Effect of dietary amino acids on behavior and serum levels of amino acids in stress loaded rats

Interrelationships between behavior and concentrations of serum amino acids in stressed rats with immobilization and water immersion were investigated. Rats were subjected to 7 h of immobilization with water-immersion stress (IWS) in each sequential day, and serum amino acids were then determined. On the first day, serum taurine, threonine, valine, leucine, isoleucine, phenylalanine, lysine, and histidine (increased-type of amino acids) were significantly increased, but alanine (decreased-type) was significantly decreased. On days 3 and 7, the increase was retained, except for threonine, histidine, and lysine. Spontaneous activities (locomotion, rearing behavior, hole-poking) under loading water-immersion stress were significantly decreased, but a supplementation of branched-chain amino acids (BCAA) led to recovery. We suggest that pretreatment with some kind of increased type of amino acid, such as BCAA, might effectively prevent decline in spontaneous activities evoked by water-immersion stress.     

Amino Acid Deficiencies In Casein

A new order of growth limiting amino acids is indicated for casein as follows: methionine; threonine; tryptophan, isoleucine, leucine, histidine, valine and phenylalanine; lysine; nonessential amino acids.     

Histidine and arginine are associated with inflammation and oxidative stress in obese women

The aims of the present study were to examine the serum amino acid profiles in obese and non-obese women and investigate the relationships between the serum amino acids and inflammation and oxidative stress in a human case-control study. Serum amino acids, inflammatory biomarkers (C-reactive protein and IL-6) and oxidative biomarkers (superoxide dismutase, malondialdehyde and glutathione peroxidase) were measured and compared in 235 obese women and 217 non-obese controls. The relationships between serum amino acids and inflammatory and oxidative biomarkers were examined using multiple linear regression. Among the amino acids determined, serum histidine, arginine, threonine, glycine, lysine and serine were found to be significantly lower in obese women as compared to non-obese controls (P < 0·001). The difference was the greatest for histidine (P < 0·001). In obese women, both histidine and arginine were negatively associated with inflammation and oxidative stress. In non-obese controls, histidine was negatively associated with oxidative stress. The findings in this study indicate that the metabolism of amino acids is abnormal in obese women in whom histidine and arginine have close relationships with inflammation and oxidative stress.     

Induction of threonine imbalance by dispensable amino acids: relation to competition for amino acid transport into brain.

The ability of low protein diets containing small neutral, dispensable amino acids to induce threonine imbalance has been examined. Diets containing amino acids which compete for threonine transport in vitro (serine, alanine, alpha-amino-n-butyrate) caused depressions of growth and food intake which could be corrected to varying degrees by adding threonine to the diet. Large neutral, indispensable amino acids, moderately inhibitory of threonine transport, also induced the imbalance. Some amino acids that had little or no effect on threonine transport in vitro (acidic amino acids and proline) did not cause growth and food intake depressions. Other non-inhibitory amino acids (arginine and lysine) caused growth depressions which were not satisfactorily corrected by additional threonine alone, but were prevented by supplements of all the indispensable amino acids including threonine. Ornithine which was also not inhibitory of threonine transport was an exception. It induced a moderate growth depression which was corrected by additional threonine. Similar studies showed that histidine or tryptophan imbalance could be induced by feeding diets containing only those large neutral amino acids which compete for histidine or tryptophan transport in vitro. These experiments show that, based on the results of transport competition experiments, it is generally possible to devise amino acid supplements which can induce a dietary imbalance of a given amino acid.     

Gastric inhibitory polypeptide and insulin responses to orally administered amino acids in genetically obese hyperglycemic (ob/ob) mice

The effects of oral administration of eight L-amino acids (alanine, arginine, cysteine, glycine, histidine, hydroxyproline, lysine and threonine) individually or as an amino acid mixture on plasma gastric inhibitory polypeptide (GIP), insulin and glucose concentrations were examined in 18-h fasted obese hyperglycemic (ob/ob) mice. At a dose of 5.4 mmol/kg body weight, arginine, cysteine, histidine and the amino acid mixture were equipotent in terms of increasing plasma GIP and insulin concentrations. Alanine, hydroxyproline and lysine also increased plasma GIP, but insulin concentrations were unchanged. In contrast, threonine failed to affect either GIP or insulin concentrations. There was no correlation between either the incremental or integrated GIP and insulin responses, and none of the amino acids administered affected circulating glucose concentrations. The results indicate that a range of essential and nonessential neutral and basic amino acids stimulate the release of GIP in ob/ob mice. However, GIP made only a modest contribution to the stimulation of insulin secretion following administration of amino acids in the presence of basal glycemia.     

