Low baseline ribosome-related gene expression and resistance training-induced declines in ribosome-related gene expression are associated with skeletal muscle hypertrophy in young men and women.

Ribosomes are essential cellular machinery for protein synthesis. It is hypothesised that ribosome content supports muscle growth and that individuals with more ribosomes have greater increases in muscle size following resistance training (RT). Aerobic conditioning (AC) also elicits distinct physiological adaptations; however, no measures of ribosome content following AC have been conducted. We used ribosome-related gene expression as a proxy measure for ribosome content and hypothesised that AC and RT would increase ribosome-related gene expression. Fourteen young men and women performed 6 weeks of single-legged AC followed by 10 weeks of double-legged RT. Muscle biopsies were taken following AC and following RT in the aerobically conditioned (AC+RT) and unconditioned (RT) legs. No differences in regulatory genes (Ubf, Cyclin D1, Tif-1a and Polr-1b) involved in ribosomal biogenesis or ribosomal RNA (45S, 5.8S, 18S and 28S rRNAs) expression were observed following AC and RT, except for c-Myc (RT > AC+RT) and 5S rRNA (RT < AC+RT at pre-RT) with 18S external transcribed spacer and 5.8S internal transcribed spacer expression decreasing from pre-RT to post-RT in the RT leg only. When divided for change in leg-lean soft tissue mass (ΔLLSTM) following RT, legs with the greatest ΔLLSTM had lower expression in 11/13 measured ribosome-related genes before RT and decreased expression in 9/13 genes following RT. These results indicate that AC and RT did not increase ribosome-related gene expression. Contrary to previous research, the greatest increase in muscle mass was associated with lower changes in ribosome-related gene expression over the course of the 10-week training programme. This may point to the importance of translational efficiency rather than translational capacity (i.e. ribosome content) in mediating long-term exercise-induced adaptations in skeletal muscle.

Does perceived caloric and nutrient intake influence the acute effect of beverage consumption on cardiovascular function?

Our objective was to explore whether consuming the same high-fat/sugar beverage affects endothelial function differently depending on whether it is presented as "unhealthy" [accurate high calorie (kcal), fat, and sugar information displayed] versus "healthy" (inaccurate low kcal, fat, and sugar information displayed). Twenty-five, young (21 ± 2 yr), healthy, food-stress/shame-prone women completed three conditions: milkshake consumption (540 kcal, 80 g sugar, and 14 g fat) where correct, "unhealthy" nutritional information was shown to participants (milkshake condition), consumption of the same milkshake but with incorrect, "healthy" information shown to participants (100 kcal, 3 g sugar, and 4 g fat; sham-nutrishake condition), and water consumption (control condition). Pre- and postbeverage we assessed 1) endothelial function via standard brachial artery flow-mediated dilation (FMD); 2) perceived shame, stress, beverage healthiness, and harm; and 3) blood (plasma) glucose, insulin, triglycerides and oral fluid cortisol, and tumor necrosis factor-alpha (TNFα) receptor binding. Glucose, triglycerides, and insulin increased in the milkshake and sham-nutrishake conditions (P < 0.05). The milkshake was perceived as less healthy (P < 0.001) and more harmful (P < 0.001) than the sham-nutrishake. Shame, stress, oral fluid cortisol and TNFα receptor binding did not increase postconsumption. FMD decreased after the milkshake condition (pre: 7.4 ± 3.3%; post-60 min: 4.9 ± 2.9%; post-90 min: 4.5 ± 3.1%, P < 0.001) but not the sham-nutrishake (pre: 5.7 ± 2.2%; post-60 min: 5.5 ± 2.6%; post-90 min: 5.0 ± 2.4%, P = 0.43) or control conditions (pre: 7.0 ± 2.6%; post-60 min: 6.6 ± 4.1%; post-90 min: 6.0 ± 3.2%, P = 0.29). Shear rate stimulus covariation did not alter FMD results. Lower perceived beverage healthiness was significantly associated with a greater reduction in FMD (ρ = 0.36, P = 0.002). In conclusion, a high-fat/sugar milkshake reduced FMD only when presented as high in fat, sugar, and calories. This suggests that perceptions about nutritional information contribute to the impact of food intake on endothelial function and that nocebo effects could be involved in cardiovascular disease etiology.NEW & NOTEWORTHY This was the first study to investigate how perceived nutritional content influences the impact of a high-sugar/fat beverage on endothelial function. We found that a high-sugar/fat beverage only reduced endothelial function when it was presented to participants as high in calories, fat, and sugar. This suggests that perceived nutritional information contributes to the impact of high sugar and fat intake on endothelial function.

Does Aerobic Exercise Increase Skeletal Muscle Mass in Female and Male Adults?

