Three months of bed rest induce a residual transcriptomic signature resilient to resistance exercise countermeasures.

This study explored the muscle genome-wide response to long-term unloading (84-day bed rest) in 21 men. We hypothesized that a part of the bed rest-induced gene expression signature would be resilient to a concurrent flywheel resistance exercise (RE) countermeasure. Using DNA microarray technology analyzing 35 345 gene-level probe-sets, we identified 335 annotated probe-sets that were downregulated, and 315 that were upregulated after bed rest (P < .01). Besides a predictable differential expression of genes and pathways related to mitochondria (downregulation; false-discovery rates (FDR) <1E-04), ubiquitin system (upregulation; FDR = 3E-02), and skeletal muscle energy metabolism and structure (downregulation; FDR ≤ 3E-03), 84-day bed rest also altered circadian rhythm regulation (upregulation; FDR = 3E-02). While most of the bed rest-induced changes were counteracted by RE, 209 transcripts were resilient to the exercise countermeasure. Genes upregulated after bed rest were particularly resistant to training (P < .001 vs downregulated, non-reversed genes). Specifically, "Translation Factors," "Proteasome Degradation," "Cell Cycle," and "Nucleotide Metabolism" pathways were not normalized by RE. This study provides an unbiased high-throughput transcriptomic signature of one of the longest unloading periods in humans to date. Classical disuse-related changes in structural and metabolic genes/pathways were identified, together with a novel upregulation of circadian rhythm transcripts. In the context of previous bed rest campaigns, the latter seemed to be related to the duration of unloading, suggesting the transcriptomic machinery continues to adapt throughout extended disuse periods. Despite that the RE training offset most of the bed rest-induced muscle-phenotypic and transcriptomic alterations, we contend that the human skeletal muscle also displays a residual transcriptomic signature of unloading that is resistant to an established exercise countermeasure.

Early accentuated muscle hypertrophy is strongly associated with myonuclear accretion.

The current study explored whether the marked hypertrophic response noted with a short-term unilateral concurrent exercise paradigm was associated with more prominent changes in myonuclei accretion, ribosome biogenesis, and capillarization compared with resistance exercise alone (RE). Ten men (age 25 ± 4 yr) performed aerobic and resistance exercise (AE + RE) for one leg while the other leg did RE. Muscle biopsies were obtained before and after 5 wk of training and subjected to fiber-type specific immunohistochemical analysis, and quantification of total RNA content and mRNA/rRNA transcript abundance. Type II fiber cross-sectional area (CSA) increased with both AE + RE (22%) and RE (16%), while type I fiber CSA increased mainly with AE + RE (16%). The change score tended to differ between legs for type I CSA (P = 0.099), and the increase in smallest fiber diameter was greater in AE + RE than RE (P = 0.029). The number of nuclei per fiber increased after AE + RE in both fiber types, and this increase was greater (P = 0.027) than after RE. A strong correlation was observed between changes in number of nuclei per fiber and fiber CSA in both fiber types, for both AE + RE and RE (r > 0.8, P < 0.004). RNA content increased after AE + RE (24%, P = 0.019), but the change-scores did not differ across legs. The capillary variables generally increased in both fiber types, with no difference across legs. In conclusion, the accentuated hypertrophic response to AE + RE was associated with more pronounced myonuclear accretion, which was strongly correlated with the degree of fiber hypertrophy. This suggests that myonuclear accretion could play a role in facilitating muscle hypertrophy also during very short training periods.

Aerobic exercise augments muscle transcriptome profile of resistance exercise.

