Muscle Protein Synthesis Rates Research Articles

Neuromuscular electrical stimulation prior to presleep protein feeding stimulates the use of protein-derived amino acids for overnight muscle protein synthesis.

Neuromuscular electrical stimulation (NMES) as well as presleep dietary protein ingestion represent effective strategies to stimulate muscle protein synthesis rates. Here we demonstrate that in older men after a day of bed rest, the application of NMES prior to presleep protein feeding stimulates the use of dietary protein-derived amino acids for overnight muscle protein synthesis by 18% compared with presleep protein feeding only.

Anabolic sensitivity of postprandial muscle protein synthesis to the ingestion of a protein-dense food is reduced in overweight and obese young adults

Excess body fat diminishes muscle protein synthesis rates in response to hyperinsulinemic-hyperaminoacidemic clamps. However, muscle protein synthetic responses after the ingestion of a protein-dense food source across a range of body mass indexes (BMIs) have not been compared. We compared the myofibrillar protein synthetic response and underlying nutrient-sensing mechanisms after the ingestion of lean pork between obese, overweight, and healthy-weight adults. Ten healthy-weight [HW; BMI (in kg/m2): 22.7 ± 0.4], 10 overweight (OW; BMI: 27.1 ± 0.5), and 10 obese (OB; BMI: 35.9 ± 1.3) adults received primed continuous l-[ring-13C6]phenylalanine infusions. Blood and muscle biopsy samples were collected before and after the ingestion of 170 g pork (36 g protein and 3 g fat) to assess skeletal muscle anabolic signaling, amino acid transporters [large neutral and small neutral amino acid transporters (LAT1, SNAT2) and CD98], and myofibrillar protein synthesis. At baseline, OW and OB groups showed greater relative amounts of mammalian target of rapamycin complex 1 (mTORC1) protein than the HW group. Pork ingestion increased mTORC1 phosphorylation only in the HW group (P = 0.001). LAT1 and SNAT2 protein content increased during the postprandial period in all groups (time effect, P < 0.05). Basal myofibrillar protein synthetic responses were similar between groups (P = 0.43). However, myofibrillar protein synthetic responses (0-300 min) were greater in the HW group (1.6-fold; P = 0.005) after pork ingestion than in the OW and OB groups. There is a diminished myofibrillar protein synthetic response to the ingestion of protein-dense food in overweight and obese adults compared with healthy-weight controls. These data indicate that impaired postprandial myofibrillar protein synthetic response may be an early defect with increasing fat mass, potentially dependent on altered anabolic signals, that reduces muscle sensitivity to food ingestion. This trial was registered at clinicaltrials.gov as NCT02613767.

Both basal and post-prandial muscle protein synthesis rates, following the ingestion of a leucine-enriched whey protein supplement, are not impaired in sarcopenic older males

Studying the muscle protein synthetic response to food intake in elderly is important, as it aids the development of interventions to combat sarcopenia. Although sarcopenic elderly are the target group for many of these nutritional interventions, no studies have assessed basal or post-prandial muscle protein synthesis rates in this population. To assess the basal and post-prandial muscle protein synthesis rates between healthy and sarcopenic older men. A total of 15 healthy (69±1y) and 15 sarcopenic (81±1y) older men ingested a leucine-enriched whey protein nutritional supplement containing 21g of protein, 9g of carbohydrate, and 3g of fat. Stable isotope methodology combined with frequent collection of blood and muscle samples was applied to assess basal and post-prandial muscle protein fractional synthetic rates. Handgrip strength, muscle mass, and gait speed were assessed to identify sarcopenia, according to international criteria. Basal mixed muscle protein fractional synthetic rates (FSR) averaged 0.040±0.005 and 0.032±0.003%/h (mean±SEM) in the sarcopenic and healthy group, respectively (P=0.14). Following protein ingestion, FSR increased significantly to 0.055±0.004 and 0.053±0.004%/h in the post-prandial period in the sarcopenic (P=0.003) and healthy groups (P<0.001), respectively, with no differences between groups (P=0.45). Furthermore, no differences were observed between groups in muscle protein synthesis rates during the early (0.058±0.007 vs 0.060±0.008%/h, sarcopenic vs healthy, respectively) and late (0.052±0.004 vs 0.048±0.003%/h) stages of the post-prandial period (P=0.93 and P=0.34, respectively). Basal muscle protein synthesis rates are not lower in sarcopenic older men compared to healthy older men. The ingestion of 21g of a leucine-enriched whey protein effectively increases muscle protein synthesis rates in both sarcopenic and healthy older men. Public trial registry number: NTR3047.

