The myonuclear domain in adult skeletal muscle fibres: past, present and future.

Most cells in the body are mononuclear whereas skeletal muscle fibres are uniquely multinuclear. The nuclei of muscle fibres (myonuclei) are usually situated peripherally which complicates the equitable distribution of gene products. Myonuclear abundance can also change under conditions such as hypertrophy and atrophy. Specialised zones in muscle fibres have different functions and thus distinct synthetic demands from myonuclei. The complex structure and regulatory requirements of multinuclear muscle cells understandably led to the hypothesis that myonuclei govern defined 'domains' to maintain homeostasis and facilitate adaptation. The purpose of this review is to provide historical context for the myonuclear domain and evaluate its veracity with respect to mRNA and protein distribution resulting from myonuclear transcription. We synthesise insights from past and current in vitro and in vivo genetically modified models for studying the myonuclear domain under dynamic conditions. We also cover the most contemporary knowledge on mRNA and protein transport in muscle cells. Insights from emerging technologies such as single myonuclear RNA-sequencing further inform our discussion of the myonuclear domain. We broadly conclude: (1) the myonuclear domain can be flexible during muscle fibre growth and atrophy, (2) the mechanisms and role of myonuclear loss and motility deserve further consideration, (3) mRNA in muscle is actively transported via microtubules and locally restricted, but proteins may travel far from a myonucleus of origin and (4) myonuclear transcriptional specialisation extends beyond the classic neuromuscular and myotendinous populations. A deeper understanding of the myonuclear domain in muscle may promote effective therapies for ageing and disease.

Gut check: Unveiling the influence of acute exercise on the gut microbiota.

The human gastrointestinal microbiota and its unique metabolites regulate a diverse array of physiological processes with substantial implications for human health and performance. Chronic exercise training positively modulates the gut microbiota and its metabolic output. The benefits of chronic exercise for the gut microbiota may be influenced by acute changes in microbial community structure and function that follow a single exercise bout (i.e., acute exercise). Thus, an improved understanding of changes in the gut microbiota that occur with acute exercise could aid in the development of evidence-based exercise training strategies to target the gut microbiota more effectively. In this review, we provide a comprehensive summary of the existing literature on the acute and very short-term (<3 weeks) exercise responses of the gut microbiota and faecal metabolites in humans. We conclude by highlighting gaps in the literature and providing recommendations for future research in this area. NEW FINDINGS: What is the topic of this review? The chronic benefits of exercise for the gut microbiota are likely influenced by acute changes in microbial community structure and function that follow a single exercise bout. This review provides a summary of the existing literature on acute exercise responses of the gut microbiota and its metabolic output in humans. What advances does it highlight? Acute aerobic exercise appears to have limited effects on diversity of the gut microbiota, variable effects on specific microbial taxa, and numerous effects on the metabolic activity of gut microbes with possible implications for host health and performance.

Myosin Heavy Chain Composition, Creatine Analogues, and the Relationship of Muscle Creatine Content and Fast-Twitch Proportion to Wilks Coefficient in Powerlifters.

