Availability Of Essential Amino Acids Research Articles

Plant Protein Blend Ingestion Stimulates Post-Exercise Myofibrillar Protein Synthesis Rates Equivalently to Whey in Resistance-Trained Adults.

Whey protein ingestion is typically considered an optimal dietary strategy to maximize myofibrillar protein synthesis (MyoPS) following resistance exercise. While single source plant protein ingestion is typically less effective, at least partly, due to less favorable amino acid profiles, this could theoretically be overcome by blending plant-based proteins with complementary amino acid profiles. We compared the post-exercise MyoPS response following the ingestion of a novel plant-derived protein blend with an isonitrogenous bolus of whey protein. Ten healthy, resistance trained, young adults (male/female: 8/2; age: 26 ± 6 y; BMI: 24 ± 3 kg·m-2) received a primed continuous infusion of L-[ring-2H5]-phenylalanine and completed a bout of bilateral leg resistance exercise before ingesting 32 g protein from whey (WHEY) or a plant protein blend (BLEND; 39.5% pea, 39.5% brown rice, 21.0% canola) in a randomized, double-blind crossover fashion. Blood and muscle samples were collected at rest, and 2 and 4 h after exercise and protein ingestion, to assess plasma amino acid concentrations, and postabsorptive and post-exercise MyoPS rates. Plasma essential amino acid availability over the 4 h postprandial post-exercise period was ~44% higher in WHEY compared with BLEND (P = 0.04). From equivalent postabsorptive values (WHEY, 0.042 ± 0.020%·h-1; BLEND, 0.043 ± 0.015%·h-1) MyoPS rates increased following exercise and protein ingestion (time effect; P < 0.001) over a 0-2 h (WHEY, 0.085 ± 0.037%·h-1; BLEND, 0.080 ± 0.037%·h-1) and 2-4 h (WHEY, 0.085 ± 0.036%·h-1; BLEND, 0.086 ± 0.034%·h-1) period, with no differences between conditions during either period or throughout the entire (0-4 h) postprandial period (time × condition interactions; all P > 0.05). Ingestion of a novel plant-based protein blend stimulates post-exercise MyoPS to an equivalent extent as a whey protein, demonstrating the utility of plant protein blends to optimize post-exercise skeletal muscle reconditioning.

Effects of Individual Essential Amino Acids on Growth Rates of Young Rats Fed a Low-Protein Diet.

To investigate the effects of individual essential amino acids (EAA) on growth and the underlying mechanisms, EAA individually supplemented a low-protein (LP) diet fed to young rats in the present study. Treatments were an LP diet that contained 6% crude protein (CP), a high-protein (HP) diet that contained 18% CP, and 10 LP diets supplemented with individual EAA to achieve an EAA supply equal to that of the HP diet. The CP concentration of the LP diet was ascertained from the results of the first experiment, which examined the effects of dietary CP concentrations on growth rates, with CP ranging from 2% to 26%. Weight gain was increased with the supplementation of His, Ile, Lys, Thr, or Trp as compared to the LP diet (p < 0.05). Feed intake was greater for the His-, Lys-, and Thr-supplemented treatments as compared to the LP group (p < 0.05). Protein utilization efficiency was lower for the HP group than other groups (p < 0.01). The supplementation of Leu, Lys, and Val led to reduced protein utilization efficiency (p < 0.05), but the supplementation of Thr and Trp led to greater efficiency than the LP group (p < 0.05). Compared to the LP group, plasma urea concentrations were elevated with individual EAA supplementation, with the exception of the Thr addition. The added EAA resulted in increased concentrations of the corresponding EAA in plasma, except for Arg and Phe supplementation. The supplementation of Arg, His, Leu, Lys, and Met individually stimulated mTORC1 pathway activity (p < 0.05), and all EAA resulted in the decreased expression of ATF4 (p < 0.05). In summary, the supplementation of His, Ile, Lys, Thr, or Trp to an LP diet improved the growth performance of young rats. Responses to His and Lys additions were related to the activated mTORC1 pathway and feed intake increases. The improved growth performance resulting from the addition of a single EAA is not solely attributed to the increased plasma availability of EAA. Rather, it may be the consequence of a confluence of factors encompassing signaling pathways, the availability of amino acids, and other associated elements. The additivity of these factors results in independent responses to several EAA with no order of limitation, as is universally encoded in growth models for all production animal species.

Coingestion of Collagen With Whey Protein Prevents Postexercise Decline in Plasma Glycine Availability in Recreationally Active Men.