Protein coingestion with alcohol following strenuous exercise attenuates alcohol-induced intramyocellular apoptosis and inhibition of autophagy.

Alcohol ingestion decreases postexercise rates of muscle protein synthesis, but the mechanism(s) (e.g., increased protein breakdown) underlying this observation is unknown. Autophagy is an intracellular "recycling" system required for homeostatic substrate and organelle turnover; its dysregulation may provoke apoptosis and lead to muscle atrophy. We investigated the acute effects of alcohol ingestion on autophagic cell signaling responses to a bout of concurrent (combined resistance- and endurance-based) exercise. In a randomized crossover design, eight physically active males completed three experimental trials of concurrent exercise with either postexercise ingestion of alcohol and carbohydrate (12 ± 2 standard drinks; ALC-CHO), energy-matched alcohol and protein (ALC-PRO), or protein (PRO) only. Muscle biopsies were taken at rest and 2 and 8 h postexercise. Select autophagy-related gene (Atg) proteins decreased compared with rest with ALC-CHO (P < 0.05) but not ALC-PRO. There were parallel increases (P < 0.05) in p62 and PINK1 commensurate with a reduction in BNIP3 content, indicating a diminished capacity for mitochondria-specific autophagy (mitophagy) when alcohol and carbohydrate were coingested. DNA fragmentation increased in both alcohol conditions (P < 0.05); however, nuclear AIF accumulation preceded this apoptotic response with ALC-CHO only (P < 0.05). In contrast, increases in the nuclear content of p53, TFEB, and PGC-1α in ALC-PRO were accompanied by markers of mitochondrial biogenesis at the transcriptional (Tfam, SCO2, and NRF-1) and translational (COX-IV, ATPAF1, and VDAC1) level (P < 0.05). We conclude that alcohol ingestion following exercise triggers apoptosis, whereas the anabolic properties of protein coingestion may stimulate mitochondrial biogenesis to protect cellular homeostasis.

Ingestion of Wheat Protein Increases In Vivo Muscle Protein Synthesis Rates in Healthy Older Men in a Randomized Trial

Background: Muscle mass maintenance is largely regulated by basal muscle protein synthesis and the capacity to stimulate muscle protein synthesis after food intake. The postprandial muscle protein synthetic response is modulated by the amount, source, and type of protein consumed. It has been suggested that plant-based proteins are less potent in stimulating postprandial muscle protein synthesis than animal-derived proteins. However, few data support this contention.Objective: We aimed to assess postprandial plasma amino acid concentrations and muscle protein synthesis rates after the ingestion of a substantial 35-g bolus of wheat protein hydrolysate compared with casein and whey protein.Methods: Sixty healthy older men [mean ± SEM age: 71 ± 1 y; body mass index (in kg/m2): 25.3 ± 0.3] received a primed continuous infusion of L-[ring-13C6]-phenylalanine and ingested 35 g wheat protein (n = 12), 35 g wheat protein hydrolysate (WPH-35; n = 12), 35 g micellar casein (MCas-35; n = 12), 35 g whey protein (Whey-35; n = 12), or 60 g wheat protein hydrolysate (WPH-60; n = 12). Plasma and muscle samples were collected at regular intervals.Results: The postprandial increase in plasma essential amino acid concentrations was greater after ingesting Whey-35 (2.23 ± 0.07 mM) than after MCas-35 (1.53 ± 0.08 mM) and WPH-35 (1.50 ± 0.04 mM) (P < 0.01). Myofibrillar protein synthesis rates increased after ingesting MCas-35 (P < 0.01) and were higher after ingesting MCas-35 (0.050% ± 0.005%/h) than after WPH-35 (0.032% ± 0.004%/h) (P = 0.03). The postprandial increase in plasma leucine concentrations was greater after ingesting Whey-35 than after WPH-60 (peak value: 580 ± 18 compared with 378 ± 10 μM, respectively; P < 0.01), despite similar leucine contents (4.4 g leucine). Nevertheless, the ingestion of WPH-60 increased myofibrillar protein synthesis rates above basal rates (0.049% ± 0.007%/h; P = 0.02).Conclusions: The myofibrillar protein synthetic response to the ingestion of 35 g casein is greater than after an equal amount of wheat protein. Ingesting a larger amount of wheat protein (i.e., 60 g) substantially increases myofibrillar protein synthesis rates in healthy older men. This trial was registered at clinicaltrials.gov as NCT01952639.

Is Cancer Cachexia Attributed to Impairments in Basal or Postprandial Muscle Protein Metabolism?

