An observational human study investigating the effect of anabolic androgenic steroid use on the transcriptome of skeletal muscle and whole blood using RNA-Seq.

BACKGROUND: The effects of Anabolic Androgenic Steroids (AAS) are largely illustrated through Androgen Receptor induced gene transcription, yet RNA-Seq has yet to be conducted on human whole blood and skeletal muscle. Investigating the transcriptional signature of AAS in blood may aid AAS detection and in muscle further understanding of AAS induced hypertrophy. METHODS: Males aged 20-42 were recruited and sampled once: sedentary controls (C), resistance trained lifters (RT) and resistance trained current AAS users (RT-AS) who ceased exposure ≤ 2 or ≥ 10 weeks prior to sampling. RT-AS were sampled twice as Returning Participants (RP) if AAS usage ceased for ≥ 18 weeks. RNA was extracted from whole blood and trapezius muscle samples. RNA libraries were sequenced twice, for validation purposes, on the DNBSEQ-G400RS with either standard or CoolMPS PE100 reagents following MGI protocols. Genes were considered differentially expressed with FDR < 0.05 and a 1.2- fold change. RESULTS: Cross-comparison of both standard reagent whole blood (N = 55: C = 7, RT = 20, RT-AS ≤ 2 = 14, RT-AS ≥ 10 = 10, RP = 4; N = 46: C = 6, RT = 17, RT-AS ≤ 2 = 12, RT-AS ≥ 10 = 8, RP = 3) sequencing datasets, showed that no genes or gene sets/pathways were differentially expressed between time points for RP or between group comparisons of RT-AS ≤ 2 vs. C, RT, or RT-AS ≥ 10. Cross-comparison of both muscle (N = 51, C = 5, RT = 17, RT-AS ≤ 2 = 15, RT-AS ≥ 10 = 11, RP = 3) sequencing (one standard & one CoolMPS reagent) datasets, showed one gene, CHRDL1, which has atrophying potential, was upregulated in RP visit two. In both muscle sequencing datasets, nine differentially expressed genes, overlapped with RT-AS ≤ 2 vs. RT and RT-AS ≤ 2 vs. C, but were not differentially expressed with RT vs. C, possibly suggesting they are from acute doping alone. No genes seemed to be differentially expressed in muscle after the long-term cessation of AAS, whereas a previous study found long term proteomic changes. CONCLUSION: A whole blood transcriptional signature of AAS doping was not identified. However, RNA-Seq of muscle has identified numerous differentially expressed genes with known impacts on hypertrophic processes that may further our understanding on AAS induced hypertrophy. Differences in training regimens in participant groupings may have influenced results. Future studies should focus on longitudinal sampling pre, during and post-AAS exposure to better control for confounding variables.

The MMAAS Project: An Observational Human Study Investigating the Effect of Anabolic Androgenic Steroid Use on Gene Expression and the Molecular Mechanism of Muscle Memory.

OBJECTIVE: It remains unknown whether myonuclei remain elevated post anabolic-androgenic steroid (AAS) usage in humans. Limited data exist on AAS-induced changes in gene expression. DESIGN: Cross-sectional/longitudinal. SETTING: University. PARTICIPANTS: Fifty-six men aged 20 to 42 years. INDEPENDENT VARIABLES: Non-resistance-trained (C) or resistance-trained (RT), RT currently using AAS (RT-AS), of which if AAS usage ceased for ≥18 weeks resampled as Returning Participants (RP) or RT previously using AAS (PREV). MAIN OUTCOME MEASURES: Myonuclei per fiber and cross-sectional area (CSA) of trapezius muscle fibers. RESULTS: There were no significant differences between C (n = 5), RT (n = 15), RT-AS (n = 17), and PREV (n = 6) for myonuclei per fiber. Three of 5 returning participants (RP1-3) were biopsied twice. Before visit 1, RP1 ceased AAS usage 34 weeks before, RP2 and RP3 ceased AAS usage ≤2 weeks before, and all had 28 weeks between visits. Fiber CSA decreased for RP1 and RP2 between visits (7566 vs 6629 µm 2 ; 7854 vs 5677 µm 2 ) while myonuclei per fiber remained similar (3.5 vs 3.4; 2.5 vs 2.6). Respectively, these values increased for RP3 between visits (7167 vs 7889 µm 2 ; 2.6 vs 3.3). CONCLUSIONS: This cohort of past AAS users did not have elevated myonuclei per fiber values, unlike previous research, but reported AAS usage was much lower. Training and AAS usage history also varied widely among participants. Comparable myonuclei per fiber numbers despite decrements in fiber CSA postexposure adheres with the muscle memory mechanism, but there is variation in usage relative to sampling date and low numbers of returning participants.

Thermoregulation is not impaired in breast cancer survivors during moderate-intensity exercise performed in warm and hot environments.