INTRODUCTION: It is uncertain whether aerobic exercise in the form of walking contributes to the preservation or increase in total or regional skeletal muscle mass (SMM). PURPOSE: This study aimed to determine the effects of aerobic exercise on total and regional (upper body verses leg SMM) in male ( n = 105) and female ( n = 133) adults with overweight and obesity. METHODS: A retrospective analysis of data from four randomized controlled trials. Participants included those who completed the given trial (control, n = 63; intervention, n = 175) and with complete magnetic resonance imaging (MRI) measured adipose tissue and SMM pre- and postintervention. Macronutrient intake was assessed for a subsample of participants. Supervised exercise was performed by walking on a treadmill for durations ranging from 12 to 24 wk at intensities between 50% and 75% of V̇O 2peak . RESULTS: All MRI-measured adipose tissue depots were reduced, and cardiorespiratory fitness was increased by aerobic exercise compared with controls ( P < 0.001). Independent of baseline SMM, aerobic exercise was associated with a small reduction (estimated mean difference ± standard error) in whole-body SMM (-0.310 ± 0.150 kg, P = 0.039) and upper body SMM (-0.273 ± 0.121 kg, P = 0.025) compared with control. No between-group difference was observed for change in leg SMM ( P > 0.10). A negative association was observed between the relative change in body weight and change in total ( R2 = 0.37, P < 0.001), upper body ( R2 = 0.21, P < 0.001), and leg SMM ( R2 = 0.09, P = 0.701). The SMM-to-adipose tissue ratio increased in response to aerobic exercise and was positively associated with weight loss ( P < 0.001). Change in SMM was not associated with dietary protein intake ( P > 0.10). CONCLUSIONS: Aerobic exercise performed while walking preserves, but does not increase, SMM in exercising muscle of adults. SMM not directly targeted by aerobic exercise may not be maintained.

Lipidomic studies reveal two specific circulating phosphatidylcholines as surrogate biomarkers of the omega-3 index.

Optimal dietary intake of omega-3 long-chain polyunsaturated fatty acids (n3-LCPUFAs) is critical to human health across the lifespan. However, omega-3 index (O3I) determination is not routinely assessed due to complicated procedures for n3-LCPUFA analysis from the phospholipid (PL) fraction of erythrocytes. Herein, a high-throughput method for lipidomics based on multisegment injection-nonaqueous capillary electrophoresis-mass spectrometry was applied to identify circulating PLs as surrogate biomarkers of O3I in two randomized placebo-controlled trials. An untargeted lipidomic data workflow using a subgroup analysis of serum extracts from sunflower oil versus high-dose fish oil (FO)-supplemented participants revealed that ingested n3-LCPUFAs were primarily distributed as their phosphatidylcholines (PCs) relative to other PL classes. In both high-dose FO (5.0 g/day) and EPA-only trials (3.0 g/day), PC (16:0_20:5) was the most responsive PL, whereas PC (16:0_22:6) was selective to DHA-only supplementation. We also demonstrated that the sum concentration of both these PCs in fasting serum or plasma samples was positively correlated to the O3I following FO (r = 0.708, P = 1.02 × 10-11, n = 69) and EPA- or DHA-only supplementation (r = 0.768, P = 1.01 × 10-33, n = 167). Overall, DHA was more effective in improving the O3I (ΔO3I = 4.90 ± 1.33%) compared to EPA (ΔO3I = 2.99 ± 1.19%) in young Canadian adults who had a poor nutritional status with an O3I (3.50 ± 0.68%) at baseline. Our method enables the rapid assessment of the O3I by directly measuring two circulating PC species in small volumes of blood, which may facilitate screening applications for population and precision health.

The effects of single-leg disuse on skeletal muscle strength and size in the nonimmobilized leg of uninjured adults: a meta-analysis.

The effects of single-leg immobilization on changes in skeletal muscle strength and size in the nonimmobilized leg remain controversial. Some studies have shown decreases, or even increases, in skeletal muscle strength and size of the nonimmobilized leg, thus challenging its role as an internal control. Here, we meta-analyze changes in knee extensor strength and size in the nonimmobilized leg of noninjured adults who participated in single-leg disuse studies. We extracted data from the nonimmobilized leg of participants from 15 of 40 studies included in our previous meta-analysis on single-leg disuse. Single-leg disuse had a trivial effect on knee extensor strength (Hedges' gav = -0.13 [-0.23, -0.03], P < 0.01, -3.6 ± 5.6%, N = 13 studies, n = 194 participants) and no impact on knee extensor size (0.06 [-0.06, 0.19], P = 0.21, 0.8 ± 2.9%, N = 9, n = 107) in the nonimmobilized leg. By comparison, single-leg disuse had a large effect on knee extensor strength (-0.85 [-1.01, -0.69], P < 0.01, -20.4 ± 6.4%; mean difference between legs = 16.8 ± 7.8% [12.8, 20.8], P < 0.001) and a medium effect on knee extensor size (-0.40 [-0.55, -0.25], P < 0.01, -7.0 ± 4%; mean difference = 7.8 ± 5.6% [11.6, 4.0], P < 0.002) in the immobilized leg. These results highlight the utility of the nonimmobilized leg to act as an internal control in single-leg immobilization studies.NEW & NOTEWORTHY Our meta-analyses show a trivial effect of single-leg immobilization on leg extensor strength and no effect on leg extensor size in the nonimmobilized leg in uninjured adults. Thus, the nonimmobilized leg in single-leg immobilization studies can serve as useful internal control when examining changes in knee extensor strength and size.