The optimal amino acid supplementation of barley for the growing pig. 1. Response of nitrogen metabolism to progressive supplementation.

1. In five experiments, involving 142 female pigs weighing on average 33 kg, estimates were made of the amounts of essential amino acids which minimized urinary N excretion when diets with barley as the only source of protein were given at the rate of 120 g/kg0.75 per d. 2. With additions of lysine (4.0 g/kg diet) and threonine, (1.2 g/kg diet) to barley urinary N excretion decreased from 0.91 to 0.36 g/kg0.75 per d, corresponding to an increase in biological value (bv) from 0.51 to 0.86. 3. With these additions of lysine and threonine, there were no responses to additions of tryptophan, methionine or isoleucine, or to further additions of lysine or threonine, but addition of histidine significantly reduced N excretion. 4. No optimal addition of histidine could be determined; the mean rate of N excretion after addition of histidine (not less than 0.3 g/kg diet) was 0.27 g/kg0.75 per d, corresponding to a BV of 0.93. 5. There was a variation between pigs from different litters in their responses to added histidine. Those with low rates of N excretion on the unsupplemented diet did not respond to additions of histidine, but those with high rates did. 6. It is concluded that additions of only three amino acids can greatly improve the nutritive value of barley protein for the growing pig and that the amino acid composition of the supplemented protein closely approaches the ideal; it is also similar to whole-body tissue protein.     

Human milk nonprotein nitrogen components: changing patterns of free amino acids and urea in the course of early lactation

Free amino acids and urea were analyzed in 78 human milk samples obtained during the first 5 wk of lactation from 10 mothers delivering at term. Significant differences (p less than 0.05) in the concentrations between colostral and mature milk were found for glutamic acid, glutamine, alanine, glycine, cystine, and phosphoethanolamine which increased, and with serine, phosphoserine, aspartic acid + asparagine, arginine, lysine, isoleucine, phenylalanine, proline, methionine, tryptophan, and beta-alanine which decreased. Some of these changes occurred within the first 5 days of lactation, so that differences between transitional and mature milk became negligible (glutamic acid, alanine, and serine, aspartic acid + asparagine, lysine, isoleucine, methionine, tryptophan, respectively). No significant differences between any of the three stages of lactation were found regarding the concentrations of total free amino acids, urea, taurine, threonine, valine, leucine, histidine, and tyrosine. Possible relevances for free amino acids, including nonprotein ones, in human milk are discussed.     

L-leucine, L-methionine, and L-lysine are involved in the regulation of intermediary metabolism-related gene expression in rainbow trout hepatocytes

Using rainbow trout hepatocytes stimulated with l-leucine, l-methionine, or l-lysine in the presence or absence of bovine insulin, we investigated the ability of these amino acids to mimic the effects of a pool of amino acids on protein kinase B (Akt)/target of rapamycin (TOR) signaling pathways and expression of 6 genes known to be subjected to insulin and/or amino acid regulation [glucose-6-phosphatase (G6Pase), phosphoenolpyruvate carboxykinase (PEPCK), glucokinase (GK), pyruvate kinase (PK), fatty acid synthase (FAS), and serine dehydratase (SDH)]. Emphasis was placed on leucine, known to be a signaling molecule in mammals, and methionine and lysine that are essential amino acids limiting in plant-based diets for fish. In the presence of insulin, leucine (but not methionine or lysine) phosphorylated Akt and ribosomal protein S6 as previously observed with a pool of amino acids, suggesting that leucine might participate in the activation of the TOR pathway by amino acids in fish, as in mammals. G6Pase, PEPCK, GK, and SDH gene expression were higher in leucine-treated cells compared with control cells. Leucine combined with insulin reduced G6Pase gene expression by 90% and increased FAS gene expression > 4-fold compared with the control treatment. Methionine weakly decreased G6Pase, GK, and SDH gene expression and lysine weakly but significantly decreased the mRNA level of PEPCK. Thus, leucine regulated gluconeogenesis and lipogenesis, but not glycolysis, in the same way as a pool of amino acids. Methionine appeared to be involved in the regulation of specific genes, whereas lysine only had limited effects. These findings are particularly relevant regarding the involvement of amino acids in the regulation of metabolism-related gene expression.     