Recent reports suggest that aerobic exercise may boost the hypertrophic response to short-term resistance training. This study explored the effects of an acute aerobic exercise bout on the transcriptional response to subsequent resistance exercise. Ten moderately trained men performed ∼45 min cycling on one leg followed by 4 × 7 maximal knee extensions for each leg, 15 min later. Thus, one limb performed aerobic and resistance exercise (AE + RE) while the opposing leg did resistance exercise only (RE). Biopsies were obtained from the vastus lateralis muscle of each leg 3 h after the resistance exercise bout. Using DNA microarray, we analyzed differences [≥1.5-fold, false discovery rate (FDR) ≤10%] in gene expression profiles for the two modes of exercise. There were 176 genes up (127)- or downregulated (49) by AE + RE compared with RE. Among the most significant differentially expressed genes were established markers for muscle growth and oxidative capacity, novel cytokines, transcription factors, and micro-RNAs (miRNAs). The most enriched functional categories were those linked to carbohydrate metabolism and transcriptional regulation. Upstream analysis revealed that vascular endothelial growth factor, cAMP-response element-binding protein, Tet methylcytosine dioxygenase, and mammalian target of rapamycin were regulators highly activated by AE + RE, whereas JnK, NF-κß, MAPK, and several miRNAs were inhibited. Thus, aerobic exercise alters the skeletal muscle transcriptional signature of resistance exercise to initiate important gene programs promoting both myofiber growth and improved oxidative capacity. These results provide novel insight into human muscle adaptations to diverse exercise modes and offer the very first genomic basis explaining how aerobic exercise may augment, rather than compromise, muscle growth induced by resistance exercise.

Muscle, functional and cognitive adaptations after flywheel resistance training in stroke patients: a pilot randomized controlled trial.

BACKGROUND: Resistance exercise (RE) improves neuromuscular function and physical performance after stroke. Yet, the effects of RE emphasizing eccentric (ECC; lengthening) actions on muscle hypertrophy and cognitive function in stroke patients are currently unknown. Thus, this study explored the effects of ECC-overload RE training on skeletal muscle size and function, and cognitive performance in individuals with stroke. METHODS: Thirty-two individuals with chronic stroke (≥6 months post-stroke) were randomly assigned into a training group (TG; n = 16) performing ECC-overload flywheel RE of the more-affected lower limb (12 weeks, 2 times/week; 4 sets of 7 maximal closed-chain knee extensions; <2 min of contractile activity per session) or a control group (CG; n = 16), maintaining daily routines. Before and after the intervention, quadriceps femoris volume, maximal force and power for each leg were assessed, and functional and dual task performance, and cognitive functions were measured. RESULTS: Quadriceps femoris volume of the more-affected leg increased by 9.4 % in TG. Muscle power of the more-affected, trained (48.2 %), and the less-affected, untrained limb (28.1 %) increased after training. TG showed enhanced balance (8.9 %), gait performance (10.6 %), dual-task performance, executive functions (working memory, verbal fluency tasks), attention, and speed of information processing. CG showed no changes. CONCLUSION: ECC-overload flywheel resistance exercise comprising 4 min of contractile activity per week offers a powerful aid to regain muscle mass and function, and functional performance in individuals with stroke. While the current intervention improved cognitive functions, the cause-effect relationship, if any, with the concomitant neuromuscular adaptations remains to be explored. TRIAL REGISTRATION: Clinical Trials NCT02120846.

Effects of In-Season Inertial Resistance Training With Eccentric Overload in a Sports Population at Risk for Patellar Tendinopathy.

Gual, G, Fort-Vanmeerhaeghe, A, Romero-Rodríguez, D, and Tesch, PA. Effects of in-season inertial resistance training with eccentric overload in a sports population at risk for patellar tendinopathy. J Strength Cond Res 30(7): 1834-1842, 2016-Volleyball and basketball players can be considered as a population at risk for patellar tendinopathy. Given the paradox that eccentric training elicits therapeutic benefits yet might provoke such injury, we investigated the influence of a weekly bout of inertial squat resistance exercise offering eccentric overload on lower limb muscle power and patellar tendon complaints. Players of 8 (4 basketball and 4 volleyball) teams (38 women and 43 men) were randomly assigned to either the intervention (IG) or control (CG) group. Although IG and CG maintained scheduled in-season training routines over 24 weeks, IG, in addition, performed 1 weekly session of eccentric overload by 4 sets of 8 repetitions of the squat using flywheel inertial resistance. Victorian Institute of Sports Assessment patellar tendinopathy questionnaire (VISA-p), vertical countermovement jump, and squat power, both concentric (Squat-Con) and eccentric (Squat-Ecc), tests were performed before (T1), during (T2), and after (T3) the 24 weeks of intervention. Neither group suffered from patellar tendinopathy during the study period. VISA-p displayed no differences across groups at any measurement period. Countermovement jump scores significantly (p ≤ 0.05) differed between groups in favor of the IG. Both Squat-Con and Squat-Ecc mean scores from the IG were significantly (p < 0.01) higher than the CG. Adding a weekly eccentric overload squat training bout to a regular basketball and volleyball exercise routine enhances lower limb muscle power without triggering patellar tendon complaints. Future studies, using the current exercise paradigm, aim to explore its efficacy to prevent or combat patellar tendinopathy in sports calling for frequent explosive jumps.

