Short-term high-calorie high-fat feeding induces hyperinsulinemia and blunts skeletal muscle microvascular blood flow in healthy humans.

Skeletal muscle microvascular blood flow (MBF) plays an important role in glucose disposal in muscle. Impairments in muscle MBF contribute to insulin resistance and prediabetes. Animal studies show that short-term (3 day) high-fat feeding blunts skeletal muscle MBF before impairing insulin-stimulated glucose disposal. It is not known whether this occurs in humans. We investigated the temporal impact of a 7-day high-calorie high-fat (HCHF) diet intervention (+52% kJ; 41% fat) on fasting and postprandial cardiometabolic outcomes in 14 healthy adults (18-37 yr). Metabolic health and vascular responses to a mixed-meal challenge (MMC) were measured at pre (day 0)-, mid (day 4)- and post (day 8)-intervention. There were no significant differences in body weight, body fat %, fasting blood glucose, and fasting plasma insulin concentrations at pre-, mid- and postintervention. Compared with preintervention there was a significant increase in insulin (but not glucose) total area under the curve in response to the MMC at midintervention (P = 0.041) and at postintervention (P = 0.028). Unlike at pre- and midintervention, at postintervention muscle MBF decreased at 60 min (P = 0.024) and 120 min (P = 0.023) after the MMC. However, macrovascular blood flow was significantly increased from 0 to 60 min (P < 0.001) and 120 min (P < 0.001) after the MMC at pre-, mid- and postintervention. Therefore, short-term HCHF feeding in healthy individuals leads to elevated postprandial insulin but not glucose levels and a blunting of meal-induced skeletal muscle MBF responses but not macrovascular blood flow responses.NEW & NOTEWORTHY This is the first study to investigate skeletal muscle microvascular blood flow (MBF) responses in humans after short-term high-calorie high-fat (HCHF) diet. The main findings were that HCHF diet causes elevated postprandial insulin in healthy individuals within 3 days and blunts meal-induced muscle MBF within 7 days, despite no impairments in postprandial glucose or macrovascular blood flow.

Similar muscle hypertrophy following eight weeks of resistance training to momentary muscular failure or with repetitions-in-reserve in resistance-trained individuals.

This study examined the influence of resistance training (RT) proximity-to-failure, determined by repetitions-in-reserve (RIR), on quadriceps hypertrophy and neuromuscular fatigue. Resistance-trained males (n = 12) and females (n = 6) completed an 8-week intervention involving two RT sessions per week. Lower limbs were randomised to perform the leg press and leg extension exercises either to i) momentary muscular failure (FAIL), or ii) a perceived 2-RIR and 1-RIR, respectively (RIR). Muscle thickness of the quadriceps [rectus femoris (RF) and vastus lateralis (VL)] and acute neuromuscular fatigue (i.e., repetition and lifting velocity loss) were assessed. Data was analysed with Bayesian linear mixed-effect models. Increases in quadriceps thickness (average of RF and VL) from pre- to post-intervention were similar for FAIL [0.181 cm (HDI: 0.119 to 0.243)] and RIR [0.182 cm (HDI: 0.115 to 0.247)]. Between-protocol differences in RF thickness slightly favoured RIR [-0.036 cm (HDI: -0.113 to 0.047)], but VL thickness slightly favoured FAIL [0.033 cm (HDI: -0.046 to 0.116)]. Mean volume was similar across the RT intervention between FAIL and RIR. Lifting velocity and repetition loss were consistently greater for FAIL versus RIR, with the magnitude of difference influenced by the exercise and the stage of the RT intervention.

Terminating RT sets with a close proximity-to-failure (e.g., 1- to 2-RIR) can be sufficient to promote similar hypertrophy of the quadriceps as reaching momentary muscular failure in resistance-trained individuals over eight weeks, but the overall influence of proximity-to-failure on muscle-specific hypertrophy may also depend on other factors (e.g., exercise selection, order, and subsequent musculature targeted).Due to high repetition loss (from the first to final set) when sets are terminated at momentary muscular failure, performing RT with 1- to 2-RIR allows for similar volume load and repetition volume accumulation as reaching momentary muscular failure across eight weeks, possibly influencing the overall RT stimulus achieved.Performing RT to momentary muscular failure consistently induces higher levels of acute neuromuscular fatigue versus RT performed with 1- to 2-RIR; however, improved fatigue resistance overtime may attenuate acute neuromuscular fatigue and subsequent repetition loss (but may depend on the exercise performed).

Accuracy of Intraset Repetitions-in-Reserve Predictions During the Bench Press Exercise in Resistance-Trained Male and Female Subjects.

