Correction to: Impact of whole dairy matrix on musculoskeletal health and aging-current knowledge and research gaps.

The article Impact of whole dairy matrix on musculoskeletal health and aging-current knowledge and research gaps written by N.R.W. Geiker, C. Mølgaard, S. Iuliano, R. Rizzoli,Y. Manios, L.J.C. van Loon, J.-M. Lecerf, G. Moschonis, J.-Y. Reginster, I. Givens, A. Astrup.

Impact of whole dairy matrix on musculoskeletal health and aging-current knowledge and research gaps.

Dairy products are included in dietary guidelines worldwide, as milk, yoghurt, and cheese are good sources of calcium and protein, vital nutrients for bones and muscle mass maintenance. Bone growth and mineralization occur during infancy and childhood, peak bone mass being attained after early adulthood. A low peak bone mass has consequences later in life, including increased risk of osteoporosis and fractures. Currently, more than 200 million people worldwide suffer from osteoporosis, with approximately 9 million fractures yearly. This poses a tremendous economic burden on health care. Between 5% and 10% of the elderly suffer from sarcopenia, the loss of muscle mass and strength, further increasing the risk of fractures due to falls. Evidence from interventional and observational studies support that fermented dairy products in particular exert beneficial effects on bone growth and mineralization, attenuation of bone loss, and reduce fracture risk. The effect cannot be explained by single nutrients in dairy, which suggests that a combined or matrix effect may be responsible similar to the matrix effects of foods on cardiometabolic health. Recently, several plant-based beverages and products have become available and marketed as substitutes for dairy products, even though their nutrient content differs substantially from dairy. Some of these products have been fortified, in efforts to mimic the nutritional profile of milk, but it is unknown whether the additives have the same bioavailability and beneficial effect as dairy. We conclude that the dairy matrix exerts an effect on bone and muscle health that is more than the sum of its nutrients, and we suggest that whole foods, not only single nutrients, need to be assessed in future observational and intervention studies of health outcomes. Furthermore, the importance of the matrix effect on health outcomes argues in favor of making future dietary guidelines food based.

Beetroot juice supplementation reduces whole body oxygen consumption but does not improve indices of mitochondrial efficiency in human skeletal muscle.

KEY POINTS: Oral consumption of nitrate (NO3(-)) in beetroot juice has been shown to decrease the oxygen cost of submaximal exercise; however, the mechanism of action remains unresolved. We supplemented recreationally active males with beetroot juice to determine if this altered mitochondrial bioenergetics. Despite reduced submaximal exercise oxygen consumption, measures of mitochondrial coupling and respiratory efficiency were not altered in muscle. In contrast, rates of mitochondrial hydrogen peroxide (H2O2) emission were increased in the absence of markers of lipid or protein oxidative damage. These results suggest that improvements in mitochondrial oxidative metabolism are not the cause of beetroot juice-mediated improvements in whole body oxygen consumption. ABSTRACT: Ingestion of sodium nitrate (NO3(-)) simultaneously reduces whole body oxygen consumption (V̇O2) during submaximal exercise while improving mitochondrial efficiency, suggesting a causal link. Consumption of beetroot juice (BRJ) elicits similar decreases in V̇O2 but potential effects on the mitochondria remain unknown. Therefore we examined the effects of 7-day supplementation with BRJ (280 ml day(-1), ∼26 mmol NO3(-)) in young active males (n = 10) who had muscle biopsies taken before and after supplementation for assessments of mitochondrial bioenergetics. Subjects performed 20 min of cycling (10 min at 50% and 70% V̇O2 peak) 48 h before 'Pre' (baseline) and 'Post' (day 5 of supplementation) biopsies. Whole body V̇O2 decreased (P < 0.05) by ∼3% at 70% V̇O2 peak following supplementation. Mitochondrial respiration in permeabilized muscle fibres showed no change in leak respiration, the content of proteins associated with uncoupling (UCP3, ANT1, ANT2), maximal substrate-supported respiration, or ADP sensitivity (apparent Km). In addition, isolated subsarcolemmal and intermyofibrillar mitochondria showed unaltered assessments of mitochondrial efficiency, including ADP consumed/oxygen consumed (P/O ratio), respiratory control ratios and membrane potential determined fluorometrically using Safranine-O. In contrast, rates of mitochondrial hydrogen peroxide (H2O2) emission were increased following BRJ. Therefore, in contrast to sodium nitrate, BRJ supplementation does not alter key parameters of mitochondrial efficiency. This occurred despite a decrease in exercise V̇O2, suggesting that the ergogenic effects of BRJ ingestion are not due to a change in mitochondrial coupling or efficiency. It remains to be determined if increased mitochondrial H2O2 contributes to this response.

