Effects of Sulfur-containing Amino Acids and Glycine on Plasma Cholesterol Level in Rats Fed on a High Cholesterol Diet

The effects of the addition of individual amino acids on methionine-induced hypercholesterolemia (experiment 1), and the interacting effects of dietary protein level and sulfur-containing amino acids and glycine on plasma cholesterol concentration (experiment 2) were studied in growing rats fed on a high cholesterol diet. In experiment 1, rats were fed on a 25% casein-0.75% methionine (25CM) diet containing 2.5% of individual amino acids for 2 weeks. Methionine-induced hypercholesterolemia was prevented by the concurrent addition of glycine or serine, but the other amino acids tested (alanine, threonine, leucine, phenylalanine, lysine, arginine, and glutamic acid) had no effect. Histidine rather enhanced the hypercholesterolemia. In experiment 2, rats were fed on a 10%, 25%, or 50% casein diet containing 0.75% methionine, 0.60% cystine, 0.63% taurine, 2.5% glycine, or 0.75% methionine +2.5% glycine for 3 weeks. Dietary addition of 0.75% methionine increased the plasma cholesterol concentration for the 25% and 50% casein diets, but it decreased the plasma cholesterol for the 10% casein diet. When the addition level of methionine was doubled in the 10% casein diet, the plasma cholesterol concentration was significantly higher for the 1.5% methionine-added diet than for the 0.75% methionine-added diet. Cystine and taurine lowered plasma cholesterol for all dietary casein levels. Methionine-induced hypercholesterolemia with 25% and 50% casein diets was prevented by the glycine supplementation. These data suggest that sulfur-containing amino acids and glycine are important in plasma cholesterol regulation.     

Regulation of polyamine synthesis by dietary alpha-aminoisobutyric acid and ornithine

An experiment was conducted to determine the effect of feeding ornithine in combination with alpha-aminoisobutyric acid (AIB), an inhibitor of arginase, on the regulation of polyamine synthesis in chicks. A total of 48 chicks with genetically elevated renal arginase activity was fed diets containing crystalline amino acids and 1% AIB with or without 2% ornithine. Feeding AIB reduced renal arginase activity, while renal and hepatic ornithine decarboxylase (ODC) activity increased. Feeding AIB plus ornithine caused no further reduction in renal arginase activity compared with that in chicks fed the AIB-supplemented diet. Renal and hepatic ODC activities, however, fell to below control levels. Renal, hepatic, and breast muscle ornithine concentrations increased substantially when ornithine was fed. AIB plus ornithine increased renal putrescine and spermidine concentrations. It was concluded that AIB could partially overcome the ornithine-induced inhibition of ODC activity. These findings support the hypothesis that dietary manipulation of precursor amino acids of polyamines in the presence of metabolites that induce ODC activity can influence tissue polyamine concentrations.     

Cysteine supplementation decreases plasma homocysteine concentration in rats fed on a low-casein diet in rats

The effect of dietary supplementation with cysteine on the plasma homocysteine concentration was investigated in rats fed on 10% casein (10C) and 30% casein (30C) diets. The 10C diet significantly increased the plasma homocysteine concentration as compared with the 30C diet. The hyperhomocysteinemia induced by the 10C diet was significantly suppressed by cysteine supplementation even at a 0.3% level, whereas cysteine did not decrease the plasma homocysteine concentration when added to the 30C diet. In contrast, 0.3% methionine supplementation of the 10C diet tended to increase the plasma homocysteine concentration. Cysteine supplementation to rats fed on the 10C diet did not alter the plasma cysteine concentration and the hepatic activities of cystathionine beta-synthase and betaine:homocysteine S-methyltransferase, whereas it significantly decreased the hepatic concentrations of S-adenosylmethionine and betaine. These results suggest that cysteine supplementation might be effective for suppressing the hyperhomocysteinemia induced by a low-protein diet.     