Whey protein ingestion during recovery from exercise increases myofibrillar but not muscle connective protein synthesis rates. It has been speculated that whey protein does not provide sufficient glycine to maximize postexercise muscle connective protein synthesis rates. In the present study, we assessed the impact of coingesting different amounts of collagen with whey protein as a nutritional strategy to increase plasma glycine availability during recovery from exercise. In a randomized, double-blind, crossover design, 14 recreationally active men (age: 26 ± 5years; body mass index: 23.8 ± 2.1kg·m-2) ingested in total 30g protein, provided as whey protein with 0g (WHEY), 5g (WC05); 10g (WC10), and 15g (WC15) of collagen protein immediately after a single bout of resistance exercise. Blood samples were collected frequently over 6hr of postexercise recovery to assess postprandial plasma amino acid kinetics and availability. Protein ingestion strongly increased plasma amino acid concentrations (p < .001) with no differences in plasma total amino acid availability between treatments (p > .05). The postprandial rise in plasma leucine and essential amino acid availability was greater in WHEY compared with the WC10 and WC15 treatments (p < .05). Plasma glycine and nonessential amino acid concentrations declined following whey protein ingestion but increased following collagen coingestion (p < .05). Postprandial plasma glycine availability averaged -8.9 ± 5.8, 9.2 ± 3.7, 23.1 ± 6.5, and 39.8 ± 11.0mmol·360min/L in WHEY, WC05, WC10, and WC15, respectively (incremental area under curve values, p < .05). Coingestion of a small amount of collagen (5g) with whey protein (25g) is sufficient to prevent the decline in plasma glycine availability during recovery from lower body resistance-type exercise in recreationally active men.

Nutritional value of torula yeast as an alternative protein feedstuff for hybrid Striped Bass (White Bass × Striped Bass)

AbstractObjectiveTwo separate trials were conducted to evaluate the digestibility of a novel alternative ingredient, enhanced torula yeast (TY) Cyberlindnera jadinii, in diets for hybrid Striped Bass (HSB; White Bass Morone chrysops × Striped Bass M. saxatilis) and to evaluate graded inclusion levels of this product when replacing soybean products.MethodsIn trial 1, the digestibility of TY was determined by testing a reference diet and a TY diet (reference and TY ingredient at an 80:20 ratio). A total of 90 fish were fed the experimental digestibility diets for 21 days, and fecal samples were collected using the stripping technique. In trial 2, a comparative feeding trial evaluated the replacement of soybean meal by TY on a crude protein basis at 50, 100, 200, and 300 g of TY/kg of experimental diet. A total of 150 HSB (18.6 ± 0.21 g/fish [mean + SE]) were distributed among fifteen 110‐L glass aquaria. Each diet was fed to triplicate groups of fish for 9 weeks.ResultIn trial 1, apparent digestibility coefficients for protein, lipid, organic matter, and energy of TY were 0.97, 0.98, 0.75, and 0.75, respectively. The availability of essential amino acids also was generally high, ranging from 0.74 to 0.99. For trial 2, a second‐order polynomial regression indicated that the optimal inclusion levels of TY in diets were 102, 112, and 109 g/kg for weight gain, feed efficiency, and protein conversion efficiency, respectively. However, the muscle yield linearly decreased with the graded inclusion levels of TY. Linear increases in intraperitoneal fat, hepatosomatic index, and whole‐body lipid were observed as the inclusion of TY increased in the diet.ConclusionIn conclusion, nutrients from TY were well utilized by HSB, and it can replace soybean products without affecting growth performance, with the optimal TY inclusion level being 102–112 g/kg of diet.

Pallidin function in Drosophila surface glia regulates sleep and is dependent on amino acid availability

The Pallidin protein is a central subunit of a multimeric complex called biogenesis of lysosome-related organelles complex 1 (BLOC1) that regulates specific endosomal functions and has been linked to schizophrenia. We show here that downregulation of Pallidin and other members of BLOC1 in the surface glia, the Drosophila equivalent of the blood-brain barrier, reduces and delays nighttime sleep in a circadian-clock-dependent manner. In agreement with BLOC1 involvement in amino acid transport, downregulation of the large neutral amino acid transporter 1 (LAT1)-like transporters JhI-21 and mnd, as well as of TOR (target of rapamycin) amino acid signaling, phenocopy Pallidin knockdown. Furthermore, supplementing food with leucine normalizes the sleep/wake phenotypes of Pallidin downregulation, and we identify a role for Pallidin in the subcellular trafficking of JhI-21. Finally, we provide evidence that Pallidin in surface glia is required for GABAergic neuronal activity. These data identify a BLOC1 function linking essential amino acid availability and GABAergic sleep/wake regulation.

Responses of the Macroalga Ulva prolifera Müller to Ocean Acidification Revealed by Complementary NMR- and MS-Based Omics Approaches.