Protein intake and lean body mass preservation during energy intake restriction in overweight older adults.

BACKGROUND: Dietary-induced weight loss is generally accompanied by a decline in skeletal muscle mass. The loss of muscle mass leads to a decline in muscle strength and impairs physical performance. A high dietary protein intake has been suggested to allow muscle mass preservation during energy intake restriction. OBJECTIVE: To investigate the impact of increasing dietary protein intake on lean body mass, strength and physical performance during 12 weeks of energy intake restriction in overweight older adults. DESIGN: Sixty-one overweight and obese men and women (63±5 years) were randomly assigned to either a high protein diet (HP; 1.7 g kg(-1) per day; n=31) or normal protein diet (NP; 0.9 g kg(-1) per day; n=30) during a 12-week 25% energy intake restriction. During this controlled dietary intervention, 90% of the diet was provided by the university. At baseline and after the intervention, body weight, lean body mass (dual-energy X-ray absorptiometry), leg strength (1-repetition maximum), physical performance (Short Physical Performance Battery, 400 m) and habitual physical activity (actigraph) were assessed. RESULTS: Body weight declined in both groups with no differences between the HP and NP groups (-8.9±2.9 versus -9.1±3.4 kg, respectively; P=0.584). Lean body mass declined by 1.8±2.2 and 2.1±1.4 kg, respectively, with no significant differences between groups (P=0.213). Leg strength had decreased during the intervention by 8.8±14.0 and 8.9±12.8 kg, with no differences between groups (P=0.689). Physical performance as measured by 400 m walking speed improved in both groups, with no differences between groups (P=0.219). CONCLUSIONS: Increasing protein intake above habitual intake levels (0.9 g kg(-1) per day) does not preserve lean body mass, strength or physical performance during prolonged energy intake restriction in overweight older adults.

In vivo postprandial lipid partitioning in liver and skeletal muscle in prediabetic and diabetic rats.

AIMS/HYPOTHESIS: Insulin resistance and type 2 diabetes have been associated with ectopic lipid deposition. This study investigates the derangements in postprandial lipid handling in liver and skeletal muscle tissue at different stages during the pathogenesis of type 2 diabetes in a rat model. METHODS: Four groups (n = 6) of male Zucker diabetic fatty rats were used for this study: prediabetic fa/fa rats and healthy fa/+ littermates at the age of 6 weeks, and diabetic fa/fa rats and healthy fa/+ littermates at the age of 12 weeks. In vivo (1)H-[(13)C] magnetic resonance spectroscopy measurements were performed in liver and tibialis anterior muscle at baseline and 4, 24 and 48 h after oral administration of 1.5 g [U-(13)C]algal lipid mixture per kilogram body weight. Total and (13)C-labelled intracellular lipid contents were determined from the magnetic resonance spectra. RESULTS: In both prediabetic and diabetic rats, total lipid contents in muscle and liver were substantially higher than in healthy controls and this was accompanied by a 2.3-fold greater postprandial lipid uptake in the liver (p < 0.001). Interestingly, in prediabetic rats, skeletal muscle appeared to be protected from excess lipid uptake whereas after developing overt diabetes muscle lipid uptake was 3.4-fold higher than in controls (p < 0.05). Muscle lipid use was significantly lower in prediabetic and diabetic muscle, indicative of impairments in lipid oxidation. CONCLUSIONS/INTERPRETATION: In vivo postprandial lipid handling is disturbed in both liver and skeletal muscle tissue in prediabetic and diabetic rats, but the uptake of dietary lipids in muscle is only increased after the development of overt diabetes.

Disturbed energy metabolism and muscular dystrophy caused by pure creatine deficiency are reversible by creatine intake.

