Protein Intake at Breakfast Promotes a Positive Whole-Body Protein Balance in a Dose-Response Manner in Healthy Children: A Randomized Trial.

Background: Protein ingestion promotes whole-body net protein balance (NB) in children, which is a prerequisite for growth. Determining how much protein is required at breakfast to promote a positive NB, which may be negative after the traditional overnight fast in children, has yet to be determined. Objective: We determined the impact of incremental doses of milk protein at breakfast as well as the impact of daily dietary protein distribution on NB in children. Methods: A total of 28 children [14 boys, 14 girls; age range: 7-11 y; body mass index (mean ± SD, in kg/m2): 16.0 ± 1.9] completed 2 intervention trials. During the breakfast meal, participants consumed an isoenergetic beverage with different amounts of protein (0, 7, 14, or 21 g for Groups A-D, respectively) and [15N]-glycine to measure whole body protein metabolism. Whole-body nitrogen turnover, protein synthesis (PS), protein breakdown, and NB were measured over 9 and 24 h. Results: Following an overnight fast, children were in negative NB (-64.5 mg · kg-1 · h-1). Protein ingestion at breakfast induced a stepwise increase in NB over 9 h [Groups A (6.2 mg · kg-1 · h-1) < B (27.9 mg · kg-1 · h-1) < C (46.9 mg · kg-1 · h-1) < D (66.0 mg · kg-1 · h-1)] with all conditions different from each other (all P < 0.01). PS was 42% greater in Group D than in Group A over 9 h (P < 0.05). Conclusions: Consuming ≥7 g of the total daily protein intake at breakfast attenuates the observed overnight protein losses in children during the subsequent 9 h following breakfast consumption. The dose-dependent increase in NB over a daytime fed period, inclusive of breakfast and lunch, highlights the importance of breakfast protein intake on acute anabolism in healthy active children. This trial was registered at clinicaltrials.gov as NCT02465151.

Postexercise Dietary Protein Ingestion Increases Whole-Body Leucine Balance in a Dose-Dependent Manner in Healthy Children.

Background: Protein ingestion is important in enhancing whole-body protein balance in children. The effect of discrete bolus protein ingestion on acute postexercise recovery has yet to be determined.Objective: This study determined the effect of increasing doses of ingested protein on postexercise whole-body leucine balance in healthy, active children.Methods: Thirty-five children (26 boys, 9 girls; age range: 9-13 y; weight mean ± SD: 44.9 ± 10.6 kg) underwent a 5-d adaptation diet (0.95 g protein ⋅ kg-1 ⋅ d-1) before performing 20 min of cycling 3 times with a concurrent, primed, constant infusion of [13C]leucine. After exercise, participants consumed an isoenergetic beverage (140 kcal) containing variable amounts of bovine skim-milk protein and carbohydrates (sucrose) (0, 5, 10, and 15 g protein made up with 35, 30, 25, and 20 g carbohydrates, respectively). Blood and breath samples were taken over the 3 h of recovery to determine non-steady state whole-body leucine oxidation (LeuOX) and net leucine balance (LeuBAL).Results: LeuOX (secondary outcome) peaked 60 min after beverage ingestion and demonstrated a relative dose-response over the 3 h of recovery (15 g = 10 > 5 > 0 g; P < 0.001). LeuBAL (primary outcome) demonstrated a dose-response over the 3 h [15 g (24.2 ± 8.2 mg/kg) > 10 g (11.6 ± 4.3 mg/kg) > 5 g (5.7 ± 1.9 mg/kg) > 0 g (-3.0 ± 1.7 mg/kg); all P < 0.01] with all conditions different from zero (all P < 0.001).Conclusions: Over the 3-h postexercise period, LeuBAL was negative with carbohydrate ingestion alone; however, the co-ingestion of carbohydrates and 5 g high-quality dietary protein was sufficient to promote a positive postexercise whole-body protein balance in healthy, active children. Moreover, LeuBAL increased in a dose-dependent manner within the protein range studied. Children should consider consuming a source of dietary protein after physical activity to enhance whole-body anabolism. This trial was registered at clinicaltrials.gov as NCT01598935.