Machek, SB, Hwang, PS, Cardaci, TD, Wilburn, DT, Bagley, JR, Blake, DT, Galpin, AJ, and Willoughby, DS. Myosin heavy chain composition, creatine analogues, and the relationship of muscle creatine content and fast-twitch proportion to Wilks coefficient in powerlifters. J Strength Cond Res 34(11): 3022-3030, 2020-Little data exist on powerlifting-specific skeletal muscle adaptations, and none elucidate sex differences in powerlifters. Powerlifters tend to display higher fast-twitch fiber content and phosphagen system dependence. Nevertheless, it is unknown whether fast-twitch fiber or muscle creatine content are predictive of competitive powerlifting performance (via Wilks coefficient). Twelve actively competing powerlifters (PL; n = 6M/6F; age = 21.3 ± 1.0; 3.0 ± 1.8 year competing; 7.3 ± 6.6 meets attended) and 10 sedentary controls (CON; n = 5M/5F; age = 19.4 ± 2.0 year) underwent vastus lateralis muscle biopsies and venipuncture to compare the myosin heavy chain (MHC) fiber type and creatine analogue profiles between groups of both sexes, and determine whether MHC IIa and muscle total creatine (MTC) composition predict powerlifting performance. Samples were analyzed for specific MHC isoform (I, IIa, and IIx) content via mixed homogenate SDS-PAGE, and creatine analogues (MTC, muscle creatine transporter [SLC6A8], serum total creatine [STC], and serum creatinine [CRT]). Furthermore, MHC IIa and MTC content were compared with Wilks coefficient using Pearson correlation coefficients. Male PL MHC content was 50 ± 6% I, 45 ± 6% IIa, and 5 ± 11% IIx, versus 46 ± 6% I, 53 ± 6 IIa, and 0% IIx in female PL. Conversely, male CON MHC content was 33 ± 5% I, 38 ± 7% IIa, and 30 ± 8% IIx, vs. 35 ± 9% I, 44 ± 8% IIa, and 21 ± 17% IIx in female CON. Muscle total creatine, SLC6A8, STC, and CRT did not significantly differ between groups nor sexes. Finally, neither MHC IIa content (r = -0.288; p = 0.364) nor MTC (r = 0.488; p = 0.108) significantly predicted Wilks coefficient, suggesting these characteristics alone do not determine powerlifting skill variation.

Epigenetic Responses to Acute Resistance Exercise in Trained vs. Sedentary Men.

Bagley, JR, Burghardt, KJ, McManus, R, Howlett, B, Costa, PB, Coburn, JW, Arevalo, JA, Malek, MH, and Galpin, AJ. Epigenetic responses to acute resistance exercise in trained vs. sedentary men. J Strength Cond Res 34(6): 1574-1580, 2020-Acute resistance exercise (RE) alters DNA methylation, an epigenetic process that influences gene expression and regulates skeletal muscle adaptation. This aspect of cellular remodeling is poorly understood, especially in resistance-trained (RT) individuals. The study purpose was to examine DNA methylation in response to acute RE in RT and sedentary (SED) young men, specifically targeting genes responsible for metabolic, inflammatory, and hypertrophic muscle adaptations. Vastus lateralis biopsies were performed before (baseline), 30 minutes after, and 4 hours after an acute RE bout (3 × 10 repetitions at 70% 1 repetition maximum [1RM] leg press and leg extension) in 11 RT (mean ± SEM: age = 26.1 ± 1.0 years; body mass = 84.3 ± 0.2 kg; leg press 1RM = 412.6 ± 25.9 kg) and 8 SED (age = 22.9 ± 1.1 years; body mass = 75.6 ± 0.3 kg; leg press 1RM = 164.8 ± 22.5 kg) men. DNA methylation was analyzed through methylation sensitive high-resolution melting using real-time polymerase chain reaction. Separate 2 (group) × 3 (time) repeated-measures analyses of variance and analyses of covariance were performed to examine changes in DNA methylation for each target gene. Results showed that acute RE (a) hypomethylated LINE-1 (measure of global methylation) in RT but not SED, (b) hypermethylated metabolic genes (GPAM and SREBF2) in RT, while lowering SREBF2 methylation in SED, and (c) did not affect methylation of genes associated with inflammation (IL-6 and TNF-α) or hypertrophy (mTOR and AKT1). However, basal IL-6 and TNF-α were lower in SED compared with RT. These findings indicate the same RE stimulus can illicit different epigenetic responses in RT vs. SED men and provides a molecular mechanism underpinning the need for differential training stimuli based on subject training backgrounds.

Gut Microbiota Composition Is Related to Cardiorespiratory Fitness in Healthy Young Adults.