Cachexia is a significant clinical problem associated with very poor quality of life, reduced treatment tolerance and outcomes, and a high mortality rate. Mechanistically, any sizeable loss of skeletal muscle mass must be underpinned by a structural imbalance between muscle protein synthesis and breakdown rates. Recent data indicate that the loss of muscle mass with aging is, at least partly, attributed to a blunted muscle protein synthetic response to protein feeding. Whether such anabolic resistance is also evident in conditions where cachexia is present remains to be addressed. Only few data are available on muscle protein synthesis and breakdown rates in vivo in cachectic cancer patients. When calculating the theoretical changes in basal or postprandial fractional muscle protein synthesis and breakdown rates that would be required to lose 5% of body weight within a six-month period, we can define the changes that would need to occur to explain the muscle mass loss observed in cachectic patients. If changes in both post-absorptive and postprandial muscle protein synthesis and breakdown rates contribute to the loss of muscle mass, it would take alterations as small as 1%–2% to induce a more than 5% decline in body weight. Therefore, when trying to define impairments in basal and/or postprandial muscle protein synthesis or breakdown rates using contemporary stable isotope methodology in cancer cachexia, we need to select large homogenous groups of cancer patients (>40 patients) to allow us to measure physiological and clinically relevant differences in muscle protein synthesis and/or breakdown rates. Insight into impairments in basal or postprandial muscle protein synthesis and breakdown rates in cancer cachexia is needed to design more targeted nutritional, pharmaceutical and/or physical activity interventions to preserve skeletal muscle mass and, as such, to reduce the risk of complications, improve quality of life, and lower mortality rates during the various stages of the disease.

Insulin does not stimulate muscle protein synthesis during increased plasma branched-chain amino acids alone but still decreases whole body proteolysis in humans.

Insulin stimulates muscle protein synthesis when the levels of total amino acids, or at least the essential amino acids, are at or above their postabsorptive concentrations. Among the essential amino acids, branched-chain amino acids (BCAA) have the primary role in stimulating muscle protein synthesis and are commonly sought alone to stimulate muscle protein synthesis in humans. Fourteen healthy young subjects were studied before and after insulin infusion to examine whether insulin stimulates muscle protein synthesis in relation to the availability of BCAA alone. One half of the subjects were studied in the presence of postabsorptive BCAA concentrations (control) and the other half in the presence of increased plasma BCAA (BCAA). Compared with that prior to the initiation of the insulin infusion, fractional synthesis rate of muscle protein (%/h) did not change (P > 0.05) during insulin in either the control (0.04 ± 0.01 vs 0.05 ± 0.01) or the BCAA (0.05 ± 0.02 vs. 0.05 ± 0.01) experiments. Insulin decreased (P < 0.01) whole body phenylalanine rate of appearance (μmol·kg-1·min-1), indicating suppression of muscle proteolysis, in both the control (1.02 ± 0.04 vs 0.76 ± 0.04) and the BCAA (0.89 ± 0.07 vs 0.61 ± 0.03) experiments, but the change was not different between the two experiments (P > 0.05). In conclusion, insulin does not stimulate muscle protein synthesis in the presence of increased circulating levels of plasma BCAA alone. Insulin's suppressive effect on proteolysis is observed independently of the levels of circulating plasma BCAA.

Exercise strategies to protect against the impact of short-term reduced physical activity on muscle function and markers of health in older men: study protocol for a randomised controlled trial.

BackgroundMuscles get smaller and weaker as we age and become more vulnerable to atrophy when physical activity is reduced or removed. This research is designed to investigate the potentially protective effects of two separate exercise strategies against loss in skeletal muscle function and size, and other key indices of health, following 14 days of reduced physical activity in older men.MethodsThree groups of 10 older men (aged 65–80 years) will undertake 2 weeks of reduced activity by decreasing daily steps from more than 3500 to less than 1500 (using pedometers to record step count). Two of the three groups will then undertake additional exercise interventions, either: 4 weeks of progressive resistance training prior to the step-reduction intervention (PT-group), or home-based ‘exercise snacking’ three times per day during the step-reduction intervention (ES-group). The third group undertaking only the step-reduction intervention (control) will provide a comparison against which to assess the effectiveness of the protective exercise strategies. Pre and post step-reduction assessments of muscle function, standing balance, anthropometry and muscle architecture will be taken. Pre and post step-reduction in postprandial metabolic control, resting systemic inflammation, adipose inflammation, oxidative stress, immune function, sleep quality, dietary habits, and quality of life will be measured. The stress response to exercise, and signalling protein and gene expression for muscle protein synthesis and breakdown following an acute bout of exercise will also be assessed pre and post step-reduction. Rates of muscle protein synthesis and adipose triglyceride turnover during the step-reduction intervention will be measured using stable isotope methodology. All participants will then undertake 2 weeks of supervised resistance training with the aim of regaining any deficit from baseline in muscle function and size.DiscussionThis study aims to identify exercise strategies that could be implemented to protect against loss of muscle power during 2 weeks of reduced activity in older men, and to improve understanding of the way in which a short-term reduction in physical activity impacts upon muscle function and health.Trial registrationClinicalTrials.gov: NCT02495727 (Initial registration: 25 June 2015)