Creatine (Cr) plays an important role in muscle energy homeostasis by its participation in the ATP-phosphocreatine phosphoryl exchange reaction mediated by creatine kinase. Given that the consequences of Cr depletion are incompletely understood, we assessed the morphological, metabolic and functional consequences of systemic depletion on skeletal muscle in a mouse model with deficiency of l-arginine:glycine amidinotransferase (AGAT(-/-)), which catalyses the first step of Cr biosynthesis. In vivo magnetic resonance spectroscopy showed a near-complete absence of Cr and phosphocreatine in resting hindlimb muscle of AGAT(-/-) mice. Compared with wild-type, the inorganic phosphate/ß-ATP ratio was increased fourfold, while ATP levels were reduced by nearly half. Activities of proton-pumping respiratory chain enzymes were reduced, whereas F(1)F(0)-ATPase activity and overall mitochondrial content were increased. The Cr-deficient AGAT(-/-) mice had a reduced grip strength and suffered from severe muscle atrophy. Electron microscopy revealed increased amounts of intramyocellular lipid droplets and crystal formation within mitochondria of AGAT(-/-) muscle fibres. Ischaemia resulted in exacerbation of the decrease of pH and increased glycolytic ATP synthesis. Oral Cr administration led to rapid accumulation in skeletal muscle (faster than in brain) and reversed all the muscle abnormalities, revealing that the condition of the AGAT(-/-) mice can be switched between Cr deficient and normal simply by dietary manipulation. Systemic creatine depletion results in mitochondrial dysfunction and intracellular energy deficiency, as well as structural and physiological abnormalities. The consequences of AGAT deficiency are more pronounced than those of muscle-specific creatine kinase deficiency, which suggests a multifaceted involvement of creatine in muscle energy homeostasis in addition to its role in the phosphocreatine-creatine kinase system.

Both resistance- and endurance-type exercise reduce the prevalence of hyperglycaemia in individuals with impaired glucose tolerance and in insulin-treated and non-insulin-treated type 2 diabetic patients.

AIMS/HYPOTHESIS: The present study compares the impact of endurance- vs resistance-type exercise on subsequent 24 h blood glucose homeostasis in individuals with impaired glucose tolerance (IGT) and type 2 diabetes. METHODS: Fifteen individuals with IGT, 15 type 2 diabetic patients treated with exogenous insulin (INS), and 15 type 2 diabetic patients treated with oral glucose-lowering medication (OGLM) participated in a randomised crossover experiment. Participants were studied on three occasions for 3 days under strict dietary standardisation, but otherwise free-living conditions. Blood glucose homeostasis was assessed by ambulatory continuous glucose monitoring over the 24 h period following a 45 min session of resistance-type exercise (75% one repetition maximum), endurance-type exercise (50% maximum workload capacity) or no exercise at all. RESULTS: Average 24 h blood glucose concentrations were reduced from 7.4 ± 0.2, 9.6 ± 0.5 and 9.2 ± 0.7 mmol/l during the control experiment to 6.9 ± 0.2, 8.6 ± 0.4 and 8.1 ± 0.5 mmol/l (resistance-type exercise) and 6.8 ± 0.2, 8.6 ± 0.5 and 8.5 ± 0.5 mmol/l (endurance-type exercise) over the 24 h period following a single bout of exercise in the IGT, OGLM and INS groups, respectively (p < 0.001 for both treatments). The prevalence of hyperglycaemia (blood glucose >10 mmol/l) was reduced by 35 ± 7 and 33 ± 11% over the 24 h period following a single session of resistance- and endurance-type exercise, respectively (p < 0.001 for both treatments). CONCLUSIONS/INTERPRETATION: A single session of resistance- or endurance-type exercise substantially reduces the prevalence of hyperglycaemia during the subsequent 24 h period in individuals with IGT, and in insulin-treated and non-insulin-treated type 2 diabetic patients. Both resistance- and endurance-type exercise can be integrated in exercise intervention programmes designed to improve glycaemic control. TRIAL REGISTRATION: Clinicaltrials.gov NCT00945165. FUNDING: The Netherlands Organization for Health Research and Development (ZonMw, the Netherlands).

Prolonged exercise training increases intramuscular lipid content and perilipin 2 expression in type I muscle fibers of patients with type 2 diabetes.

The aim of the present study was to investigate changes in intramuscular triglyceride (IMTG) content and perilipin 2 expression in skeletal muscle tissue following 6 mo of endurance-type exercise training in type 2 diabetes patients. Ten obese male type 2 diabetes patients (age 62 ± 1 yr, body mass index BMI 31 ± 1 kg/m²) completed three exercise sessions/week consisting of 40 min of continuous endurance-type exercise at 75% V(O2 peak) for a period of 6 mo. Muscle biopsies collected at baseline and after 2 and 6 mo of intervention were analyzed for IMTG content and perilipin 2 expression using fiber type-specific immunofluorescence microscopy. Endurance-type exercise training reduced trunk body fat by 6 ± 2% and increased whole body oxygen uptake capacity by 13 ± 7% (P < 0.05). IMTG content increased twofold in response to the 6 mo of exercise training in both type I and type II muscle fibers (P < 0.05). A threefold increase in perilipin 2 expression was observed from baseline to 2 and 6 mo of intervention in the type I muscle fibers only (1.1 ± 0.3, 3.4 ± 0.6, and 3.6 ± 0.6% of fibers stained, respectively, P < 0.05). Exercise training induced a 1.6-fold increase in mitochondrial content after 6 mo of training in both type I and type II muscle fibers (P < 0.05). In conclusion, this is the first study to report that prolonged endurance-type exercise training increases the expression of perilipin 2 alongside increases in IMTG content in a type I muscle fiber-type specific manner in type 2 diabetes patients.