Ocean acidification (OA) is a dramatic perturbation of seawater environments due to increasing anthropogenic emissions of CO2. Several studies indicated that OA frequently induces marine biota stress and a reduction of biodiversity. Here, we adopted the macroalga Ulva prolifera as a model and applied a complementary multi-omics approach to investigate the metabolic profiles under normal and acidified conditions. Our results show that U. prolifera grows at higher rates in acidified environments. Consistently, we observed lower sucrose and phosphocreatine concentrations in response to a higher demand of energy for growth and a higher availability of essential amino acids, likely related to increased protein biosynthesis. In addition, pathways leading to signaling and deterrent compounds appeared perturbed. Finally, a remarkable shift was observed here for the first time in the fatty acid composition of triglycerides, with a decrease in the relative abundance of PUFAs towards an appreciable increase of palmitic acid, thus suggesting a remodeling in lipid biosynthesis. Overall, our studies revealed modulation of several biosynthetic pathways under OA conditions in which, besides the possible effects on the marine ecosystem, the metabolic changes of the alga should be taken into account considering its potential nutraceutical applications.

Protein Matrix Impacts Essential Amino Acid Response To Feeding

It is well established that muscle protein turnover is highly response to increases in plasma essential amino acids (EAA). Digestion and amino acid absorption kinetics appear to strongly modulate post-prandial muscle protein synthesis. Despite this relationship, little is known about the effects of the protein matrix on plasma EAA appearance. PURPOSE: To assess if protein matrix alters plasma EAA pharmacokinetics (PK). METHODS: 24 healthy young individuals (24.7 ± 4.7y; BMI = 24.6 ± 3.2 kg/m2) completed this study. Participants consumed one of the following protein matrices: whey protein isolate (W; 6.8 g of EAA), beef sirloin (B; 7.0 g of EAA), or free form EAA enriched W (FW; 5.6 g of EAA). Blood samples were collected before and four hours following the ingestion of W, B, and FW. Plasma EAA were analyzed via LC-MS/MS. Standard pK values were calculated: area under the curve above baseline (AUCi), max EAA concentration (Cmax), EAA concentration change from baseline to EAACmax (ΔEAA), rate to Cmax, and time to Cmax. To examine group differences in EAA pK a one-way analysis of variance with Tukey post hoc test was performed with significance accepted at P ≤ 0.05. RESULTS: Significant group differences were noted for all measured pK (p ≤ 0.005). Post hoc comparisons indicated AUCi was significantly lower in W (50705.6 ± 13483.3 μmol/L/min) as compared to FW (84793.8 ± 15989.7 μmol/L/min; p = 0.004) and B (77084.6 ± 25076.0 μmol/L/min; p = 0.028). Cmax, ΔEAA, and rate to Cmax was significantly higher in FW (Cmax = 1,920 ± 242.6 μmol/L; ΔEAA = 1012.4 ± 200.8 μmol/L; rate to Cmax = 30.1 ± 13.0 μmol/L/min) as compared to W (Cmax = 1343.1 ± 144.4 μmol/L, p ≤ 0.001; ΔEAA = 556.9 ± 160.1 μmol/L, p ≤ 0.001; rate to Cmax = 13.6 ± 7.6 μmol/L/min, p = 0.003) and B (Cmax = 1339.9 ± 213.5 μmol/L, p ≤ 0.001; ΔEAA = 543.8 ± 164.2 μmol/L, p ≤ 0.001; rate to Cmax = 4.9 ± 2.7 μmol/L/min, p ≤ 0.001). Lastly, the time to Cmax was slowest in B (135.0 ± 62.1 mins) as compared to W (47.5 ± 14.9 mins; p ≤ 0.001) and FW (36.3 ± 7.4 mins; p ≤ 0.001). CONCLUSION: Despite similar EAA content, pharmacokinetics were significantly different between treatments. Despite the lowest EAA content, FW resulted in the greatest plasma EAA availability and quickest time to peak. These results indicate that protein/amino acid digestion is a primary determinant of plasma EAA availability.

Effects of Consuming Ounce-Equivalents Portions of Animal and Plant Protein Foods, As Defined by the Dietary Guidelines for Americans on Essential Amino Acid Availability