The Effect of Postexercise Milk Protein Intake on Rehydration of Children.

PURPOSE: In adults, rehydration after exercise in the heat can be enhanced with a protein-containing beverage; however, whether this applies to children remains unknown. This study examined the effect of milk protein intake on postexercise rehydration in children. METHOD: Fifteen children (10-12 years) performed three exercise trials in the heat (34.4 ± 0.2 °C, 47.9 ± 1.1% relative humidity). In a randomized, counterbalanced crossover design, participants consumed iso-caloric and electrolyte-matched beverages containing 0 g (CONT), 0.76 g (Lo-PRO) or 1.5 g (Hi-PRO) of milk protein/100 mL in a volume equal to 150% of their body mass (BM) loss during exercise. BM was then assessed over 4 h of recovery. RESULTS: Fluid balance demonstrated a significant condition × time interaction (p = .012) throughout recovery; Hi-PRO was less negative than CONT at 2 hr (p = .01) and tended to be less negative at 3 h (p = .07). Compared with CONT, beverage retention was enhanced by Hi-PRO at 2 h (p < .05). CONCLUSION: A postexercise beverage containing milk protein can favorably affect fluid retention in children. Further research is needed to determine the optimal volume and composition of a rehydration beverage for complete restoration of fluid balance.

Chronic α-hydroxyisocaproic acid treatment improves muscle recovery after immobilization-induced atrophy.

Muscle disuse atrophy is observed routinely in patients recovering from traumatic injury and can be either generalized resulting from extended bed rest or localized resulting from single-limb immobilization. The present study addressed the hypothesis that a diet containing 5% α-hydroxyisocaproic acid (α-HICA), a leucine (Leu) metabolite, will slow the loss and/or improve recovery of muscle mass in response to disuse. Adult 14-wk-old male Wistar rats were provided a control diet or an isonitrogenous isocaloric diet containing either 5% α-HICA or Leu. Disuse atrophy was produced by unilateral hindlimb immobilization ("casting") for 7 days and the contralateral muscle used as control. Rats were also casted for 7 days and permitted to recover for 7 or 14 days. Casting decreased gastrocnemius mass, which was associated with both a reduction in protein synthesis and S6K1 phosphorylation as well as enhanced proteasome activity and increased atrogin-1 and MuRF1 mRNA. Although neither α-HICA nor Leu prevented the casting-induced muscle atrophy, the decreased muscle protein synthesis was not observed in α-HICA-treated rats. Neither α-HICA nor Leu altered the increased proteasome activity and atrogene expression observed with immobilization. After 14 days of recovery, muscle mass had returned to control values only in the rats fed α-HICA, and this was associated with a sustained increase in protein synthesis and phosphorylation of S6K1 and 4E-BP1 of previously immobilized muscle. Proteasome activity and atrogene mRNA content were at control levels after 14 days and not affected by either treatment. These data suggest that whereas α-HICA does not slow the loss of muscle produced by disuse, it does speed recovery at least in part by maintaining an increased rate of protein synthesis.

Phytonutrients for bone health during ageing.

Osteoporosis is a skeletal disease characterized by a decrease in bone mass and bone quality that predispose an individual to an increased risk of fragility fractures. Evidence demonstrating a positive link between certain dietary patterns (e.g. Mediterranean diet or high consumption of fruits and vegetables) and bone health highlights an opportunity to investigate their potential to protect against the deterioration of bone tissue during ageing. While the list of these phytonutrients is extensive, this review summarizes evidence on some which are commonly consumed and have gained increasing attention over recent years, including lycopene and various polyphenols (e.g. polyphenols from tea, grape seed, citrus fruit, olive and dried plum). Evidence to define a clear link between these phytonutrients and bone health is currently insufficient to generate precise dietary recommendations, owing to mixed findings or a scarcity in clinical data. Moreover, their consumption typically occurs within the context of a diet consisting of a mix of phytonutrients and other nutrients rather than in isolation. Future clinical trials that can apply a robust set of outcome measurements, including the determinants of bone strength, such as bone quantity (i.e. bone mineral density) and bone quality (i.e. bone turnover and bone microarchitecture), will help to provide a more comprehensive outlook on how bone responds to these various phytonutrients. Moreover, future trials that combine these phytonutrients with established bone nutrients (i.e. calcium and vitamin D) are needed to determine whether combined strategies can produce more robust effects on skeletal health.