Chronic stroke patients show early and robust improvements in muscle and functional performance in response to eccentric-overload flywheel resistance training: a pilot study.

BACKGROUND: Resistance exercise comprising eccentric (ECC) muscle actions enhances muscle strength and function to aid stroke patients in conducting daily tasks. The purpose of this study was to assess the efficacy of a novel ECC-overload flywheel resistance exercise paradigm to induce muscle and functional performance adaptations in chronic stroke patients. METHODS: Twelve patients (~8 years after stroke onset) performed 4 sets of 7 coupled concentric (CON) and ECC actions using the affected limb on a flywheel leg press (LP) device twice weekly for 8 weeks. Maximal CON and ECC isokinetic torque at 30, 60 and 90°/s, isometric knee extension and LP force, and CON and ECC peak power in LP were measured before and after training. Balance (Berg Balance Scale, BBS), gait (6-Min Walk test, 6MWT; Timed-Up-and-Go, TUG), functional performance (30-s Chair-Stand Test, 30CST), spasticity (Modified Ashworth Scale) and perceived participation (Stroke Impact Scale, SIS) were also determined. RESULTS: CON and ECC peak power increased in both the trained affected (34 and 44%; P < 0.01), and the untrained, non-affected leg (25 and 34%; P < 0.02). Power gains were greater (P = 0.008) for ECC than CON actions. ECC isokinetic torque at 60 and 90°/s increased in the affected leg (P < 0.04). The increase in isometric LP force for the trained, affected leg across tests ranged 10-20% (P < 0.05). BBS (P = 0.004), TUG (P = 0.018), 30CST (P = 0.024) and SIS (P = 0.058) scores improved after training. 6MWT and spasticity remained unchanged. CONCLUSIONS: This novel, short-term ECC-overload flywheel RE training regime emerges as a valid, safe and viable method to improve muscle function, balance, gait and functional performance in men and women suffering from chronic stroke.

Flywheel resistance exercise to maintain muscle oxidative potential during unloading.

BACKGROUND: As spaceflight compromises skeletal muscle oxidative and aerobic work capacity, this study assessed the efficacy of resistance exercise (RE) to counteract muscle metabolic perturbations induced by 5 wk unilateral lower limb unloading (UL). METHODS: There were 21 men and women (30-56 yr) who were randomly assigned to either UL with (Group, Grp; UL+RE; N = 10) or without (Grp UL; N = 11) concurrent RE. Iso-inertial RE comprised four sets of seven maximal coupled concentric-eccentric knee extensions executed 2-3 times per week. Percutaneous biopsies were obtained from m. vastus lateralis before and after either intervention. Levels of mRNA expression of factors regulating skeletal muscle oxidative capacity i.e., peroxisome proliferator-activated receptor gamma coactivator-1 (PGC-1alpha) and vascular endothelial growth factor (VEGF), and glycolytic capacity, i.e., phosphofructokinase (PFK), glycogen phosphorylase and synthase, hexokinase, and phosphorylase kinase alpha1, were subsequently analyzed. RESULTS: Grp UL showed decreased (36%) PGC-1alpha expression, increased (1.5-fold) PFK expression, and a trend toward decreased VEGF post-intervention. Grp UL+RE showed no changes. DISCUSSION: These results suggest that 5 wk unloading reduces skeletal muscle oxidative capacity and increases glycolytic enzyme activity. More importantly, only 12 bouts of high-force, low-volume resistance exercise attenuated these responses. Thus, the current resistance exercise paradigm emphasizing eccentric overload effectively counteracts unwarranted metabolic alterations induced by 5 wk unloading and may, therefore, aid in maintaining skeletal muscle integrity and endurance, and hence astronaut health and fitness during spaceflight.

