Jamie I. Baum

Mechanisms involved in the coordinate regulation of mTORC1 by insulin and amino acids.

In this study, we explored the coordinate regulation of mTORC1 by insulin and amino acids. Rat livers were perfused with medium containing various concentrations of insulin and/or amino acids. At fasting (1×) or 2× (2×AA) concentrations of amino acids, insulin maximally stimulated Akt phosphorylation but had no effect on global rates of protein synthesis. In the absence of insulin, 4×AA produced a moderate stimulation of protein synthesis and activation of mTORC1. The combination of 4×AA and insulin produced a maximal stimulation of protein synthesis and activation of mTORC1. These effects were accompanied by decreases in raptor and PRAS40 and an increase in RagC associated with mTOR (mammalian target of rapamycin). The studies were extended to a cell culture model in which mTORC1 activity was repressed by deprivation of leucine and serum, and resupplementation with the amino acid and insulin acted in an additive manner to restore mTORC1 activation. In deprived cells, mTORC1 was activated by expressing either constitutively active (ca) Rheb or a caRagB·caRagC complex, and coexpression of the constructs had an additive effect. Notably, resupplementation with leucine in cells expressing caRheb or with insulin in cells expressing the caRagB·caRagC complex was as effective as resupplementation with both leucine and insulin in non-transfected cells. Moreover, changes in mTORC1 activity correlated directly with altered association of mTOR with RagB/RagC, Rheb, raptor, and PRAS40. Overall, the results suggest that amino acids signal through the Rag complex and insulin through Rheb to achieve coordinate activation of mTORC1.

Protein Consumption and the Elderly: What Is the Optimal Level of Intake?

Maintaining independence, quality of life, and health is crucial for elderly adults. One of the major threats to living independently is the loss of muscle mass, strength, and function that progressively occurs with aging, known as sarcopenia. Several studies have identified protein (especially the essential amino acids) as a key nutrient for muscle health in elderly adults. Elderly adults are less responsive to the anabolic stimulus of low doses of amino acid intake compared to younger individuals. However, this lack of responsiveness in elderly adults can be overcome with higher levels of protein (or essential amino acid) consumption. The requirement for a larger dose of protein to generate responses in elderly adults similar to the responses in younger adults provides the support for a beneficial effect of increased protein in older populations. The purpose of this review is to present the current evidence related to dietary protein intake and muscle health in elderly adults.

Diet-induced obesity alters anabolic signalling in mice at the onset of skeletal muscle regeneration

Obesity is classified as a metabolic disorder that is associated with delayed muscle regeneration following damage. For optimal skeletal muscle regeneration, inflammation along with extracellular matrix remodelling and muscle growth must be tightly regulated. Moreover, the regenerative process is dependent on the activation of myogenic regulatory factors (MRFs) for myoblast proliferation and differentiation. The purpose of this study was to determine how obesity alters inflammatory and protein synthetic signalling and MRF expression at the onset of muscle regeneration in mice.

Breakfasts Higher in Protein Increase Postprandial Energy Expenditure, Increase Fat Oxidation, and Reduce Hunger in Overweight Children from 8 to 12 Years of Age

BACKGROUND Currently 1 in every 3 children aged 2-19 y is overweight or obese. Breakfast is a key component of a healthy diet and has the potential to affect childrens health. OBJECTIVE The objective of this study was to determine whether consumption of a protein-based breakfast (PRO) increases postprandial energy metabolism and substrate oxidation, reduces hunger, and reduces food intake at lunch compared with a carbohydrate-based breakfast (CHO) in normal weight (NW) vs. overweight/obese (OW) children. METHODS A randomized, crossover-design study was conducted in NW (n = 16; 33 ± 1 kg) and OW (n = 13; 46 ± 2 kg) children (10 ± 1 y). Participants were served either a PRO [344 kcal, 21% protein (18 g), 52% carbohydrate, and 27% fat] or CHO [327 kcal, 4% protein (3 g), 67% carbohydrate, and 29% fat]. Energy expenditure (EE), substrate oxidation, appetite, and blood glucose were measured over a 4 h period. Four hour postprandial participants were provided with access to a lunch buffet and food intake was recorded. RESULTS After breakfast, OW children in the PRO group had higher (P < 0.0001) EEs and fat oxidation over the 4 h period than did the NW children in the CHO and PRO groups. There was no difference in postprandial EE or carbohydrate oxidation between the CHO and PRO groups over the 4 h period; however, fat oxidation was 16% higher (P < 0.05) after the PRO than the CHO and postprandial carbohydrate oxidation at 4 h was 32% higher after the PRO than the CHO (P < 0.01), independent of weight group. All participants had decreased feelings of hunger (-14%; P < 0.01) and increased fullness (+32%; P < 0.05) after the PRO than the CHO. Finally, there was no difference in food intake within the NW and OW groups. CONCLUSION This study indicates that breakfast macronutrient composition affects postprandial responses in both NW and OW children. A PRO increases postprandial EE and fat oxidation, reduces hunger, and increases satiety when compared with a carbohydrate-based breakfast.