ABSTRACT: Refalo, MC, Remmert, JF, Pelland, JC, Robinson, ZP, Zourdos, MC, Hamilton, DL, Fyfe, JJ, and Helms, ER. Accuracy of intraset repetitions-in-reserve predictions during the bench press exercise in resistance-trained male and female subjects. J Strength Cond Res 38(3): e78-e85, 2024-This study assessed the accuracy of intraset repetitions-in-reserve (RIR) predictions to provide evidence for the efficacy of RIR prescription as a set termination method to inform proximity to failure during resistance training (RT). Twenty-four resistance trained male ( n = 12) and female ( n = 12) subjects completed 2 experimental sessions involving 2 sets performed to momentary muscular failure (barbell bench press exercise) with 75% of 1 repetition maximum (1RM), whereby subjects verbally indicated when they perceived to had reached either 1 RIR or 3 RIR. The difference between the predicted RIR and the actual RIR was defined as the "RIR accuracy" and was quantified as both raw (i.e., direction of error) and absolute (i.e., magnitude of error) values. High raw and absolute mean RIR accuracy (-0.17 ± 1.00 and 0.65 ± 0.78 repetitions, respectively) for 1-RIR and 3-RIR predictions were observed (including all sets and sessions completed). We identified statistical equivalence (equivalence range of ±1 repetition, thus no level of statistical significance was set) in raw and absolute RIR accuracy between (a) 1-RIR and 3-RIR predictions, (b) set 1 and set 2, and (c) session 1 and session 2. No evidence of a relationship was found between RIR accuracy and biological sex, years of RT experience, or relative bench press strength. Overall, resistance-trained individuals are capable of high absolute RIR accuracy when predicting 1 and 3 RIR on the barbell bench press exercise, with a minor tendency for underprediction. Thus, RIR prescriptions may be used in research and practice to inform the proximity to failure achieved upon set termination.

Enhanced skeletal muscle contractile function and corticospinal excitability precede strength and architectural adaptations during lower-limb resistance training.

PURPOSE: Evolving investigative techniques are providing greater understanding about the early neuromuscular responses to resistance training among novice exercisers. The aim of this study was to investigate the time-course of changes in muscle contractile mechanics, architecture, neuromuscular, and strength adaptation during the first 6-weeks of lower-limb resistance training. METHODS: Forty participants: 22 intervention (10 males/12 females; 173.48 ± 5.20 cm; 74.01 ± 13.13 kg) completed 6-week resistance training, and 18 control (10 males/8 females; 175.52 ± 7.64 cm; 70.92 ± 12.73 kg) performed no resistance training and maintained their habitual activity. Radial muscle displacement (Dm) assessed via tensiomyography, knee extension maximal voluntary contraction (MVC), voluntary activation (VA), corticospinal excitability and inhibition via transcranial magnetic stimulation, motor unit (MU) firing rate, and muscle thickness and pennation angle via ultrasonography were assessed before and after 2, 4, and 6-weeks of dynamic lower-limb resistance training or control. RESULTS: After 2-weeks training, Dm reduced by 19-25% in the intervention group; this was before any changes in neural or morphological measures. After 4-weeks training, MVC increased by 15% along with corticospinal excitability by 16%; however, there was no change in VA, corticospinal inhibition, or MU firing rate. After 6-weeks training there was further MVC increase by 6% along with muscle thickness by 13-16% and pennation angle by 13-14%. CONCLUSION: Enhanced contractile properties and corticospinal excitability occurred before any muscle architecture, neural, and strength adaptation. Later increases in muscular strength can be accounted for by architectural adaptation.

Towards an improved understanding of proximity-to-failure in resistance training and its influence on skeletal muscle hypertrophy, neuromuscular fatigue, muscle damage, and perceived discomfort: A scoping review.

