulated by a host of biomarkers involved in the dynamic balance of bone tissue. In contrast, resistance training can increase BMD; however, the efficacy of resistance training in protecting BMD in distance runners has not been elucidated. The aim of this study was to investigate the relationship between resistance training, testosterone and bone metabolism biomarker concentrations, and BMD in young adult male distance runners. Twenty-five apparently healthy men (23-32 years; mean ± SD: 25.9 ± 2.9 years; 1.77 ± 0.04 m; 75.4 ± 8.5 kg) were categorized into 1 of 3 groups: untrained control participants (CON; n = 8); nonresistance-trained runners (NRT; n = 8); or resistance-trained runners (RT; n = 9). Blood was collected and analyzed for concentrations of free and total testosterone and 14 bone metabolism biomarkers. Bone mineral density was assessed using dual-energy X-ray absorptiometry. At all measured sites, BMD was greater (p ≤ 0.05) for RT compared with NRT and CON. Vitamin D concentration was greater (p ≤ 0.05) in RT and NRT compared with CON. Concentrations of testosterone and remaining bone biomarkers did not differ between groups (p > 0.05). Resistance-trained runners had greater BMD than nonresistance-trained runners and untrained peers. This difference did not seem to be modulated by biomarkers that contribute to bone formation or resorption, indicating that differences in BMD are associated with habitual load-bearing exercise using external resistance. Runners should perform resistance exercise at least once per week because this is associated with greater BMD.

Adding Resistance Training to the Standard of Care for Inpatient Substance Abuse Treatment in Men With Human Immunodeficiency Virus Improves Skeletal Muscle Health Without Altering Cytokine Concentrations.

Vingren, JL, Curtis, JH, Levitt, DE, Duplanty, AA, Lee, EC, McFarlin, BK, and Hill, DW. Adding resistance training to the standard of care for inpatient substance abuse treatment in men with human immunodeficiency virus improves skeletal muscle health without altering cytokine concentrations. J Strength Cond Res 32(1): 76-82, 2018-Substance abuse and human immunodeficiency virus (HIV) infection can independently lead to myopathy and related inflammatory alterations; importantly, these effects seem to be additive. Resistance training (RT) can improve muscle health in people living with HIV (PLWH), but the efficacy of this intervention has not been examined for PLWH recovering from substance abuse. The purpose of this study was to determine the effect of RT on muscle health markers (mass, strength, and power) and basal circulating biomarkers for men living with HIV undergoing substance abuse treatment. Men living with HIV undergoing 60-day inpatient substance abuse treatment completed either RT (3×/wk) or no exercise training (control) for 6 weeks. Muscle mass, strength, and power, and fasting circulating cytokines (interferon γ, tumor necrosis factor-α, interleukin (IL)-1ß, IL-2, IL-4, IL-6, and IL-10), vascular cellular adhesion molecule-1, and cortisol were measured before (PRE) and after (POST) the 6-week period. Both groups received the standard of care for HIV and substance abuse treatment determined by the inpatient facility. Muscle mass, strength, and power increased (p ≤ 0.05) from PRE to POST for RT but were unchanged for control. No differences were found for circulating biomarkers. Adding RT to the standard of care for substance abuse treatment improved aspects of muscle health (mass, strength, and power) in men living with HIV. These improvements are associated with a lower risk of a number of health conditions. Therefore, practitioners should consider implementing RT interventions as part of substance abuse treatment programs in this population to help manage long-term health.

Synergistic Impact of d-δ-Tocotrienol and Geranylgeraniol on the Growth and HMG CoA Reductase of Human DU145 Prostate Carcinoma Cells.