Milk beverages can reduce nutrient inadequacy among Brazilian pre-school children: a dietary modelling study.

BACKGROUND: Previous studies found high prevalence of inadequate intakes of vitamins E, D and K, calcium and potassium among Brazilian pre-school children, with suboptimal consumption of dairy products. Dietary modelling was applied to determine the theoretical impact of improving dairy products consumption on nutrient adequacy in 4-5-year-old Brazilian children. METHODS: Adherence to the dairy recommendation of two servings/day was calculated using data from the Brazil Kids Nutrition and Health Study (KNHS) (n = 228). Two modelling scenarios were applied to test the impact on nutrient intakes of (1) adding one or two servings of a frequently consumed cow's milk or a widely available fortified alternative: pre-school children milk (PCM), and of (2) substituting the current milk consumed by PCM. Mean nutrient intakes and percentage of children adhering to the nutrient recommendations were determined at baseline and after applying modelling scenarios. RESULTS: Seventy-six percent (n = 174) of children did not meet the recommended daily two servings of dairy products, 56% had less than one serving of dairy products on the day of recall. The mean consumption of whole milk (fortified and unfortified) was 147 g/d, yoghurt 114 g/d and cheese 34 g/d. The addition of one serving of cow's milk demonstrated a 17% reduction in calcium inadequacy, 18% reduction in vitamin A and 3% reduction in zinc inadequacy. Adding one serving of PCM further reduced calcium inadequacy from 87 to 41%, vitamin E from 81 to 37%, and zinc inadequacy by 10%. Replacing the child's current milk with a PCM resulted in further reduction of micronutrient inadequacies, including calcium, vitamin D and vitamin E. CONCLUSIONS: Dairy products consumption in pre-school children should be encouraged to reduce nutrient inadequacies. In particular, consumption of PCM would help to reduce calcium, vitamin D and vitamin E inadequacy, nutrients of concern in this population.

Impact of moderate dietary protein restriction on glucose homeostasis in a model of estrogen deficiency.

The need to consume adequate dietary protein to preserve physical function during ageing is well recognized. However, the effect of protein intakes on glucose metabolism is still intensively debated. During age-related estrogen withdrawal at the time of the menopause, it is known that glucose homeostasis may be impaired but the influence of dietary protein levels in this context is unknown. The aim of the present study is to elucidate the individual and interactive effects of estrogen deficiency and suboptimal protein intake on glucose homeostasis in a preclinical model involving ovariectomy (OVX) and a 13 week period of a moderately reduced protein intake in 7 month-old ageing rats. To investigate mechanisms of action acting via the pancreas-liver-muscle axis, fasting circulating levels of insulin, glucagon, IGF-1, FGF21 and glycemia were measured. The hepatic lipid infiltration and the protein expression of GLUT4 in the gastrocnemius were analyzed. The gene expression of some hepatokines, myokines and lipid storage/oxidation related transcription factors were quantified in the liver and the gastrocnemius. We show that, regardless of the estrogen status, moderate dietary protein restriction increases fasting glycemia without modifying insulinemia, body weight gain and composition. This fasting hyperglycemia is associated with estrogen status-specific metabolic alterations in the muscle and liver. In estrogen-replete (SHAM) rats, GLUT4 was down-regulated in skeletal muscle while in estrogen-deficient (OVX) rats, hepatic stress-associated hyperglucagonemia and high serum FGF21 were observed. These findings highlight the importance of meeting dietary protein needs to avoid disturbances in glucose homeostasis in ageing female rats with or without estrogen withdrawal.

Absorption, conjugation and excretion of the flavanones, naringenin and hesperetin from alpha-rhamnosidase-treated orange juice in human subjects.