Sodium nitrate co-ingestion with protein does not augment postprandial muscle protein synthesis rates in older, type 2 diabetes patients.

The age-related anabolic resistance to protein ingestion is suggested to be associated with impairments in insulin-mediated capillary recruitment and postprandial muscle tissue perfusion. The present study investigated whether dietary nitrate co-ingestion with protein improves muscle protein synthesis in older, type 2 diabetes patients. Twenty-four men with type 2 diabetes (72 ± 1 yr, 26.7 ± 1.4 m/kg(2) body mass index, 7.3 ± 0.4% HbA1C) received a primed continuous infusion of l-[ring-(2)H5]phenylalanine and l-[1-(13)C]leucine and ingested 20 g of intrinsically l-[1-(13)C]phenylalanine- and l-[1-(13)C]leucine-labeled protein with (PRONO3) or without (PRO) sodium nitrate (0.15 mmol/kg). Blood and muscle samples were collected to assess protein digestion and absorption kinetics and postprandial muscle protein synthesis rates. Upon protein ingestion, exogenous phenylalanine appearance rates increased in both groups (P < 0.001), resulting in 55 ± 2% and 53 ± 2% of dietary protein-derived amino acids becoming available in the circulation over the 5h postprandial period in the PRO and PRONO3 groups, respectively. Postprandial myofibrillar protein synthesis rates based on l-[ring-(2)H5]phenylalanine did not differ between groups (0.025 ± 0.004 and 0.021 ± 0.007%/h over 0-2 h and 0.032 ± 0.004 and 0.030 ± 0.003%/h over 2-5 h in PRO and PRONO3, respectively, P = 0.7). No differences in incorporation of dietary protein-derived l-[1-(13)C]phenylalanine into de novo myofibrillar protein were observed at 5 h (0.016 ± 0.002 and 0.014 ± 0.002 mole percent excess in PRO and PRONO3, respectively, P = 0.8). Dietary nitrate co-ingestion with protein does not modulate protein digestion and absorption kinetics, nor does it further increase postprandial muscle protein synthesis rates or the incorporation of dietary protein-derived amino acids into de novo myofibrillar protein in older, type 2 diabetes patients.

Increasing Insulin Availability Does Not Augment Postprandial Muscle Protein Synthesis Rates in Healthy Young and Older Men.

Skeletal muscle protein synthesis is highly responsive to food intake. It has been suggested that the postprandial increase in circulating insulin modulates the muscle protein synthetic response to feeding. The objective of the study was to investigate whether a greater postprandial rise in circulating insulin level increases amino acid uptake in muscle and augments postprandial muscle protein synthesis rates. Forty-eight healthy young (age 22 ± 1 y; body mass index 22.0 ± 0.3 kg/m2) and older males (age 68 ± 1 y; body mass index 26.3 ± 0.4 kg/m2) ingested 20 g intrinsically L-[1-13C]-leucine- and L-[1-13C]-phenylalanine-labeled casein protein with or without local insulin infusion. Primed continuous infusions of L-[1-13C]-leucine and L-[ring-2H5]-phenylalanine were applied, with arterial and venous blood samples and muscle biopsies being collected during a 5-hour postprandial period. Insulin administration did not increase overall leg blood flow (P = .509) but increased amino acid uptake over the leg in both young and older subjects (P = .003). The greater amino acid uptake over the leg did not further increase postprandial muscle protein synthesis rates (0.050% ± 0.006% and 0.037% ± 0.004% per hour vs 0.044% ± 0.004% and 0.037% ± 0.002% per hour in the insulin-stimulated vs control condition in the young and older groups, respectively; P = .804) and did not affect postprandial deposition of dietary protein-derived amino acids in de novo muscle protein (P = .872). Greater postprandial plasma insulin availability stimulates amino acid uptake over the leg but does not further augment postprandial muscle protein synthesis rates or stimulate the postprandial deposition of protein derived amino acids into de novo muscle protein in healthy young and older men.