ObjectivesThe objective of this study was to assess the effects of consuming ounce-equivalents (oz-eq) portions of lean pork versus nuts, beans, and eggs, as defined by the Dietary Guidelines for Americans (DGA), as part of a meal, on essential amino acid (EAA) substrate availability for protein anabolism in young adults. MethodsIn this single-blinded, randomized crossover study, 30 participants (15 male, 15 female; age: 26 ± 4.9 y; BMI: 26.4 ± 4.5 kg/m2; mean ± SE) completed four testing sessions where they consumed a standardized test salad on each day with 2 oz-eq of either lean pork, whole eggs, black beans, or almonds. Blood samples were taken at baseline (prior to commencing meal consumption) and at 30, 60,120, 180,240, and 300 minutes following the initiation of meal consumption. Plasma from the blood samples were analyzed for amino acid concentrations via high-performance liquid chromatography (HPLC). ResultsThe EAAs positive incremental area under the curve (iAUCpos) over 300 minutes for lean pork, egg, black beans, and almonds was 7353 ± 397 μg/mL (least-square mean ± standard error (SE)), 4653 ± 408 μg/mL, 2100 ± 398 μg/mL, and 1220 ± 398 μg/mL, respectively. Pork resulted in signifcantly greater EAAs availability compared to egg, black beans, and almonds (Tukey adjusted, P < 0.0001 for all, while egg resulted in significantly greater EAAs availability compared to black beans and almonds (Tukey adjusted P < 0.0001 for both). No difference for EAAs iAUCpos was found between black beans and almonds. ConclusionsBased on the oz-eq concept used for the DGA, the protein sources included in this study, namely lean pork, whole egg, black beans, and almonds, are not equivalent regarding plasma EAAs availability for protein anabolism in young adults, in response to being consumed as part of a meal. More specifically on an oz-eq basis, lean pork provides a greater response than whole egg, followed by black beans and almonds, in terms of its ability to provide plasma EAAs as a substrate for protein anabolism This research serves as an important resource for future Dietary Guidelines Advisory Committees to reevaluate the appropriateness of equating Protein Foods on the current oz-eq basis. Funding SourcesNational Pork Board - Pork Checkoff and American Egg Board - Egg Nutrition Center.

Amino Acid Availability Is Not Essential for Life-Span Extension by Dietary Restriction in the Fly.

Dietary restriction (DR) is one of the most potent ways to extend health and life span. Key progress in understanding the mechanisms of DR, and aging more generally, was made when dietary protein, and more specifically essential amino acids (EAA), were identified as the dietary component to restrict to obtain DR's health and life-span benefits. This role of dietary amino acids has influenced work on aging mechanisms, especially in nutrient sensing, for example, Target of Rapamycin and insulin(-like) signaling networks. Experimental biology in Drosophila melanogaster has been instrumental in generating and confirming the hypothesis that EAA availability is important in aging. Here, we expand on previous work testing the involvement of EAA in DR through large-scale (N = 6 238) supplementation experiments across 4 diets and 2 genotypes in female flies. Surprisingly, we find that EAA are not essential to DR's life-span benefits. Importantly, we do identify the fecundity benefits of EAA supplementation suggesting the supplemented EAA were bioavailable. Furthermore, we find that the effects of amino acids on life span vary by diet and genetic line studied and that at our most restricted diet fecundity is constrained by other nutrients than EAA. We suggest that DR for optimal health is a concert of nutritional effects, orchestrated by genetic, dietary, and other environmental interactions. Our results question the universal importance of amino acid availability in the biology of aging and DR.

Različitost aminokiselinskog sastava u žitaricama

The quality of protein is based on their amino acid composition, especially on the content and availability of essential amino acids. Cereals are important sources of protein for human nutrition, but are limited in the amounts of essential amino acids, notably lysine. The aim of this study was to analyze the chemical composition and amino acid profiles of different cereals that are important for nutritional purposes in human diet. The content of protein, moisture and crude fat in cereals varied significantly from 7.83 to 13.22%, 11.45 to 13.80%, and from 1.67 to 6.35%, respectively. The obtained results showed that oat had the highest contents of crude protein (13.22%), crude fat (6.35%) and crude cellulose (9.42%) compared to other cereals. Significant (p &lt; 0.05) variation existed in the content of essential and nonessential amino acids among samples with the highest level in oat and wheat. Essential amino acids accounted for one-third of the total amino acids in the tested cereals. Glutamic acid was found to be the most abundant amino acid. It could be concluded that the amino acid composition of oat is the most favorable among cereals due to its high protein content and the content of lysine which can be found in limited amounts in most of the cereals.

Različitost aminokiselinskog sastava u žitaricama

The quality of protein is based on their amino acid composition, especially on the content and availability of essential amino acids. Cereals are important sources of protein for human nutrition, but are limited in the amounts of essential amino acids, notably lysine. The aim of this study was to analyze the chemical composition and amino acid profiles of different cereals that are important for nutritional purposes in human diet. The content of protein, moisture and crude fat in cereals varied significantly from 7.83 to 13.22%, 11.45 to 13.80%, and from 1.67 to 6.35%, respectively. The obtained results showed that oat had the highest contents of crude protein (13.22%), crude fat (6.35%) and crude cellulose (9.42%) compared to other cereals. Significant (p &lt; 0.05) variation existed in the content of essential and nonessential amino acids among samples with the highest level in oat and wheat. Essential amino acids accounted for one-third of the total amino acids in the tested cereals. Glutamic acid was found to be the most abundant amino acid. It could be concluded that the amino acid composition of oat is the most favorable among cereals due to its high protein content and the content of lysine which can be found in limited amounts in most of the cereals.