We have determined the absorption, conjugation and excretion of naringenin-7-O-rutinoside (narirutin) compared to the corresponding glucoside in an orange juice matrix in human subjects. Healthy volunteers (eight men and eight women), in a double blind, randomised, crossover study, consumed orange juice with (1) natural content of naringenin-7-O-rutinoside; (2) alpha-rhamnosidase-treated to yield naringenin-7-O-glucoside. Blood was sampled at twelve time points and three fractions of urine were collected over 24 h. The area under the plasma-time curve of naringenin from (2) alpha-rhamnosidase-treated orange juice was increased about 4-fold (P < 0.0001), peak plasma concentration (Cmax) was 5.4-fold higher (P < 0.0001) and Tmax was decreased from 311 to 92 min (P = 0.002) compared to untreated orange juice (1), indicating a change in absorption site from the colon to the small intestine. Furthermore, the amount in urine was increased from 7 to 47 % (P < 0.0001) of the dose after consumption of the alpha-rhamnosidase-treated orange juice (2). All urine samples contained both naringenin-7- and -4'-O-glucuronides. In addition, to examine the effect of dose and alpha-rhamnosidase treatment on hesperetin conjugate profiles, a further treatment where (3) orange juice fortified with three times the original content of hesperetin-7-O-rutinoside was used. Five hesperetin metabolites (3'-O-glucuronide; 7-O-glucuronide; 5,7-O-diglucuronide; 3',7-O-diglucuronide; 3'-O-sulphate) were present after all treatments (1-3), with the same profile of the conjugates. The present data show that bioavailability of naringenin is increased by conversion from rutinoside to glucoside, but the profile of the conjugates of flavanones formed and excreted in urine is neither affected by the absorption site nor by a 3-fold change in dose.

Global Landscape of Nutrient Inadequacies in Toddlers and Young Children.

Toddlers and young children need an adequate and diverse diet to provide all of the nutrients required for optimal growth and development. Unfortunately, inadequate intake of vitamins and minerals is still identified by the World Health Organization (WHO) as a major public health threat for young children. Organizations like the WHO and the World Bank focus primarily on iron, zinc, vitamin A, and iodine for children ≤5 years of age in low-income countries. In addition to the data from these organizations, individual-level food consumption surveys are needed to provide a fuller picture of food and nutrient intakes. Where studies are available, intakes of dietary fiber and vitamin D are generally below recommendations for toddlers and young children. Other nutrient gaps differ by country and are related to food availability and local dietary habits. For example, young children in the US regularly consume dairy products, and <10% fall below recommendations for calcium intake compared to 2- to 4-year-old toddlers in the Philippines where dairy food consumption is low, and 66-84% fall below calcium recommendations. Dietary intake studies can help to identify the foods and beverages most relevant to alleviate nutrient gaps and improve dietary intakes of toddlers and young children around the world.

Contribution of Milk Beverages to Nutrient Adequacy of Young Children and Preschool Children in the Philippines.

Malnutrition is a major public health concern in the Philippines. Milk and dairy products are important sources of energy, protein, and micronutrients for normal growth and development in children. This study aims to assess the contribution of different types of milk to nutrient intakes and nutrient adequacy among young and preschool children in the Philippines. Filipino children aged one to four years (n = 2992) were analysed while using dietary intake data from the 8th National Nutrition Survey 2013. Children were stratified by age (one to two years and three to four years) and by milk beverage consumption type: young children milk (YCM) and preschool children milk (PCM), other milks (mostly powdered milk with different degrees of fortification of micronutrients), and non-dairy consumers (no milks or dairy products). The mean nutrient intakes and the odds of meeting nutrient adequacy by consumer groups were compared, percentage of children with inadequate intakes were calculated. Half (51%) of Filipino children (all ages) did not consume any dairy on a given day, 15% consumed YCM or PCM, and 34% consumed other milks. Among children one to two years, those who consumed YCM had higher mean intakes of iron, magnesium, potassium, zinc, B vitamins, folate, and vitamins C, D, and E (all p < 0.001) when compared to other milk consumers. Non-dairy consumers had mean intakes of energy, total fat, fibre, calcium, phosphorus, iron, potassium, zinc, folate, and vitamins D and E that were far below the recommendations. Children who consumed YCM or PCM had the highest odds in meeting adequacy of iron, zinc, thiamin, vitamin B6, folate, and vitamins C, D, and E as compared to other milks or non-dairy consumers, after adjusting for covariates. This study supports the hypothesis that dairy consumers had higher intakes of micronutrients and higher nutrient adequacy than children who consumed no milk or dairy products. Secondly, YCM or PCM have demonstrated to be good dairy options to achieve nutrient adequacy in Filipino children.