The growth-suppressive effect of d-δ-tocotrienol and geranylgeraniol is at least partially attributed to their impact on 3-hydroxy-3-methylglutaryl coenzyme A (HMG CoA) reductase, the rate-limiting enzyme in the mevalonate pathway that provides essential intermediates for the posttranslational modification of growth-related proteins including RAS. We hypothesize that these agents synergistically impact cell growth based on their complementary mechanisms of action with HMG CoA reductase. d-δ-tocotrienol (0-40 µmol/L; half maximal inhibitory concentration [IC50] = 15 µmol/L) and geranylgeraniol (0-100 µmol/L; IC50 = 60 µmol/L) each induced concentration-dependent suppression of the growth of human DU145 prostate carcinoma cells. Blends of the two agents synergistically suppressed the growth of DU145 cells, with combination index values ranging 0.67-0.75. While 7.5 µmol/L d-δ-tocotrienol and 30 µmol/L geranylgeraniol individually had no impact on cell cycle distribution in DU145 cells, a blend of the agents induced cell cycle arrest at the G1 phase. The synergistic downregulation of the expression of HMG CoA reductase by 7.5 µmol/L d-δ-tocotrienol and 30 µmol/L geranylgeraniol was accompanied by a reduction in membrane K-RAS protein. Our finding supports the cancer chemopreventive action of plant-based diets and their isoprenoid constituents. Properly formulated isoprenoids and derivatives may provide novel approaches in prostate cancer prevention and therapy.

Effect of alcohol after muscle-damaging resistance exercise on muscular performance recovery and inflammatory capacity in women.

PURPOSE: To investigate the effect of acute alcohol consumption on muscular performance recovery, assessed by maximal torque production, and on inflammatory capacity, assessed by lipopolysaccharide (LPS)-stimulated cytokine production, following muscle-damaging resistance exercise in women. METHODS: Thirteen recreationally resistance-trained women completed two identical exercise bouts (300 maximal single-leg eccentric leg extensions) followed by alcohol (1.09 g ethanol kg-1 fat-free body mass) or placebo ingestion. Blood was collected before (PRE), and 5 (5 h-POST), 24 (24 h-POST), and 48 (48 h-POST) hours after exercise and analyzed for LPS-stimulated cytokine production (TNF-α, IL-1ß, IL-6, and IL-8 and IL-10). Maximal torque production (concentric, eccentric, isometric) was measured for each leg at PRE, 24 h-POST, and 48 h-POST. RESULTS: Although the exercise bout increased LPS-stimulated production of TNF-α (%change from PRE: 5 h-POST 109%; 24 h-POST 49%; 48 h-POST 40%) and decreased LPS-stimulated production of IL-8 (5 h-POST -40%; 24 h-POST -50%; 48 h-POST: -43%) and IL-10 (5 h-POST: -37%; 24 h-POST -32%; 48 h-POST -31%), consuming alcohol after exercise did not affect this response. Regardless of drink condition, concentric, eccentric, and isometric torque produced by the exercised leg were lower at 24 h-POST (concentric 106 ± 6 Nm, eccentric 144 ± 9 Nm, isometric 128 ± 8 Nm; M ± SE) compared to PRE (concentric 127 ± 7 Nm, eccentric 175 ± 11 Nm, isometric 148 ± 8 Nm). Eccentric torque production was partially recovered and isometric torque production was fully recovered by 48 h-POST. CONCLUSIONS: Alcohol consumed after muscle-damaging resistance exercise does not appear to affect inflammatory capacity or muscular performance recovery in resistance-trained women. Combined with previous findings in men, these results suggest a gender difference regarding effects of alcohol on exercise recovery.

Oral Consumption of Vitamin K2 for 8 Weeks Associated With Increased Maximal Cardiac Output During Exercise.