Single-leg disuse decreases skeletal muscle strength, size, and power in uninjured adults: A systematic review and meta-analysis.

We aimed to quantify declines from baseline in lower limb skeletal muscle size and strength of uninjured adults following single-leg disuse. We searched EMBASE, Medline, CINAHL, and CCRCT up to 30 January 2022. Studies were included in the systematic review if they (1) recruited uninjured participants; (2) were an original experimental study; (3) employed a single-leg disuse model; and (4) reported muscle strength, size, or power data following a period of single-leg disuse for at least one group without a countermeasure. Studies were excluded if they (1) did not meet all inclusion criteria; (2) were not in English; (3) reported previously published muscle strength, size, or power data; or (4) could not be sourced from two different libraries, repeated online searches, and the authors. We used the Cochrane Risk of Bias Assessment Tool to assess risk of bias. We then performed random-effects meta-analyses on studies reporting measures of leg extension strength and extensor size. Our search revealed 6548 studies, and 86 were included in our systematic review. Data from 35 and 20 studies were then included in the meta-analyses for measures of leg extensor strength and size, respectively (40 different studies). No meta-analysis for muscle power was performed due to insufficient homogenous data. Effect sizes (Hedges' gav ) with 95% confidence intervals for leg extensor strength were all durations = -0.80 [-0.92, -0.68] (n = 429 participants; n = 68 aged 40 years or older; n ≥ 78 females); ≤7 days of disuse = -0.57 [-0.75, -0.40] (n = 151); >7 days and ≤14 days = -0.93 [-1.12, -0.74] (n = 206); and >14 days = -0.95 [-1.20, -0.70] (n = 72). Effect sizes for measures of leg extensor size were all durations = -0.41 [-0.51, -0.31] (n = 233; n = 32 aged 40 years or older; n ≥ 42 females); ≤7 days = -0.26 [-0.36, -0.16] (n = 84); >7 days and ≤14 days = -0.49 [-0.67, -0.30] (n = 102); and >14 days = -0.52 [-0.74, -0.30] (n = 47). Decreases in leg extensor strength (cast: -0.94 [-1.30, -0.59] (n = 73); brace: -0.90 [-1.18, -0.63] (n = 106)) and size (cast: -0.61[-0.87, -0.35] (n = 41); brace: (-0.48 [-1.04, 0.07] (n = 41)) following 14 days of disuse did not differ for cast and brace disuse models. Single-leg disuse in adults resulted in a decline in leg extensor strength and size that reached a nadir beyond 14 days. Bracing and casting led to similar declines in leg extensor strength and size following 14 days of disuse. Studies including females and males and adults over 40 years of age are lacking.

Does initial skeletal muscle size or sex affect the magnitude of muscle loss in response to 14 days immobilization?

We aimed to determine whether there was a relationship between pre-immobilization skeletal muscle size and the magnitude of muscle atrophy following 14 days of unilateral lower limb immobilization. Our findings (n = 30) show that pre-immobilization leg fat-free mass and quadriceps cross-sectional area (CSA) were unrelated to the magnitude of muscle atrophy. However, sex-based differences may be present, but confirmatory work is required. In women, pre-immobilization leg fat-free mass and CSA were associated with changes in quadriceps CSA after immobilization (n = 9, r2 = 0.54-0.68; P < 0.05). The extent of muscle atrophy is not affected by initial muscle mass, but there is potential for sex-based differences.

Aerobic conditioning alters the satellite cell and ribosome response to acute eccentric contractions in young men and women.

