Total Plasma Amino Acid Concentrations Research Articles

Time Courses of Gastric Volume and Content after Different Types of Casein Ingestion in Healthy Men: A Randomized Crossover Study

Few studies have evaluated differences in the curd-forming ability of casein on gastric volume and content directly after ingestion in humans. This study evaluated the time course of gastric volume and curd conditions in the stomach after protein ingestion. This was an open-labeled, randomized crossover trial. Ten healthy men [age: 33.4 ± 7.3 y; BMI (kg/m2): 21.9 ± 0.9] received 350 g of 3 isonitrogenous and isocaloric protein drinks containing 30 g micellar casein (MCN), sodium caseinate (SCN), or whey protein concentrate (WPC). The gastric antrum cross-sectional area (CSA) and curd in the stomach were measured using ultrasonography within 5 h after ingestion. The differences between test foods were tested using the MIXED model and post hoc tests using Fisher's protected least significant difference. The incremental AUC of the gastric antrum CSA after MCN ingestion was 1.3-fold and 1.5-fold higher than that after the ingestion of SCN and WPC, respectively (both P < 0.05), but not different between SCN and WPC. The number of participants with curds ≥20 mm with a high echogenicity clot observed in the stomach within 5 h after MCN ingestion was significantly greater than that after the ingestion of other proteins (n = 9 for MCN, n = 2 for SCN, and n = 0 for WPC; bothP < 0.01). The regression line slopes on total plasma amino acid concentration and gastric antrum CSA were significantly different between the participants with and without curds. In contrast to SCN and WPC, MCN ingestion resulted in slow kinetics of gastric antrum CSA. Differences in curd formation of casein in the stomach affect gastric emptying and plasma amino acid absorption kinetics after ingestion in healthy men. This trial was registered at University Hospital Medical Information Network Clinical Trials Registry as UMIN000038388 (https://center6.umin.ac.jp/cgi-open-bin/ctr_e/ctr_view.cgi?recptno=R000043746).

Four weeks treatment with the GLP-1 receptor analogue liraglutide lowers liver fat and concomitantly circulating glucagon in individuals with overweight.

We investigated the effect of pharmacologically induced weight loss on markers of glucagon resistance in individuals with overweight during treatment with the glucagon-like peptide-1 receptor agonist liraglutide. We performed an open-label study in 14 men with overweight (age 38 ± 11 years, BMI 32 ± 4 kg/m2) without simultaneously diabetes. Subjects were treated with liraglutide, initiated and titrated with 0.6 mg/day/week to reach the final dose of 3.0 mg/day. Subjects were examined at baseline, during titration (Week 4), after 2 weeks of steady state (Week 6) of final dosing of liraglutide and 3 weeks after discontinuation of liraglutide (follow-up). Study participants lost 3.3 ± 1.9 kg (3%) total body weight during the first 4 weeks of treatment with liraglutide. Simultaneously, liver fat content decreased from 12.4 ± 11.6% to 10.2 ± 11.1%, p = 0.025, whereas fat content in the spleen and subcutaneous tissue was unaltered. Markers of glucagon resistance, including plasma glucagon and the glucagon-alanine-index, also decreased significantly during treatment, but total and individual plasma amino acid concentrations did not. Insulin resistance (HOMA-IR) was unchanged during treatment, whereas insulin clearance increased. Treatment with the GLP-1 receptor analogue liraglutide decreased liver fat content, and simultaneously attenuated glucagon concentrations and the glucagon-alanine index in individuals with overweight without diabetes.

Ingestion of an ample amount of meat substitute based on a lysine-enriched, plant-based protein blend stimulates postprandial muscle protein synthesis to a similar extent as an isonitrogenous amount of chicken in healthy, young men.