Assessment of the essentiality of methionine, threonine, tryptophan, histidine and isoleucine in immature dogs.

A completely purified L-amino acid diet was used to evaluate the indispensability of methionine, threonine, tryptophan, histidine and isoleucine in the immature dog. Immature dogs fed diets devoid of methionine, threonine, tryptophan, histidine or isoleucine had severely depressed food intakes, accompanying weight loss, and a distinctly negative nitrogen balance. Partial or total removal of these amino acids resulted in a marked increase in blood and urinary urea. Estimates are made for the dietary requirements for these amino acids in the immature dog.     

The amino acid requirements of the preruminant calf.

1. Ten calves (50--58 kg live weight) were given a diet consisting of diluted whole milk, wheat gluten and supplemented with appropriate nutrients including amino acids but deficient in lysine. The lysine reequirements of these calves, which were growing at approximately 0.25 kg/d, were estimated from responses to lysine supplementation of this diet. From plasma urea, plasma lysine, nitrogen retention and apparent digestibility of N responses the estimated lysine requirements were 8.5, 7.5, 7.2 and 7.6 g/d respectively. 2. From the mean lysine requirement (7.8 g/d) and the ratio, lysine: otheressential amino acids in carcasses of similar calves the estimated requirements were (g/d): methionine 2.1, cystine 1.6, threonine 4.9, valine 4.8, isoleucine 3.4, leucine 8.4, tyrosine 3.0, phenylalanine 4.4, histidine 3.0, arginine 8.5, tryptophan 1.0.     

Newer concepts of the indispensable amino acids

In healthy adult humans, eight amino acids (isoleucine, leucine, lysine, methionine, phenylalanine, threonine, tryptophan, and valine) were shown classically by nitrogen balance studies to be indispensable. Subsequent studies classifying histidine as indispensable are reviewed in this article. We also review the evidence that in certain nutritional or disease states or in certain stages of development otherwise dispensable amino acids may become indispensable. Arginine, citrulline, ornithine, cysteine, and tyrosine thus may be considered as acquired indispensable amino acids. Evidence for the indispensability of taurine is also considered. We propose a classification of the indispensability of amino acids based on clinical and therapeutic considerations.     

Effect of acid hydrolysis time on amino acid determination in casein and processed cheeses with different fat content

This work deals with hydrolysis curves for 15 amino acids (aspartic acid, threonine, serine, glutamic acid, proline, glycine, alanine, valine, isoleucine, leucine, phenylalanine, tyrosine, histidine, lysine and arginine) in bovine casein and in model-processed cheeses with 30, 45 and 60% (w/w) fat in dry matter. The effect of a different fat content on hydrolysis curves of the amino acids was observed. The hydrolysis curves were obtained by means of nonlinear regression using 13 hydrolysis intervals (range, 0–144 h). The amino acid content in hydrolysates was measured using ion exchange chromatography. Correction factors were calculated with the aim of increasing the accuracy of the amino acid determinations. With the rising fat content in samples, the correction factors increased for the majority of amino acids observed. The highest correction factors were determined for threonine, serine and tyrosine.     

Oxidation of an indicator amino acid by young pigs receiving diets with varying levels of lysine or threonine, and an assessment of amino acid requirements

The catabolism of [14C]phenylalanine was used to indicate the effects of varying the dietary level of lysine and threonine on the retention of dietary amino acids by 2-week-old pigs receiving diets containing skim milk and a mixture of free amino acids. Reducing the dietary level of lysine from 16 to 12 g/kg had no influence on phenylalanine oxidation, reducing the lysine level from 12 to 11 then to 10 g/kg caused an almost linear increase in phenylalanine oxidation whereas further reduction to 9 or 8 g/kg resulted in a less-marked increase in phenylalanine oxidation. This showed that 12 g lysine/kg was required to maximize amino acid retention and indicated that lysine was conserved more effectively at low dietary concentrations than at dietary concentrations approaching the requirement. Reducing the dietary level of threonine from 8 to 6 g/kg had no influence on phenylalanine oxidation, whereas further reduction to 4 g/kg caused a linear increase in phenylalanine catabolism showing that 6 g threonine/kg was required to maximize amino acid retention. Reduction of the levels of lysine, threonine and methionine from the generous levels characteristic of a diet containing 240 g protein from skim milk/kg, to the requirement levels determined separately in the presence of the generous levels of all the other amino acids, resulted in a twofold increase in phenylalanine catabolism. This shows that the pig seems able to conserve limiting intakes of a single amino acid, but not if the intakes of two or three amino acids are limiting.     