Satellite cells (SCs) and ribosomes are key determinants of the skeletal muscle adaptive response. Both are thought to increase acutely after resistance exercise and chronically with resistance training. However, the acute SC and ribosome exercise response with prior aerobic conditioning is unknown. Fourteen young men and women underwent 6 wk of single-legged aerobic conditioning followed by an acute bout of 300 eccentric contractions on each leg. Muscle biopsies were taken from the vastus lateralis of the aerobically conditioned (AC) and the control (CTL) legs before (Pre), 24 (24 h), and 48 (48 h) h post-contractions. Pre-eccentric contractions, 45S pre-rRNA and 5.8S internal transcribed spacer (ITS) expression were lower in the AC leg compared with the CTL leg. SC content (PAX7+ cells/100 fibers) in type I and mixed fibers showed a main effect of condition, where values were greater in the AC leg compared with the CTL. A main effect of condition for Pax7 and MyoD1 mRNA expression was observed where expression was greater in the AC leg compared with the CTL. AC had greater RNA concentration and mRNA expression of Ubf and Tif-1a compared with CTL. Only the AC leg increased (Pre-24h) 45S pre-rRNA, 5.8S ITS, and 28S ITS following eccentric contractions. We discovered that aerobic conditioning increased type-I SC abundance and the acute increase in ribosome content following eccentric contractions.

Increased protein intake derived from leucine-enriched protein enhances the integrated myofibrillar protein synthetic response to short-term resistance training in untrained men and women: a 4-day randomized controlled trial.

Leucine is a critical amino acid stimulating myofibrillar protein synthesis (MyoPS). The consumption of higher leucine-containing drinks stimulates MyoPS, but we know less about higher leucine solid foods. Here, we examined the effect of short-term resistance exercise training (STRT) combined with supplementation of a protein and leucine-enriched bar, compared with STRT alone, on integrated (%/day) rates of MyoPS and anabolic protein signaling. In a nonblinded, randomized crossover trial, eight young adults performed four sessions of STRT without or while consuming the study bar (STRT+Leu, 16 g of protein containing ∼3 g of leucine) for two 4-day phases, separated by 2 days nonexercise (Rest) washout. In combination with serial muscle biopsies, deuterated water permitted the measurement of MyoPS and protein signaling phosphorylation. MyoPS during STRT (1.43 ± 0.06%/day) and STRT+Leu (1.53 ± 0.06%/day) were greater than Rest (1.31 ± 0.05%/day), and MyoPS during STRT+Leu (1.53 ± 0.06%/day) was greater than STRT alone (1.43 ± 0.06%/day). STRT+Leu increased the ratio of phosphorylated to total mechanistic target of rapamycin and 4EBP1 compared to Rest. Engaging in STRT increased integrated MyoPS and protein signaling in young adults and was enhanced with increased protein intake derived from a leucine-enriched protein bar. This study was registered at clinicaltrials.gov as NCT03796897.

Short-term aerobic conditioning prior to resistance training augments muscle hypertrophy and satellite cell content in healthy young men and women.

Factors influencing inter-individual variability of responses to resistance training (RT) remain to be fully elucidated. We have proposed the importance of capillarization in skeletal muscle for the satellite cell (SC) response to RT-induced muscle hypertrophy, and hypothesized that aerobic conditioning (AC) would augment RT-induced adaptations. Fourteen healthy young (22 ± 2 years) men and women underwent AC via 6 weeks of unilateral cycling followed by 10 weeks of bilateral RT to investigate how AC alters SC content, activity, and muscle hypertrophy following RT. Muscle biopsies were taken at baseline (unilateral), post AC (bilateral), and post RT (bilateral) in the aerobically conditioned (AC + RT) and unconditioned (RT) legs. Immunofluorescence was used to determine muscle capillarization, fiber size, SC content, and activity. Type I and type II fiber cross-sectional area (CSA) increased following RT, and when legs were analyzed independently, AC + RT increased type I, type II, and mixed-fiber CSA, where the RT leg tended to increase type II (p = .05), but not type I or mixed-fiber CSA. SC content, activation, and differentiation increased with RT, where type I total and quiescent SC content was greater in AC + RT compared to the RT leg. Those with the greatest capillary-to-fiber perimeter exchange index before RT had the greatest change in CSA following RT and a significant relationship was observed between type II fiber capillarization and the change in type II-fiber CSA with RT (r = 0.35). This study demonstrates that AC prior to RT can augment RT-induced muscle adaptions and that these differences are associated with increases in capillarization.

Sex-Based Differences in the Myogenic Response and Inflammatory Gene Expression Following Eccentric Contractions in Humans.