Plant-based proteins are considered to be less effective in their capacity to stimulate muscle protein synthesis when compared with animal-based protein sources, likely due to differences in amino acid contents. We compared the postprandial muscle protein synthetic response following the ingestion of a lysine-enriched plant-based protein product with an isonitrogenous amount of chicken. Twenty-four men (age 24 ± 5 years; BMI 22·9 ± 2·6 kg·m-2) participated in this parallel, double-blind, randomised controlled trial and consumed 40 g of protein as a lysine-enriched wheat and chickpea protein product (Plant, n 12) or chicken breast fillet (Chicken, n 12). Primed, continuous intravenous l-(ring-13C6)-phenylalanine infusions were applied while repeated blood and muscle samples were collected over a 5-h postprandial period to assess plasma amino acid responses, muscle protein synthesis rates and muscle anabolic signalling responses. Postprandial plasma leucine and essential amino acid concentrations were higher following Chicken (P < 0·001), while plasma lysine concentrations were higher throughout in Plant (P < 0·001). Total plasma amino acid concentrations did not differ between interventions (P = 0·181). Ingestion of both Plant and Chicken increased muscle protein synthesis rates from post-absorptive: 0·031 ± 0·011 and 0·031 ± 0·013 to postprandial: 0·046 ± 0·010 and 0·055 ± 0·015 % h-1, respectively (P-time < 0·001), with no differences between Plant and Chicken (time x treatment P = 0·068). Ingestion of 40 g of protein in the form of a lysine-enriched plant-based protein product increases muscle protein synthesis rates to a similar extent as an isonitrogenous amount of chicken in healthy, young men. Plant-based protein products sold as meat replacers may be as effective as animal-based protein sources to stimulate postprandial muscle protein synthesis rates in healthy, young individuals.

Evaluation of dietary crude protein concentrations, fishmeal, and sorghum inclusions in broiler chickens offered wheat-based diet via Box-Behnken response surface design.

The objective of this study was to investigate the impacts of dietary crude protein (CP), fishmeal and sorghum on nutrient utilisation, digestibility coefficients and disappearance rates of starch and protein, amino acid concentrations in systemic plasma and their relevance to growth performance of broiler chickens using the Box-Behnken response surface design. The design consisted of three factors at three levels including dietary CP (190, 210, 230 g/kg), fishmeal (0, 50, 100 g/kg), and sorghum (0, 150, 300 g/kg). A total of 390 male, off-sex Ross 308 chicks were offered experimental diets from 14 to 35 days post-hatch. Growth performance, nutrient utilisation, starch and protein digestibilities and plasma free amino acids were determined. Dietary CP had a negative linear impact on weight gain where the transition from 230 to 190 g/kg CP increased weight gain by 9.43% (1835 versus 2008 g/bird, P = 0.006). Moreover, dietary CP linearly depressed feed intake (r = -0.486. P < 0.001). Fishmeal inclusions had negative linear impacts on weight gain (r = -0.751, P < 0.001) and feed intake (r = -0.495, P < 0.001). There was an interaction between dietary CP and fishmeal for FCR. However, growth performance was not influenced by dietary inclusions of sorghum. Total plasma amino acid concentrations were negatively related to weight gain (r = -0.519, P < 0.0001). The dietary transition from 0 to 100 g/kg fishmeal increased total amino acid concentrations in systemic plasma by 35% (771 versus 1037 μg/mL, P < 0.001). It may be deduced that optimal weight gain (2157 g/bird), optimal feed intake (3330 g/bird) and minimal FCR (1.544) were found in birds offered 190 g/kg CP diets without fishmeal inclusion, irrespective of sorghum inclusions. Both fishmeal and sorghum inclusions did not alter protein and starch digestion rate in broiler chickens; however, moderate reductions in dietary CP could advantage broiler growth performance.

Amino Acid Profile in Malnourished Patients with Liver Cirrhosis and Its Modification with Oral Nutritional Supplements: Implications on Minimal Hepatic Encephalopathy.

Low plasma levels of branched chain amino acids (BCAA) in liver cirrhosis are associated with hepatic encephalopathy (HE). We aimed to identify a metabolic signature of minimal hepatic encephalopathy (MHE) in malnourished cirrhotic patients and evaluate its modification with oral nutritional supplements (ONS) enriched with ß-Hydroxy-ß-methylbutyrate (HMB), a derivative of the BCAA leucine. Post hoc analysis was conducted on a double-blind placebo-controlled trial of 43 individuals with cirrhosis and malnutrition, who were randomized to receive, for 12 weeks, oral supplementation twice a day with either 220 mL of Ensure® Plus Advance (HMB group, n = 22) or with 220 mL of Ensure® Plus High Protein (HP group, n = 21). MHE evaluation was by psychometric hepatic encephalopathy score (PHES). Compared to the HP group, an HMB-specific treatment effect led to a larger increase in Val, Leu, Phe, Trp and BCAA fasting plasma levels. Both treatments increased Fischer’s ratio and urea without an increase in Gln or ammonia fasting plasma levels. MHE was associated with a reduced total plasma amino acid concentration, a reduced BCAA and Fischer´s ratio, and an increased Gln/Glu ratio. HMB-enriched ONS increased Fischer´s ratio without varying Gln or ammonia plasma levels in liver cirrhosis and malnutrition, a protective amino acid profile that can help prevent MHE.