While proximity-to-failure is considered an important resistance training (RT) prescription variable, its influence on physiological adaptations and short-term responses to RT is uncertain. Given the ambiguity in the literature, a scoping review was undertaken to summarise evidence for the influence of proximity-to-failure on muscle hypertrophy, neuromuscular fatigue, muscle damage and perceived discomfort. Literature searching was performed according to PRISMA-ScR guidelines and identified three themes of studies comparing either: i) RT performed to momentary muscular failure versus non-failure, ii) RT performed to set failure (defined as anything other than momentary muscular failure) versus non-failure, and iii) RT performed to different velocity loss thresholds. The findings highlight that no consensus definition for "failure" exists in the literature, and the proximity-to-failure achieved in "non-failure" conditions is often ambiguous and variable across studies. This poses challenges when deriving practical recommendations for manipulating proximity-to-failure in RT to achieve desired outcomes. Based on the limited available evidence, RT to set failure is likely not superior to non-failure RT for inducing muscle hypertrophy, but may exacerbate neuromuscular fatigue, muscle damage, and post-set perceived discomfort versus non-failure RT. Together, these factors may impair post-exercise recovery and subsequent performance, and may also negatively influence long-term adherence to RT.KEY POINTS This scoping review identified three broad themes of studies investigating proximity-to-failure in RT, based on the specific definition of set failure used (and therefore the research question being examined), to improve the validity of study comparisons and interpretations.There is no consensus definition for set failure in RT, and the proximity-to-failure achieved during non-failure RT is often unclear and varies both within and between studies, which together poses challenges when interpreting study findings and deriving practical recommendations regarding the influence of RT proximity-to-failure on muscle hypertrophy and other short-term responses.Based on the limited available evidence, performing RT to set failure is likely not superior to non-failure RT to maximise muscle hypertrophy, but the optimal proximity to failure in RT for muscle hypertrophy is unclear and may be moderated by other RT variables (e.g., load, volume-load). Also, RT performed to set failure likely induces greater neuromuscular fatigue, muscle damage, and perceived discomfort than non-failure RT, which may negatively influence RT performance, post-RT recovery, and long-term adherence.

Bayesian analysis of changes in standing horizontal and vertical jump after different modes of resistance training.

Training interventions often have small effects and are tested in small samples. We used a Bayesian approach to examine the change in jump distance after different resistance training programmes. Thirty-three 18- to 45-year-old males completed one of three lower limb resistance training programmes: deadlift (DL), hip thrust (HT) or back squat (BS). Horizontal and vertical jump performance was assessed over the training intervention. Examination of Bayesian posterior distributions for jump distance estimated that the probability of a change above a horizontal jump smallest worthwhile change (SWC) of 4.7 cm for the DL group was ~12%. For the HT and BS groups, the probability of a change above the SWC was ~87%. The probability of a change above a vertical jump SWC of 1.3 cm for the DL group was ~31%. For the HT and BS groups, the probability of a change above the vertical jump SWC was ~62% and ~67%, respectively. Our study illustrates that a Bayesian approach provides a rich inferential interpretation for small sample training studies with small effects. The extra information from such a Bayesian approach is useful to practitioners in Sport and Exercise Science where small effects are expected and sample size is often constrained.

Influence of resistance training load on measures of skeletal muscle hypertrophy and improvements in maximal strength and neuromuscular task performance: A systematic review and meta-analysis.

This systematic review and meta-analysis determined resistance training (RT) load effects on various muscle hypertrophy, strength, and neuromuscular performance task [e.g., countermovement jump (CMJ)] outcomes. Relevent studies comparing higher-load [>60% 1-repetition maximum (RM) or <15-RM] and lower-load (≤60% 1-RM or ≥ 15-RM) RT were identified, with 45 studies (from 4713 total) included in the meta-analysis. Higher- and lower-load RT induced similar muscle hypertrophy at the whole-body (lean/fat-free mass; [ES (95% CI) = 0.05 (-0.20 to 0.29), P = 0.70]), whole-muscle [ES = 0.06 (-0.11 to 0.24), P = 0.47], and muscle fibre [ES = 0.29 (-0.09 to 0.66), P = 0.13] levels. Higher-load RT further improved 1-RM [ES = 0.34 (0.15 to 0.52), P = 0.0003] and isometric [ES = 0.41 (0.07 to 0.76), P = 0.02] strength. The superiority of higher-load RT on 1-RM strength was greater in younger [ES = 0.34 (0.12 to 0.55), P = 0.002] versus older [ES = 0.20 (-0.00 to 0.41), P = 0.05] participants. Higher- and lower-load RT therefore induce similar muscle hypertrophy (at multiple physiological levels), while higher-load RT elicits superior 1-RM and isometric strength. The influence of RT loads on neuromuscular task performance is however unclear.

Protein-carbohydrate ingestion alters Vps34 cellular localization independent of changes in kinase activity in human skeletal muscle.