The effect of transport system A and N amino acids and of nerve and epidermal growth factors on the induction of ornithine decarboxylase activity

The induction of ornithine decarboxylase (EC 4.1.1.17) (ODC) by amino acids and by the peptide hormones nerve growth factor (NGF) and epidermal growth factor (EGF) in salts-glucose media has been studied. Only those neutral amino acids taken into the cell via one of the Na+ dependent transport systems stimulate ODC activity. Asparagine and the nonmetabolizable alpha-amino-isobutyric acid (AIB) were used as representatives of this class of inducing amino acids, and their intracellular concentrations were related to the levels of ODC induced. A threshold intracellular concentration of asparagine or AIB has to be attained before ODC can be induced. Further slight increases in intracellular concentrations of asparagine or AIB produce disproportionately large increases of ODC, resulting in a sigmoidal curve of ODC induction. These results, and the fact that the decrease in ODC levels caused by valine is associated with a concurrent decrease in the intracellular level of the inducing amino acid, suggest that the intracellular amino acid level is causally related to the induction of ornithine decarboxylase. Glutamic acid, EGF, and NGF do not induce ODC except in the presence of an inducing amino acid. They act synergistically with the inducing amino acid and produce higher ODC levels at the same intracellular concentration of the inducing amino acid.     

Suppression of methionine-induced hyperhomocysteinemia by glycine and serine in rats

The hyperhomocysteinemia induced by a dietary addition of 1% methionine was significantly suppressed by the concurrent addition of 1% glycine or 1.4% serine to the same degree. The methionine-induced increase in the hepatic concentration of methionine metabolites was significantly suppressed by glycine and serine, but the hepatic cystathionine beta-synthase activity was not enhanced by these amino acids. When the methionine-supplemented diet was changed to the methionine plus glycine or serine diet, the plasma homocysteine concentration rapidly decreased during and after the first day. The hyperhomocysteinemia induced by an intraperitoneal injection with methionine was also suppressed by concurrent injection with glycine or serine, although the effect of serine was significantly greater than that of glycine. These results indicate that glycine and serine were effective for suppressing methionine-induced hyperhomocysteinemia: serine and its precursor glycine are considered to have elicited their effects mainly by stimulating cystathionine synthesis by supplying serine, another substrate for cystathionine synthesis.     

Effects of a high soy protein diet on intestinal polyamines and ornithine decarboxylase activity in rats

This study was performed to determine whether intestinal luminal polyamine concentrations are affected by a high soy protein diet when compared with a high casein diet or a normoprotein casein diet. We also determined the effects of these diets, with differences in polyamines content, on mucosal polyamines and ornithine decarboxylase (ODC) activity to assess cell proliferation. Three groups of eight male Wistar rats were fed either a 50% soy protein diet, a 50% casein diet, or an 18% casein diet as a control. After 4 weeks of feeding, both intestinal content and mucosa were recovered. Polyamines were assayed by high performance liquid chromatography. ODC activity was measured by the release of (14)CO(2) from (14)C-L-ornithine. Luminal putrescine and cadaverine concentrations were higher in the jejunum than in the ileum, suggesting an absorptive process. The highest concentrations of intestinal polyamines were observed in rats fed the soy protein diet (P < 0.05). Only minor differences were observed in mucosal polyamines according to the diets. ODC activity was also higher in the intestinal mucosa of rats fed the high soy protein diet (P < 0.05). These results suggest that intestinal luminal polyamine concentrations and ODC activity are modulated by the dietary protein source.     

Effects of cysteine on amino acid concentrations and transsulfuration enzyme activities in rat liver with protein-calorie malnutrition

The changes in amino acid concentrations and transsulfuration enzyme activities in liver were investigated after 4-week fed on 23% casein diet (control group) and 5% casein diet without (protein-calorie malnutrition, PCM group) or with (PCMC group) oral administration of cysteine, 250 mg/kg (twice daily, starting from the fourth week) using rats as an animal model. By supplementation with cysteine in PCM rats (PCMC group), cysteine level was elevated almost close to the control level, and glutathione (GSH), aspartic acid and serine levels were restored greater than the control levels. The measurement of transsulfuration enzyme activities exhibited that gamma-glutamylcysteine ligase (gamma-GCL) activity was up-regulated in rats with protein restriction (PCM group), and cysteine supplementation (PCMC group) down-regulated to the control level. One-week supplementation of cysteine (PCMC group) significantly down-regulated the cysteine sulfinate decarboxylase activity. These results indicate that the availability of sulfur amino acid(s) especially cysteine appears to play a role in determining the flux of cysteine between cysteine catabolism and GSH synthesis.     