Bacteria residing in the human gastrointestinal tract has a symbiotic relationship with its host. Animal models have demonstrated a relationship between exercise and gut microbiota composition. This was the first study to explore the relationship between cardiorespiratory fitness (maximal oxygen consumption, VO2max) and relative gut microbiota composition (Firmicutes to Bacteroidetes ratio [F/B]) in healthy young adults in a free-living environment. Twenty males and 17 females (25.7 ± 2.2 years), who did not take antibiotics in the last 6 months, volunteered for this study. VO2max was measured using a symptom-limited graded treadmill test. Relative microbiota composition was determined by analyzing DNA extracted from stool samples using a quantitative polymerase chain reaction that specifically measured the quantity of a target gene (16S rRNA) found in Firmicutes and Bacteroidetes. Relationships between F/B and potentially related dietary, anthropometric, and fitness variables were assessed using correlation analyses with an appropriate Bonferroni adjustment (p < .004). The average F/B ratio in all participants was 0.94 ± 0.03. The F/B ratio was significantly correlated to VO2max (r = .48, p < .003), but no other fitness, nutritional intake, or anthropometric variables (p > .004). VO2max explained ∼22% of the variance of an individual's relative gut bacteria as determined by the F/B ratio. These data support animal findings, demonstrating a relationship between relative human gut microbiota composition and cardiorespiratory fitness in healthy young adults. Gastrointestinal bacteria is integral in regulating a myriad of physiological processes, and greater insight regarding ramifications of exercise and nutrition on gut microbial composition may help guide therapies to promote human health.

Muscle health and performance in monozygotic twins with 30 years of discordant exercise habits.

INTRODUCTION: Physical health and function depend upon both genetic inheritance and environmental factors (e.g., exercise training). PURPOSE: To enhance the understanding of heritability/adaptability, we explored the skeletal muscle health and physiological performance of monozygotic (MZ) twins with > 30 years of chronic endurance training vs. no specific/consistent exercise. METHODS: One pair of male MZ twins (age = 52 years; Trained Twin, TT; Untrained Twin, UT) underwent analyses of: (1) anthropometric characteristics and blood profiles, (2) markers of cardiovascular and pulmonary health, and (3) skeletal muscle size, strength, and power and molecular markers of muscle health. RESULTS: This case study represents the most comprehensive physiological comparison of MZ twins with this length and magnitude of differing exercise history. TT exhibited: (1) lower body mass, body fat%, resting heart rate, blood pressure, cholesterol, triglycerides, and plasma glucose, (2) greater relative cycling power, anaerobic endurance, and aerobic capacity (VO2max), but lower muscle size/strength and poorer muscle quality, (3) more MHC I (slow-twitch) and fewer MHC IIa (fast-twitch) fibers, (4) greater AMPK protein expression, and (5) greater PAX7, IGF1Ec, IGF1Ea, and FN14 mRNA expression than UT. CONCLUSIONS: Several measured differences are the largest reported between MZ twins (TT expressed 55% more MHC I fibers, 12.4 ml/kg/min greater VO2max, and 8.6% lower body fat% vs. UT). These data collectively (a) support utilizing chronic endurance training to improve body composition and cardiovascular health and (b) suggest the cardiovascular and skeletal muscle systems exhibit greater plasticity than previously thought, further highlighting the importance of studying MZ twins with large (long-term) differences in exposomes.

Skeletal Muscle Fatigability and Myosin Heavy Chain Fiber Type in Resistance Trained Men.

Bagley, JR, McLeland, KA, Arevalo, JA, Brown, LE, Coburn, JW, and Galpin, AJ. Skeletal muscle fatigability and myosin heavy chain fiber type in resistance trained men. J Strength Cond Res 31(3): 602-607, 2017-Forty years ago, Thorstensson and Karlsson in 1976 described the link between muscle fatigability and fiber type, finding that more fast-twitch fibers were associated with a quicker onset of quadriceps fatigue. This provided the foundation for the Classic Thorstensson Test of fatigability and subsequent noninvasive fiber type prediction equation. This equation was developed with data from recreationally active (REC) men but has been implemented in participants with heterogeneous physical activity/exercise backgrounds. The accuracy of this approach in resistance trained (RET) men has not been established. Moreover, muscle fiber typing techniques have evolved considerably since this seminal work. Therefore, we reexamined this relationship using RET men and a more sensitive fiber typing method (single fiber myosin heavy chain [MHC] isoform classification). Fifteen RET men (age = 24.8 ± 1.3 years) performed maximal knee extensions (via isokinetic dynamometry) to determine peak torque (PT) and quadriceps fatigue percentage (FP) after 30 and 50 repetitions. Vastus lateralis (VL) single fiber MHC type was determined and fibers were grouped as %Fast (expressing MHC IIa, IIa/IIX, or IIx; no MHC I containing fibers). Resistance trained men exhibited 46% greater PT (RET = 207 ± 28 N·m vs. REC = 130 ± 8 N·m) and 28% more %Fast (RET = 61 ± 4% vs. REC = 44 ± 4%) than REC men. Additionally, RET men had a relatively homogeneous FP (64 ± 1%) ranging from 53 to 72%. No relationship was found between FP and MHC fiber type (R = 0.01, p > 0.05). The Classic Thorstensson Test may not accurately estimate VL fiber type composition in RET men, highlighting the (a) unique phenotypical/functional adaptations induced by chronic RET and (b) the need for more sensitive cellular/molecular analyses in RET muscle.