DIGESTIBILITY OF NUTRIENTS AND BROILER CHICKEN PRODUCTIVITY UNDER THE ACTION OF PROBIOTIC ENTERO-ACTIVE

The probiotic “Entero-active”, due to the formation of lactic and acetic acids, creates an unfavourable pH environment for pathogenic microflora, stimulates the growth of intestinal normal flora, which has a positive effect on the composition of the microbiocenosis. In addition, probiotic microorganisms produce biologically active substances, enzymes and amino acids. The aim of the research was to investigate the digestion of feed nutrients by broiler chickens fed by the probiotic supplement Entero-active. Optimal doses of probiotic feeding are determined on the basis of obtained results. The influence of probiotic supplement on digestibility of feed nutrients, availability of amino acids and retention of mineral elements of mixed fodder is established. Tendencies of changes in productivity and safety of poultry under the effect of probiotic are revealed. The broiler chickens fed by additive Entero-active have increased the digestibility of dry matter by 2.4% (P &lt;0.01), protein by 3.4% (P &lt;0.001), fiber by 31.1% (P &lt;0.001) and NFE by 4.0% (P &lt;0.001) comparing with the control group. According to our research, the application of probiotic supplement Entero-active for the broilers’ feeding increases the availability of essential amino acids, i.e. lysine by 4.8%, histidine by 3.8%, arginine by 4.9%, threonine by 7.5%, valine by 4.1%, methionine by 2.6%, isoleucine by 7.6% compared with the control indicator. It was found the absorption of Ca, P, Mg, Mn, and increases respectively, by 23.5%, 6.9%, 12.2%, and by 33.1% under the action of probiotics. The Entero-active probiotic application in the diet of broiler chickens increases the pre-slaughter live weight by 16.7%, the weight of ungutted carcasses by 15.0% and gutted carcasses by 17.3%, feed costs per 1 kg increase by 12.9%, relative to control. Our studies have made it possible to identify certain patterns in the digestion of nutrients by poultry, depending on the different doses of probiotic Entero-active used in their diets.

Invasive Plants Are a Valuable Alternate Protein Source and Can Contribute to Meeting Climate Change Targets

Agriculture has come under pressure to meet global food demands, whilst having to meet economic and ecological targets. This has opened newer avenues for investigation in unconventional protein sources. Current agricultural practises manage marginal lands mostly through animal husbandry, which; although effective in land utilisation for food production, largely contributes to global green-house gas (GHG) emissions. Assessing the revalorisation potential of invasive plant species growing on these lands may help encourage their utilisation as an alternate protein source and partially shift the burden from livestock production; the current dominant source of dietary protein, and offer alternate means of income from such lands. Six globally recognised invasive plant species found extensively on marginal lands; Gorse (Ulex europaeus), Vetch (Vicia sativa), Broom (Cytisus scoparius), Fireweed (Chamaenerion angustifolium), Bracken (Pteridium aquilinum), and Buddleia (Buddleja davidii) were collected and characterised to assess their potential as alternate protein sources. Amino acid profiling revealed appreciable levels of essential amino acids totalling 33.05 ± 0.04 41.43 ± 0.05, 33.05 ± 0.11, 32.63 ± 0.04, 48.71 ± 0.02 and 21.48 ± 0.05 mg/g dry plant mass for Gorse, Vetch, Broom Fireweed, Bracken, and Buddleia, respectively. The availability of essential amino acids was limited by protein solubility, and Gorse was found to have the highest soluble protein content. It was also high in bioactive phenolic compounds including cinnamic- phenyl-, pyruvic-, and benzoic acid derivatives. Databases generated using satellite imagery were used to locate the spread of invasive plants. Total biomass was estimated to be roughly 52 Tg with a protein content of 5.2 Tg with a total essential amino acid content of 1.25 Tg (~24%). Globally, Fabaceae was the second most abundant family of invasive plants. Much of the spread was found within marginal lands and shrublands. Analysis of intrinsic agricultural factors revealed economic status as the emergent factor, driven predominantly by land use allocation, with shrublands playing a pivotal role in the model. Diverting resources from invasive plant removal through herbicides and burning to leaf protein extraction may contribute toward sustainable protein, effective land use, and achieving emission targets, while simultaneously maintaining conservation of native plant species.