Cardiovascular responses to rowing on a novel ergometer designed for both resistance and aerobic training in space.

BACKGROUND: Astronauts are required to perform both resistance and aerobic exercise while in orbit. This study assessed the aerobic energy yield and related physiological measurements using a nongravity dependent flywheel device designed for both resistance and aerobic exercise (RAD) in space. METHODS: Eight physically active men and women performed all-out rowing on the RAD. For comparison, exercise was also carried out employing a commercially available rowing ergometer (C2). RESULTS: Peak oxygen uptake during exercise using RAD and C2 averaged 3.11 +/- 0.49 and 3.18 +/- 0.50 L x min(-1), respectively. Similarly, peak plasma lactate concentration (9.6 vs. 11.2 mmol x L(-1)), heart rate (183 vs. 184 bpm), and rate of perceived exertion (15.8 vs. 16.0) were comparable across exercise using the two devices. DISCUSSION: Collectively, the results suggest that this novel exercise modality offers cardiovascular and metabolic responses, and thus aerobic exercise stimulus that is equally effective as that evoked by established technology for indoor rowing. Given the need for physiologically sound and highly effective exercise countermeasures that features small mass and envelope, and allows for resistance and aerobic exercise in a single apparatus, we believe this novel hardware should be considered for use in space.

Microgravity-induced skeletal muscle atrophy in women and men: implications for long-duration spaceflights to the Moon and Mars.

The inclusion of women on spaceflights has historically been limited. Recently, the first woman who will travel to the Moon was selected, and more women are participating in long-duration spaceflights. However, physiological data from real and simulated microgravity exposure are limited in women. This investigation studied women (n = 8, 34 ± 1 yr) and men (n = 9, 32 ± 1 yr) who underwent 2 (women) or 3 (men) mo of simulated microgravity (6° head-down tilt bed rest). Quadriceps and triceps surae muscle volumes were assessed via MRI before bed rest, bed rest day 29 (BR29, women and men), bed rest day 57 (BR57, women), and bed rest day 89 (BR89, men). Volume of both muscle groups decreased (P < 0.05) in women and men at all bed rest timepoints. Quadriceps muscle volume loss in women was greater than men at 1 mo (BR29: -17% vs. -10%, P < 0.05) and this 1-mo loss for women was similar to men at 3 mo (BR89: -18%, P > 0.05). In addition, the loss in women at 2 mo (BR57: -21%) exceeded men at 3 mo (P < 0.05). For the triceps surae, there was a trend for greater muscle volume loss in women compared with men at 1 mo (BR29: -18% vs. -16%, P = 0.08), and loss in women at 2 mo was similar to men at 3 mo (BR57: -29%, BR89: -29%, P > 0.05). The collective evidence suggests that women experience greater lower limb muscle atrophy than men at least through the first 4 mo of microgravity exposure. More sex-specific microgravity studies are needed to help protect the health of women traveling on long-duration orbital and interplanetary spaceflights.NEW & NOTEWORTHY This study adds to the limited evidence regarding sex-specific responses to real or simulated microgravity exposure, which collectively suggests a sex-specific muscle atrophy profile, with women losing more than men at least through the first 4 mo of weightlessness. Considering the increase in women being selected for space missions, including the first women to travel to the Moon, more physiological data on women in response to microgravity are needed.

Skeletal muscle morphology, satellite cells, and oxidative profile in relation to physical function and lifelong endurance training in very old men.