The Nutritional Impact of Milk Beverages in Reducing Nutrient Inadequacy among Children Aged One to Five Years in the Philippines: A Dietary Modelling Study.

Around half of Filipino children are not consuming any dairy products on a given day, which has shown to be associated with increased risk of inadequate nutrient intakes. The current study applies dietary modelling to assess the nutritional impact of meeting dairy recommendations in reducing nutrient inadequacy in children aged one to five years in the Philippines. Dietary intake data of Filipino children aged one to five years (n = 3864) were analyzed from the 8th National Nutrition Survey 2013. Children who did not meet national dairy recommendations were identified. Two scenarios were applied, based on two types of commonly consumed milk products by the survey participants. In scenario one, one serving of powdered milk was added to the diet of these children. In scenario two, one serving of a young children milk (YCM) or preschool children milk (PCM) was added to the diet of children aged one to two years and three to five years, respectively. Mean nutrient intakes and percentages of children with inadequate intakes were estimated before and after applying modelling scenarios. Scenario one demonstrated improvement in calcium, phosphorus, sodium, vitamin A and riboflavin intakes, while in scenario two, further improvement of intakes of a wider range of nutrients including iron, selenium, zinc, magnesium, potassium, vitamins C, D, E, thiamin, niacin, vitamins B6, and B12 was observed. In both scenarios, if all children would meet their dairy recommendations, theoretical reductions in population nutrient inadequacy would be observed for all micronutrients, for example, only 20% of children aged one to two years would be inadequate in vitamin A instead of the current 60%, iron inadequacy would see a 5% reduction, and approximately 10% reduction for calcium and 20% reduction for folate. The present study is the first to apply dietary modelling to assess the theoretical impact of meeting dairy recommendations on nutrient inadequacy in children in the Philippines. Dairy consumption should be encouraged as part of the strategy to reduce nutrient inadequacies. Calcium, iron, vitamins D, E, and folate are of concern in the Philippines as the level of inadequacies are extremely high in early years, YCM and PCM can help increase the intake of these nutrients.

Increased bioavailability of hesperetin-7-glucoside compared with hesperidin results in more efficient prevention of bone loss in adult ovariectomised rats.

Hesperidin (Hp), a citrus flavonoid predominantly found in oranges, shows bone-sparing effects in ovariectomised (OVX) animals. In human subjects, the bioavailability of Hp can be improved by the removal of the rhamnose group to yield hesperetin-7-glucoside (H-7-glc). The aim of the present work was to test whether H-7-glc was more bioavailable and therefore more effective than Hp in the prevention of bone loss in the OVX rat. Adult 6-month-old female Wistar rats were sham operated or OVX, then pair fed for 90 d a casein-based diet supplemented or not with freeze-dried orange juice enriched with Hp or H-7-glc at two dose equivalents of the hesperetin aglycone (0.25 and 0.5 %). In the rats fed 0.5 %, a reduction in OVX-induced bone loss was observed regarding total bone mineral density (BMD):+7.0 % in OVX rats treated with Hp (HpOVX) and +6.6 % in OVX rats treated with H-7-glc (H-7-glcOVX) v. OVX controls (P < 0.05). In the rats fed 0.25 % hesperetin equivalents, the H-7-glcOVX group showed a 6.6 % improvement in total femoral BMD v. the OVX controls (P < 0.05), whereas the Hp diet had no effect at this dose. The BMD of rats fed 0.25 % H-7-glc was equal to that of those given 0.5 % Hp, but was not further increased at 0.5 % H-7-glc. Plasma hesperetin levels and relative urinary excretion were significantly enhanced in the H-7-glc v. Hp groups, and the metabolite profile showed the absence of eriodictyol metabolites and increased levels of hesperetin sulphates. Taken together, improved bioavailability of H-7-glc may explain the more efficient bone protection of this compound.