Improving human skeletal muscle myosin heavy chain fiber typing efficiency.

Single muscle fiber sodium dodecyl sulfate polyacrylamide gel-electrophoresis (SDS-PAGE) is a sensitive technique for determining skeletal muscle myosin heavy chain (MHC) composition of human biopsy samples. However, the number of fibers suitable to represent fiber type distribution via this method is undefined. Muscle biopsies were obtained from the vastus lateralis (VL) of nine resistance-trained males (25 ± 1 year, height = 179 ± 5 cm, mass = 82 ± 8 kg). Single fiber MHC composition was determined via SDS-PAGE. VL fiber type distribution [percent MHC I, I/IIa, IIa, IIa/IIx, and total "hybrids" (i.e. I/IIa + IIa/IIx)] was evaluated according to number of fibers analyzed per person (25 vs. 125). VL fiber type distribution did not differ according to number of fibers analyzed (P > 0.05). VL biopsy fiber type distribution of nine subjects is represented by analyzing 25 fibers per person. These data may help minimize cost, personnel-time, and materials associated with this technique, thereby improving fiber typing efficiency in humans.

Cardiovascular and Perceptual Responses to an Ultraendurance Channel Swim: A Case Study.

Ultraendurance open water swimming presents unique physiological challenges. This case study aimed to describe cardiovascular and perceptual responses during a successful solo channel swim. Investigators followed a female swimmer's Catalina Channel (32.2 km) crossing, monitoring water temperature (T(water)) and air temperature (T(air)), distance remaining (DR), average velocity, and heart rate (HR(swim)) at regular intervals. Every 24 minutes, the swimmer reported perceived pain (on a scale of 0-10), rating of perceived exertion (RPE [scale of 6-20]), perceived thermal sensation (scale 0-8), and thirst (scale 1-9). Data are presented as mean ± SD where applicable. The participant finished in 9 hours, 2 minutes, and 48 seconds; T(water) averaged 19.1 ± 0.4ºC, and T(air) averaged 18.6 ± 0.9ºC. Her HR(swim) ranged from 148 to 155 beats/min, and thermal sensation ranged from 3 to 4. Pain inconsistently varied from 0 to 5 during the swim. The RPE remained between 12 and 14 for the first 8 hours, but increased dramatically near the end (reaching 18). Thirst sensation steadily increased throughout the swim, again reaching maximal values on completion. Physiologically and statistically significant correlations existed between thirst and DR (r = -0.905), RPE and HR(swim) (r = 0.741), RPE and DR (r = -0.694), and pain and DR (r = -0.671). The primary findings were that, despite fluctuations in perceptual stressors, the swimmer maintained a consistent exercise intensity as indicated by HR(swim); and during ultraendurance swimming, pain, RPE, and thirst positively correlated with distance swum. We hope these findings aid in the preparation and performance of future athletes by providing information on what swimmers may expect during an ultraendurance attempt and by increasing the understanding of physiological and perceptual responses during open water swimming.

Actual vs. perceived exertion during active virtual reality game exercise.