The production of intrinsically labeled milk and meat protein is feasible and provides functional tools for human nutrition research.

Administration of labeled, free amino acids does not allow direct assessment of in vivo dietary protein digestion and absorption kinetics. Consequently, dietary protein sources with labeled amino acids incorporated within their protein matrix are required. The aim of the present study was to produce intrinsically L-[1-(13)C]phenylalanine-labeled milk and meat protein that would permit in vivo assessment of postprandial protein digestion and absorption kinetics in humans. One lactating dairy cow was continuously infused with 420 µmol of L-[1-(13)C]phenylalanine/min for 96 h, with plasma and milk being collected before, during, and after isotope infusion. Twenty-four hours after infusion, the cow was slaughtered to produce intrinsically labeled meat. Levels of L-[1-(13)C]phenylalanine enrichment as high as 40 mole percent excess (MPE) in milk and 1.5 MPE in meat protein were achieved. In a subsequent human proof-of-principle experiment, 2 healthy young males (25±1 yr; 66.2±5.2 kg) each ingested 135 g of L-[1-(13)C]phenylalanine intrinsically labeled minced beef, after which plasma samples were collected at regular time intervals. Plasma L-[1-(13)C]phenylalanine enrichments increased during the first 90 min following beef ingestion, reaching peak plasma enrichment levels of 0.61±0.04 MPE. Whole-body net protein balance, assessed by continuous infusion of L-[ring-(2)H(5)]phenylalanine and L-[ring-(2)H(2)]tyrosine, was higher in the postprandial period compared with basal values (6.4±0.1 vs. -4.5±0.1 µmol/kg per h). In conclusion, the production of intrinsically L-[1-(13)C]phenylalanine-labeled milk and meat protein is feasible and provides functional tools to investigate in vivo protein digestion and absorption kinetics in humans.

Endocrine responses during overnight recovery from exercise: impact of nutrition and relationships with muscle protein synthesis.

Nocturnal endocrine responses to exercise performed in the evening and the potential role of nutrition are poorly understood. To gain novel insight, 10 healthy men ingested carbohydrate with (C+P) and without (C) protein in a randomized order and double-blind manner during 2 hr of interval cycling followed by resistance-type exercise and into early postexercise recovery. Blood samples were obtained hourly throughout 9 hr of postexercise overnight recovery for analysis of key hormones. Muscle samples were taken from the vastus lateralis before and after exercise and then again the next morning (7 a.m.) to calculate mixed-muscle protein fractional synthetic rate (FSR). Overnight plasma hormone concentrations were converted into overall responses (expressed as area under the concentration curve) and did not differ between treatments for either growth hormone (1,464 ± 257 vs. 1,432 ± 164 pg/ml · 540 min) or total testosterone (18.3 ± 1.2 vs. 17.9 ± 1.2 nmol/L · 540 min, C and C+P, respectively). In contrast, the overnight cortisol response was higher with C+P (102 ± 11 nmol/L · 540 min) than with C (81 ± 8 nmol/L · 540 min; p = .02). Mixed-muscle FSR did not differ between C and C+P during overnight recovery (0.062% ± 0.006% and 0.062% ± 0.009%/hr, respectively) and correlated significantly with the plasma total testosterone response (r = .7, p < .01). No correlations with FSR were apparent for the response of growth hormone (r = -.2, p = .4), cortisol (r = .1, p = .6), or the ratio of testosterone to cortisol (r = .2, p = .5). In conclusion, protein ingestion during and shortly after exercise does not modulate the endocrine response or muscle protein synthesis during overnight recovery.

Muscle fiber capillarization is associated with various indices of skeletal muscle mass in healthy, older men.