Effects of Casein Hydrolysate Ingestion on Thermoregulatory Responses in Healthy Adults during Exercise in Heated Conditions: A Randomized Crossover Trial.

Food ingestion has been shown to affect thermoregulation during exercise, while the impact of protein degradant consumption remains unclear. We investigated the effects of casein hydrolysate ingestion on thermoregulatory responses during exercise in the heat. In a randomized, placebo-controlled, double-blind, crossover trial, five men and five women consumed either 5 g of casein hydrolysate or placebo. Thirty minutes after ingestion, participants cycled at 60% VO2max until voluntary exhaustion wearing a hot-water (43 °C) circulation suit. Exercise time to exhaustion, body core temperature, forearm sweat rate, and forearm cutaneous vascular conductance did not differ different between the conditions. However, chest sweat rate and mean skin temperature increased upon casein hydrolysate ingestion compared with placebo during exercise. Increased chest sweat rate upon casein hydrolysate ingestion was associated with elevated sudomotor sensitivity to increasing body core temperature, but not the temperature threshold for initiating sweating. A positive correlation was found between chest sweat rate and plasma total amino acid concentration during exercise. These results suggest that casein hydrolysate ingestion enhances sweating heterogeneously by increasing peripheral sensitivity of the chest’s sweating mechanism and elevating skin temperature during exercise in the heat. However, the physiological link between plasma amino acid concentration and sweat rate remains unclear.

Postprandial Amino Acid Kinetics of Milk Protein Mixtures are Affected by Composition, But Not Denaturation, in Neonatal Piglets

Multiple studies have indicated that formula-fed infants show a different growth trajectory compared with breastfed infants. The observed growth rates are suggested to be linked to higher postprandial levels of branched chain amino acids (BCAAs) and insulin related to differences in protein quality. We evaluated the effects of milk protein denaturation and milk protein composition on postprandial plasma and hormone concentrations. Neonatal piglets were bolus-fed randomly, in an incomplete crossover design, 2 of 3 milk protein solutions: native whey protein isolate (NWPI), denatured whey protein isolate (DWPI), or protein base ingredient, comprising whey and casein (PBI). Postprandial plasma amino acids (AAs), insulin, glucagon-like peptide 1, glucose, and paracetamol concentrations were assayed. Plasma responses were fitted with a model of first-order absorption with linear elimination. DWPI (91% denatured protein) compared with NWPI (91% native protein) showed lower essential amino acids (EAAs) (∼10%) and BCAA (13-19%) concentrations in the first 30-60 min. However, total amino acid (TAA) concentration per time-point and area under the curve (AUC), as well as EAA and BCAA AUC were not different. PBI induced a ∼30% lower postprandial insulin spike than NWPI, yet plasma TAA concentration at several time-points and AUC was higher in PBI than in NWPI. The TAA rate constant for absorption (k a) was twofold higher in PBI than in NWPI. Plasma BCAA levels from 60 to 180 min and AUC were higher in PBI than in NWPI. Plasma EAA concentrations and AUCs in PBI and NWPI were not different. Denaturation of WPI had a minimal effect on postprandial plasma AA concentration. The differences between PBI and NWPI were partly explained by the difference in AA composition, but more likely differences in protein digestion and absorption kinetics. We conclude that modifying protein composition, but not denaturation, of milk protein solutions impacts the postprandial amino acid availability in neonatal piglets.

Plasma amino acid profiles of dairy heifers grazing pasture, chicory and plantain

ABSTRACTThe objective of this study was to quantify amino acid (AA) profiles in plasma collected from dairy heifers grazing swards containing ryegrass/clover, chicory and plantain in autumn and spring. The study comprised a 28-day autumn trial and a 21-day spring trial using 56 Friesian x Jersey heifers. Total AA concentrations ranged from 2849 to 3165 μM in autumn and from 3195 to 3454 μM in spring, with no treatment effect. Season affected essential AA (EAA) and total AA concentrations. Total plasma AA concentrations were lower in autumn (2965 μM) than in spring (3303 μM). Based on these results, feeding dairy heifer’s either pure chicory, binary mixture or pure plantain does not affect EAA and total AA of Friesian x Jersey heifers. Further, no relationship was established between AA and weight gain of heifers in this study.