In the current study, we compared muscle morphology in three advanced aging cohorts that differed in physical function, including a unique cohort of lifelong endurance athletes. Biopsies from the vastus lateralis muscle of seven lifelong endurance athletes (EAs) aged 82-92 yr, and 19 subjects from the Uppsala Longitudinal Study of Adult Men (ULSAM) aged 87-91 yr were analyzed. ULSAM subjects were divided into high- (n = 9, HF) and low- (n = 10, LF) function groups based on strength and physical function tests. The analysis included general morphology, fiber type and cross-sectional area, capillarization, deficient cytochrome c oxidase (COX) activity, number of myonuclei and satellite cells, and markers of regeneration and denervation. Fibers with central nuclei and/or nuclear clumps were observed in all groups. EA differed from LF and HF by having a higher proportion of type I fibers, 52% more capillaries in relation to fiber area, fewer COX-negative fibers, and less variation in fiber sizes (all P < 0.05). There were no differences between the groups in the number of myonuclei and satellite cells per fiber, and no significant differences between LF and HF (P > 0.05). In conclusion, signs of aging were evident in the muscle morphology of all groups, but neither endurance training status nor physical function influenced signs of regeneration and denervation processes. Lifelong endurance training, but not higher physical function, was associated with higher muscle oxidative capacity, even beyond the age of 80.NEW & NOTEWORTHY Here we show that lifelong endurance training, but not physical function, is associated with higher muscle oxidative capacity, even beyond the age of 80 yr. Neither endurance training status nor physical function was significantly associated with satellite cells or markers of regeneration and denervation in muscle biopsies from these very old men.

Quadriceps muscle use in the flywheel and barbell squat.

BACKGROUND: Resistance exercise has been proposed as an aid to counteract quadriceps muscle atrophy in astronauts during extended missions in orbit. While space authorities have advocated the squat exercise should be prescribed, no exercise system suitable for in-flight use has been validated with regard to quadriceps muscle use. We compared muscle involvement in the terrestrial "gold standard" squat using free weights and a nongravity dependent flywheel resistance exercise device designed for use in space. METHODS: The subjects were 10 strength-trained men who performed 5 sets of 10 repetitions using the barbell squat (BS; 10 repetition maximum) or flywheel squat (FS; each repetition maximal), respectively. Functional magnetic resonance imaging (MRI) and surface electromyography (EMG) techniques assessed quadriceps muscle use. Exercise-induced contrast shift of MR images was measured by means of transverse relaxation time (T2). EMG root mean square (RMS) was measured during concentric (CON) and eccentric (ECC) actions and normalized to EMG RMS determined during maximal voluntary contraction. RESULTS: The quadriceps muscle group showed greater exercise-induced T2 increase following FS compared with BS. Among individual muscles, the rectus femoris displayed greater T2 increase with FS (+24 +/- 14%) than BS (+8 +/- 4%). Normalized quadriceps EMG showed no difference across exercise modes. DISCUSSION: Collectively, the results of this study suggest that quadriceps muscle use in the squat is comparable, if not greater, with flywheel compared with free weight resistance exercise. Data appear to provide support for use of flywheel squat resistance exercise as a countermeasures adjunct during spaceflight.

Flywheel resistance training calls for greater eccentric muscle activation than weight training.

Changes in muscle activation and performance were studied in healthy men in response to 5 weeks of resistance training with or without "eccentric overload". Subjects, assigned to either weight stack (grp WS; n = 8) or iso-inertial "eccentric overload" flywheel (grp FW; n = 9) knee extensor resistance training, completed 12 sessions of four sets of seven concentric-eccentric actions. Pre- and post-measurements comprised maximal voluntary contraction (MVC), rate of force development (RFD) and training mode-specific force. Root mean square electromyographic (EMG(RMS)) activity of mm. vastus lateralis and medialis was assessed during MVC and used to normalize EMG(RMS) for training mode-specific concentric (EMG(CON)) and eccentric (EMG(ECC)) actions at 90°, 120° and 150° knee joint angles. Grp FW showed greater (p < 0.05) overall normalized angle-specific EMG(ECC) of vastii muscles compared with grp WS. Grp FW showed near maximal normalized EMG(CON) both pre- and post-training. EMG(CON) for Grp WS was near maximal only post-training. While RFD was unchanged following training (p > 0.05), MVC and training-specific strength increased (p < 0.05) in both groups. We believe the higher EMG(ECC) activity noted with FW exercise compared to standard weight lifting could be attributed to its unique iso-inertial loading features. Hence, the resulting greater mechanical stress may explain the robust muscle hypertrophy reported earlier in response to flywheel resistance training.

Strength, power, fiber types, and mRNA expression in trained men and women with different ACTN3 R577X genotypes.