After muscle injury, the interaction between muscle satellite cells (SC) and the immune response is instrumental for the repair and regeneration of skeletal muscle tissue. Studies have reported sex-based differences in the skeletal muscle inflammatory and regenerative response following injury. However, many of these studies investigated such differences by manipulating the concentration of estradiol, in rodents and humans, without directly comparing males to females. We sought to explore differences in the myogenic and inflammatory response following unaccustomed eccentric exercise in males and females. We hypothesized that females would have a blunted myogenic and inflammatory response as compared to males. Methods: 26 (13 male, 13 female) healthy young adults (22 ± 0.4 years [mean ± SEM]) performed 300 maximal eccentric contractions (180°/s) of the knee extensors. Muscle biopsies were taken before (pre) and 48 h (post) following eccentric damage. SC content and activation were determined by immunohistochemical and real time-polymerase chain reaction (rt-PCR) analysis. Inflammatory markers were analyzed using rt-PCR. Results: Following eccentric damage, males had a greater expansion of type I-associated SC (p < 0.05), and there was a trend for a greater expansion in total SC (type I + II fibers) (p = 0.06) compared to females. There was a trend for a greater increase in Pax7 and CCL2 gene expression in males compared to females (p = 0.09). Conclusion: We conclude that there are sex-based differences in the myogenic and inflammatory response, where females have a blunted SC and inflammatory response.

Declines in muscle protein synthesis account for short-term muscle disuse atrophy in humans in the absence of increased muscle protein breakdown.

BACKGROUND: We determined the short-term (i.e. 4 days) impacts of disuse atrophy in relation to muscle protein turnover [acute fasted-fed muscle protein synthesis (MPS)/muscle protein breakdown (MPB) and integrated MPS/estimated MPB]. METHODS: Healthy men (N = 9, 22 ± 2 years, body mass index 24 ± 3 kg m-2 ) underwent 4 day unilateral leg immobilization. Vastus lateralis (VL) muscle thickness (MT) and extensor strength and thigh lean mass (TLM) were measured. Bilateral VL muscle biopsies were collected on Day 4 at t = -120, 0, 90, and 180 min to determine integrated MPS, estimated MPB, acute fasted-fed MPS (l-[ring-13 C6 ]-phe), and acute fasted tracer decay rate representative of MPB (l-[15 N]-phe and l-[2 H8 ]-phe). Protein turnover cell signalling was measured by immunoblotting. RESULTS: Immobilization decreased TLM [pre: 7477 ± 1196 g, post: 7352 ± 1209 g (P < 0.01)], MT [pre: 2.67 ± 0.50 cm, post: 2.55 ± 0.51 cm (P < 0.05)], and strength [pre: 260 ± 43 N m, post: 229 ± 37 N m (P < 0.05)] with no change in control legs. Integrated MPS decreased in immob vs. control legs [control: 1.55 ± 0.21% day-1 , immob: 1.29 ± 0.17% day-1 (P < 0.01)], while tracer decay rate (i.e. MPB) (control: 0.02 ± 0.006, immob: 0.015 ± 0.015) and fractional breakdown rate (FBR) remained unchanged [control: 1.44 ± 0.51% day-1 , immob: 1.73 ± 0.35% day-1 (P = 0.21)]. Changes in MT correlated with those in MPS but not FBR. MPS increased in the control leg following feeding [fasted: 0.043 ± 0.012% h-1 , fed: 0.065 ± 0.017% h-1 (P < 0.05)] but not in immob [fasted: 0.034 ± 0.014% h-1 , fed: 0.049 ± 0.023% h-1 (P = 0.09)]. There were no changes in markers of MPB with immob (P > 0.05). CONCLUSIONS: Human skeletal muscle disuse atrophy is driven by declines in MPS, not increases in MPB. Pro-anabolic therapies to mitigate disuse atrophy would likely be more effective than therapies aimed at attenuating protein degradation.

Dairy and Dairy Alternative Supplementation Increase Integrated Myofibrillar Protein Synthesis Rates, and Are Further Increased when Combined with Walking in Healthy Older Women.

BACKGROUND: The stimulation of muscle protein synthesis (MPS) by dietary protein is reduced with age. We hypothesized that twice-daily milk consumption would increase daily rates of MPS in older women relative to a nondairy milk alternative and that MPS would be enhanced by increased physical activity (PA). METHODS: Twenty-two older women were randomly assigned to 1 of 3 experimental groups: whole milk (WM; n = 7, 69 ± 3 y), skim milk (SM; n = 7, 68 ± 3 y), or an almond beverage (AB; n = 8, 63 ± 3 y). From days 1 to 3, participants consumed a standardized diet (0.8 g protein⋅kg-1 ⋅d-1) and performed their habitual PA (Phase 1, Baseline). From days 4 to 6, participants continued to perform habitual PA, but consumed an intervention diet consisting of the standardized diet plus twice-daily beverages (250 mL each) of either WM, SM, or AB (Phase 2, Diet Intervention). Finally, from days 7 to 9, the intervention diet was consumed, and PA via daily steps was increased to ∼150% of habitual daily steps (Phase 3, Intervention Diet + PA). Deuterated water was ingested throughout the study, and muscle biopsies were taken on days 1, 4, 7, and 10 to measure MPS. RESULTS: Daily MPS rates were not differentially affected by the addition of WM, SM, or AB to a standardized diet. There was, however, a significant effect of study phase such that, when collapsed across conditions, MPS was significantly increased from Phase 1 to Phase 2 (+0.133%⋅d-1; 95% CI: 0.035-0.231; P < 0.01) and further increased from Phase 2 to Phase 3 (+0.156%⋅d-1; 95% CI: 0.063-0.250; P < 0.01). CONCLUSIONS: Increasing PA through walking was sufficient to increase daily MPS rates in older women, irrespective of whether dietary protein intake is increased beyond the recommended intake of 0.8 g⋅kg-1 ⋅d-1. The trial was registered at clinicaltrials.gov as NCT04981652.