The optimum dietary amino acid pattern for growing pigs. 1. Experiments by amino acid deletion

A series of four nitrogen-balance experiments was carried out with growing pigs to determine the optimum balance amongst the amino acids in the diet. The reduction in N retention when 20% of a single amino acid was removed from the diet was used to calculate a dietary amino acid pattern in which each amino acid would be equally limiting. A mixture of amino acids simulating the amino acid pattern of casein was used with the same efficiency as casein. From two successive deletion experiments an optimum balance amongst the essential amino acids was derived. Expressed relative to lysine = 100 this had threonine 72, valine 75, methionine + cystine 63, isoleucine 60, leucine 110, phenylalanine + tyrosine 120, tryptophan 18. No estimate was made for histidine. Essential amino acids in this pattern were mixed with non-essential amino acids in ratios of 36:64 up to 57:43. The highest efficiency of N retention was achieved with diets having a ratio of at least 45:55. This included (g/16 g N) lysine 6.5, threonine 4.7, valine 4.9, methionine + cystine 4.1, isoleucine 3.9, leucine 7.2, phenylalanine + tyrosine 7.8, tryptophan 12. The N of diets with this amino acid pattern was utilized significantly better than when the pattern proposed by the Agricultural Research Council (1981) was used. The flow of amino acids past the terminal ileum of pigs given the semi-synthetic diet with this amino acid pattern was no greater than that observed with protein-free diets. The proposed pattern thus describes the intrinsic requirements of the growing pig for absorbed amino acids.     

Effects of leptin on intake of specific micro- and macronutrients in a woman with leptin gene deficiency studied off and on leptin at stable body weight

In this report, we examine the effects of leptin on the intake of specific macro- and micronutrients in a female patient with leptin gene deficiency. The patient was studied off and on leptin at stable body weight, within the normal to mildly overweight range. The data were obtained by detailed dietary assessments, measuring dietary intakes by weighed food and fluid consumption records, and analyzed using nutrition analysis software. Overall, significant differences were found in the off versus on leptin treatment periods in the following categories: (i) macronutrients: kilocalories, protein, carbohydrates, monounsaturated fats, MFA 18:1 oleic and total fiber; (ii) vitamins: vitamin C, pyridoxine and pantothenic acid; (iii) minerals: potassium, magnesium, copper and chromium; and amino acids: threonine, lysine and histidine. The nutritional data from this study indicates a direct link between the effects of leptin and ingestion of several specific micronutrients. The mechanisms underlying these effects warrant further investigation and study.     

Threonine, tryptophan and histidine requirements of immature beagle dogs

The threonine, tryptophan and histidine requirements of growing male Beagle dogs were determined using diets containing the equivalent of 14% protein as crystalline L-amino acids. For each amino acid the requirement was taken to be the minimum dietary quantity required for optimal growth, feed efficiency and nitrogen retention. In experiment 1 the threonine requirement was found to be provided by 0.52% or more dietary threonine. In experiments 2 and 3 the minimal dietary tryptophan and histidine requirements of immature dogs were estimated as 0.17 and 0.21%, respectively. The requirements for threonine and tryptophan are similar to those for growing rats. However, the histidine requirement of Beagles appears to be lower than that of rats.     

Response of rats to diets of equal chemical score: effect of lysine or threonine as the limiting amino acid and of an amino acid excess

The effect of lysine or threonine deficiency with or without excesses of all other amino acids was evaluated in a 21-d feeding study with male rats. Four amino acid mixtures were designed to be first limiting for the rat in lysine or threonine and contained either 0 or 50% excess of nonlimiting amino acids. These mixtures were incorporated into purified diets to provide seven levels [20-140% of the National Research Council (NRC) requirement] of the limiting amino acid. Food intake, body weight gain and carcass composition were measured for each rat to determine the effects of the identity of the limiting amino acid and of amino acid excess on the response to dietary chemical score. Significant effects and/or interactions of the identity of the limiting amino acid (i.e., Lys or Thr) and the presence of excess amino acids were seen for each of the measured responses. At equivalent dietary percentages of the NRC requirement, threonine deficiency supported greater body weight gain than did lysine deficiency. At equivalent deficiencies (Lys vs. Thr) threonine-deficient rats were more susceptible to adverse effects of excess amino acids. When the limiting amino acids were incorporated into the diet through incremental addition of the deficient amino acid mixture, rats responded to levels of lysine or threonine in excess of the NRC requirement. These results suggest that the current NRC requirements for these amino acids are too low and that aspects of the dietary amino acid composition other than the percentage deficit of the limiting amino acid can be important determinants of animal response.     