Alpha-actinins are structural proteins of the Z-line. Human skeletal muscle expresses two alpha-actinin isoforms, alpha-actinin-2 and alpha-actinin-3, encoded by their respective genes ACTN2 and ACTN3. ACTN2 is expressed in all muscle fiber types, while only type II fibers, and particularly the type IIb fibers, express ACTN3. ACTN3 (R577X) polymorphism results in loss of alpha-actinin-3 and has been suggested to influence skeletal muscle function. The X allele is less common in elite sprint and power athletes than in the general population and has been suggested to be detrimental for performance requiring high power. The present study investigated the association of ACTN3 genotype with muscle power during 30-s Wingate cycling in 120 moderately to well-trained men and women and with knee extensor strength and fatigability in a subset of 21 men performing isokinetic exercise. Muscle biopsies were obtained from the vastus lateralis muscle to determine fiber-type composition and ACTN2 and ACTN3 mRNA levels. Peak and mean power and the torque-velocity relationship and fatigability output showed no difference across ACTN3 genotypes. Thus this study suggests that R577X polymorphism in ACTN3 is not associated with differences in power output, fatigability, or force-velocity characteristics in moderately trained individuals. However, repeated exercise bouts prompted an increase in peak torque in RR but not in XX genotypes, suggesting that ACTN3 genotype may modulate responsiveness to training. Our data further suggest that alpha-actinins do not play a significant role in determining muscle fiber-type composition. Finally, we show that ACTN2 expression is affected by the content of alpha-actinin-3, which implies that alpha-actinin-2 may compensate for the lack of alpha-actinin-3 and hence counteract the phenotypic consequences of the deficiency.

Muscle2View, a CellProfiler pipeline for detection of the capillary-to-muscle fiber interface and high-content quantification of fiber type-specific histology.

Because manual immunohistochemical analysis of features such as skeletal muscle fiber typing, capillaries, myonuclei, and fiber size-related parameters is time consuming and prone to user subjectivity, automatic computational methods could allow for faster and more objective evaluation. Here, we developed Muscle2View, a free CellProfiler-based pipeline that integrates all key fiber-morphological variables, including the novel quantification of the capillary-to-fiber interface, in one single tool. Provided that the images are of sufficient quality and the settings are configured for the specific study, the pipeline allows for automatic and unsupervised analysis of fiber borders, myonuclei, capillaries, and morphometric parameters in a fiber type-specific manner from large batches of images in <10 min/tissue sample. The novel identification of the capillary-to-fiber interface allowed for the calculation of microvascular factors such as capillary contacts (CC), individual capillary-to-fiber ratio (C/Fi), and capillary-to-fiber perimeter exchange (CFPE) index. When comparing the Muscle2View pipeline to manual or semiautomatic analysis, overall the results revealed strong correlations. For several variables, however, there were differences (5-15%) between values computed by manual counting and Muscle2View, suggesting that the methods should not necessarily be used interchangeably. Collectively, we demonstrate that the Muscle2View pipeline can provide unbiased and high-content analysis of muscle cross-sectional immunohistochemistry images. In addition to the classical morphological measurements, the Muscle2View can identify the complex capillary-to-fiber network and myonuclear density in a fiber type-specific manner. This robust analysis is done in one single run within a user-friendly and flexible environment based on the free and widely used image software CellProfiler.NEW & NOTEWORTHY Here, we developed a freely available CellProfiler-based pipeline termed Muscle2View, which provides unbiased, high-content analysis of muscle cross-sectional immunohistochemistry images. In addition to fiber typing, myonuclei counting, and the quantification of fiber type-specific morphological measurements, the Muscle2View pipeline can identify the complex capillary-to-fiber network from a batch of images within minutes. Thus, the Muscle2View is a viable tool for researchers aiming to quantify immunohistochemical variables from skeletal muscle biopsies.

Altered regulation of the PINK1 locus: a link between type 2 diabetes and neurodegeneration?