Fish Oil for Healthy Aging: Potential Application to Master Athletes.

Master athletes perform high volumes of exercise training yet display lower levels of physical functioning and exercise performance when compared with younger athletes. Several reports in the clinical literature show that long chain n-3 polyunsaturated fatty acid (LC n-3 PUFA) ingestion promotes skeletal muscle anabolism and strength in untrained older persons. There is also evidence that LC n-3 PUFA ingestion improves indices of muscle recovery following damaging exercise in younger persons. These findings suggest that LC n-3 PUFA intake could have an ergogenic effect in master athletes. However, the beneficial effect of LC n-3 PUFA intake on skeletal muscle in response to exercise training in both older and younger persons is inconsistent and, in some cases, generated from low-quality studies or those with a high risk of bias. Other factors such as the choice of placebo and health status of participants also confound interpretation of existing reports. As such, when considered on balance, the available evidence does not indicate that ingestion of LC n-3 PUFAs above current population recommendations (250-500 mg/day; 2 portions of oily fish per week) enhances exercise performance or recovery from exercise training in master athletes. Further work is now needed related to how the dose, duration, and co-ingestion of LC n-3 PUFAs with other nutrients such as amino acids impact the adaptive response to exercise training. This work should also consider how LC n-3 PUFA supplementation may differentially alter the lipid profile of cellular membranes of key regulatory sites such as the sarcolemma, mitochondria, and sarcoplasmic reticulum.

Transcriptomic links to muscle mass loss and declines in cumulative muscle protein synthesis during short-term disuse in healthy younger humans.

Muscle disuse leads to a rapid decline in muscle mass, with reduced muscle protein synthesis (MPS) considered the primary physiological mechanism. Here, we employed a systems biology approach to uncover molecular networks and key molecular candidates that quantitatively link to the degree of muscle atrophy and/or extent of decline in MPS during short-term disuse in humans. After consuming a bolus dose of deuterium oxide (D2 O; 3 mL.kg-1 ), eight healthy males (22 ± 2 years) underwent 4 days of unilateral lower-limb immobilization. Bilateral muscle biopsies were obtained post-intervention for RNA sequencing and D2 O-derived measurement of MPS, with thigh lean mass quantified using dual-energy X-ray absorptiometry. Application of weighted gene co-expression network analysis identified 15 distinct gene clusters ("modules") with an expression profile regulated by disuse and/or quantitatively connected to disuse-induced muscle mass or MPS changes. Module scans for candidate targets established an experimentally tractable set of candidate regulatory molecules (242 hub genes, 31 transcriptional regulators) associated with disuse-induced maladaptation, many themselves potently tied to disuse-induced reductions in muscle mass and/or MPS and, therefore, strong physiologically relevant candidates. Notably, we implicate a putative role for muscle protein breakdown-related molecular networks in impairing MPS during short-term disuse, and further establish DEPTOR (a potent mTOR inhibitor) as a critical mechanistic candidate of disuse driven MPS suppression in humans. Overall, these findings offer a strong benchmark for accelerating mechanistic understanding of short-term muscle disuse atrophy that may help expedite development of therapeutic interventions.

An intron variant of the GLI family zinc finger 3 (GLI3) gene differentiates resistance training-induced muscle fiber hypertrophy in younger men.