Effects of protein quantity and type on diet induced thermogenesis in overweight adults: A randomized controlled trial

Protein content of a meal is hypothesized to drive DIT dose-dependently. However, no single meal study exists comparing two different doses of protein on DIT. In addition, the source of protein, particularly whey protein, was shown to have a higher DIT than casein and soy in the acute setting, however the mechanism behind this difference is not yet clear. The aim of the present work is therefore to evaluate the efficacy of two different doses and types of protein (whey protein and casein) on DIT in overweight adults. Randomized, double blind crossover including seventeen overweight men and women assigned to four isocaloric study treatments where protein and carbohydrate were exchanged: control, 30g of whey protein microgels (WPM30), 50g WPM (WPM50) or 50g micellar casein (MC50). Energy expenditure was measured by indirect calorimetry. Blood, breath and urine samples were collected in order to measure substrate oxidation, amino acid profile, glucose and insulin, protein turnover and other metabolic parameters. DIT was 6.7±3.7%, 13.0±5.0%, 18.0±5.0% and 16.0±5.0% for control, WPM30, WPM50 and MC50, respectively. There was a significant difference between WPM50 and WPM30 (p<0.005) and a trend was observed between WPM50 and MC50 (p=0.06). WPM50 resulted in the highest total, essential, and branched-chain plasma amino acid concentrations when compared with the other study treatments (p<0.005) and a higher insulin concentration than MC50 (p<0.005). Protein oxidation was higher for WPM50 than MC50. Protein turnover was significantly correlated with DIT through total leucine oxidation (r=0.52, p=0.005). Our findings show that DIT does increase at a dose beyond 30g of WPM and that the type of dairy protein may have an effect on DIT with WPM tending towards a higher DIT than casein. Although further research is required to understand the mechanism behind the effect of different protein sources on thermogenesis, we suggest that amongst the components of protein turnover, protein oxidation maybe an important driver of thermogenesis at doses higher than 30g. These results have concrete implications when choosing a dose of protein to optimize its thermogenic effect. NCT02303080 www.clinicaltrials.gov.

Soya milk exerts different effects on plasma amino acid responses and incretin hormone secretion compared with cows’ milk in healthy, young men

Apart from the well-known action of insulin, the mechanism by which soya and cows' milk improve postprandial glycaemia control was examined. In total, twelve healthy, young, Chinese men were studied on three separate occasions, in random order with isovolumetric (322 ml) control water, soya milk and cows' milk. Plasma total amino acid concentrations increased 30 min after test meals consumption and were higher after soya milk (230 %) and cow milk (240 %) consumption compared with water. Cows' milk ingestion induced higher branched-chain amino acids (BCAA) (40 %) than soya milk. Postprandial incretin concentrations increased after meal consumption. Cows' milk meal was accompanied by higher incremental AUC (iAUC) (170 %) for glucagon-like peptide-1 (GLP-1) compared with soya milk and control (P=0·06). However, glucose-dependent insulinotropic polypeptide (GIP) concentrations increased to significantly greater levels after soya milk consumption (iAUC 60 % higher) compared with cows' milk and control. Consumption of both soya and cows' milk with carbohydrates induced a similar reduction in glycaemic response through a different mechanism, beyond insulin action. Plasma amino acids (alanine and arginine), and incretins in particular (GIP was stimulated), may be involved in the hyperinsulinaemia after soya milk meals. However, BCAA and GLP-1 release may be responsible for the reduced glycaemia after cows' milk consumption by delaying gastric emptying. This could be the result of different milk protein/amino acid composition, but also differences in milk carbohydrate composition (i.e. lactose v. sucrose). It can be concluded that soya milk is a good alternative to cows' milk with regard to glycaemic regulation, with different mechanisms involved.

Distinct physiological, plasma amino acid, and liver transcriptome responses to purified dietary beef, chicken, fish, and pork proteins in young rats.

We report on the impact of purified dietary meat proteins from four species on plasma insulin, lipid and amino acid (AA) concentrations, and hepatic transcriptome (RNA-sequencing). Young rats received semi-synthetic diets for 1 wk that differed only regarding protein source; casein (reference) was replaced by beef, chicken, fish, or pork proteins. Compared to casein, all proteins, except pork, increased total plasma AA concentrations. Pork protein reduced adipose tissue mass and liver triacylglycerol, which was accompanied by increased plasma triacylglycerol concentrations. Plasma cholesterol was reduced by fish protein. The number of differentially expressed genes ranged between 609 (pork) and 1258 (chicken); on average one-third of the changes were specific for each meat protein. Pathway responses were most similar for beef and chicken, followed by pork and fish. Although the extent varied, all meat proteins induced mRNA translation, antigen processing/presentation, intracellular vesicular trafficking, and oxidoreductive-transformation pathways, and suppressed signal-transduction (Notch, TGFB/SMAD, insulin) and mitochondrial biogenesis pathways. Lipid- and AA-metabolic pathways were repressed, except by pork. AA-transport pathways were induced by beef and fish only, and complement/coagulation-pathways were suppressed by chicken and beef. Fish suppressed nuclear-transport and cofactor metabolism. To conclude, short-term feeding of different meat proteins resulted in distinct physiological and transcriptome changes in young rats.