INTRODUCTION: Muscle fiber capillarization plays a fundamental role in the regulation of skeletal muscle mass maintenance. However, it remains unclear to what extent capillarization is related to various other skeletal muscle characteristics. In this study we determined whether muscle fiber capillarization is independently associated with measures of skeletal muscle mass, both on a whole-body and cellular level, and post-absorptive muscle protein synthesis rates in healthy older men. METHODS: Forty-six healthy older (70 ± 4 y) men participated in a trial during which basal muscle protein synthesis rates were assessed using stable isotope tracer methodology. Blood and muscle biopsy samples were collected to assess post-absorptive muscle protein synthesis rates over a 3-hour period. Immunohistochemistry was performed to determine various indices of muscle fiber capillarization, size, type distribution, and myonuclear content/domain size. Dual energy x-ray absorptiometry scans were performed to determine whole-body and appendicular lean tissue mass. RESULTS: Capillary-to-fiber ratio (C/Fi) and perimeter exchange (CFPE) index correlated with whole-body lean tissue mass (r = 0.43, P < 0.01 and r = 0.25, P < 0.10, respectively), appendicular lean tissue mass (r = 0.52, P < 0.001 and r = 0.37, P < 0.05, respectively) as well as appendicular lean tissue mass divided by body mass index (r = 0.65, P < 0.001 and r = 0.62, P < 0.001, respectively). Muscle fiber size correlated with C/Fi (r = 0.45, P < 0.01), but not with CFPE index. No associations were observed between different indices of muscle fiber capillarization and post-absorptive muscle protein synthesis rates in healthy, older men. CONCLUSION: The present study provides further evidence that muscle fiber capillarization may be a critical factor in the regulation of skeletal muscle maintenance in healthy older men.

Higher Levels of Physical Activity Are Associated with Greater Fruit and Vegetable intake in Older Adults.

OBJECTIVE: High levels of physical activity (PA) and optimal nutrition independently improve healthy aging, but few data are available about how PA may influence food preferences in older populations. Therefore, the aim of our study was to establish if there is an association between habitual PA and intake of nutrient-dense foods (i.e. fruits and vegetables). DESIGN: A cross-sectional survey was conducted. SETTING: The Netherlands. PARTICIPANTS: 2466 older adults (56% male, age 62±9 yr). MEASUREMENTS: PA was assessed using the short questionnaire to assess health (SQUASH) and participants were classified into quintiles of weekly PA (MET-h/wk). Total fruit and vegetable intake was assessed using a validated food frequency questionnaire (FFQ) and were corrected for energy intake (g/kcal/d) in the analyses. Multiple regression analyses were performed to determine the association between PA and fruit and vegetable intake, including covariates. RESULTS: Being in the higher quintiles of PA (Q3, Q4 and Q5) was positively associated with more daily fruit and vegetable consumption, even after correction for total energy intake (Q3; ß=0.089, P<0.001, Q4; ß=0.047, P=0.024, Q5; ß=0.098, P<0.001). CONCLUSIONS: Older adults who are moderately to highly physically active tend to consume more fruit and vegetable compared to less active peers, when corrected for total energy intake. Female gender, under- and overreporting dietary intake (Goldberg score), non-smoking, high level of education, less alcohol consumption and a lower body mass index positivity affected this relationship. Our data will help health-care professionals to accelerate their efforts to treat and prevent chronic diseases.

Continuous low- to moderate-intensity exercise training is as effective as moderate- to high-intensity exercise training at lowering blood HbA(1c) in obese type 2 diabetes patients.

AIMS/HYPOTHESIS: Exercise represents an effective interventional strategy to improve glycaemic control in type 2 diabetes patients. However, the impact of exercise intensity on the benefits of exercise training remains to be established. In the present study, we compared the clinical benefits of 6 months of continuous low- to moderate-intensity exercise training with those of continuous moderate- to high-intensity exercise training, matched for energy expenditure, in obese type 2 diabetes patients. METHODS: Fifty male obese type 2 diabetes patients (age 59 +/- 8 years, BMI 32 +/- 4 kg/m(2)) participated in a 6 month continuous endurance-type exercise training programme. All participants performed three supervised exercise sessions per week, either 55 min at 50% of whole body peak oxygen uptake (VO(2)peak (low to moderate intensity) or 40 min at 75% of VO(2)peak (moderate to high intensity). Oral glucose tolerance, blood glycated haemoglobin, lipid profile, body composition, maximal workload capacity, whole body and skeletal muscle oxidative capacity and skeletal muscle fibre type composition were assessed before and after 2 and 6 months of intervention. RESULTS: The entire 6 month intervention programme was completed by 37 participants. Continuous endurance-type exercise training reduced blood glycated haemoglobin levels, LDL-cholesterol concentrations, body weight and leg fat mass, and increased VO(2)peak, lean muscle mass and skeletal muscle cytochrome c oxidase and citrate synthase activity (p < 0.05). No differences were observed between the groups training at low to moderate or moderate to high intensity. CONCLUSIONS/INTERPRETATION: When matched for energy cost, prolonged continuous low- to moderate-intensity endurance-type exercise training is equally effective as continuous moderate- to high-intensity training in lowering blood glycated haemoglobin and increasing whole body and skeletal muscle oxidative capacity in obese type 2 diabetes patients. TRIAL REGISTRATION: ISRCTN32206301 FUNDING: None.