Background: Virtual exercise has become more common as emerging and converging technologies make active virtual reality games (AVRGs) a viable mode of exercise for health and fitness. Our lab has previously shown that AVRGs can elicit moderate to vigorous exercise intensities that meet recommended health benefit guidelines. Dissociative attentional focuses during AVRG gameplay have the potential to widen the gap between participants' perception of exertion and actual exertion. Objective: The aim of this study was to determine actual exertion (AEx) vs. perceived exertion (PEx) levels during AVRGs by measuring heart rate (HR) and ratings of perceived exertion (RPE) in two different settings. Materials and methods: HR and RPE were collected on participants (N = 32; age 22.6 ± 2.6) during 10 min of gameplay in LabS and GymS using the HTC VIVE with the following games played: Fruit Ninja VR (FNVR), Beat Saber (BS), and Holopoint (HP). Results: Participants exhibited significantly higher levels of AEx compared to reported PEx for all three AVRGs (Intensity): FNVR [AEx = 11.6 ± 1.8 (Light), PEx = 9.0 ± 2.0 (Very Light)], BS [AEx = 11.3 ± 1.7 (Light), PEx = 10.3 ± 2.1 (Very Light)], HP [AEx = 13.1 ± 2.3 (Somewhat Hard), PEx = 12.3 ± 2.4 (Light-Somewhat Hard)]. Additionally, participants playing in the GymS experienced significantly higher levels of AEx [12.4 ± 2.3 (Light-Somewhat Hard)] and PEx [10.8 ± 2.5 (Very Light-Light)] compared to the LabS [AEx = 11.6 ± 1.8 (Light), PEx = 10.3 ± 2.6 (Very Light-Light)]. Conclusion: Perceptions of exertion may be lower than actual exertion during AVRG gameplay, and exertion levels can be influenced by the setting in which AVRGs are played. This may inform VR developers and health clinicians who aim to incorporate exercise/fitness regimens into upcoming 'virtual worlds' currently being developed at large scales (i.e., the "metaverse").

Validity of field expedient devices to assess core temperature during exercise in the cold.

INTRODUCTION: Exposure to cold environments affects human performance and physiological function. Major medical organizations recommend rectal temperature (TREC) to evaluate core body temperature (TcORE) during exercise in the cold; however, other field expedient devices claim to measure TCORE. The purpose of this study was to determine if field expedient devices provide valid measures of TcRE during rest and exercise in the cold. METHODS: Participants included 13 men and 12 women (age = 24 +/- 3 yr, height = 170.7 +/- 10.6 cm, mass = 73.4 +/- 16.7 kg, body fat = 18 +/- 7%) who reported being healthy and at least recreationally active. During 150 min of cold exposure, subjects sequentially rested for 30 min, cycled for 90 min (heart rate = 120-140 bpm), and rested for an additional 30 min. Investigators compared aural (T(AUR)), expensive axillary (T(AXLe)), inexpensive axillary (T(AXLi)), forehead (T(FOR)), gastrointestinal (T(GI)), expensive oral (T(ORLe)), inexpensive oral (T(ORLi)), and temporal (T(TEM)) temperatures to T(REc) every 15 min. Researchers used mean difference between each device and T(REC) (i.e., mean bias) as the primary criterion for validity. RESULTS: T(AUR), T(AXLe), T(AXLi), T(FOR), TORLe, T(ORLi), and TTEM provided significantly lower measures compared to T(REC) and fell below our validity criterion. T(GI) significantly exceeded T(REC) at three of eleven time points, but no significant difference existed between mean T(REC) and T(GI) across time. Only T(GI) achieved our validity criterion and compared favorably to T(REC). CONCLUSION: T(GI) offers a valid measurement with which to assess T(CORE) during rest and exercise in the cold; athletic trainers, mountain rescuers, and military medical personnel should avoid other field expedient devices in similar conditions.

Rapid gut microbiome changes in a world-class ultramarathon runner.

The human gut microbiome is a dynamic ecosystem with prolific health connotations. Physical activity is emerging as a potent regulator of human microbiome composition. This study examined changes in the gut microbiome of a world-class ultramarathon runner before and after competing in the Western States Endurance Run (WSER), a 163 km mountain footrace. Anthropometrics and body composition were assessed and the ultramarathoner's submaximal and maximal performance profiles were evaluated. Gut microbiome analyses were performed at four time-points: 21 weeks and 2 weeks before and 2 hours and 10 days after WSER. Aerobic power (VO2 max) was 4.24 L/min (66.7 ml kg-1  min-1 ), and running economy (51.1 ml kg-1  min-1 at 268 m/min) and lactate threshold (~83% VO2 max) values were comparable to that of highly trained distance runners. Two hours post-race, considerable changes in the ultrarunners' gut microbiome were observed. Alpha diversity (Shannon Diversity Index) increased from 2.73 to 2.80 and phylum-level bacterial composition (Firmicutes/Bacteroidetes ratio) rose from 4.4 to 14.2. Underlying these macro-level microbial alterations were demonstrable increases in select bacterial genera such as Veillonella (+14,229%) and Streptococcus (+438%) concomitant with reductions in Alloprevotella (-79%) and Subdolingranulum (-50%). To our knowledge, this case study shows the most rapid and pronounced shifts in human gut microbiome composition after acute exercise in the human literature. These findings provide yet another example of how exercise can be a powerful modulator of human health.