We examined the association between genotype and resistance training-induced changes (12 wk) in dual x-ray energy absorptiometry (DXA)-derived lean soft tissue mass (LSTM) as well as muscle fiber cross-sectional area (fCSA; vastus lateralis; n = 109; age = 22 ± 2 y, BMI = 24.7 ± 3.1 kg/m2 ). Over 315 000 genetic polymorphisms were interrogated from muscle using DNA microarrays. First, a targeted investigation was performed where single nucleotide polymorphisms (SNP) identified from a systematic literature review were related to changes in LSTM and fCSA. Next, genome-wide association (GWA) studies were performed to reveal associations between novel SNP targets with pre- to post-training change scores in mean fCSA and LSTM. Our targeted investigation revealed no genotype-by-time interactions for 12 common polymorphisms regarding the change in mean fCSA or change in LSTM. Our first GWA study indicated no SNP were associated with the change in LSTM. However, the second GWA study indicated two SNP exceeded the significance level with the change in mean fCSA (P = 6.9 × 10-7 for rs4675569, 1.7 × 10-6 for rs10263647). While the former target is not annotated (chr2:205936846 (GRCh38.p12)), the latter target (chr7:41971865 (GRCh38.p12)) is an intron variant of the GLI Family Zinc Finger 3 (GLI3) gene. Follow-up analyses indicated fCSA increases were greater in the T/C and C/C GLI3 genotypes than the T/T GLI3 genotype (P < .05). Data from the Auburn cohort also revealed participants with the T/C and C/C genotypes exhibited increases in satellite cell number with training (P < .05), whereas T/T participants did not. Additionally, those with the T/C and C/C genotypes achieved myonuclear addition in response to training (P < .05), whereas the T/T participants did not. In summary, this is the first GWA study to examine how polymorphisms associate with the change in hypertrophy measures following resistance training. Future studies are needed to determine if the GLI3 variant differentiates hypertrophic responses to resistance training given the potential link between this gene and satellite cell physiology.

The Effect of a Multi-ingredient Supplement on Resistance Training-induced Adaptations.

INTRODUCTION: Resistance exercise training (RET) induces muscle hypertrophy that, when combined with co-temporal protein ingestion, is enhanced. However, fewer studies have been conducted when RET is combined with multi-ingredient supplements. PURPOSE: We aimed to determine the effect of a high-quality multi-ingredient nutritional supplement (SUPP) versus an isonitrogenous (lower protein quality), isoenergetic placebo (PL) on RET-induced gains in lean body mass (LBM), muscle thickness, and muscle cross-sectional area (CSA). We hypothesized that RET-induced gains in LBM and muscle CSA would be greater in SUPP versus PL. METHODS: In a double-blind randomized controlled trial, 26 (13 male, 13 female) healthy young adults (mean ± SD, 22 ± 2 yr) were randomized to either the SUPP group (n = 13; 20 g whey protein, 2 g leucine, 2.5 g creatine monohydrate, 300 mg calcium citrate, 1000 IU vitamin D) or the PL group (n = 13; 20 g collagen peptides, 1.4 g alanine, 0.6 g glycine) groups, ingesting their respective supplements twice daily. Measurements were obtained before and after a 10-wk linear progressive RET program. RESULTS: Greater increases in LBM were observed for SUPP versus PL (SUPP: +4.1 ± 1.3 kg, PL: +2.8 ± 1.7 kg, P < 0.05). No additive effect of the supplement could be detected on vastus lateralis muscle CSA, but SUPP did result in increased biceps brachii muscle CSA and thickness (P < 0.05). CONCLUSIONS: We conclude that when combined with RET, the consumption of SUPP increased LBM and upper-body CSA and thickness to a greater extent than to that observed in the PL group of healthy young adults.

Brief Vigorous Stair Climbing Effectively Improves Cardiorespiratory Fitness in Patients With Coronary Artery Disease: A Randomized Trial.

Background: Cardiac rehabilitation exercise reduces the risk of secondary cardiovascular disease. Interval training is a time-efficient alternative to traditional cardiac rehabilitation exercise and stair climbing is an accessible means. We aimed to assess the effectiveness of a high-intensity interval stair climbing intervention on improving cardiorespiratory fitness ( V ˙ O 2 peak ) compared to standard cardiac rehabilitation care. Methods: Twenty participants with coronary artery disease (61 ± 7 years, 18 males, two females) were randomly assigned to either traditional moderate-intensity exercise (TRAD) or high-intensity interval stair climbing (STAIR). V ˙ O 2 peak was assessed at baseline, following 4 weeks of six supervised exercise sessions and after 8 weeks of ~24 unsupervised exercise sessions. TRAD involved a minimum of 30 min at 60-80%HRpeak, and STAIR consisted of three bouts of six flights of 12 stairs at a self-selected vigorous intensity (~90 s/bout) separated by recovery periods of walking (~90 s). This study was registered as a clinical trial at clinicaltrials.gov (NCT03235674). Results: Two participants could not complete the trial due to the time commitment of the testing visits, leaving n = 9 in each group who completed the interventions without any adverse events. V ˙ O 2 peak increased after supervised and unsupervised training in comparison to baseline for both TRAD [baseline: 22.9 ± 2.5, 4 weeks (supervised): 25.3 ± 4.4, and 12 weeks (unsupervised): 26.5 ± 4.8 mL/kg/min] and STAIR [baseline: 21.4 ± 4.5, 4 weeks (supervised): 23.4 ± 5.6, and 12 weeks (unsupervised): 25 ± 6.2 mL/kg/min; p (time) = 0.03]. During the first 4 weeks of training (supervised) the STAIR vs. TRAD group had a higher %HRpeak (101 ± 1 vs. 89 ± 1%; p ≤ 0.001), across a shorter total exercise time (7.1 ± 0.1 vs. 36.7 ± 1.1 min; p = 0.009). During the subsequent 8 weeks of unsupervised training, %HRpeak was not different (87 ± 8 vs. 96 ± 8%; p = 0.055, mean ± SD) between groups, however, the STAIR group continued to exercise for less time per session (10.0 ± 3.2 vs. 24.2 ± 17.0 min; p = 0.036). Conclusions: Both brief, vigorous stair climbing, and traditional moderate-intensity exercise are effective in increasing V ˙ O 2 peak , in cardiac rehabilitation exercise programmes.