Post-prandial amino acid and leucine responses to serum albumin concentrate compared to whey protein concentrate in healthy subjects

Background Supplemental protein sources vary in composition in regards to size, structure, and amino acid composition of the varying individual proteins. Serum albumin concentrate (SA) is a new protein source extracted from edible-grade bovine plasma, which was recently made available on commercial scale. As compared to the low MW proteins in whey protein concentrate (WPC), SA consists of higher MW protein constituents such as albumin (66kD, >60%) and globulin proteins (130 kD, >10%), which contain a high quality amino acid composition. These differences in the protein profile of SA may both delay and sustain amino acid absorption in comparison to WPC. The objective of this study was to determine the post-prandial amino acid response to oral ingestion of SA and to compare the response to that of WPC. Methods Eight healthy, 20-30 year old, male (n=4, 80.3 kg BW) and female (n=4, 63.7 kg BW) volunteers participated in an open label, clinical study utilizing a cross-over design (Clinicaltrials.gov: NCT01643265). The volunteers received each of the two protein sources in a random order with a 7 d washout period between treatments. After a 12 h overnight fast, 7 mL of blood was collected from a catheter inserted into a forearm vein. Baseline amino acid values at 30 and 15 min were determined prior to administration of the experimental protein sources. Twenty grams of either WPC or SA was provided in 335 mL of water. Subsequent blood samples were collected at 15, 30, 45, 60, 90, 120, 180, and 210 min after ingestion for analysis of individual amino acid concentrations. An EZ:faastTM amino acid analysis kit was used for sample pretreatment and chromatographic separation was conducted using a Agilent 6460 triple quadrupole LC/MS system. Data were analyzed using repeated measures and General Linear Models procedures from SAS (Version 9.2) including treatment, gender, and test period as independent variables in the statistical model. Significance is reported as P<0.05 and P<0.01. Consent to publish the results was obtained from all participants. Results Total plasma amino acid concentration at baseline was found to be higher (P<0.01) at baseline in male subjects (2,633 nmoles/mL) compared to female subjects (2,153 nmoles/mL). In contrast, plasma leucine concentration was found to be higher (P<0.05) in female subjects (156 nmoles/mL) compared to male subjects (103 nmoles/mL). Rises in post-prandial individual amino acid concentration were observed for both WPC and SA (P<0.01). The rises reflect the amino acid pattern of the protein source ingested. The ingestion of whey protein concentrate increased plasma total amino acid concentration 30 min post-prandially compared to SA (P<0.05) by 10%, without differences in leucine. In contrast, the ingestion of SA promoted 14% higher leucine concentrations 120 min post-prandially than did WPC (P<0.05). Conclusion Individual post-prandial amino acids reflect the amino acid pattern and characteristics of the protein source ingested. The digestion and absorption of the protein and amino acid composition of serum albumin concentrate

Effects of dietary amylose/amylopectin ratio on growth performance, feed utilization, digestive enzymes, and postprandial metabolic responses in juvenile obscure puffer Takifugu obscurus

The effect of dietary amylose/amylopectin (AM/AP) ratio on growth, feed utilization, digestive enzyme activities, plasma parameters, and postprandial blood glucose responses was evaluated in juvenile obscure puffer, Takifugu obscurus. Five isonitrogenous (430 g kg(-1) crude protein) and isolipidic (90 g kg(-1) crude lipid) diets containing an equal starch level (250 g kg(-1) starch) with different AM/AP ratio diets of 0/25, 3/22, 6/19, 9/16 and 12/13 were formulated. Each experimental diet was fed to triplicate groups (25 fish per tank), twice daily during a period of 60 days. After the growth trial, a postprandial blood response test was carried out. Fish fed diet 6/19 showed best growth, feed efficiency and protein efficiency ratio. Hepatosomatic index, plasma total cholesterol concentration, liver glycogen and lipid content, and gluconokinase, pyruvate kinase and fructose-1,6-bisphosphatase activities were lower in fish fed highest AM/AP diet (12/13) than in fish fed the low-amylose diets. Activities of liver and intestinal trypsin in fish fed diet 3/22 and diet 6/19 were higher than in fish fed diet 9/16 and diet 12/13. Activities of liver and intestinal amylase and intestinal lipase, and starch digestibility were negatively correlated with dietary AM/AP ratio. Fish fed diet 3/22 and diet 6/19 showed higher plasma total amino acid concentration than fish fed the other diets, while plasma urea nitrogen concentration and activities of alanine aminotransferase and aspartate aminotransferase showed the opposite trend. Equal values were found for viscerosomatic index and condition factor, whole body and muscle composition, plasma high-density and low-density lipoprotein cholesterol concentrations, and activities of lipase and hexokinase and glucose-6-phosphatase in liver. Postprandial plasma glucose and triglyceride peak value of fish fed diet 12/13 were lower than in fish fed the low-amylose diets, and the peak time of plasma glucose was later than in fish fed the other diets. Plasma glucose and triglyceride concentrations showed a significant difference at 2 and 4 h after a meal and varied between dietary treatments. According to regression analysis of weight gain against dietary AM/AP ratio, the optimum dietary AM/AP ratio for maximum growth of obscure puffer was 0.25. The present result indicates that dietary AM/AP ratio could affect growth performance and feed utilization, some plasma parameters, digestive enzyme as well as hepatic glucose metabolic enzyme activities in juvenile obscure puffer.