This study aimed to assess how female breast cancer survivors (BCS) respond physiologically, hematologically, and perceptually to exercise under heat stress compared to females with no history of breast cancer (CON). Twenty-one females (9 BCS and 12 CON [age; 54 ± 7 years, stature; 167 ± 6 cm, body mass; 68.1 ± 7.62 kg, and body fat; 30.9 ± 3.8%]) completed a warm (25℃, 50% relative humidity, RH) and hot (35℃, 50%RH) trial in a repeated-measures crossover design. Trials consisted of 30 min of rest, 30 min of walking at 4 metabolic equivalents, and a 6-minute walk test (6MWT). Physiological measurements (core temperature (Tre ), skin temperature (Tskin ), heart rate (HR), and sweat analysis) and perceptual rating scales (ratings of perceived exertion, thermal sensation [whole body and localized], and thermal comfort) were taken at 5- and 10-min intervals throughout, respectively. Venous blood samples were taken before and after to assess; IL-6, IL-10, CRP, IFN-γ, and TGF-ß1 . All physiological markers were higher during the 35 versus 25℃ trial; Tre (~0.25℃, p = 0.002), Tskin (~3.8℃, p < 0.001), HR (~12 beats·min-1 , p = 0.023), and whole-body sweat rate (~0.4 L·hr-1 , p < 0.001), with no difference observed between groups in either condition (p > 0.05). Both groups covered a greater 6MWT distance in 25 versus 35℃ (by ~200 m; p = 0.003). Nevertheless, the control group covered more distance than BCS, regardless of environmental temperature (by ~400 m, p = 0.03). Thermoregulation was not disadvantaged in BCS compared to controls during moderate-intensity exercise under heat stress. However, self-paced exercise performance was reduced for BCS regardless of environmental temperature.

Exercise heat acclimation and post-exercise hot water immersion improve resting and exercise responses to heat stress in the elderly.

OBJECTIVES: To investigate the efficacy of heat acclimation (HA) in the young (YEX) and elderly (EEX) following exercise-HA, and the elderly utilising post-exercise hot water immersion HA (EHWI). DESIGN: Cross-sectional study. METHOD: Twenty-six participants (YEX: n = 11 aged 22 ±â€¯2 years, EEX:n = 8 aged 68 ±â€¯3 years, EHWI: n = 7 aged 73 ±â€¯3 years) completed two pre-/post-tests, separated by five intervention days. YEX and EEX exercised in hot conditions to raise rectal temperature (Trec) ≥38.5 °C within 60 min, with this increase maintained for a further 60 min. EHWI completed 30 min of cycling in temperate conditions, then 30 min of HWI (40 °C), followed by 30 min seated blanket wrap. Pre- and post-testing comprised 30 min rest, followed by 30 min of cycling exercise (3.5 W·kg-1 Hprod), and a six-minute walk test (6MWT), all in 35 °C, 50% RH. RESULTS: The HA protocols did not elicit different mean heart rate (HR), Trec, and duration Trec ≥ 38.5 °C (p > 0.05) between YEX, EEX, and EHWI groups. Resting Trec, peak skin temperature, systolic and mean arterial pressure, perceived exertion and thermal sensation decreased, and 6MWT distance increased pre- to post-HA (p < 0.05), with no difference between groups. YEX also demonstrated a reduction in resting HR (p < 0.05). No change was observed in peak Trec or HR, vascular conductance, sweat rate, or thermal comfort in any group (p > 0.05). CONCLUSIONS: Irrespective of age or intervention, HA induced thermoregulatory, perceptual and exercise performance improvements. Both exercise-HA (EEX), and post-exercise HWI (EHWI) are considered viable interventions to prepare the elderly for heat stress.

Validity of a wearable sweat rate monitor and routine sweat analysis techniques using heat acclimation.

INTRODUCTION: the aim of this study was to assess the validity of a novel wearable sweat rate monitor against an array of sweat analysis techniques which determine sudomotor function when exercising moderately under heat stress. Construct validity was determined utilising a 5-day short-term heat acclimation (STHA) intervention. METHODS: Nineteen healthy individuals (age: 41 ± 23 years, body mass: 74.0 ± 12.2 kg, height: 174.9 ± 6.9 cm) [male; n = 15, female; n = 4] completed nine trials over a three-week period, in a controlled chamber set to 35 °C, 50% relative humidity for all sessions. The pre and post-trials were separated by five consecutive controlled hyperthermia HA sessions. Sweat analysis was compared from pre and post-trial, whereby whole body sweat rate (WBSR) was assessed via pre and post nude body mass. Local sweat rate (LSR) was determined via technical absorbent patches (TA) (weighed pre and post) and a novel wearable KuduSmart® (SMART) monitor which was placed on the left arm during the 30-min of exercise. Tegaderm patches, used to measure sweat sodium chloride conductivity (SC), and TA patches were placed on the back, chest and forearm for the 30-min cycling. RESULTS: Sudomotor function significantly adapted via STHA (p < 0.05); demonstrated by a WBSR increase of 24%, LSR increase via the TA method (back: 26%, chest: 45% and arm: 48%) and LSR increase by the SMART monitor (35%). Finally, SC decreased (back: -21%, chest: -25% and arm: -24%, p < 0.05). CONCLUSION: All sweat techniques were sensitive to sudomotor function adaptation following STHA, reinforcing their validity. The real time data given by the wearable KuduSmart® monitor provides coaches and athletes instant comparable sudomotor function feedback to traditional routinely used sweat analysis techniques.