Background • Vitamin K1 and K2 are not typically common in a Western diet because they are found in a variety of fermented foods. Vitamin K2 in particular has been demonstrated to restore mitochondrial function and has a key role in production of mitochondrial adenosine triphosphate. Thus, it is reasonable to speculate that dietary supplementation with vitamin K2 could increase the function of muscle with high mitochondrial content (ie, skeletal and cardiac muscle). Objective • The purpose of this study was to determine if 8 wk of dietary supplementation with Vitamin K2 could alter cardiovascular responses to a graded cycle ergometer test. Design • The study was a randomized controlled trial. Setting • The study took place in the Applied Physiology Laboratory of the Department of Biological Sciences at the University of North Texas (Denton, TX, USA). Participants • Participants were aerobically trained males and female athletes (N = 26). Intervention • Participants were randomly assigned either to a control group that received a rice flour placebo or to an intervention group that received vitamin K2. For weeks 1 to 4, participants received 300 mg/d; for weeks 5 to 8, they received 150 mg/d. Subjects assigned to the control group received similar doses to mirror the intervention group. Subjects consumed the supplements during an 8-wk period while they maintained their typical exercise habits. Outcome Measures • At baseline and postintervention, participants completed a standard, graded exercise test on an electronically braked cycle ergometer. Before the test, participants were fitted with a mouth piece, and their oxygen consumption, carbon dioxide production, respiratory rate, and respiratory exchange ratio were measured. In addition, participants were fitted with skin-mounted electrodes that measured noninvasive cardiac output, stroke volume, and heart rate. To assess the cumulative exercise change, an area-under-the-curve (AUC) value was calculated separately for each outcome variable at each treatment time point. Results • Vitamin K2 supplementation was associated with a 12% increase in maximal cardiac output, with P = .031, with a trend toward an increase in heart-rate AUC, with P = .070. No significant changes occurred in stroke volume. Conclusions • Although vitamin K2 supplementation has previously been reported to improve cardiovascular function in diseased patients, to the research team's knowledge, the current study is the first to report its potential in active individuals. More research is needed to fully evaluate the potential effects of the observed effects.

Effect of Acute Alcohol Ingestion on Resistance Exercise-Induced mTORC1 Signaling in Human Muscle.

Duplanty, AA, Budnar, RG, Luk, HY, Levitt, DE, Hill, DW, McFarlin, BK, Huggett, DB, and Vingren, JL. Effect of acute alcohol ingestion on resistance exercise-induced mTORC1 signaling in human muscle. J Strength Cond Res 31(1): 54-61, 2017-The purpose of this project was to further elucidate the effects postexercise alcohol ingestion. This project had many novel aspects including using a resistance exercise (RE) only exercise design and the inclusion of women. Ten resistance-trained males and 9 resistance-trained females completed 2 identical acute heavy RE trials (6 sets of Smith machine squats) followed by ingestion of either alcohol or placebo. All participants completed both conditions. Before exercise (PRE) and 3 (+3 hours) and 5 (+5 hours) hours postexercise, muscle tissue samples were obtained from the vastus lateralis by biopsies. Muscle samples were analyzed for phosphorylated mTOR, S6K1, and 4E-BP1. For men, there was a significant interaction effect for mTOR and S6K1 phosphorylation. At +3 hours, mTOR and S6K1 phosphorylation was higher for placebo than for alcohol. For women, there was a significant main effect for time. mTOR phosphorylation was higher at +3 hours than at PRE and at +5 hours. There were no significant effects found for 4E-BP1 phosphorylation in men or women. The major findings of this study was that although RE elicited similar mTORC1 signaling both in men and in women, alcohol ingestion seemed to only attenuate RE-induced phosphorylation of the mTORC1 signaling pathway in men. This study provides evidence that alcohol should not be ingested after RE as this ingestion could potentially hamper the desired muscular adaptations to RE by reducing anabolic signaling, at least in men.

The effect of post­resistance exercise alcohol ingestion on lipopolysaccharide­stimulated cytokines.