NEW FINDINGS: What is the central question of the study? Is Vps34 a nutrient-sensitive activator of mTORC1 in human skeletal muscle? What is the main finding and its importance? We show that altering nutrient availability, via protein-carbohydrate feeding, does not increase Vps34 kinase activity in human skeletal muscle. Instead, feeding increased Vps34-mTORC1 co-localization in parallel to increased mTORC1 activity. These findings may have important implications in the understanding nutrient-induced mTORC1 activation in skeletal muscle via interaction with Vps34. ABSTRACT: The Class III PI3Kinase, Vps34, has recently been proposed as a nutrient sensor, essential for activation of the mechanistic target of rapamycin (mTOR) complex 1 (mTORC1). We therefore investigated the effects of increasing nutrient availability through protein-carbohydrate (PRO-CHO) feeding on Vps34 kinase activity and cellular localization in human skeletal muscle. Eight young, healthy males (21 ± 0.5 yrs, 77.7 ± 9.9 kg, 25.9 ± 2.7 kg/m2 , mean ± SD) ingested a PRO-CHO beverage containing 20/44/1 g PRO/CHO/FAT respectively, with skeletal muscle biopsies obtained at baseline and 1 h and 3 h post-feeding. PRO-CHO feeding did not alter Vps34 kinase activity, but did stimulate Vps34 translocation toward the cell periphery (PRE (mean ± SD) - 0.273 ± 0.040, 1 h - 0.348 ± 0.061, Pearson's Coefficient (r)) where it co-localized with mTOR (PRE - 0.312 ± 0.040, 1 h - 0.348 ± 0.069, Pearson's Coefficient (r)). These alterations occurred in parallel to an increase in S6K1 kinase activity (941 ± 466% of PRE at 1 h post-feeding). Subsequent in vitro experiments in C2C12 and human primary myotubes displayed no effect of the Vps34-specific inhibitor SAR405 on mTORC1 signalling responses to elevated nutrient availability. Therefore, in summary, PRO-CHO ingestion does not increase Vps34 activity in human skeletal muscle, whilst pharmacological inhibition of Vps34 does not prevent nutrient stimulation of mTORC1 in vitro. However, PRO-CHO ingestion promotes Vps34 translocation to the cell periphery, enabling Vps34 to associate with mTOR. Therefore, our data suggests that interaction between Vps34 and mTOR, rather than changes in Vps34 activity per se may be involved in PRO-CHO activation of mTORC1 in human skeletal muscle.

Skipping Breakfast Before Exercise Creates a More Negative 24-hour Energy Balance: A Randomized Controlled Trial in Healthy Physically Active Young Men.

BACKGROUND: At rest, omission of breakfast lowers daily energy intake, but also lowers energy expenditure, attenuating any effect on energy balance. The effect of breakfast omission on energy balance when exercise is prescribed is unclear. OBJECTIVES: The aim of this study was to assess the effect on 24-h energy balance of omitting compared with consuming breakfast prior to exercise. METHODS: Twelve healthy physically active young men (age 23 ± 3 y, body mass index 23.6 ± 2.0 kg/m2) completed 3 trials in a randomized order (separated by >1 week): a breakfast of oats and milk (431 kcal; 65 g carbohydrate, 11 g fat, 19 g protein) followed by rest (BR); breakfast before exercise (BE; 60 min cycling at 50 % peak power output); and overnight fasting before exercise (FE). The 24-h energy intake was calculated based on the food consumed for breakfast, followed by an ad libitum lunch, snacks, and dinner. Indirect calorimetry with heart-rate accelerometry was used to measure substrate utilization and 24-h energy expenditure. A [6,6-2H2]glucose infusion was used to investigate tissue-specific carbohydrate utilization. RESULTS: The 24-h energy balance was -400 kcal (normalized 95% CI: -230, -571 kcal) for the FE trial; this was significantly lower than both the BR trial (492 kcal; normalized 95% CI: 332, 652 kcal) and the BE trial (7 kcal; normalized 95% CI: -153, 177 kcal; both P < 0.01 compared with FE). Plasma glucose utilization in FE (mainly representing liver glucose utilization) was positively correlated with energy intake compensation at lunch (r = 0.62, P = 0.03), suggesting liver carbohydrate plays a role in postexercise energy-balance regulation. CONCLUSIONS: Neither exercise energy expenditure nor restricted energy intake via breakfast omission were completely compensated for postexercise. In healthy men, pre-exercise breakfast omission creates a more negative daily energy balance and could therefore be a useful strategy to induce a short-term energy deficit. This trial was registered at clinicaltrials.gov as NCT02258399.

Influence of the "Slingshot" Bench Press Training Aid on Bench Press Kinematics and Neuromuscular Activity in Competitive Powerlifters.