Extraordinary fast-twitch fiber abundance in elite weightlifters.

Human skeletal muscle fibers exist across a continuum of slow → fast-twitch. The amount of each fiber type (FT) influences muscle performance but remains largely unexplored in elite athletes, particularly from strength/power sports. To address this nescience, vastus lateralis (VL) biopsies were performed on World/Olympic (female, n = 6, "WCF") and National-caliber (female, n = 9, "NCF"; and male, n = 6, "NCM") American weightlifters. Participant accolades included 3 Olympic Games, 19 World Championships, 25 National records, and >170 National/International medals. Samples were analyzed for myosin heavy chain (MHC) content via SDS-PAGE using two distinct techniques: single fiber (SF) distribution (%) and homogenate (HG) composition. The main finding was that these athletes displayed the highest pure MHC IIa concentrations ever reported in healthy VL (23±9% I, 5±3% I/IIa, 67±13% IIa, and 6±10% IIa/IIx), with WCF expressing a notable 71±17% (NCF = 67±8%, NCM = 63±16%). No pure MHC IIx were found with SF. Secondary analysis revealed the heavyweights accounted for 91% of the MHC IIa/IIx fibers, which caused a correlation between this FT and body mass. Additionally, when compared to SF, HG overestimated MHC I (23±9 vs. 31±9%) and IIx (0±0 vs. 3±6%) by misclassifying I/IIa fibers as I and IIa/IIx fibers as IIx, highlighting the limitation of HG as a measure of isoform distribution. These results collectively suggest that athlete caliber (World vs. National) and/or years competing in the sport determine FT% more than sex, particularly for MHC IIa. The extreme fast-twitch myofiber abundance likely explains how elite weightlifters generate high forces in rapid time-frames.

Metabolic Cost and Exercise Intensity During Active Virtual Reality Gaming.

OBJECTIVE: The aim of this study was to investigate the exercise intensity of active virtual reality games (AVRGs) by oxygen consumption (VO2), heart rate (HR), and rating of perceived exertion (RPE). A second aim was to compare the AVRG intensities to current American College of Sports Medicine exercise guidelines using metabolic equivalents (METs) and %VO2 reserve (%VO2R). MATERIALS & METHODS: HR, VO2, and RPE were collected on participants (N = 41; age: 25.2 ± 4.4 years) during 10-minutes of supine rest and while the participants played each of the following AVRGs for 10 minutes: Thrill of the Fight (TOF), Audioshield (AS), and Holopoint (HP). RESULTS: Compared to resting values of HR (63 ± 10 bpm) and VO2 (4.9 ± 0.6 mL/[kg·min]), there were significant elevations in these variables during TOF (149 ± 16 bpm and 32.5 ± 7.1 mL/[kg·min]), AS (131 ± 24 bpm and 19.1 ± 5.9 mL/[kg·min]), and HP (135 ± 22 bpm and 24.8 ± 6.6 mL/[kg·min]). Based on 95% confidence intervals (CI) of %VO2R, TOF was classified vigorous (68.6% ± 2.8%), HP moderate (49.7% ± 2.7%), and AS light intensity (35.7% ± 2.4%). The 95% CI of METs indicated that TOF was classified vigorous (9.3 ± 0.3 METs), HP moderate to vigorous (7.1 ± 0.3 METs), and AS moderate intensity (5.5 ± 0.3 METs). Lastly, 95% CI of RPE led to TOF being classified as moderate (12.7 ± 0.4), whereas HP (10.5 ± 0.4) and AS (9.3 ± 0.3) were light intensity. CONCLUSIONS: These data suggest that these AVRGs can elicit significant increases in VO2 that are game-dependent, indicating increased energy expenditure. Furthermore, each game had a lower intensity categorization based on RPE compared to %VO2R or METs. These data provide information on the metabolic cost of movement-specific games and may aid consumers and fitness specialists in developing exercise programs with AVRGs.