Dose-dependent effects of leucine supplementation on preservation of muscle mass in cancer cachectic mice

Cancer cachexia, which is characterized by muscle wasting, is associated with increased morbidity and mortality. Because muscle protein synthesis may be increased and protein breakdown reduced by leucine supplementation, we used the C26 tumor-bearing cachectic mouse model to assess the effects of dietary supplementation with leucine on muscle weight and the markers of muscle protein breakdown (mRNA of atrogin and murf). Male CD2F1 mice were subcutaneously inoculated with tumor cells (tumor-bearing mice; TB) or were sham injected (control; C). They were fed standard diets or diets supplemented with leucine [1 gr (TB1Leu) or 8 gr (TB8Leu) supplemented leucine per kg feed]; TB and C received 8.7% Leu/g protein, TB1Leu received 9.6% Leu/g protein and TB8Leu received 14.6 Leu/g protein. After 21 days, the following were determined: body weights, plasma amino-acid concentrations, tumor size and muscle mass of the gastrocnemius (mG), tibialis anterior (mTA), extensor digitorum longus (mEDL) and soleus (mS) muscles. In tumor-bearing (TB) mice, carcass and skeletal muscle masses decreased, and levels of atrogin and murf mRNA in the mEDL increased. Muscle-mass loss was counteracted dose-dependently by leucine supplementation: relative to TB, the mass of the mG was +23% in TB8Leu, and +22% in mTA (p<0.05). However, leucine supplementation did not change atrogin and murf mRNA levels. Total plasma amino acid concentrations increased in TB, especially for taurine, lysine, arginine and alanine (p<0.05). Leucine supplementation attenuated the increase in total plasma amino-acid concentrations (p<0.05). Irrespective of changes in muscle protein breakdown markers, leucine supplementation reduced muscle wasting in tumor-bearing cachectic mice and attenuated changes in plasma amino acids.

Lactobacillus plantarum Consumption Increases PepT1-Mediated Amino Acid Absorption by Enhancing Protein Kinase C Activity in Spontaneously Colitic Mice1–3

Although probiotic consumption has generally been shown to have many beneficial effects for the prevention and treatment of inflammatory bowel disease, the effects of Lactobacillus plantarum (LP) on intestinal nutrient absorption, particularly oligopeptide transporter 1 (PepT1)-mediated absorption of dietary protein under inflammatory conditions, has not yet been characterized. In this study, we first investigated the effects of LP consumption on plasma amino acid concentrations and PepT1-mediated absorption of cephalexin in the small intestine of wild-type (WT) mice and interleukin-10 knockout (IL-10−/−) mice, a model of spontaneous colitis. We then analyzed expression and distribution of PepT1 and protein kinase C (PKC) activity in the jejunum of these mice. LP consumption (109 colony-forming units/0.5 mL) delivered by gavage once per day for 4 wk increased the total plasma amino acid concentration and the concentration of plasma cephalexin through enhancement of PepT1-mediated uptake in LP treated IL-10−/− mice compared with IL-10−/− mice. However, Western blotting and quantitative PCR analysis revealed no significant differences in PepT1 protein and mRNA expression between LP-treated and untreated mice. Additionally, immunofluorescence analysis showed that PepT1 did not appear to be mislocalized in IL-10−/− mice. Interestingly, IL-10−/− mice had significantly lower PKC activity and expression of phosphorylated PKC compared with WT mice, and these decreases could be prevented by LP treatment. These data suggest that consumption of LP enhances PepT1-mediated amino acid absorption, likely through alterations in PKC activity, as opposed to changes in expression or distribution of PepT1 in the small intestine of IL-10−/− mice.