Timing and pattern of postexercise protein ingestion affects whole-body protein balance in healthy children: a randomized trial.

The dose and timing of postexercise protein ingestion can influence whole-body protein balance (WBPB) in adults, although comparable data from children are scarce. This study investigated how protein intake (both amount and distribution) postexercise can affect WBPB in physically active children. Thirty-five children (26 males; 9-13 years old) underwent a 5-day adaptation diet, maintaining a protein intake of 0.95 g·kg-1·day-1. Participants consumed [15N]glycine (2 mg·kg-1) before performing 3 × 20 min of variable-intensity cycling, and whole-body protein kinetics were assessed over 6 and 24 h of recovery. Fifteen grams of protein was distributed across 2 isoenergetic carbohydrate-containing beverages (15 and 240 min postexercise) containing reciprocal amounts of protein (i.e., 0 + 15 g, 5 + 10 g, 10 + 5 g, and 15 + 0 g for Groups A-D, respectively). Over the 6 h that included the exercise bout and consumption of the first beverage at 15 min postexercise, WBPB (i.e., synthesis - breakdown) demonstrated a linear increase of 0.647 g·kg-1·day-1 per 1 g protein intake (P < 0.001). Over 24 h, robust regression revealed that WBPB was best modeled by a parabola (P < 0.05), suggesting that a maximum in WBPB was achieved between groups B and C. In conclusion, despite a dose response early in recovery, a periodized protein intake with multiple smaller doses after physical activity may be more beneficial than a single bolus dose in promoting daily WBPB in healthy active children.

Carnosol Inhibits Pro-Inflammatory and Catabolic Mediators of Cartilage Breakdown in Human Osteoarthritic Chondrocytes and Mediates Cross-Talk between Subchondral Bone Osteoblasts and Chondrocytes.

AIM: The aim of this work was to evaluate the effects of carnosol, a rosemary polyphenol, on pro-inflammatory and catabolic mediators of cartilage breakdown in chondrocytes and via bone-cartilage crosstalk. MATERIALS AND METHODS: Osteoarthritic (OA) human chondrocytes were cultured in alginate beads for 4 days in presence or absence of carnosol (6 nM to 9 µM). The production of aggrecan, matrix metalloproteinase (MMP)-3, tissue inhibitor of metalloproteinase (TIMP)-1, interleukin (IL)-6 and nitric oxide (NO) and the expression of type II collagen and ADAMTS-4 and -5 were analyzed. Human osteoblasts from sclerotic (SC) or non-sclerotic (NSC) subchondral bone were cultured for 3 days in presence or absence of carnosol before co-culture with chondrocytes. Chondrocyte gene expression was analyzed after 4 days of co-culture. RESULTS: In chondrocytes, type II collagen expression was significantly enhanced in the presence of 3 µM carnosol (p = 0.008). MMP-3, IL-6, NO production and ADAMTS-4 expression were down-regulated in a concentration-dependent manner by carnosol (p<0.01). TIMP-1 production was slightly increased at 3 µM (p = 0.02) and ADAMTS-5 expression was decreased from 0.2 to 9 µM carnosol (p<0.05). IL-6 and PGE2 production was reduced in the presence of carnosol in both SC and NSC osteoblasts while alkaline phosphatase activity was not changed. In co-culture experiments preincubation of NSC and SC osteoblasts wih carnosol resulted in similar effects to incubation with anti-IL-6 antibody, namely a significant increase in aggrecan and decrease in MMP-3, ADAMTS-4 and -5 gene expression by chondrocytes. CONCLUSIONS: Carnosol showed potent inhibition of pro-inflammatory and catabolic mediators of cartilage breakdown in chondrocytes. Inhibition of matrix degradation and enhancement of formation was observed in chondrocytes cocultured with subchondral osteoblasts preincubated with carnosol indicating a cross-talk between these two cellular compartments, potentially mediated via inhibition of IL-6 in osteoblasts as similar results were obtained with anti-IL-6 antibody.

Low-load resistance training during step-reduction attenuates declines in muscle mass and strength and enhances anabolic sensitivity in older men.