Monocarboxylate transporter expression at the onset of skeletal muscle regeneration.

The onset of skeletal muscle regeneration is characterized by proliferating myoblasts. Proliferating myoblasts have an increased energy demand and lactate exchange across the sarcolemma can be used to address this increased demand. Monocarboxylate transporters (MCTs) are involved in lactate transport across the sarcolemma and are known to be affected by various physiological stimuli. However, MCT expression at the onset of skeletal muscle regeneration has not been determined. The purpose of this study was to determine if skeletal muscle regeneration altered MCT expression in regenerating tibialis anterior (TA) muscle. Male C57/BL6 mice were randomly assigned to either a control (uninjured) or bupivacaine (injured) group. Three days post injection, the TA was extracted for determination of protein and gene expression. A 21% decrease in muscle mass to tibia length (2.4 ± 0.1 mg/mm vs. 1.9 ± 0.2 mg/mm, P < 0.02) was observed. IGF‐1 and MyoD gene expression increased 5.0‐fold (P < 0.05) and 3.5‐fold (P < 0.05), respectively, 3 days post bupivacaine injection. MCT‐1 protein was decreased 32% (P < 0.03); however, MCT‐1 gene expression was not altered. There was no difference in MCT4 protein or gene expression. Lactate dehydrogenase (LDH)‐A protein expression increased 71% (P < 0.0004). Protein levels of LDH‐B and mitochondrial enzyme cytochrome C oxidase subunit decreased 3 days post bupivacaine injection. CD147 and PKC‐θ protein increased 64% (P < 0.03) and 79% (P < 0.02), respectively. MCT1 but not MCT4 expression is altered at the onset of skeletal muscle regeneration possibly in an attempt to regulate lactate uptake and use by skeletal muscle cells.

Glucagon acts in a dominant manner to repress insulin-induced mammalian target of rapamycin complex 1 signaling in perfused rat liver

The opposing actions of insulin and glucagon on hepatic carbohydrate metabolism are well documented. In contrast, relatively little is known about how the two hormones interact to regulate hepatic protein metabolism. Previously, we reported that glucagon in the absence of insulin represses signaling through the mammalian target of rapamycin complex 1 (mTORC1). In the present study, we sought to determine whether or not the action of one hormone would dominate over the other in the regulation of mTORC1 signaling. Livers were perfused in situ with medium containing either no added hormones (control), 10 nM insulin, 100 nM glucagon, or a combination of the hormones. Compared with control livers, insulin stimulated Akt phosphorylation and mTORC1 signaling, as assessed by increased phosphorylation of the mTORC1 targets eIF4E-binding protein (4E-BP)1 and ribosomal protein S6 kinase (S6K)1, and promoted assembly of the eIF4G x eIF4E complex. Glucagon alone had no effect on mTORC1 signaling but stimulated the activity of protein kinase A (PKA). In the presence of a combination of insulin and glucagon, Akt and TSC2 phosphorylation and PKA activity were all increased compared with controls. However, mTORC1 signaling was repressed compared with livers perfused with medium containing insulin alone, and this effect was associated with reduced assembly of the mTORC1 x eIF3 complex. Overall, the results suggest that glucagon acts in a dominant manner to repress insulin-induced mTORC1 signaling, which is in contrast to previous studies showing a dominant action of insulin in the control of hepatic gluconeogenesis.

Effect of Aronia melanocarpa (Black Chokeberry) supplementation on the development of obesity in mice fed a high-fat diet