Determination of muscle protein synthesis rates in fish using 2H2O and 2H NMR analysis of alanine

Following administration of deuterated water (2H2O), the fractional synthetic rate (FSR) of a given endogenous protein can be estimated by 2H-enrichment quantification of its alanine residues. Currently, this is measured by mass spectrometry following a derivatization procedure. Muscle FSR was measured by 1H/2H NMR analysis of alanine from seabass kept for 6 days in 5% 2H-enriched saltwater, following acid hydrolysis and amino acid isolation by cation-exchange chromatography of muscle tissue. The analysis is simple and robust, and provides precise measurements of excess alanine 2H-enrichment in the 0.1–0.4% range from 50 mmol of alanine recovered from muscle protein.

Erythropoietin Does Not Enhance Skeletal Muscle Protein Synthesis Following Exercise in Young and Older Adults.

Purpose: Erythropoietin (EPO) is a renal cytokine that is primarily involved in hematopoiesis while also playing a role in non-hematopoietic tissues expressing the EPO-receptor (EPOR). The EPOR is present in human skeletal muscle. In mouse skeletal muscle, EPO stimulation can activate the AKT serine/threonine kinase 1 (AKT) signaling pathway, the main positive regulator of muscle protein synthesis. We hypothesized that a single intravenous EPO injection combined with acute resistance exercise would have a synergistic effect on skeletal muscle protein synthesis via activation of the AKT pathway.Methods: Ten young (24.2 ± 0.9 years) and 10 older (66.6 ± 1.1 years) healthy subjects received a primed, constant infusion of [ring-13C6] L-phenylalanine and a single injection of 10,000 IU epoetin-beta or placebo in a double-blind randomized, cross-over design. 2 h after the injection, the subjects completed an acute bout of leg extension resistance exercise to stimulate skeletal muscle protein synthesis.Results: Significant interaction effects in the phosphorylation levels of the members of the AKT signaling pathway indicated a differential activation of protein synthesis signaling in older subjects when compared to young subjects. However, EPO offered no synergistic effect on vastus lateralis mixed muscle protein synthesis rate in young or older subjects.Conclusions: Despite its ability to activate the AKT pathway in skeletal muscle, an acute EPO injection had no additive or synergistic effect on the exercise-induced activation of muscle protein synthesis or muscle protein synthesis signaling pathways.

A single session of neuromuscular electrical stimulation does not augment postprandial muscle protein accretion.

The loss of muscle mass and strength that occurs with aging, termed sarcopenia, has been (at least partly) attributed to an impaired muscle protein synthetic response to food intake. Previously, we showed that neuromuscular electrical stimulation (NMES) can stimulate fasting muscle protein synthesis rates and prevent muscle atrophy during disuse. We hypothesized that NMES prior to protein ingestion would increase postprandial muscle protein accretion. Eighteen healthy elderly (69 ± 1 yr) males participated in this study. After a 70-min unilateral NMES protocol was performed, subjects ingested 20 g of intrinsically l-[1-(13)C]phenylalanine-labeled casein. Plasma samples and muscle biopsies were collected to assess postprandial mixed muscle and myofibrillar protein accretion as well as associated myocellular signaling during a 4-h postprandial period in both the control (CON) and stimulated (NMES) leg. Protein ingestion resulted in rapid increases in both plasma phenylalanine concentrations and l-[1-(13)C]phenylalanine enrichments, which remained elevated during the entire 4-h postprandial period (P < 0.05). Mixed-muscle protein-bound l-[1-(13)C]phenylalanine enrichments increased significantly over time following protein ingestion, with no differences between the CON (0.0164 ± 0.0019 MPE) and NMES (0.0164 ± 0.0019 MPE) leg (P > 0.05). In agreement, no differences were observed in the postprandial rise in myofibrillar protein bound l-[1-(13)C]phenylalanine enrichments between the CON and NMES legs (0.0115 ± 0.0014 vs. 0.0133 ± 0.0013 MPE, respectively, P > 0.05). Significant increases in mTOR and P70S6K phosphorylation status were observed in the NMES-stimulated leg only (P < 0.05). We conclude that a single session of NMES prior to food intake does not augment postprandial muscle protein accretion in healthy older men.

Enteral β-hydroxy-β-methylbutyrate supplementation increases protein synthesis in skeletal muscle of neonatal pigs.