Mutations in PINK1 cause the mitochondrial-related neurodegenerative disease Parkinson's. Here we investigate whether obesity, type 2 diabetes, or inactivity alters transcription from the PINK1 locus. We utilized a cDNA-array and quantitative real-time PCR for gene expression analysis of muscle from healthy volunteers following physical inactivity, and muscle and adipose tissue from nonobese or obese subjects with normal glucose tolerance or type 2 diabetes. Functional studies of PINK1 were performed utilizing RNA interference in cell culture models. Following inactivity, the PINK1 locus had an opposing regulation pattern (PINK1 was down-regulated while natural antisense PINK1 was up-regulated). In type 2 diabetes skeletal muscle, all transcripts from the PINK1 locus were suppressed and gene expression correlated with diabetes status. RNA interference of PINK1 in human neuronal cell lines impaired basal glucose uptake. In adipose tissue, mitochondrial gene expression correlated with PINK1 expression although remained unaltered following siRNA knockdown of Pink1 in primary cultures of brown preadipocytes. In conclusion, regulation of the PINK1 locus, previously linked to neurodegenerative disease, is altered in obesity, type 2 diabetes and inactivity, while the combination of RNAi experiments and clinical data suggests a role for PINK1 in cell energetics rather than in mitochondrial biogenesis.

Skeletal muscle proteolysis in response to short-term unloading in humans.

Skeletal muscle atrophy is evident after muscle disuse, unloading, or spaceflight and results from decreased protein content as a consequence of decreased protein synthesis, increased protein breakdown or both. At this time, there are essentially no human data describing proteolysis in skeletal muscle undergoing atrophy on Earth or in space, primarily due to lack of valid and accurate methodology. This particular study aimed at assessing the effects of short-term unloading on the muscle contractile proteolysis rate. Eight men were subjected to 72-h unilateral lower limb suspension (ULLS) and intramuscular interstitial levels of the naturally occurring proteolytic tracer 3-methylhistidine (3MH) were measured by means of microdialysis before and on completion of this intervention. The 3MH concentration following 72-h ULLS (2.01 +/- 0.22 nmol/ml) was 44% higher (P < 0.05) than before ULLS (1.56 +/- 0.20 nmol/ml). The present experimental model and the employed method determining 3MH in microdialysates present a promising tool for monitoring skeletal muscle proteolysis or metabolism of specific muscles during conditions resulting in atrophy caused by, e.g., disuse and real or simulated microgravity. This study provides evidence that the atrophic processes are evoked rapidly and within 72 h of unloading and suggests that countermeasures should be employed in the early stages of space missions to offset or prevent muscle loss during the period when the rate of muscle atrophy is the highest.

Muscle fluid shift does not alter EMG global variables during sustained isometric actions.

Body fluid redistribution occurs in astronauts traveling in space, potentially altering interstitial water content and hence impedance. This in turn may impact the features of electromyographic (EMG) signals measured to compare in-flight muscle function with pre- and post-flight conditions. Thus, the current study aimed at investigating the influence of similar fluid shifts on EMG spectral variables during muscle contractile activity. Ten men performed sustained isometric actions (120 s) at 20% and 60% of maximum voluntary contraction (MVC) following 1-h rest in the vertical or supine position. From single differential EMG signals, recorded from the soleus (SOL), the medial (MG) and lateral (LG) gastrocnemius muscles, initial value and rate of change over time (slope) of mean power frequency (MNF) and average rectified value (ARV) were assessed. MNF initial value showed dependence on muscle (P<0.01), but was unaffected by body tilt. MNF rate of change increased (P<0.001) with increased force and differed across muscles (P<0.05), but was not influenced (P=0.85) by altered body position. Thus, fluid shift resulting from vertical to supine tilt had no impact on myoelectrical manifestations of muscle fatigue. Furthermore, since such alteration of body fluid distribution resembles that occurring in microgravity, our findings suggest this may not be a methodological limitation, when comparing EMG fatigue indices on Earth versus in space.

Clinical Applications of Iso-Inertial, Eccentric-Overload (YoYo™) Resistance Exercise.