Acute effects of breakfasts containing α-lactalbumin, or gelatin with or without added tryptophan, on hunger, ‘satiety’ hormones and amino acid profiles

Proteins are the most satiating macronutrients. Tryptophan (TRP) may contribute to the satiating effect, as it serves as a precursor for the anorexigenic neurotransmitter serotonin. To address the role of TRP in the satiating properties of dietary protein, we compared three different breakfasts, containing either alpha-lactalbumin (high in TRP), gelatin (low in TRP) or gelatin with added TRP (gelatin+TRP, high in TRP), on appetite. Twenty-four subjects (22-29 kg/m2; aged 19-37 years) received a subject-specific breakfast at t = 0 with 10, 55 and 35 % energy from protein, carbohydrate and fat respectively in a randomised, single-blind design. Hunger, glucagon-like peptide (GLP)-1, ghrelin, amino acid concentrations and energy intake during a subsequent lunch were determined. Suppression of hunger was stronger 240 min after the breakfast with alpha-lactalbumin compared with gelatin and gelatin+TRP. Total plasma amino acid concentrations were lower with alpha-lactalbumin compared with gelatin with or without TRP (from t = 180-240 min). TRP concentrations were higher after alpha-lactalbumin than after gelatin with or without TRP from t = 0-100 min, whereas from t = 100-240 min, TRP concentrations were lower after gelatin than after alpha-lactalbumin and gelatin+TRP. The plasma ratio of TRP to other large neutral amino acids (LNAA) was, only at t = 100 min, lower after gelatin+TRP than after the other breakfasts. Plasma amino acid responses, TRP concentrations and TRP:LNAA ratios were not correlated with hunger. GLP-1 and ghrelin concentrations were similar for all diets. Energy intake during a subsequent lunch was similar for all diets. Summarised, an alpha-lactalbumin breakfast suppresses hunger more than a gelatin or gelatin+TRP breakfast. This cannot be explained by (possible) differences found in TRP concentrations and TRP:LNAA ratios in the breakfasts and in plasma, as well as in circulating total amino acids, GLP-1 and ghrelin.

Plasma Amino Acid Concentrations During Late Rehabilitation in Patients With Traumatic Brain Injury

Børsheim E, Bui Q-UT, Wolfe RR. Plasma amino acid concentrations during late rehabilitation in patients with traumatic brain injury. Objectives To investigate whether the basal plasma amino acid concentrations in patients with traumatic brain injury (TBI) have returned to levels found in healthy controls at about 17 months postinjury and to determine the effect of intake of a mixture of essential amino acids (EAA) on plasma amino acid concentrations in TBI versus healthy controls. Design Peripheral venous amino acid concentrations in subjects with TBI were compared with concentrations in healthy controls both at rest and for 1 hour after intake of 7g of EAA. Setting Postacute brain injury rehabilitation center. Participants Six men with TBI (age ± standard deviation, 27±6y; months postinjury, 17±4) and 6 healthy men (age, 43±7y). Intervention Intake of a drink consisting of 7g of EAA. Main Outcome Measures Individual and total plasma amino acid concentrations. Results Total amino acid concentration was about 12% lower in TBI versus controls ( P=.022). Valine was reduced by 33% in the TBI group versus controls ( P=.003), whereas the other EAA did not differ between groups. After intake of the EAA drink, plasma non-EAA increased to a significantly higher level in controls versus TBI subjects ( P=.017). Conclusions Plasma total amino acid concentration is still reduced 17 months postinjury in patients with TBI versus healthy controls, mainly because of a lower valine level. This may be of importance for both brain and muscle metabolic functions, and warrant further study. Further, ingested EAA are apparently not as readily converted to non-EAA in TBI patients as in healthy controls, suggesting that in recovery from TBI, certain non-EAA may become provisionally essential.