BACKGROUND: Products derived from black chokeberry are claimed to be beneficial in treating chronic diseases, such as obesity and diabetes. OBJECTIVE: The objective of this study was to determine if supplementation with Aronia melanocarpa (black chokeberry) juice concentrate (AJC) has anti-obesity properties in mice fed obesogenic diets. METHODS: Male C57BL/6J mice (n = 10/dietary treatment) were placed on either a low-fat, high-sucrose (LFHS; 5% fat), LFHS+AJC (1.44 g AJC/kg diet), high-fat (HF; 30% fat), or HF+AJC for 12-weeks. RESULTS: Final body weight was lower in LFHS+AJC compared to LFHS, HF and HF+AJC (∼14, 20% and ∼16%, respectively; P < 0.05). Mice receiving LFHS and LFHS+AJC had significantly higher (P = 0.001) energy intake than HF and HF+AJC. LFHS-fed mice had less (–30%) epididymal fat (p < 0.05) than HF-fed mice, however mice on the LFHS+AJC had less epididymal fat per gram body weight than LFHS controls. There was no effect of diet or AJC on adipose tissue gene expression. There was no difference in plasma insulin, glucose or triglycerides between groups, however there was a positive effect of AJC on adiponectin (P = 0.059). There was also a significant effect of diet (LFHS versus HF) on HOMA-IR (P = 0.004) and HOMA-BCF (P = 0.002). CONCLUSIONS: The results from this study demonstrate that AJC supplementation has the potential to prevent weight gain and markers of obesity. Further research is needed to determine mechanisms of action.

Breakfast Macronutrient Composition Influences Thermic Effect of Feeding and Fat Oxidation in Young Women Who Habitually Skip Breakfast

The purpose of this study was to determine if breakfast macronutrient composition improved thermic effect of feeding (TEF) and appetite after a one-week adaptation in young women who habitually skip breakfast. A randomized, controlled study was conducted in females (24.1 ± 2 years), who skip breakfast (≥5 times/week). Participants were placed into one of three groups for eight days (n = 8 per group): breakfast skipping (SKP; no breakfast), carbohydrate (CHO; 351 kcal; 59 g CHO, 10 g PRO, 8 g fat) or protein (PRO; 350 kcal; 39 g CHO, 30 g PRO, 8 g fat). On days 1 (D1) and 8 (D8), TEF, substrate oxidation, appetite and blood glucose were measured. PRO had higher (p < 0.05) TEF compared to SKP and CHO on D1 and D8, with PRO having 29% higher TEF than CHO on D8. On D1, PRO had 30.6% higher fat oxidation than CHO and on D8, PRO had 40.6% higher fat oxidation than CHO. SKP had higher (p < 0.05) fat oxidation on D1 and D8 compared to PRO and CHO. There was an interaction (p < 0.0001) of time and breakfast on appetite response. In addition, CHO had a significant increase (p < 0.05) in PP hunger response on D8 vs. D1. CHO and PRO had similar PP (postprandial) glucose responses on D1 and D8. Consumption of PRO breakfast for 8 days increased TEF compared to CHO and SKP, while consumption of CHO for one week increased PP hunger response.

Differential effects of leucine supplementation in young and aged mice at the onset of skeletal muscle regeneration

Aging decreases the ability of skeletal muscle to respond to injury. Leucine has been demonstrated to target protein synthetic pathways in skeletal muscle thereby enhancing this response. However, the effect of aging on leucine-induced alterations in protein synthesis at the onset of skeletal muscle regeneration has not been fully elucidated. The purpose of this study was to determine if aging alters skeletal muscle regeneration and leucine-induced alterations in markers of protein synthesis. The tibialis anterior of young (3 months) and aged (24 months) female C57BL/6J mice were injected with either bupivacaine or PBS, and the mice were given ad libitum access to leucine-supplemented or normal drinking water. Protein and gene expression of markers of protein synthesis and degradation, respectively, were analyzed at three days post-injection. Following injury in young mice, leucine supplementation was observed to elevate only p-p70S6K. In aged mice, leucine was shown to elicit higher p-mTOR content with and without injury, and p-4EBP-1 content post-injury. Additionally in aged mice, leucine was shown to elicit higher content of relative p70S6K post-injury. Our study shows that leucine supplementation affects markers of protein synthesis at the onset of skeletal muscle regeneration differentially in young and aged mice.

The Link between Dietary Protein Intake, Skeletal Muscle Function and Health in Older Adults

Skeletal muscle mass and function are progressively lost with age, a condition referred to as sarcopenia. By the age of 60, many older adults begin to be affected by muscle loss. There is a link between decreased muscle mass and strength and adverse health outcomes such as obesity, diabetes and cardiovascular disease. Data suggest that increasing dietary protein intake at meals may counterbalance muscle loss in older individuals due to the increased availability of amino acids, which stimulate muscle protein synthesis by activating the mammalian target of rapamycin (mTORC1). Increased muscle protein synthesis can lead to increased muscle mass, strength and function over time. This review aims to address the current recommended dietary allowance (RDA) for protein and whether or not this value meets the needs for older adults based upon current scientific evidence. The current RDA for protein is 0.8 g/kg body weight/day. However, literature suggests that consuming protein in amounts greater than the RDA can improve muscle mass, strength and function in older adults.