Many low-birth weight infants are at risk for poor growth due to an inability to achieve adequate protein intake. Administration of the amino acid leucine stimulates protein synthesis in skeletal muscle of neonates. To determine the effects of enteral supplementation of the leucine metabolite β-hydroxy-β-methylbutyrate (HMB) on protein synthesis and the regulation of translation initiation and degradation pathways, overnight-fasted neonatal pigs were studied immediately (F) or fed one of five diets for 24 h: low-protein (LP), high-protein (HP), or LP diet supplemented with 4 (HMB4), 40 (HMB40), or 80 (HMB80) μmol HMB·kg body wt(-1)·day(-1) Cell replication was assessed from nuclear incorporation of BrdU in the longissimus dorsi (LD) muscle and jejunum crypt cells. Protein synthesis rates in LD, gastrocnemius, rhomboideus, and diaphragm muscles, lung, and brain were greater in HMB80 and HP and in brain were greater in HMB40 compared with LP and F groups. Formation of the eIF4E·eIF4G complex and S6K1 and 4E-BP1 phosphorylation in LD, gastrocnemius, and rhomboideus muscles were greater in HMB80 and HP than in LP and F groups. Phosphorylation of eIF2α and eEF2 and expression of SNAT2, LAT1, MuRF1, atrogin-1, and LC3-II were unchanged. Numbers of BrdU-positive myonuclei in the LD were greater in HMB80 and HP than in the LP and F groups; there were no differences in jejunum. The results suggest that enteral supplementation with HMB increases skeletal muscle protein anabolism in neonates by stimulation of protein synthesis and satellite cell proliferation.

Diminished Postprandial Muscle Protein Synthetic Response To Protein Ingestion In Obese Adults

Excess body fat blunts muscle protein synthesis rates under hyperinsulinemic, hyperaminoacidemic clamp conditions. However, the efficacy of the ingestion of single meal containing ample amounts of protein to augment postprandial muscle protein synthesis rates has not been studied in obese and overweight adults. PURPOSE: We aimed to compare myofibrillar protein synthesis after protein ingestion in healthy weight, overweight, and obese adults. METHODS: 10 healthy-weight (24±1 y, BMI 22.7±0.4 kg/m2, HOMA-IR 1.4±0.2), 10 overweight (26±2 y, BMI 27.1±0.5 kg/m2, HOMA-IR 1.25±0.11), and 10 obese (27±3 y, BMI 35.9±1.3 kg/m2, HOMA-IR 5.8±0.8) men and women underwent a primed continuous L-[ring-13C6]phenylalanine infusion. Blood and muscle biopsy samples were collected at rest and after ingestion of 36 g of protein to assess plasma amino acid and insulin concentrations and myofibrillar protein synthesis rates. RESULTS: Protein ingestion increased plasma essential amino acid concentrations similarly in all participants (time effect: P<0.05) with concentrations peaked at 2 h and returning to baseline values at 5 h. The obese participants had a greater peak postprandial plasma insulin response to protein ingestion than the overweight and healthy-weight participants (obese 56.7±8.0 compared to overweight 11.9±1.6 and healthy-weight 14.3±1.7 μIU/L, P<0.001). Protein ingestion increased myofibrillar protein synthesis in both the healthy-weight (rest: 0.057±0.006 %/h, fed: 0.084±0.014 %/h) and overweight groups (rest: 0.061±0.007, fed: 0.087±0.019 %/h) (P<0.05) with no increase in the obese group (rest: 0.055±0.005 %/h, fed: 0.067±0.005 %/h, P=0.45). CONCLUSIONS: Increased adiposity led to an impaired postprandial muscle protein synthetic response to protein ingestion in obese adults when compared to healthy-weight and overweight individuals. These data suggest that poor skeletal muscle remodeling may underlie early metabolic impairments in apparently “healthy” obese adults. Funding provided by the National Pork Board

Exercise Enhances The Overnight Muscle Protein Synthetic Response To Pre-sleep Protein Feeding In Older Males