Dugdale, JH, Hunter, AM, Di Virgilio, TG, Macgregor, LJ, and Hamilton, DL. Influence of the "Slingshot" bench press training aid on bench press kinematics and neuromuscular activity in competitive powerlifters. J Strength Cond Res 33(2): 327-336, 2019-This study examined the acute effects of the "Slingshot" (SS) on bench press performance, prime mover surface electromyographic (sEMG) amplitude, and barbell velocity during maximal and submaximal bench pressing in competitive male powerlifters. Fifteen male powerlifters (mean ± SD; age: 27.05 ± 5.94 years; mass: 94.15 ± 13.43 kg; 1 repetition maximum [1RM] bench press: 139.7 ± 16.79 kg) participated in the study. Bench press strength, average barbell velocity, and sEMG amplitude of the prime mover muscles (triceps brachii, pectoralis major, and anterior deltoid) were measured during 2 conditions; "Raw" (without use of any assistance) and "SS" (using the "Slingshot" to perform both the weight achieved during "Raw" 1RM testing [Raw max/SS], and absolute 1RM using the "SS"). The results showed that the "SS" significantly increased bench press 1RM performance by a mean ± SD of 20.67 ± 3.4 kg. Barbell velocity and stick point analysis indicate that this improvement is likely driven by an increase in peak and prestick barbell velocity as triceps root mean square (RMS) was lower throughout all rep max phases with the "SS." The "SS" also caused reductions in RMS, specifically of the triceps at all rep ranges but barbell velocity was better maintained in the last reps of all sets. These data indicate that the "SS" specifically deloaded the triceps muscle throughout all rep ranges and provide assistance to maintaining barbell velocity under fatigue during later repetitions of multiple repetition sets. The "SS" training aid could therefore be used in deload phases of bench press training or as an overreaching and velocity training aid.

Preexercise breakfast ingestion versus extended overnight fasting increases postprandial glucose flux after exercise in healthy men.

The aim of this study was to characterize postprandial glucose flux after exercise in the fed versus overnight fasted state and to investigate the potential underlying mechanisms. In a randomized order, twelve men underwent breakfast-rest [(BR) 3 h semirecumbent], breakfast-exercise [(BE) 2 h semirecumbent before 60 min of cycling (50% peak power output)], and overnight fasted exercise [(FE) as per BE omitting breakfast] trials. An oral glucose tolerance test (OGTT) was completed after exercise (after rest on BR). Dual stable isotope tracers ([U-13C] glucose ingestion and [6,6-2H2] glucose infusion) and muscle biopsies were combined to assess postprandial plasma glucose kinetics and intramuscular signaling, respectively. Plasma intestinal fatty acid binding (I-FABP) concentrations were determined as a marker of intestinal damage. Breakfast before exercise increased postexercise plasma glucose disposal rates during the OGTT, from 44 g/120 min in FE {35 to 53 g/120 min [mean (normalized 95% confidence interval)] to 73 g/120 min in BE [55 to 90 g/120 min; P = 0.01]}. This higher plasma glucose disposal rate was, however, offset by increased plasma glucose appearance rates (principally OGTT-derived), resulting in a glycemic response that did not differ between BE and FE ( P = 0.11). Plasma I-FABP concentrations during exercise were 264 pg/ml (196 to 332 pg/ml) lower in BE versus FE ( P = 0.01). Breakfast before exercise increases postexercise postprandial plasma glucose disposal, which is offset (primarily) by increased appearance rates of orally ingested glucose. Therefore, metabolic responses to fed-state exercise cannot be readily inferred from studies conducted in a fasted state.

Lipid remodeling and an altered membrane-associated proteome may drive the differential effects of EPA and DHA treatment on skeletal muscle glucose uptake and protein accretion.

In striated muscle, eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) have differential effects on the metabolism of glucose and differential effects on the metabolism of protein. We have shown that, despite similar incorporation, treatment of C2C12 myotubes (CM) with EPA but not DHA improves glucose uptake and protein accretion. We hypothesized that these differential effects of EPA and DHA may be due to divergent shifts in lipidomic profiles leading to altered proteomic profiles. We therefore carried out an assessment of the impact of treating CM with EPA and DHA on lipidomic and proteomic profiles. Fatty acid methyl esters (FAME) analysis revealed that both EPA and DHA led to similar but substantials changes in fatty acid profiles with the exception of arachidonic acid, which was decreased only by DHA, and docosapentanoic acid (DPA), which was increased only by EPA treatment. Global lipidomic analysis showed that EPA and DHA induced large alterations in the cellular lipid profiles and in particular, the phospholipid classes. Subsequent targeted analysis confirmed that the most differentially regulated species were phosphatidylcholines and phosphatidylethanolamines containing long-chain fatty acids with five (EPA treatment) or six (DHA treatment) double bonds. As these are typically membrane-associated lipid species we hypothesized that these treatments differentially altered the membrane-associated proteome. Stable isotope labeling by amino acids in cell culture (SILAC)-based proteomics of the membrane fraction revealed significant divergence in the effects of EPA and DHA on the membrane-associated proteome. We conclude that the EPA-specific increase in polyunsaturated long-chain fatty acids in the phospholipid fraction is associated with an altered membrane-associated proteome and these may be critical events in the metabolic remodeling induced by EPA treatment.