Slowly digestible carbohydrate sources can be used to attenuate the postprandial glycemic response to the ingestion of diabetes-specific enteral formulas.

PURPOSE: The purpose of this study is to compare the glycemic and insulinemic responses following the ingestion of recently developed diabetes-specific enteral formulas versus a standard and a high-fat formula. METHODS: Fifteen type 2 diabetes patients were selected to participate in a randomized, double-blind, crossover study. Two enteral formulas (47 energy percent [En%] carbohydrate, 34En% fat, and 4 g fiber/200 mL) were defined with either isomaltulose (formula 1) or sucromalt (formula 2) as the main carbohydrate source. For comparison, an isoenergetic diabetes-specific, high-fat (33En% carbohydrate, 50En% fat, 2.9 g fiber/200 mL) and a standard formula (55En% carbohydrate, 30En% fat, 2.8 g fiber/200 mL) were tested. RESULTS: Ingestion of formulas 1 and 2 and the high-fat formula resulted in an attenuated blood glucose response when compared with the standard formula (P < .05). In accordance, peak plasma glucose concentrations were significantly lower when compared with the standard formula (189 +/- 3.6 mg/dL [10.5 +/- 0.2 mmol/L], 196.2 +/- 3.6 mg/dL [10.9 +/- 0.2 mmol/L], 187.2 +/- 3.6 mg/dL [10.4 +/- 0.2 mmol/L], and 237.6 +/- 3.6 mg/dL [13.2 +/- 0.2 mmol/L], respectively). Plasma insulin responses were lower after consumption of the newly developed and high-fat formulas. Ingestion of the high-fat formula resulted in a greater postprandial triglyceride response (P < .05). CONCLUSIONS: Diabetes-specific enteral formulas rich in slowly digestible carbohydrate sources can be equally effective in attenuating the postprandial blood glucose response as low-carbohydrate, high-fat enteral formulas without elevating the plasma triglyceride response.

The production of intrinsically labeled milk protein provides a functional tool for human nutrition research.

Oral or intravenous administration of labeled, free amino acids does not allow the direct assessment of protein digestion and absorption kinetics following dietary protein intake. Consequently, dietary protein sources with labeled amino acids incorporated within the protein are required. The aim of this study was to produce milk proteins intrinsically labeled with l-[1-(13)C]phenylalanine that would allow the assessment of protein digestion and absorption kinetics and the subsequent muscle protein synthetic response to dietary protein intake in vivo in humans. Two Holstein cows (body weight of 726 +/- 38 kg) were continuously infused with l-[1-(13)C]phenylalanine at 402 micromol/min for 44 to 48 h, during and after which plasma samples and milk were collected. After milk protein separation, casein was used in a subsequent human proof-of-principle experiment. Two healthy males (aged 61 +/- 1 yr; body mass index of 22.4 +/- 0.1 kg/m(2)) ingested 35 g of casein highly enriched with [1-(13)C] phenylalanine. Plasma samples were collected at regular intervals, and skeletal muscle biopsies were collected before and 6 h after casein ingestion. In the initial experiment, a total of 5.83 kg of l-[1-(13)C]phenylalanine-enriched milk protein (casein enrichment was 29.4 molar percent excess) was collected during stable isotope infusion in the cows. In the proof-of-principle study, ingestion of 35 g of intrinsically labeled casein resulted in peak plasma l-[1-(13)C]phenylalanine enrichments within 90 min after protein ingestion (9.75 +/- 1.47 molar percent excess). Skeletal muscle protein synthesis rates calculated over the entire 6-h period averaged 0.058 +/- 0.012%/h. The production of intrinsically labeled milk protein is feasible and provides dietary protein that can be used to investigate protein digestion and absorption and the subsequent muscle protein synthetic response in vivo in humans.

Muscle mass and strength gains following 6 months of resistance type exercise training are only partly preserved within one year with autonomous exercise continuation in older adults.