A method of assessing essential amino acid availability from microbial and ruminally undegraded protein in lactating dairy cows

The objective of this study was to extend a stable isotope-based assessment of AA absorption from rumen-degradable protein (RDP) sources to include determination of essential AA (EAA) availability from microbial protein (MCP). To demonstrate the technique, a study using a 2 × 2 factorial arrangement of treatments applied in a repeated 4 × 4 Latin square design was undertaken. Factors were high and low rumen-degradable protein and high and low starch. Twelve lactating cows were blocked into 3 groups according to days in milk and randomly assigned to the 4 treatment sequences. Each period was 14 d in length with 10 d of adaption followed by 4 d of ruminal infusions of 15N-labeled ammonium sulfate. On the last day of each period, a 13C-labeled AA mixture was infused into the jugular vein over a 6-h period to assess total AA entry. Rumen, blood, urine, and milk samples were collected during the infusions. Ruminal bacteria and blood samples were assessed for AA enrichment. Total plasma AA absorption rates were derived for 6 EAA from plasma 13C AA enrichment. Absorption of 6 EAA from MCP was calculated from total AA absorption based on 15N enrichment in blood and rumen bacteria. Essential AA absorption rates from total protein, MCP, and rumen-undegradable protein were derived with standard errors of the mean of 6, 14, and 14%, respectively. An average of 45% of absorbed EAA were from MCP, which varied among 6 EAA and was interactively affected by starch and RDP in diets. Microbial AA availability measured by isotope dilution method increased with the high RDP diets and was unaffected by starch level, except for Met, which decreased with high starch. Microbial protein outflow, estimated from urinary purine derivatives, increased with RDP and was not significantly affected by starch. This was consistent with measurements from the isotope dilution method. Total AA absorption rates measured from isotope dilution were similar to estimates from CNCPS (v. 6.55), but a lower proportion of absorbed AA was derived from MCP for the former method. Compared with the isotope and CNCPS estimates, the Fleming model underestimated microbial EAA and total EAA availability. An average of 58% of the absorbed EAA was converted into milk, which varied among individual AA and was interactively affected by starch and RDP in diets. The isotope dilution approach is advantageous because it provides estimates of EAA availability for individual EAA from rumen-undegradable protein and MCP directly with fewer errors of measurement than can be achieved with intestinal disappearance methods.

Dietary protein, exercise, ageing and physical inactivity: interactive influences on skeletal muscle proteostasis.

Dietary protein is a pre-requisite for the maintenance of skeletal muscle mass; stimulating increases in muscle protein synthesis (MPS), via essential amino acids (EAA), and attenuating muscle protein breakdown, via insulin. Muscles are receptive to the anabolic effects of dietary protein, and in particular the EAA leucine, for only a short period (i.e. about 2-3 h) in the rested state. Thereafter, MPS exhibits tachyphylaxis despite continued EAA availability and sustained mechanistic target of rapamycin complex 1 signalling. Other notable characteristics of this 'muscle full' phenomenon include: (i) it cannot be overcome by proximal intake of additional nutrient signals/substrates regulating MPS; meaning a refractory period exists before a next stimulation is possible, (ii) it is refractory to pharmacological/nutraceutical enhancement of muscle blood flow and thus is not induced by muscle hypo-perfusion, (iii) it manifests independently of whether protein intake occurs in a bolus or intermittent feeding pattern, and (iv) it does not appear to be dependent on protein dose per se. Instead, the main factor associated with altering muscle full is physical activity. For instance, when coupled to protein intake, resistance exercise delays the muscle full set-point to permit additional use of available EAA for MPS to promote muscle remodelling/growth. In contrast, ageing is associated with blunted MPS responses to protein/exercise (anabolic resistance), while physical inactivity (e.g. immobilisation) induces a premature muscle full, promoting muscle atrophy. It is crucial that in catabolic scenarios, anabolic strategies are sought to mitigate muscle decline. This review highlights regulatory protein turnover interactions by dietary protein, exercise, ageing and physical inactivity.

Muscle Protein Synthesis and Whole-Body Protein Turnover Responses to Ingesting Essential Amino Acids, Intact Protein, and Protein-Containing Mixed Meals with Considerations for Energy Deficit.