Step-reduction (SR) in older adults results in muscle atrophy and an attenuated rise in postprandial muscle protein synthesis (MPS): anabolic resistance. Knowing that resistance exercise (RT) can enhance MPS, we examined whether RT could enhance MPS following 2 weeks of SR. In addition, as we postulated that SR may impair feeding-induced vasodilation limiting nutrient delivery to muscle, we also examined whether citrulline (CIT), as an arginine and nitric oxide precursor, could attenuate muscle anabolic resistance accompanying SR. We used a unilateral leg model to compare older subjects' who had undergone SR to a loaded condition of SR plus RT (SR + RT). Thirty older men (70 ± 1 years) underwent 14 days of SR (<1500 steps/day) with supplementation of either 5 g/day CIT or glycine placebo. Throughout SR, subjects performed unilateral low-load RT thrice weekly. We assessed muscle protein synthesis in the postabsorptive and postprandial state (20 g whey isolate plus 15 g glycine or as micellar-whey with 5 g CIT or 15 g glycine, n = 10/group). As MPS was similar after ingestion of either whey isolate, micellar-whey, or micellar-whey + CIT data related to different dietary groups were collapsed to compare SR and SR + RT legs. Subjects' daily steps were reduced by 80 ± 2% during SR (P < 0.001) compared with baseline. Leg fat-free mass decreased with SR (-124 ± 61 g) and increased in the SR + RT (+126 ± 68 g; P = 0.003). Myofibrillar FSR was lower (P < 0.0001) in the SR as compared with the SR + RT leg in the postabsorptive (0.026 ± 0.001%/h vs. 0.045 ± 0.001%/h) and postprandial states (0.055 ± 0.002%/h vs. 0.115 ± 0.003%/h). We conclude that low-load RT, but not supplementation with CIT, can attenuate the deleterious effects of SR in aging muscle.

Photoprotective potential of lycopene, beta-carotene, vitamin E, vitamin C and carnosic acid in UVA-irradiated human skin fibroblasts.

The photoprotective potential of the dietary antioxidants vitamin C, vitamin E, lycopene, beta-carotene, and the rosemary polyphenol, carnosic acid, was tested in human dermal fibroblasts exposed to ultraviolet-A (UVA) light. The carotenoids were prepared in special nanoparticle formulations together with vitamin C and/or vitamin E. Nanoparticle formulations, in contrast to dimethylsulphoxide, stablized lycopene in the cell culture medium and allowed efficient cellular uptake. The presence of vitamin E in the formulation further increased the stability and cellular uptake of lycopene. UVA irradiation of the human skin fibroblasts led to a 10-15-fold rise in metalloproteinase 1 (MMP-1) mRNA. This rise was suppressed in the presence of low microM concentrations of vitamin E, vitamin C, or carnosic acid but not with beta-carotene or lycopene. Indeed, in the presence of 0.5-1.0 microM beta-carotene or lycopene, the UVA-induced MMP-1 mRNA was further increased by 1.5-2-fold. This increase was totally suppressed when vitamin E was included in the nanoparticle formulation. Heme-oxygenase 1 (HO-1) mRNA expression was strongly induced by UVA irradiation but none of the antioxidants inhibited this effect at the concentrations used in this study. Indeed, beta-carotene or lycopene (0.5-1.0 microM) led to a further 1.5-fold rise in the UVA-induced HO-1 mRNA levels. In conclusion, vitamin C, vitamin E, and carnosic acid showed photoprotective potential. Lycopene and beta-carotene did not protect on their own but in the presence of vitamin E, their stability in culture was improved and the rise in MMP-1 mRNA expression was suppressed, suggesting a requirement for antioxidant protection of the carotenoids against formation of oxidative derivatives that can influence the cellular and molecular responses.

Green tea extract decreases muscle necrosis in mdx mice and protects against reactive oxygen species.