Two weeks of single-leg immobilization alters intramyocellular lipid storage characteristics in healthy, young women.

Muscle disuse rapidly induces insulin resistance (IR). Despite a relationship between intramyocellular lipid (IMCL) content and IR, during muscle-disuse IR develops before IMCL accumulation, suggesting that IMCL are not related to disuse-induced IR. However, recent studies show that it is not total IMCL content, but IMCL size and location that are related to IR. Changes in these IMCL parameters may occur prior to increases in IMCL content, thus contributing to disuse-induced IR. Omega-3 fatty acids may mitigate the effects of disuse on IR by preventing a decline in insulin signaling proteins. Twenty women (age 22 ± 3 yr) received either 5 g·day-1 omega-3 fatty acid or isoenergetic sunflower oil for 4 wk prior to, throughout 2 wk of single-leg immobilization, and during 2 wk of recovery. Changes in IMCL characteristics and insulin signaling proteins were examined in vastus lateralis samples taken before supplementation and immobilization, and following immobilization and recovery. Omega-3 supplementation had no effect. IMCL area density decreased in the subsarcolemmal region during immobilization and recovery (-19% and -56%, respectively, P = 0.009). IMCL size increased in the central intermyofibrillar region during immobilization (43%, P = 0.007), returning to baseline during recovery. PLIN5 and AKT increased during immobilization (87%, P = 0.002; 30%, P = 0.007, respectively). PLIN 5 remained elevated and AKT increased further (15%) during recovery. IRS1, AS160, and GLUT4 decreased during immobilization (-35%, P = 0.001; -44%, P = 0.03; -56%, P = 0.02, respectively), returning to baseline during recovery. Immobilization alters IMCL storage characteristics while negatively affecting unstimulated insulin signaling protein content in young women.NEW & NOTEWORTHY We report that the subcellular storage location of IMCL is altered by limb immobilization, highlighting the need to evaluate IMCL storage location when assessing the effects of disuse on IMCL content. We also found that AKT content increased during immobilization in our female population, contrary to studies in males finding that AKT decreases during disuse, highlighting that men and women may respond differently to disuse and the necessity to include women in all research.

Methodological considerations for and validation of the ultrasonographic determination of human skeletal muscle hypertrophy and atrophy.

Magnetic resonance imaging (MRI) is the current gold standard for measuring changes in muscle size (cross-sectional area [CSA] and volume) but can be cost-prohibitive and resource-intensive. We evaluated the validity of B-mode ultrasonography (US) as a low-cost alternative to MRI for measuring muscle hypertrophy and atrophy in response to resistance training and immobilization, respectively. Fourteen young men performed 10wk of unilateral resistance training (RT) to induce muscle hypertrophy. In the final two weeks of the 10wk, the subjects' contralateral leg was immobilized (IMB). The cross-sectional area of the vastus lateralis (VLCSA) was measured at the mid-thigh before and after each intervention using MRI (VLCSAMRI ) and US (VLCSAUS ). The relationship and agreement between methods were assessed. Reliability of US measurements ranged from good to excellent in all comparisons (ICC >0.67). VLCSA significantly increased after 10 weeks of RT (VLCSAUS : 7.9 ± 3.8%; VLCSAMRI : 7.8 ± 4.5%) and decreased after 2 weeks of IMB (VLCSAUS : -8.2%±5.8%; VLCSAMRI : -8.7 ± 6.1%). Significant correlations were identified between MRI and US at each time point measured (all r > 0.85) and, importantly, between MRI- and US-derived changes in VLCSA. Bland-Altman analysis revealed minimal bias in US measurements relative to the MRI (-0.5 ± 3.0%) and all measurements were within the upper and lower limits of agreement. Our data suggest that B-mode ultrasonography can be a suitable alternative to MRI for measuring changes in muscle size in response to increased and decreased muscle loading in young men.