Fiber type-specific analysis of AMPK isoforms in human skeletal muscle: advancement in methods via capillary nanoimmunoassay.

Human skeletal muscle is a heterogeneous mixture of multiple fiber types (FT). Unfortunately, present methods for FT-specific study are constrained by limits of protein detection in single-fiber samples. These limitations beget compensatory resource-intensive procedures, ultimately dissuading investigators from pursuing FT-specific research. Additionally, previous studies neglected hybrid FT, confining their analyses to only pure FT. Here we present novel methods of protein detection across a wider spectrum of human skeletal muscle FT using fully automated capillary nanoimmunoassay (CNIA) technology. CNIA allowed a ~20-fold-lower limit of 5'-AMP-activated protein kinase (AMPK) detection compared with Western blotting. We then performed FT-specific assessment of AMPK expression as a proof of concept. Individual human muscle fibers were mechanically isolated, dissolved, and myosin heavy chain (MHC) fiber typed via SDS-PAGE. Single-fiber samples were combined in pairs and grouped into MHC I, MHC I/IIa, MHC IIa, and MHC IIa/IIx for expression analysis of AMPK isoforms α1, α2, ß1, ß2, γ2, and γ3 with a tubulin loading control. Significant FT-specific differences were found for α2 (1.7-fold higher in MHC IIa and MHC IIa/IIx vs. others), γ2 (2.5-fold higher in MHC IIa vs. others), and γ3 (2-fold higher in MHC IIa and 4-fold higher in MHC IIa/IIx vs. others). Development of a protocol that combines the efficient and sensitive CNIA technology with comprehensive SDS-PAGE fiber typing marks an important advancement in FT-specific research because it allows more precise study of the molecular mechanisms governing metabolism, adaptation, and regulation in human muscle. NEW & NOTEWORTHY We demonstrate the viability of applying capillary nanoimmunoassay technology to the study of fiber type-specific protein analysis in human muscle fibers. This novel technique enables a ~20-fold-lower limit of protein detection compared with traditional Western blotting methods. Combined with SDS-PAGE methods of fiber typing, we apply this technique to compare 5'-AMP-activated protein kinase isoform expression in myosin heavy chain (MHC) I, MHC I/IIa, MHC IIa, and MHC IIa/IIx fiber types.

Lower-Limb Dominance, Performance, and Fiber Type in Resistance-trained Men.

INTRODUCTION: Large imbalances between limbs are common and potentially dangerous, yet few studies have simultaneously examined performance and physiological asymmetries. The current study examined the associations between lower-limb dominance, drop-jumping kinematics, maximal strength, and myosin heavy-chain (MHC) fiber type in the vastus lateralis. METHODS: Thirteen resistance-trained men (age, 24.3 ± 2.7 yr; height, 181.4 ± 6.6 cm; mass, 87.7 ± 11.3 kg) identified their dominant (DOM) and nondominant (ND) limb, performed drop jumps (30 cm) and maximal knee extensions (1-repetition maximum, or 1RM), and provided biopsies from both vastus lateralis muscles for single-fiber (109 ± 36 per limb per person) MHC fiber-type identification (FT%). RESULTS: All participants selected "right" as the "preferred kicking limb" (DOM). DOM displayed a trend for a greater eccentric knee angular velocity (EKV; P = 0.083) and a significantly greater concentric knee angular velocity (CKVl P = 0.002) during drop jump. DOM also tended to be stronger than ND (64.3 ± 11.3 vs 61.0 ± 8.8 kg, P = 0.063). Slow-twitch (MHC I) fibers were more prevalent in DOM (P < 0.025), whereas ND contained more fast-twitch (MHC IIa; P < 0.025). No correlations existed between categories (jumping, 1RM, and FT%). Asymmetries of >5% were present in 6 of 12 participants for EKV, 2 of 12 for CKV, 6 of 13 for 1RM, 12 of 13 for MHC I, and 11 of 13 for MHC IIa. However, only a single participant expressed asymmetries of >5% in all dependent variables (EKV, CKV, 1RM, MHC I, and MHC IIa). CONCLUSIONS: Several statistically and clinically relevant asymmetries were identified. The FT% differences between lower limbs were large and common. The findings also seem to conclude that DOM was stronger, moved faster, and contained more MHC I. However, only 23% of participants actually displayed that result. This highlights the need to analyze and report both group and individual data, particularly when interpreting findings across multiple related, but not necessarily causal, measurements.