Protein intake recommendations to optimally stimulate muscle protein synthesis (MPS) are derived from dose-response studies examining the stimulatory effects of isolated intact proteins (e.g., whey, egg) on MPS in healthy individuals during energy balance. Those recommendations may not be adequate during periods of physiological stress, specifically the catabolic stress induced by energy deficit. Providing supplemental intact protein (20–25 g whey protein, 0.25–0.3 g protein/kg per meal) during strenuous military operations that elicit severe energy deficit does not stimulate MPS-associated anabolic signaling or attenuate lean mass loss. This occurs likely because a greater proportion of the dietary amino acids consumed are targeted for energy-yielding pathways, whole-body protein synthesis, and other whole-body essential amino acid (EAA)-requiring processes than the proportion targeted for MPS. Protein feeding formats that provide sufficient energy to offset whole-body energy and protein-requiring demands during energy deficit and leverage EAA content, digestion, and absorption kinetics may optimize MPS under these conditions. Understanding the effects of protein feeding format-driven alterations in EAA availability and subsequent changes in MPS and whole-body protein turnover is required to design feeding strategies that mitigate the catabolic effects of energy deficit. In this manuscript, we review the effects, advantages, disadvantages, and knowledge gaps pertaining to supplemental free-form EAA, intact protein, and protein-containing mixed meal ingestion on MPS. We discuss the fundamental role of whole-body protein balance and highlight the importance of comprehensively assessing whole-body and muscle protein kinetics when evaluating the anabolic potential of varying protein feeding formats during energy deficit.

Albumin promotes the progression of fibroblasts through late G1 into S-phase in the absence of growth factors

ABSTRACT G1 cell cycle progression is controlled largely by growth factors in early G1 indicating that it is appropriate to divide and by nutrients in late G1 indicating sufficient raw material for cell division. We previously mapped a late G1 cell cycle checkpoint for lipids upstream from a mammalian target of rapamycin complex 1 (mTORC1)-mediated checkpoint and downstream from a mid-G1 checkpoint known as the Restriction point. We therefore investigated a role for lipids in progression through late G1 into S-phase. Quiescent BJ-hTERT human fibroblasts were primed with 10% fetal bovine serum (FBS) for 3.5 h at which time, cells were treated with a mixture of lipids and carrier bovine serum albumin (BSA) along with [3 H]-thymidine deoxyribose ([3 H]-TdR) to monitor progression into S-phase. Surprisingly, BSA by itself was more effective than FBS in promoting progression to S-phase – the lipids had no impact on progression. While insulin strongly stimulated mTORC1 activity, it did not impact on [3 H]-TdR incorporation. Although BSA modestly elevated mTORC1 activity, rapamycin strongly inhibited BSA-induced progression to S-phase. BSA treatment promoted mitosis, but not progression through a second G1. Thus, after priming quiescent cells with FBS, albumin was sufficient to promote progression into S-phase. The BSA was not simply a source of amino acids in that amino acids were present in the culture media. We propose that the presence of albumin – the most abundant protein in serum – reflects a broader availability of essential amino acids needed for cell growth.

Maternal supplementation with citrulline or arginine during gestation impacts fetal amino acid availability in a model of intrauterine growth restriction (IUGR).

Supplementing maternal diet with citrulline or arginine during gestation was shown to enhance fetal growth in a model of IUGR induced by maternal dietary protein restriction in the rat. The aims of this study were to determine in the same model whether maternal supplementation with citrulline or arginine would increase 1) citrulline and arginine concentration in fetal circulation; 2) the expression of placental amino acid transporters, and 3) the fetal availability of essential amino acids. Pregnant rats (n=8 per group) were fed either an isocaloric control (20% protein, NP) or a low protein (LP, 4% protein) diet, either alone or supplemented with 2g/kg/d of l-citrulline (LP+CIT) or isonitrogenous Arginine (LP+ARG) in drinking water throughout gestation. Fetuses were extracted by C-section on the 21st day of gestation. The gene expression of system A (Slc38a1, Slc38a2, and Slc38a4) and L (Slc7a2, Slc7a5, Slc7a8) amino acid transporters was measured in placenta and amino acid concentrations determined in maternal and fetal plasma. Maternal LP diet decreased fetal (4.01±0.03 vs. 5.45±0.07g, p<0.0001) and placental weight (0.617±0.01 vs. 0.392±0.04g, p<0.001), by 26 and 36% respectively, compared with NP diet. Supplementation with either CIT or ARG increased fetal birth weight by≈5 or 11%, respectively (4.21±0.05 and 4.48±0.05g vs. 4.01±0.03g, p<0.05). CIT supplementation produced a 5- and 2-fold increase in fetal plasma citrulline and arginine, respectively, whereas ARG supplementation only increased fetal arginine concentration. LP diet led to lower placental SNAT 4 mRNA, and higher LAT2 and SNAT1 expression, compared with NP. SNAT4, 4hFC, LAT2 mRNA were up-regulated in LP+CIT and LP+ARG group compared with the un-supplemented LP group. Higher level of LAT1 mRNA was also observed in the LP+CIT group than in the LP group (p<0.01). SNAT2 expression was unchanged in response to CIT or ARG supplementation. Fetal amino acid concentrations were decreased by LP diet, and were not restored by CIT or ARG supplementation. The current findings confirm supplementation with citrulline or arginine enhances fetal growth in a rat model of IUGR. They further suggest that: 1) citrulline and arginine administered orally to the pregnant mother may reach fetal circulation; 2) citrulline effectively raises fetal arginine availability; and 3) although it failed to increase the concentrations of essential amino acids in fetal plasma, citrulline or arginine supplementation upregulates the gene expression of several placental amino acid transporters.