In the quest for a viable non-gravity dependent method to "lift weights" in space, our laboratory introduced iso-inertial resistance (YoYo™) exercise using spinning flywheel(s), more than 25 years ago. After being thoroughly tested in individuals subjected to various established spaceflight analogs, a multi-mode YoYo™ exercise apparatus was eventually installed on the International Space Station in 2009. The method, applicable to any muscle group, provides accommodated resistance and optimal muscle loading through the full range of motion of concentric actions, and brief episodes of eccentric overload. This exercise intervention has found terrestrial applications and shown success in enhancing sports performance and preventing injury and aiding neurological or orthopedic rehabilitation. Research has proven that this technique offers unique physiological responses not possible with other exercise hardware solutions. This paper provides a brief overview of research that has made use, and explored the efficacy, of this method in healthy sedentary or physically active individuals and populations suffering from muscle wasting, disease or injury. While the collective evidence to date suggests YoYo™ offers a potent stimulus to optimize the benefits of resistance exercise, systematic research to support clinical use of this method has only begun to emerge. Thus, we also offer perspectives on unresolved issues, unexplored applications for clinical conditions, and how this particular exercise paradigm could be implemented in future clinical research and eventually being prescribed. Fields of particular interest are those aimed at promoting muscle health by preventing injury or combating muscle wasting and neurological or metabolic dysfunction due to aging or illness, or those serving in rehabilitation following trauma and/or surgery.

Metabolic adaptations in skeletal muscle after 84 days of bed rest with and without concurrent flywheel resistance exercise.

As metabolic changes in human skeletal muscle after long-term (simulated) spaceflight are not well understood, this study examined the effects of long-term microgravity, with and without concurrent resistance exercise, on skeletal muscle oxidative and glycolytic capacity. Twenty-one men were subjected to 84 days head-down tilt bed rest with (BRE; n = 9) or without (BR; n = 12) concurrent flywheel resistance exercise. Activity and gene expression of glycogen synthase, glycogen phosphorylase (GPh), hexokinase, phosphofructokinase-1 (PFK-1), and citrate synthase (CS), as well as gene expression of succinate dehydrogenase (SDH), vascular endothelial growth factor (VEFG), peroxisome proliferator-activated receptor gamma coactivator-1 (PGC-1α), and myostatin, were analyzed in samples from m. vastus lateralis collected before and after bed rest. Activity and gene expression of enzymes controlling oxidative metabolism (CS, SDH) decreased in BR but were partially maintained in BRE. Activity of enzymes regulating anaerobic glycolysis (GPh, PFK-1) was unchanged in BR. Resistance exercise increased the activity of GPh. PGC-1α and VEGF expression decreased in both BR and BRE. Myostatin increased in BR but decreased in BRE after bed rest. The analyses of these unique samples indicate that long-term microgravity induces marked alterations in the oxidative, but not the glycolytic, energy system. The proposed flywheel resistance exercise was effective in counteracting some of the metabolic alterations triggered by 84-day bed rest. Given the disparity between gene expression vs. enzyme activity in several key metabolic markers, posttranscriptional mechanisms should be explored to fully evaluate metabolic adaptations to long-term microgravity with/without exercise countermeasures in human skeletal muscle.

Unilateral lower limb suspension: From subject selection to "omic" responses.

The unilateral lower limb suspension (ULLS) method was developed, introduced, and validated in the quest for a simple, effective, and highly reliable human analog to study the consequences of spaceflight on muscle size and function. Because withdrawal of weight bearing for no more than 2-3 days is sufficient to inflict disturbances in protein metabolism of postural muscles, it is imperative ULLS serves as a very powerful method to manifest skeletal muscle adaptations similar to those experienced in 0 g. Thus the rate of global muscle loss appears rather constant over the first 2 mo, amounting to about 2-3% per week. At the microscopic level, these changes are accompanied by a corresponding decrease in individual muscle fiber size. ULLS alters metabolism favoring more carbohydrate over fat substrate utilization. Altogether, these changes result in impaired work and endurance capacity of muscles being subjected to ULLS. Maximal voluntary force decreases out of proportion to the muscle loss, suggesting motor control is modified. Past reviews offer near exhaustive information on ULLS-induced responses with regard to the above changes. Hence, the current brief review describes more broadly the evolution of the ULLS model, from issues of subject recruitment and compliance control, to recent advances unraveling molecular mechanisms facilitating unloading-induced muscle wasting. Such knowledge is critical in designing future studies aimed at exploring and developing exercise countermeasures or other means to combat the debilitating effects on muscle experienced by astronauts during long-haul missions in Orbit.