PURPOSE: To examine the effect of post-resistance exercise alcohol ingestion on lipopolysaccharide (LPS)-stimulated production of IFNγ, TNF-α, IL-1ß, IL-6, IL-8, and IL-10. METHODS: Recreationally resistance-trained men (n = 10, 25 ± 3 year, 177 ± 7 cm, 83.8 ± 15.7 kg, 14.8 ± 8.5% body fat) and women (n = 8, 23 ± 2 year, 161 ± 3 cm, 59.5 ± 6.0 kg, 26.5 ± 3.0% body fat) completed two identical heavy back squat sessions (6 × 10 at 80% 1 repetition maximum) followed by ingestion of either an alcohol (ALC; 1.09 g ethanol · kg fat-free mass(-1)) or water (PLA) drink. Blood samples were collected before exercise (PRE), and at 3 h (3 h), and 5 h (5 h) after exercise, stimulated with LPS, and analyzed for IFNγ, TNF-α, IL-1ß, IL-6, IL-8, and IL-10 concentrations. RESULTS: There were no drink conditions by time effects for IFNγ, TNF-α, IL-1ß, or IL-10. Regardless of condition, resistance exercise induce an increase in IFNγ, TNF-α, and IL-1ß at 5 h compared to PRE but a decrease in IL-10 at 3 and 5 h compared to PRE. For ALC, IL-8 was reduced at 5 h compared to PLA. From PRE to 3 h, IL-6 was reduced for ALC but increased for PLA; resistance exercise induced an increase in IL-6 for both conditions at 5 h. CONCLUSIONS: Heavy resistance exercise increased production of IFNγ, TNF-α, IL-1ß, and Il-6 and decreased production of IL-10. Alcohol ingestion after resistance exercise affected aspects of inflammatory capacity (IL-6 and IL-8 production). It appears that some of the effects previously observed for alcohol ingestion alone on the LPS-stimulated cytokine production were overwhelmed by the response to resistance exercise.

Pro- and anti-inflammatory cytokine responses to a 164-km road cycle ride in a hot environment.

PURPOSE: The purpose of this study was to examine the circulating cytokine response to a recreational 164-km road cycling event in a high ambient temperature and to determine if this response was affected by self-paced exercise time to completion. METHODS: Thirty-five men and five women were divided into tertiles based on time to complete the cycling event: slowest (SLOW), moderate (MOD), and fastest (FAST) finishers. Plasma samples were obtained 1-2 h before (PRE) and immediately after (IP) the event. A high-sensitivity multiplex assay kit was used to determine the concentration of plasma anti-inflammatory cytokines (IL-4, IL-5, IL-10, IL-13) and pro-inflammatory cytokines (IL-1ß, IL-2, IL-6, IL-7, IL-8, IL-12, GM-CSF, IFN-γ, and TNF-α). RESULTS: The concentration of plasma IL-10 increased significantly (p < 0.05) in FAST and MOD groups and had no change in the SLOW group in response to a 164-km cycling event in the hot environment. Other cytokine responses were not influenced by the Time to completion. Pro-inflammatory cytokines IL-1ß, IL-2, GM-CSF, and TNF-α decreased; whereas, IL-6 and IL-8 increased from PRE to IP. Additionally, anti-inflammatory cytokines IL-4 and IL-13 decreased. CONCLUSIONS: Completion of a 164-km cycling event induced substantial changes in circulating pro- and anti-inflammatory cytokine concentrations. Time to completion appears to have a greater influence on the systemic IL-10 response than the environmental condition; however, it is possible that a threshold for absolute intensity must be reached for environmental conditions to affect the IL-10 response to exercise. Thus, cyclists from the FAST/MOD groups appear more likely to experience an acute transient immune suppression than cyclists from the SLOW group.

A shirt containing multistage phase change material and active cooling components was associated with increased exercise capacity in a hot, humid environment.

Recent advances in clothing design include the incorporation of phase change materials (PCM) and other active cooling components (ACC) to provide better body heat dissipation. The purpose of this study was to determine the effect of wearing a shirt containing multistage PCM/ACC on exercise capacity at low (5.0), moderate-high (7.5) and extreme (9.0) levels of the physiological strain index (PSI). Fourteen individuals tested two shirts (control vs. cooling) during 45-min of interval running in a hot, humid (35 ± 1 °C; 55 ± 6% RH) environment. The cooling shirt resulted in an 8% improvement in exercise capacity at a PSI of 7.5 (p < 0.05). The observed increase in exercise capacity would likely translate to a significant improvement in exercise performance. More research is needed to determine a best practice approach for the use of cooling clothing as a counter to exercise-induced heat exposure. Practitioner Summary: In this report, we demonstrate that when forced to exercise in a hot, humid environment, an individual's exercise capacity may increase by as much as 8% when wearing a shirt composed of multistage phase change material and active cooling components.