Assessing availability of amino acids from various feedstuffs in dairy cattle using a stable isotope-based approach

Nitrogen efficiency in dairy cows can be improved by more precisely supplying essential amino acids (EAA) relative to animal needs, which requires accurate estimates of the availability of individual EAA from feedstuffs. The objective of this study was to determine EAA availability for 7 feed ingredients. Seven heifers (258 ± 28 kg BW) were randomly chosen and assigned to 8 treatment sequences in a 7 × 8 incomplete Latin square design. Treatments were a basal diet (BD), and 10% (on a dry matter basis) of BD replaced by corn silage (CS), grass hay (GH), alfalfa hay (AH), dried distillers grain (DDGS), soybean hulls (SH), wet brewers grain (BG), or corn grain (CG). Total plasma AA entry rates were estimated for each EAA within each diet by fitting a 4-pool dynamic model to observed plasma, 13C AA enrichment resulting from a 2-h constant infusion of a 13C algal AA mixture. Individual EAA availability from each test ingredient was determined by regression of entry rates for that AA on crude protein intake for each ingredient. The derived plasma total EAA entry rates for corn silage, grass hay, alfalfa hay, dried distillers grain, soyhulls, brewers grain, and corn grain were 30.6 ± 3.4, 27.4 ± 3.2, 31.3 ± 3.4, 37.2 ± 3.2, 26.4 ± 3.2, 37.8 ± 3.2, and 33.5 ± 3.2% (±standard error) of EAA from each ingredient, respectively. Using the previous estimate of 8.27% EAA utilization by splanchnic tissues during first pass, total rumen-undegradable protein EAA absorbed from the gut lumen was 33.4, 29.9, 34.1, 40.6, 28.8, 41.2, and 36.5% of the EAA in each ingredient respectively.

Effects of lowering dietary fishmeal and crude protein levels on growth performance, body composition, muscle metabolic gene expression, and chronic stress response of rainbow trout (Oncorhynchus mykiss)

A study was conducted to evaluate the effects of lowering dietary fishmeal (FM) and crude protein (CP) levels, while maintaining essential amino acid levels, on growth performance, body composition, muscle metabolic gene expression, and chronic stress response of rainbow trout, Oncorhynchus mykiss, with and without handling stress. Eight experimental diets (isocaloric) with a 4 × 2 factorial design were formulated to contain two levels of FM (20%, 5%) and four levels of CP (48%, 45%, 42%, 39%). Diets were supplemented with increasing levels of amino acids to maintain dietary essential amino acid (EAA) levels. Trout (34.8 ± 0.3 g) were fed to apparent satiation twice daily for nine weeks to assess growth performance under laboratory rearing conditions, and then for an additional six weeks with and without exposure to handling stress (30 s of chasing followed by 30 s of netted air exposure) twice per week. The 9-week growth trial demonstrated that reducing dietary FM levels from 20% to 5% significantly reduced fish growth and increased feed conversion ratio (P < .05). Reducing dietary CP levels from 48% to 42% did not affect trout growth. A dietary FM level of 20% significantly increased whole-body dry matter, CP and total EAAs (P < .05) compared to 5% FM inclusion while increasing dietary CP level significantly decreased dry matter, crude fat, and gross energy but increased total EAAs (P < .05). Reducing FM and CP levels had no effect on measured stress indices of plasma cortisol, glucose and lysozyme activity (P > .05) after 6-weeks of repeated handling stress. The expression of genes in the gcn2/eif2α/atf4 pathway, triggered in response to protein or amino acid starvation, were evaluated. General control nonderepressible 2 (gcn2) decreased with increasing dietary CP level above 42% (P < .05), but there were no dietary effects (FM or CP levels) on eif2α (eukaryotic initiation factor 2α) or atf4 (activating transcription factor 4) expression. In total, gene expression results suggest amino acid limitations on muscle protein metabolism as a result of feeding diets below 42% CP, even when supplemented with synthetic EAA to meeting published dietary requirements. In conclusion, our study demonstrated that 5% dietary FM is insufficient for maximal growth performance, while diets balanced for EAAs show an opportunity to reduce CP level from 48% to 42% without any reduction in growth performance, body composition, metabolic amino acid sufficiency or tolerance to chronic stress. Below 42% CP, reduced growth indices suggest an imbalance in EAA availability.