BACKGROUND: Duchenne muscular dystrophy is a severe X-linked congenital disorder characterized by lethal muscle wasting caused by the absence of the structural protein dystrophin. OBJECTIVE: Because generation of reactive oxygen species appears to play an important role in the pathogenesis of this disease, we tested whether antioxidant green tea extract could diminish muscle necrosis in the mdx mouse dystrophy model. DESIGN: A diet supplemented with 0.01% or 0.05% green tea extract was fed to dams and neonates for 4 wk beginning on the day of birth. Muscle necrosis and regeneration were determined in stained cryosections of soleus and elongator digitorum longus muscles. Radical scavenging by green tea extract was determined in differentiated cultured C2C12 cells treated with tert-butylhydroperoxide, with the use of 2',7'-dichlorofluorescin diacetate as a radical detector. RESULTS: This feeding regimen significantly and dose-dependently reduced necrosis in the fast-twitch muscle elongator digitorum longus but at the doses tested had no effect on the slow-twitch soleus muscle. Green tea extract concentration-dependently decreased oxidative stress induced by tert-butylhydroperoxide treatment of cultured mouse C2C12 myotubes. The lower effective dose tested in mdx mice corresponds to approximately equal to 1.4 L (7 cups) green tea/d in humans. CONCLUSION: Green tea extract may improve muscle health by reducing or delaying necrosis in mdx mice by an antioxidant mechanism.

High isoflavone content and estrogenic activity of 25 year-old Glycine max tissue cultures.

Soy isoflavones are phytoestrogens which have been associated with several health benefits. In the present study, we report the production of isoflavones in a collection of 40 strains of soya cell cultures established in 1975. A large variability in the isoflavone composition was observed and high-producing strains, with an isoflavone content of up to 46.3 mg g(-1) dry wt., were found. In comparison with soybeans, many callus strains had a higher isoflavone concentration (10-40 times) and a different ratio of genistin to daidzin forms. The highest producing strain was transferred to liquid medium in an Erlenmeyer flask and in a 10 l stirred-tank bioreactor where high isoflavone content (7% dry wt.), concentration (880 mg l(-1)) and a maximum productivity estimated to 60 mg l(-1) d(-1) were obtained. We further studied the estrogenic activity of pure compounds compared to plant cell culture extracts in the estrogen-responsive human endometrial Ishikawa cell line. Estrogen was confirmed to be 1000-10,000 times more active than isoflavones. The estrogenic activity of the extracts correlated to their isoflavone content. The activity of the malonyl isoflavones, assessed here for the first time, was lower than the aglycones. Taken together, these results suggest that soya cell cultures can be used as an alternative source to soybeans to provide high concentrations of bioactive isoflavones.

Postexercise protein ingestion increases whole body net protein balance in healthy children.

Postexercise protein ingestion increases whole body and muscle protein anabolism in adults. No study has specifically investigated the combined effects of exercise and protein ingestion on protein metabolism in healthy, physically active children. Under 24-h dietary control, 13 (seven males, six females) active children (∼ 11 yr old; 39.3 ± 5.9 kg) consumed an oral dose of [(15)N]glycine prior to performing a bout of exercise. Immediately after exercise, participants consumed isoenergetic mixed macronutrient beverages containing a variable amount of protein [0, 0.75, and 1.5 g/100 ml for control (CON), low protein (LP), and high protein (HP), respectively] according to fluid losses. Whole body nitrogen turnover (Q), protein synthesis (S), protein breakdown (B), and protein balance (WBPB) were measured throughout exercise and the early acute recovery period (9 h combined) as well as over 24 h. Postexercise protein intake from the beverage was ∼ 0.18 and ∼ 0.32 g/kg body mass for LP and HP, respectively. Q, S, and B were significantly greater (main effect time, all P < 0.001) over 9 h compared with 24 h with no differences between conditions. WBPB was also greater over 9 h compared with 24 h in all conditions (main effect time, P < 0.001). Over 9 h, WBPB was greater in HP (P < 0.05) than LP and CON with a trend (P = 0.075) toward LP being greater than CON. WBPB was positive over 9 h for all conditions but only over 24 h for HP. Postexercise protein ingestion acutely increases net protein balance in healthy children early in recovery in a dose-dependent manner with larger protein intakes (∼ 0.32 g/kg) required to sustain a net anabolic environment over an entire 24 h period.