PURPOSE: The age-related decline in skeletal muscle mass is, at least partly, attributed to anabolic resistance to food intake. To compensate for anabolic resistance, we recently introduced the ingestion of dietary protein prior to sleep as a nutritional strategy to increase overnight muscle protein synthesis rates. Here we aimed to assess whether resistance-type exercise performed in the evening can further augment the overnight muscle protein synthetic response to pre-sleep protein ingestion in older males. METHODS: In a parallel group design, twenty-three healthy older men (71±1 y) were randomly assigned to ingest 40 g casein protein before going to sleep with (PRO+EX: n=11) or without (PRO: n=12) performing resistance-type exercise earlier that evening. Overnight protein digestion and absorption kinetics, whole body protein metabolism and muscle myofibrillar protein synthesis rates were assessed using primed, continuous infusions of L-[ring-2H5]-phenylalanine and L-[ring-2H2]-tyrosine with the ingestion of intrinsically L-[1-13C]-phenylalanine labeled casein protein. RESULTS: Exogenous phenylalanine appearance rates expressed over time did not differ between treatments. A total of 53±2 vs 55±2% of the ingested protein-derived phenylalanine appeared in the circulation during overnight sleep in the PRO+EX and PRO treatment, respectively (P=0.49). Myofibrillar protein synthesis rates were 31% higher during overnight sleep when exercise was performed earlier that day (0.058±0.002 vs 0.044±0.003 %·h-1 in PRO+EX and PRO, respectively; P<0.01; L-[ring-2H5]-phenylalanine). In line, 27% more L-[1-13C]-phenylalanine was incorporated into myofibrillar protein in PRO+EX compared with PRO (0.042±0.002 vs 0.033±0.002 MPE, respectively; P<0.05; L-[1-13C]-phenylalanine). CONCLUSIONS: Resistance-type exercise augments the overnight muscle protein synthetic response to pre-sleep protein ingestion and allows more of the ingested protein to be directed towards de novo muscle protein synthesis during overnight sleep in older males. Funding: Top Institute Food and Nutrition (TIFN)

The Effects of Dietary Omega-3s on Muscle Composition and Quality in Older Adults.

This review will focus on findings from the few studies performed to date in humans to examine changes in muscle protein turnover, lean or muscle mass and physical function following fish oil-derived omega-3 fatty acid treatment. Although considerable gaps in our current knowledge exist, hypertrophic responses (e.g., improvements in the rate of muscle protein synthesis and mTOR signaling during increased amino acid availability and an increase in muscle volume) have been reported in older adults following prolonged (8 to 24 weeks) of omega-3 fatty acid supplementation. There is also accumulating evidence that increased omega-3 fatty acid levels in red blood cells are positively related to strength and measures of physical function. As a result, increased omega-3 fatty acid consumption may prove to be a promising low-cost dietary approach to attenuate or prevent aging associated declines in muscle mass and function.

Long‐term Leucine and BCAA Inclusion in a 30% Protein and Energy Restricted Diet Increases mTORC1 Signaling in Skeletal Muscle of Neonatal Pigs

Suboptimal nutrient intake during the shift from parenteral to oral feeding represents a limiting factor for growth of low‐birth weight infants. Leucine has been shown to have anabolic properties in skeletal muscle and can promote lean gain when supplemented to neonatal pigs fed a milk replacement formula. The objective of this study was to test if enteral leucine supplementation alone, or in combination with isoleucine and valine, can increase muscle protein synthesis in neonate pigs who consume 30% less crude protein (CP) and metabolizable energy than their requirements, and whether this effect is mediated through changes in muscle protein synthesis and degradation pathways, and amino acid (AA) transporter expression. Piglets (4‐d‐old) were fed by gastric tube 1 of 4 diets providing (kg BW/d) 15 g CP and 215 kcal (CON), 10.5 g CP and 156 kcal (R), 10.5 g CP + 1.19 g leucine and 156 kcal (RL), or 10.5 g CP + 1.67 g branched‐chain amino acids and 156 kcal (RBCAA) at 4‐h intervals for 9 d (n = 24). Body weights (BW) were recorded on d 0, 3, 6, and 9. On d 9, plasma AA and insulin concentrations and muscle protein synthesis rate, abundance of Na+‐coupled neutral AA transporter (SNAT2) and L‐type AA transporter (LAT1), and phosphorylation and/or abundance of insulin receptor substrate 1 (IRS‐1, Ser1101), protein kinase B (PKB), S6 kinase (S6K1), 4E‐binding protein (4EBP1), eukaryotic initiation factor 4 complex (eIF4E·eIF4G), forkhead box O (FOXO1, Ser256), atrophy F‐Box/Atrogin‐1 and muscle RING‐finger protein (MuRF1) were determined post‐feeding. Plasma leucine levels were greater in RL and RBCAA compared to CON (P ≤ 0.01). BW gain and muscle protein synthesis rate, phosphorylation of S6K1 and 4EBP1, and formation of eIF4E·eIF4G were lower in R, RL, and RBCAA than CON (P &lt; 0.01). RBCAA and RL upregulated (P ≤ 0.01) S6K1 and 4EBP1 phosphorylation and eIF4E·eIF4G formation compared to R. Phosphorylation of FOXO1 was lower in R compared to CON (P ≤ 0.05). There were no differences in phosphorylation and/or abundance of IRS‐1, PKB, SNAT2, LAT1, atrogin‐1 and MURF1 among diets. In conclusion, when protein and energy intakes are restricted, leucine or BCAA supplementation for 9 d increases the activation of the mammalian target of rapamycin pathway, but does not enhance skeletal muscle protein synthesis or promote body weight gain in neonatal pigs. Furthermore, protein degradation signaling, AA transporter abundance, and insulin signaling are unaffected by leucine or BCAA supplementation of a protein and energy restricted diet suggesting that when both dietary protein and/or energy supplies are deficient, the anabolic effect of leucine or BCAA supplementation on muscle protein accretion is limited.Support or Funding InformationNIH AR444474, NIH HD072891, NIH AR46308, USDA NIFA 2013‐67015‐20438, and USDA/ARS 6250‐51000‐055