Palatability Affects the Percentage of Metabolizable Energy as Protein Selected by Adult Beagles

The percentage of protein that dogs voluntarily choose and the effect of palatability on the quantity selected were determined. Six beagles were offered a choice of two isoenergetic purified diets containing 0 vs. 25, 9 vs. 32, 18 vs. 32, 18 vs. 48 and 25 vs. 48% metabolizable energy from protein (MEp). To examine whether palatability modifies the choice, the dogs were offered 0 vs. 25% MEp, with the 0% protein diet containing 2.9 times more sucrose than the diet containing 25% MEp. To determine the effect of concentration of protein in the diet on dietary choice and plasma amino acid concentrations (PAA), dogs were adapted to 9% MEp, followed by a choice of diets containing 9 vs. 32% MEp. The choice was repeated after adaptation to a diet containing 32% MEp. Dogs selected diets to obtain 21-27% of the MEp (mean, 25% MEp; median, 27% MEp) when sucrose was kept at 6.4%. When the protein-free diet contained 25% sucrose, dogs selected 17% of MEp, but increased food intake to ingest about the same amount of protein per day. PAA did not correlate linearly with protein intake. Food intake and total PAA were the lowest after consumption of the 9% MEp diet. We conclude that when fed equally bland diets, dogs select food to ingest approximately 25% MEp. As a palatability enhancer, sucrose increases food intake and selection of the diet containing the higher sucrose concentration.

Physiological adaptations of the raccoon dog (Nyctereutes procyonoides) to seasonal fasting-fat and nitrogen metabolism and influence of continuous melatonin treatment.

The raccoon dog (Nyctereutes procyonoides) is a middle-sized canid with profound autumnal fattening followed by winter sleep. This study investigated the effects of prolonged fasting-induced winter sleep on the fat and nitrogen metabolism of the species. Half of the animals were treated with continuous-release melatonin implants to induce artificial short photoperiod. Autumnal accumulation of fat was characterized by low plasma free fatty acid (FFA), diacylglycerol (DG), and triacylglycerol (TG) levels. After transition to winter catabolism, the circulating lipid levels increased due to enhanced lipolysis. Two months of fasting resulted in a steady 3.1 kg weight loss (28% of body mass, 0.47% day(-1)). Storage fat was mobilized during the winter sleep reflected by the elevated FFA and DG concentrations. The lowered insulin levels could be a stimulator for TG hydrolysis. The plasma total amino acid concentrations, urea levels, and urea-creatinine ratios decreased due to fasting, whereas ammonia and total protein concentrations remained stable. The effects of melatonin on energy metabolism were modest. The results indicate that the raccoon dog is well adapted to long-term wintertime fasting utilizing fat as the principal metabolic fuel. The species can maintain its protein catabolism constant for at least 60 days. Decreased cortisol and thyroid hormone concentrations may contribute to protein sparing.

Alcohol impairs leucine-mediated phosphorylation of 4E-BP1, S6K1, eIF4G, and mTOR in skeletal muscle.

Acute alcohol (EtOH) intoxication impairs skeletal muscle protein synthesis. Although this impairment is not associated with a decrease in the total plasma amino acid concentration, EtOH may blunt the anabolic response to amino acids. To examine this hypothesis, rats were administered EtOH or saline (Sal) and 2.5 h thereafter were orally administered either leucine (Leu) or Sal. The gastrocnemius was removed 20 min later to assess protein synthesis and signaling components important in translational control of protein synthesis. Oral Leu increased muscle protein synthesis by the same magnitude in Sal- and EtOH-treated rats. However, the increase in the latter group was insufficient to overcome the suppressive effect of EtOH, and the rate of synthesis remained lower than that observed in rats from the Sal-Sal group. Leu markedly increased phosphorylation of Thr residues 36, 47, and 70 on 4E-binding protein (BP)1 in muscle from rats not receiving EtOH, and this response was associated with a redistribution of eukaryotic initiation factor (eIF) 4E from the inactive eIF4E. 4E-BP1 to the active eIF4E. eIF4G complex. In EtOH-treated rats, the Leu-induced phosphorylation of 4E-BP1 and changes in eIF4E availability were partially abrogated. EtOH also prevented the Leu-induced increase in phosphorylation of eIF4G, the serine/threonine protein kinase S6K1, and the ribosomal protein S6. Moreover, EtOH attenuated the Leu-induced phosphorylation of the mammalian target of rapamycin (mTOR). The ability of EtOH to blunt the anabolic effects of Leu could not be attributed to differences in the plasma concentrations of insulin, insulin-like growth factor I, or Leu. Finally, although EtOH increased the plasma corticosterone concentration, inhibition of glucocorticoid action by RU-486 was unable to prevent EtOH-induced defects in the ability of Leu to stimulate 4E-BP1, S6K1, and mTOR phosphorylation. Hence, ethanol produces a leucine resistance in skeletal muscle, as evidenced by the impaired phosphorylation of 4E-BP1, eIF4G, S6K1, and mTOR, that is independent of elevations in endogenous glucocorticoids.