INTRODUCTION: Although resistance type exercise training (RT) effectively increases muscle mass and strength in older individuals, it remains unclear whether gains in muscle mass and strength are maintained without continued supervised training. We assessed the capacity of older individuals to maintain muscle mass and strength gains one year after partaking in a successful RT program. METHODS: Fifty-three healthy older adults performed a 24-wk supervised RT program. Upon the cessation of the training program, participants were not provided with any advice or incentives to continue exercise training. One year after completion of the training program, all participants were contacted and invited back to the laboratory to assess anthropometrics, body composition (DXA), quadriceps muscle cross-sectional area (CSA) (CT-scan), muscle strength (1RM knee extension/leg press), and muscle fiber characteristics (muscle biopsy). Following primary analyses on all participants that responded to the invitation (n = 35), participants were divided into two groups: individuals who had continued to perform exercise training on an individual basis (EXER group; n = 16) and individuals who had not continued to perform any regular exercise (STOP group; n = 19) after completing the RT program. RESULTS: The initial increases in quadriceps CSA (+506 ±â€¯209 and +584 ±â€¯287 mm2) and knee extension strength (+32 ±â€¯12 vs +34 ±â€¯10 kg) after the 24-wk RT program did not differ between the STOP and EXER group (all P > 0.05). One year after discontinuation of the RT program, participants had lost muscle mass (P < 0.01), with a greater decline in quadriceps CSA in the STOP vs EXER group (-579 ±â€¯268 vs -309 ±â€¯253 mm2, respectively; P < 0.05). Muscle strength had decreased significantly compared to values after completing the RT program (P < 0.01), with no differences observed between the STOP vs EXER group (knee extension: -21 ±â€¯8 vs -18 ±â€¯8 kg, respectively; P > 0.05), yet remained higher compared with values before the RT program (P < 0.05). CONCLUSION: Though prolonged RT can effectively increase muscle mass and strength in the older population, muscle mass gains are lost and muscle strength gains are only partly preserved within one year if the supervised exercise program is not continued.

Sarcopenia Is Related to Mortality in the Acutely Hospitalized Geriatric Patient.

BACKGROUND: Sarcopenia is defined as low skeletal muscle mass with poor physical performance, representing a strong prognostic factor for mortality in older people. Although highly prevalent in hospitalized geriatric patients, it is unknown whether sarcopenia can also predict mortality in these patients. OBJECTIVE: To determine the association between sarcopenia according the criteria of the European Working Group on Sarcopenia in Older People (EWGSOP), International Working Group on Sarcopenia (IWGS), Special Interest Group of Sarcopenia, Cachexia and Wasting Disorders (SIG) and Foundation for the National Institutes of Health (FNIH) and 2-year mortality in acutely hospitalized geriatric patients. DESIGN: 81 patients (84±5 y) admitted to the acute geriatric ward participated in this study. Body composition assessment (bio-impedance, Maltron Bioscan 920-II) and physical performance tests were performed, and mortality information was retrieved through patient files. RESULTS: Prevalence rates of sarcopenia were 51% (EWGSOP), 75% (IWGS), 69% (SIG), and 27% (FNIH). Based on Cox proportional hazard ratio (HR) analysis, 2-year mortality was significantly higher in sarcopenic patients versus non-sarcopenic patients when using the EWGSOP (2-y: HR 4.310; CI-95%:2.092-8.850; P<0.001) and FNIH criteria (2-y: HR 3.571; CI-95%:1.901-6.711; P<0.001). Skeletal muscle mass index, fat mass index, body mass index, phase angle and gait speed were significantly lower in the geriatric patients who deceased after 2 years versus those who were still alive. Cox proportional HR analyses showed that higher phase angle (HR 0.678; CI-95%:0.531- 0.864; P=0.002) and higher fat mass index (HR 0.839; CI-95%:0.758-0.928; P=0.001) significantly reduced 2-y mortality probability. Combining sarcopenia criteria and separate patient characteristics finally resulted in a model in which HRs for sarcopenia (EWGSOP and FNIH) as well as phase angle significantly predicted mortality probability. CONCLUSION: Sarcopenia is prevalent in acutely hospitalized geriatric patients and is associated with significantly higher 2-year mortality according the EWGSOP and FNIH criteria.

Protein Intake Falls below 0.6 g•kg-1•d-1 in Healthy, Older Patients Admitted for Elective Hip or Knee Arthroplasty.