Long‐term Intermittent Leucine Pulses during Continuous Feeding Impact the Plasma Metabolome of Neonatal Pigs

Infants who are unable to maintain oral feeding can be nourished by continuous administration of feed via orogastric tube. We have shown in a neonatal piglet model that intermittent pulses of leucine administered during continuous feeding enhanceslean tissue growth compared to continuous feeding alone. This response was consistent with the attainment of higher rates of muscle protein synthesis in the leucine supplemented piglets. To determine the long‐term effects of intermittent leucine pulses on metabolomic markers in the plasma of neonates, piglets (n = 23, 7‐d‐old) were fed continuously a milk replacement formula via orogastric tube for 21 d with an additional parenteral infusion of either a leucine (800 μmol·kg−1·h−1, CON+LEU)or alanine (CON+ALA) supplement for 1 h every 4 h; blood samples were collected at d 21. Plasma samples were analyzed by Metabolon (Durham, NC, USA) using GC‐MS and LC‐MS. There were 483 compounds identified in the analysis matching known chemical structures. Following log transformation and imputation of missing values, Welch's two‐sample t‐test was used to identify metabolites that differed significantly between experimental groups. The results showed that in the CON+LEU compared to the CON+ALA group, the BCAA‐related changes in metabolites was consistent with increased leucine metabolism. Concentrations of leucine‐containing dipeptides, an indicator of protein synthesis, were elevated in the CON+LEU group compared to the CON+ALA group (P &lt; 0.05). Concentrations of dimethyl arginine, a marker of proteolysis, were reduced in CON+LEU compared to CON+ALA group (P &lt;0.05). These results are consistent with the observed differences in the growth responses to the two treatments. Altogether, these data support the notion that, in addition to increasing protein synthesis, leucine supplementation ameliorates muscle protein degradation, and therefore may be useful as atherapeutic agent to promote growth in neonates. Thus, plasma metabolomics profiling may be a useful tool to identify biomarkers of protein synthesis and degradation pathways that are modulated by leucine supplementation in neonates.Support or Funding InformationNIH AR444474, NIH HD072891, USDA NIFA 2013‐67015‐20438, and USDA/ARS 6250‐51000‐055

What is the Optimal Amount of Protein to Support Post-Exercise Skeletal Muscle Reconditioning in the Older Adult?

Hyperaminoacidemia following protein ingestion enhances the anabolic effect of resistance-type exercise by increasing the stimulation of muscle protein synthesis and attenuating the exercise-mediated increase in muscle protein breakdown rates. Although factors such as the source of protein ingested and the timing of intake relative to exercise can impact post-exercise muscle protein synthesis rates, the amount of protein ingested after exercise appears to be the key nutritional factor dictating the magnitude of the muscle protein synthetic response during post-exercise recovery. In younger adults, muscle protein synthesis rates after resistance-type exercise respond in a dose-dependent manner to ingested protein and are maximally stimulated following ingestion of ~20g of protein. In contrast to younger adults, older adults are less sensitive to smaller doses of ingested protein (less than ~20g) after exercise, as evidenced by an attenuated increase in muscle protein synthesis rates during post-exercise recovery. However, older muscle appears to retain the capacity to display a robust stimulation of muscle protein synthesis in response to the ingestion of greater doses of protein (~40g), and such an amount may be required for older adults to achieve a robust stimulation of muscle protein synthesis during post-exercise recovery. The aim of this article is to discuss the current state of evidence regarding the dose-dependent relationship between dietary protein ingestion and changes in skeletal muscle protein synthesis during recovery from resistance-type exercise in older adults. We provide recommendations on the amount of protein that may be required to maximize skeletal muscle reconditioning in response to resistance-type exercise in older adults.