Skeletal Muscle Hypertrophy with Concurrent Exercise Training: Contrary Evidence for an Interference Effect.

Over the last 30+ years, it has become axiomatic that performing aerobic exercise within the same training program as resistance exercise (termed concurrent exercise training) interferes with the hypertrophic adaptations associated with resistance exercise training. However, a close examination of the literature reveals that the interference effect of concurrent exercise training on muscle growth in humans is not as compelling as previously thought. Moreover, recent studies show that, under certain conditions, concurrent exercise may augment resistance exercise-induced hypertrophy in healthy human skeletal muscle. The purpose of this article is to outline the contrary evidence for an acute and chronic interference effect of concurrent exercise on skeletal muscle growth in humans and provide practical literature-based recommendations for maximizing hypertrophy when training concurrently.

Effects of Intermittent Neck Cooling During Repeated Bouts of High-Intensity Exercise.

The purpose of this investigation was to determine the influence of intermittent neck cooling during exercise bouts designed to mimic combat sport competitions. Participants (n = 13, age = 25.3 ± 5.0 year height = 176.9 ± 7.5 cm, mass = 79.3 ± 9.0 kg, body fat = 11.8% ± 3.1%) performed three trials on a cycle ergometer. Each trial consisted of two, 5-min high-intensity exercise (HEX) intervals (HEX1 and HEX2-20 s at 50% peak power, followed by 15 s of rest), and a time to exhaustion (TTE) test. One-minute rest intervals were given between each round (RI1 and RI2), during which researchers treated the participant's posterior neck with either (1) wet-ice (ICE); (2) menthol spray (SPRAY); or (3) no treatment (CON). Neck (TNECK) and chest (TCHEST) skin temperatures were significantly lower following RI1 with ICE (vs. SPRAY). Thermal sensation decreased with ICE compared to CON following RI1, RI2, TTE, and a 2-min recovery. Rating of perceived exertion was also lower with ICE following HEX2 (vs. CON) and after RI2 (vs. SPRAY). Treatment did not influence TTE (68.9 ± 18.9s). The ability of intermittent ICE to attenuate neck and chest skin temperature rises during the initial HEX stages likely explains why participants felt cooler and less exerted during equivalent HEX bouts. These data suggest intermittent ICE improves perceptual stress during short, repeated bouts of vigorous exercise.

Three-dimensional printing of human skeletal muscle cells: An interdisciplinary approach for studying biological systems.

Interdisciplinary exploration is vital to education in the 21st century. This manuscript outlines an innovative laboratory-based teaching method that combines elements of biochemistry/molecular biology, kinesiology/health science, computer science, and manufacturing engineering to give students the ability to better conceptualize complex biological systems. Here, we utilize technology available at most universities to print three-dimensional (3D) scale models of actual human muscle cells (myofibers) out of bioplastic materials. The same methodological approach could be applied to nearly any cell type or molecular structure. This advancement is significant because historically, two-dimensional (2D) myocellular images have proven insufficient for detailed analysis of organelle organization and morphology. 3D imaging fills this void by providing accurate and quantifiable myofiber structural data. Manipulating tangible 3D models combats 2D limitation and gives students new perspectives and alternative learning experiences that may assist their understanding. This approach also exposes learners to 1) human muscle cell extraction and isolation, 2) targeted fluorescence labeling, 3) confocal microscopy, 4) image processing (via open-source software), and 5) 3D printing bioplastic scale-models (×500 larger than the actual cells). Creating these physical models may further student's interest in the invisible world of molecular and cellular biology. Furthermore, this interdisciplinary laboratory project gives instructors of all biological disciplines a new teaching tool to foster integrative thinking.