Multiple AMPK activators inhibit l-carnitine uptake in C2C12 skeletal muscle myotubes.

Mutations in the gene that encodes the principal l-carnitine transporter, OCTN2, can lead to a reduced intracellular l-carnitine pool and the disease Primary Carnitine Deficiency. l-Carnitine supplementation is used therapeutically to increase intracellular l-carnitine. As AMPK and insulin regulate fat metabolism and substrate uptake, we hypothesized that AMPK-activating compounds and insulin would increase l-carnitine uptake in C2C12 myotubes. The cells express all three OCTN transporters at the mRNA level, and immunohistochemistry confirmed expression at the protein level. Contrary to our hypothesis, despite significant activation of PKB and 2DG uptake, insulin did not increase l-carnitine uptake at 100 nM. However, l-carnitine uptake was modestly increased at a dose of 150 nM insulin. A range of AMPK activators that increase intracellular calcium content [caffeine (10 mM, 5 mM, 1 mM, 0.5 mM), A23187 (10 µM)], inhibit mitochondrial function [sodium azide (75 µM), rotenone (1 µM), berberine (100 µM), DNP (500 µM)], or directly activate AMPK [AICAR (250 µM)] were assessed for their ability to regulate l-carnitine uptake. All compounds tested significantly inhibited l-carnitine uptake. Inhibition by caffeine was not dantrolene (10 µM) sensitive despite dantrolene inhibiting caffeine-mediated calcium release. Saturation curve analysis suggested that caffeine did not competitively inhibit l-carnitine transport. To assess the potential role of AMPK in this process, we assessed the ability of the AMPK inhibitor Compound C (10 µM) to rescue the effect of caffeine. Compound C offered a partial rescue of l-carnitine uptake with 0.5 mM caffeine, suggesting that AMPK may play a role in the inhibitory effects of caffeine. However, caffeine likely inhibits l-carnitine uptake by alternative mechanisms independently of calcium release. PKA activation or direct interference with transporter function may play a role.

Differential localization and anabolic responsiveness of mTOR complexes in human skeletal muscle in response to feeding and exercise.

Mechanistic target of rapamycin (mTOR) resides as two complexes within skeletal muscle. mTOR complex 1 [mTORC1-regulatory associated protein of mTOR (Raptor) positive] regulates skeletal muscle growth, whereas mTORC2 [rapamycin-insensitive companion of mTOR (Rictor) positive] regulates insulin sensitivity. To examine the regulation of these complexes in human skeletal muscle, we utilized immunohistochemical analysis to study the localization of mTOR complexes before and following protein-carbohydrate feeding (FED) and resistance exercise plus protein-carbohydrate feeding (EXFED) in a unilateral exercise model. In basal samples, mTOR and the lysosomal marker lysosomal associated membrane protein 2 (LAMP2) were highly colocalized and remained so throughout. In the FED and EXFED states, mTOR/LAMP2 complexes were redistributed to the cell periphery [wheat germ agglutinin (WGA)-positive staining] (time effect; P = 0.025), with 39% (FED) and 26% (EXFED) increases in mTOR/WGA association observed 1 h post-feeding/exercise. mTOR/WGA colocalization continued to increase in EXFED at 3 h (48% above baseline) whereas colocalization decreased in FED (21% above baseline). A significant effect of condition (P = 0.05) was noted suggesting mTOR/WGA colocalization was greater during EXFED. This pattern was replicated in Raptor/WGA association, where a significant difference between EXFED and FED was noted at 3 h post-exercise/feeding (P = 0.014). Rictor/WGA colocalization remained unaltered throughout the trial. Alterations in mTORC1 cellular location coincided with elevated S6K1 kinase activity, which rose to a greater extent in EXFED compared with FED at 1 h post-exercise/feeding (P < 0.001), and only remained elevated in EXFED at the 3 h time point (P = 0.037). Collectively these data suggest that mTORC1 redistribution within the cell is a fundamental response to resistance exercise and feeding, whereas mTORC2 is predominantly situated at the sarcolemma and does not alter localization.

Leucine does not affect mechanistic target of rapamycin complex 1 assembly but is required for maximal ribosomal protein s6 kinase 1 activity in human skeletal muscle following resistance exercise.