Degradation and outflow of amino acids from the rumen of sheep

1. In hay-fed, cannulated sheep the apparent degradation in and outflow from the rumen were determined for graded doses of mixtures of the amino acids lysine, threonine and methionine, administered intraruminally and using polyethylene glycol (PEG) as a liquid marker. The doses ranged between 2.5 and 15 g for each amino acid in the mixtures. 2. Relative rate of apparent degradation in the first 4 h was highest for lysine, and lowest for methionine. The apparent degradation in 24 h was highest for lysine and lowest for threonine. Conversely the fraction flowing out of the rumen in intact form in 24 h was highest for threonine and lowest for lysine. Rates of apparent degradation as well as outflow were dose-dependent. 3. The validity of the estimated outflow of amino acids from the rumen was corroborated by measurements of concentrations of the amino acids in duodenal contents and in blood plasma which were also dose-dependent. 4. It was concluded that part of the requirement for the essential amino acids threonine and methionine may be met, even when these amino acids are delivered in unprotected form, given as a feed supplement.     

Amino acids and indoleamines in the brain after infusion of branched-chain amino acids to rats with liver ischemia

Rats with a portacaval anastomosis and ligation of the hepatic artery 2 days later were infused for 6 hr with a 10% glucose solution (group I) or the same solution combined with 0.24 M/liter branched-chain amino acids (BCAA, group II). Control animals with portacaval anastomosis and sham-operation (group III) or two sham-operations (group IV) were infused with a 10% glucose solution. The rats were killed by decapitation and indoleamines and amino acids were determined in the brain. Rats with liver ischemia were stuporous at the end of the experiment irrespective of treatment. The concentrations in the cortex of lysine, methionine, phenylalanine, threonine, alanine, glutamine, glycine, histidine, and tyrosine were significantly increased in group I compared to group IV. Infusion of BCAA to rats with liver-ischemia (group II) resulted in significantly lower concentrations of lysine, methionine, phenylalanine, threonine, histidine and tyrosine and increased concentrations of isoleucine, leucine, valine, and arginine compared to group I. The content of serotonin in the cortex and brain stem was significantly increased in group I compared with the BCAA-treated animals (group II) and the control groups III and IV. The concentrations of 5-hydroxyindoleacetic acid (5-HIAA) in the cortex and brain stem were higher in group I than in group IV. Infusion of BCAA to rats with liver ischemia normalized the concentrations of 5-HIAA in the cortex and brain stem.     

Intracellular amino acid levels as predictors of protein synthesis.

Protein synthesis depends on a complete complement of precursor amino acids, specific acetylating enzymes, tRNA, and so forth. It has been related to metabolism of individual amino acids, eg, valine and leucine; however, the relation of protein synthesis to the ambient concentrations of amino acids in the intracellular and extracellular pools has not been defined. Using the viable, isolated granulocyte (leukocyte) as an in vitro cell model, protein synthesis (incorporation of 4,5-3H-leucine) has been related to simultaneous amino acid concentrations in the cell and plasma by multiple regression analysis. Fifty-five normal neonates and 30 normal adults were studied. Protein synthesis was higher in the infants than in the adults (3,527 vs 2,685 pmole/hr/mg DNA). The intracellular concentrations of most amino acids were higher than their concentrations in plasma, except for valine and citrulline, which were lower. The “aminograms” in the two pools also were very different. Forty-four percent of the variance (R2) in protein synthesis was accounted for by the intracellular concentrations of leucine, glycine, alanine, and taurine in neonates and 45% by a combination of threonine, valine, methionine, and histidine in adults. The intracellular concentrations of each of these predictor amino acids in adults were, in turn, related to different combinations of the plasma concentrations of threonine, phenylalanine, tryptophan, isoleucine, histidine, citrulline, ornithine, arginine, and glycine. Thus, it is possible to identify sets of intracellular amino acids that predict the level of protein synthesis and to delineate combinations of plasma amino acids whose levels account for a significant portion of the variance in the intracellular predictor amino acids in normal human infants and adults.