OBJECTIVE: Hospitalization is generally accompanied by changes in food intake. Patients typically receive hospital meals upon personal preference within the framework of the food administration services of the hospital. In the present study, we assessed food provision and actual food and snack consumption in older patients admitted for elective hip or knee arthroplasty. DESIGN: A prospective observational study. SETTING: Orthopedic nursing ward of the Maastricht University Medical Centre+. PARTICIPANTS: In the present study, n=101 patients (age: 67±10 y; hospital stay: 6.1±1.8 d) were monitored during hospitalization following elective hip or knee arthroplasty. MEASUREMENTS: Energy and protein provided by self-selected hospital meals and snacks, and actual energy and protein (amount, distribution, and source) consumed by patients was weighed and recorded throughout 1-6 days. RESULTS: Self-selected meals provided 6.5±1.5 MJ•d-1, with 16, 48, and 34 En% provided as protein, carbohydrate, and fat, respectively. Self-selected hospital meals provided 0.75±0.16 and 0.79±0.21 g•kg-1•d-1 protein in males and females, respectively. Actual protein consumption averaged merely 0.59±0.18 and 0.50±0.21 g•kg-1•d-1, respectively. Protein consumption at breakfast, lunch, and dinner averaged 16±8, 18±9, and 20±6 g per meal, respectively. CONCLUSIONS: Though self-selected hospital meals provide patients with ~0.8 g•kg-1•d-1 protein during short-term hospitalization, actual protein consumption falls well below 0.6 g•kg-1•d-1 with a large proportion (~32%) of the provided food being discarded. Alternative strategies are required to ensure maintenance of habitual protein intake in older patients admitted for elective orthopedic surgery.

Calpain-10 gene and protein expression in human skeletal muscle: effect of acute lipid-induced insulin resistance and type 2 diabetes.

OBJECTIVE: Our objective was to investigate the effect of lipid-induced insulin resistance and type 2 diabetes on skeletal muscle calpain-10 mRNA and protein levels. RESEARCH DESIGN AND METHODS: In the first part of this study, 10 healthy subjects underwent hyperinsulinemic euglycemic (4.5 mmol/liter) clamps for 6 h with iv infusion of either saline or a 20% Intralipid emulsion (Fresenius Kabi AG, Bad Homburg, Germany). Skeletal muscle biopsies were taken before and after 3- and 6-h insulin infusion and analyzed for calpain-10 mRNA and protein expression. In the second part of the study, muscle samples obtained after an overnight fast in 10 long-standing, sedentary type 2 diabetes patients, 10 sedentary, weight-matched, normoglycemic controls, and 10 age-matched, endurance-trained cyclists were analyzed for calpain-10 mRNA and protein content. RESULTS: Intralipid infusion in healthy subjects reduced whole body glucose disposal by approximately 50% (P<0.001). Calpain-10 mRNA (P=0.01) but not protein content was reduced after 6-h insulin infusion in both the saline and Intralipid emulsion trials. Skeletal muscle calpain-10 mRNA and protein content did not differ between the type 2 diabetes patients and normoglycemic controls, but there was a strong trend for total calpain-10 protein to be greater in the endurance-trained athletes (P=0.06). CONCLUSIONS: These data indicate that skeletal muscle calpain-10 expression is not modified by insulin resistance per se and suggest that hyperinsulinemia and exercise training may modulate human skeletal muscle calpain-10 expression.

Repeated-sprint performance and plasma responses following beetroot juice supplementation do not differ between recreational, competitive and elite sprint athletes.

PURPOSE: There is an ongoing debate whether highly trained athletes are less responsive to the ergogenic properties of nitrate. We assessed the effects of nitrate supplementation on plasma nitrate and nitrite concentrations and repeated-sprint performance in recreational, competitive and elite sprint athletes. METHODS: In a randomized double-blinded cross-over design, recreational cyclists (n = 20), national talent speed-skaters (n = 22) and Olympic-level track cyclists (n = 10) underwent two 6-day supplementation periods; 140 mL/d nitrate-rich (BR; ∼800 mg/d) and nitrate-depleted (PLA; ∼0.5 mg/d) beetroot juice. Blood samples were collected and three 30-s Wingate tests were performed. RESULTS: Plasma nitrate and nitrite concentrations were higher following BR vs PLA (P < .001), with no differences between sport levels (all P > .10). Peak power over the three Wingates was not different between BR and PLA (1338 ± 30 vs 1333 ± 30 W; P = .62), and there was no interaction between treatment (BR-PLA) and Wingate number (1-2-3; P = .48). Likewise, mean power did not differ between BR and PLA (P = .86). In contrast, time to peak power improved by ∼2.8% following BR vs PLA (P = .007). This improvement in BR vs PLA was not different between Wingate 1, 2 and 3. Moreover, the effects of BR vs PLA did not differ between sport levels for any Wingate parameter (all P > .30). CONCLUSION: The plasma and repeated-sprint performance responses to beetroot juice supplementation do not differ between recreational, competitive and elite sprint athletes. Beetroot juice supplementation reduces time to reach peak power, which may improve the capacity to accelerate during high-intensity and sprint tasks in recreational as well as elite athletes.