We examined how the stimulatory effect of leucine on the mechanistic target of rapamycin complex 1 (mTORC1) pathway is affected by the presence of the remaining essential amino acids (EAAs). Nine male subjects performed resistance exercise on 4 occasions and were randomly supplied EAAs with leucine, EAAs without leucine (EAA-Leu), leucine alone, or flavored water (placebo; control). Muscle biopsies were taken from the vastus lateralis before and 60 and 90 min after exercise. Biopsies were analyzed for protein phosphorylation, kinase activity, protein-protein interactions, amino acid concentrations, and tracer incorporation. Leucine alone stimulated ribosomal protein s6 kinase 1 (S6K1) phosphorylation ∼280% more than placebo and EAA-Leu after exercise. Moreover, this response was enhanced by 60-75% after intake of EAAs compared with that of leucine alone (P < 0.05). Kinase activity of S6K1 reflected that of S6K1 phosphorylation; 60 min after exercise, the activity was elevated 3.3- and 4.2-fold with intake of leucine alone and with EAAs, respectively (P < 0.05). The interaction between mammalian target of rapamycin and regulatory-associated protein of mammalian target of rapamycin was unaltered in response to both resistance exercise and amino acid provision. Leucine alone stimulates mTORC1 signaling, although this response is enhanced by other EAAs and does not appear to be caused by alterations in mTORC1 assembly.

Live strong and prosper: the importance of skeletal muscle strength for healthy ageing.

Due to improved health care, diet and infrastructure in developed countries, since 1840 life expectancy has increased by approximately 2 years per decade. Accordingly, by 2050, a quarter of Europe's population will be over 65 years, representing a 10 % rise in half a century. With this rapid rise comes an increased prevalence of diseases of ageing and associated healthcare expenditure. To address the health consequences of global ageing, research in model systems (worms, flies and mice) has indicated that reducing the rate of organ growth, via reductions in protein synthetic rates, has multi-organ health benefits that collectively lead to improvements in lifespan. In contrast, human pre-clinical, clinical and large cohort prospective studies demonstrate that ageing leads to anabolic (i.e. growth) impairments in skeletal muscle, which in turn leads to reductions in muscle mass and strength, factors directly associated with mortality rates in the elderly. As such, increasing muscle protein synthesis via exercise or protein-based nutrition maintains a strong, healthy muscle mass, which in turn leads to improved health, independence and functionality. The aim of this review is to critique current literature relating to the maintenance of muscle mass across lifespan and discuss whether maintaining or reducing protein synthesis is the most logical approach to support musculoskeletal function and by extension healthy human ageing.

Growing older with health and vitality: a nexus of physical activity, exercise and nutrition.

The preservation of skeletal muscle mass and strength with advancing age are, we propose, critical aspects of ageing with health and vitality. Physical inactivity and poor nutrition are known to accelerate the gradual age-related decline in muscle mass and strength-sarcopenia-however, both are subject to modification. The main purpose of this review is to present the latest, evidence-based recommendations for physical activity and exercise, as well as diet for older adults that would help in preserving muscle mass and strength. We take the position that future physical activity/exercise guidelines need to make specific reference to resistance exercise and highlight the benefits of higher-intensity aerobic exercise training, alongside advocating older adults perform aerobic-based physical activity and household tasks (e.g., carrying groceries). In terms of dietary recommendations, greater emphasis should be placed on optimal rather than minimum protein intakes for older adults. Indeed, guidelines that endorse a daily protein intake of 1.2-1.5 g/kg BM/day, which are levels 50-90 % greater than the current protein Recommendation Dietary Allowance (0.8 g/kg BM/day), are likely to help preserve muscle mass and strength and are safe for healthy older adults. Being cognisant of factors (e.g., reduced appetite) that may preclude older adults from increasing their total daily protein intake, we echo the viewpoint of other active researchers in advocating that protein recommendations for older adults be based on a per meal approach in order to maximize muscle protein synthesis (MPS). On this basis, assuming three meals are consumed daily, a protein dose of 0.4-0.5 g/kg BM should be contained in each meal. We are beginning to understand ways in which to increase the utilization of ingested protein for the stimulation of MPS, namely by increasing the proportion of leucine contained in a given dose of protein, co-ingesting other nutrients (e.g., carbohydrate and fat or supplementation with n-3 polyunsaturated fatty acids) or being physically active prior to protein intake. Clearly, developing simple lifestyle interventions targeted at preserving muscle mass and strength with advancing age is crucial for facilitating longer, healthier lives into older age.

Rapamycin does not prevent increases in myofibrillar or mitochondrial protein synthesis following endurance exercise.

The present study aimed to investigate the role of the mechanistic target of rapamycin complex 1 (mTORC1) in the regulation of myofibrillar (MyoPS) and mitochondrial (MitoPS) protein synthesis following endurance exercise. Forty-two female C57BL/6 mice performed 1 h of treadmill running (18 m min(-1) ; 5° grade), 1 h after i.p. administration of rapamycin (1.5 mg · kg(-1) ) or vehicle. To quantify skeletal muscle protein fractional synthesis rates, a flooding dose (50 mg · kg(-1) ) of l-[ring-(13) C6 ]phenylalanine was administered via i.p. injection. Blood and gastrocnemius muscle were collected in non-exercised control mice, as well as at 0.5, 3 and 6 h after completing exercise (n = 4 per time point). Skeletal muscle MyoPS and MitoPS were determined by measuring isotope incorporation in their respective protein pools. Activation of the mTORC1-signalling cascade was measured via direct kinase activity assay and immunoblotting, whereas genes related to mitochondrial biogenesis were measured via a quantitative RT-PCR. MyoPS increased rapidly in the vehicle group post-exercise and remained elevated for 6 h, whereas this response was transiently blunted (30 min post-exercise) by rapamycin. By contrast, MitoPS was unaffected by rapamycin, and was increased over the entire post-exercise recovery period in both groups (P < 0.05). Despite rapid increases in both MyoPS and MitoPS, mTORC1 activation was suppressed in both groups post-exercise for the entire 6 h recovery period. Peroxisome proliferator activated receptor-γ coactivator-1α, pyruvate dehydrogenase kinase 4 and mitochondrial transcription factor A mRNA increased post-exercise (P < 0.05) and this response was augmented by rapamycin (P < 0.05). Collectively, these data suggest that endurance exercise stimulates MyoPS and MitoPS in skeletal muscle independently of mTORC1 activation.

Resistance exercise-induced S6K1 kinase activity is not inhibited in human skeletal muscle despite prior activation of AMPK by high-intensity interval cycling.

Combining endurance and strength training in the same session has been reported to reduce the anabolic response to the latter form of exercise. The underlying mechanism, based primarily on results from rodent muscle, is proposed to involve AMPK-dependent inhibition of mTORC1 signaling. This hypothesis was tested in eight trained male subjects who in randomized order performed either resistance exercise only (R) or interval cycling followed by resistance exercise (ER). Biopsies taken from the vastus lateralis before and after endurance exercise and repeatedly after resistance exercise were assessed for glycogen content, kinase activity, protein phosphorylation, and gene expression. Mixed muscle fractional synthetic rate was measured at rest and during 3 h of recovery using the stable isotope technique. In ER, AMPK activity was elevated immediately after both endurance and resistance exercise (∼90%, P < 0.05) but was unchanged in R. Thr(389) phosphorylation of S6K1 was increased severalfold immediately after exercise (P < 0.05) in both trials and increased further throughout recovery. After 90 and 180 min recovery, S6K1 activity was elevated (∼55 and ∼110%, respectively, P < 0.05) and eukaryotic elongation factor 2 phosphorylation was reduced (∼55%, P < 0.05) with no difference between trials. In contrast, markers for protein catabolism were differently influenced by the two modes of exercise; ER induced a significant increase in gene and protein expression of MuRF1 (P < 0.05), which was not observed following R exercise only. In conclusion, cycling-induced elevation in AMPK activity does not inhibit mTOR complex 1 signaling after subsequent resistance exercise but may instead interfere with the hypertrophic response by influencing key components in protein breakdown.

Omega-3 Fatty Acids and Skeletal Muscle Health.

Skeletal muscle is a plastic tissue capable of adapting and mal-adapting to physical activity and diet. The response of skeletal muscle to adaptive stimuli, such as exercise, can be modified by the prior nutritional status of the muscle. The influence of nutrition on skeletal muscle has the potential to substantially impact physical function and whole body metabolism. Animal and cell based models show that omega-3 fatty acids, in particular those of marine origin, can influence skeletal muscle metabolism. Furthermore, recent human studies demonstrate that omega-3 fatty acids of marine origin can influence the exercise and nutritional response of skeletal muscle. These studies show that the prior omega-3 status influences not only the metabolic response of muscle to nutrition, but also the functional response to a period of exercise training. Omega-3 fatty acids of marine origin therefore have the potential to alter the trajectory of a number of human diseases including the physical decline associated with aging. We explore the potential molecular mechanisms by which omega-3 fatty acids may act in skeletal muscle, considering the n-3/n-6 ratio, inflammation and lipidomic remodelling as possible mechanisms of action. Finally, we suggest some avenues for further research to clarify how omega-3 fatty acids may be exerting their biological action in skeletal muscle.