Rapid suppression of protein degradation in skeletal muscle after oral feeding of leucine in rats

A diet containing adequate amounts of protein rapidly suppresses myofibrillar protein degradation in rats and mice. This study determined whether dietary amino acids inhibit postprandial protein degradation in rat skeletal muscle. When rats fed on a 20% casein diet for 1 h after 18 h starvation, the rate of myofibrillar protein degradation measured by N(tau)-methylhistidine release from the isolated extensor digitorum longus muscle was significantly (p < 0.05) decreased at 4 h after refeeding. A diet containing an amino acid mixture which is the same composition as casein also reduced myofibrillar protein degradation at 4 h after refeeding (p < 0.05). An essential amino acid mixture (15.1%, corresponding to casein composition) and a leucine (2.9%) diets reduced the rate of myofibrillar protein degradation after refeeding (p < 0.05), whereas a protein free diet did not. Administration of leucine alone (0.135 g/100 g body weight) by a feeding tube induced a decrease in the rate of myofibrillar protein degradation at 2 h after administration (p < 0.05), whereas the serum insulin concentration was constant after leucine administration. These results suggested that leucine is one of regulating factors of myofibrillar protein degradation after refeeding of a protein diet.     

Metabolic Effects of Arginine and Methionine Supplement on Rats Fed Excess Glycine Diets

The contents of glycogen, lipid, urea and amino acids, and some enzyme activities in plasma, liver muscle and urine were determined with rats fed 10 to 12 g of 100 g body weight per day of the 10% casein diet (control) and 10% casein diets containing 7% glycine with or without 1.4% l-arginine HC1 and l-methionine for 7 days.

Nine hours after the final feeding, the amount of liver glycogen was high in the order of rats fed 10% casein diet containing 7% glycine, 10% casein diet containing 7% glycine with l-arginine and l-methionine, and the control. The amount of muscle glycogen was decreased only in those fed the control diet. The amount of liver lipid was increased by the addition of l-arginine and l-methionine to the excess glycine diet. Plasma and urinary urea was increased in animals given the excess glycine diets with or without both amino acids. In plasma liver, and muscle of animals given either of both the excess glycine diets 3 and 9 hr after the feeding, in general, glycine and serine were increased, and threonine and alanine were decreased as compared with those of rats given the control diet. However, the increase of glycine in plasma, liver and muscle detected at 9 hr after feeding the excess glycine diet was slightly prevented by the supplementation of both amino acids to the excess glycine diet. The activities of liver glycine oxidase and ornithine δ-aminotransferase of rats given the excess glycine diet with both amino acids were higher than those of other dietary groups. Liver serine dehydratase and glutamate-oxalacetate transaminase activities were high in the order of the animals fed the control, the excess glycine diet and the excess glycine diet containing both amino acids. Glutamate-pyruvate transaminase activity in the liver of rats fed the excess glycine diets with or without both amino acids were markedly higher than that of those fed the control. The activities of phosphopyruvate carboxylase and aconitase in the liver of animals given the excess glycine diet were higher than those of other dietary groups. Liver pyruvate kinase and glutamate dehydrogenase activities were similar among those dietary groups.     

Effects of Dietary Glutathione on Plasma and Liver Lipid Levels in Rats Fed on a High Cholesterol Diet

The effects of dietary glutathione (GSH) on plasma and liver lipid concentrations were investigated with rats fed on a high cholesterol diet. When graded levels of GSH, 0.75 to 5.0%, were added to the 25% casein basal diet, the plasma total cholesterol level was significantly decreased and the HDL-cholesterol level was inversely increased in all addition levels without influence on the growth of animals except for the 5% addition level; the dietary addition of 5% GSH markedly depressed the growth and food consumption of rats and caused a slight diarrhea. Plasma triglyceride and phospholipid levels were decreased by the dietary addition of GSH. The contents of cholesterol and triglyceride in the liver were decreased as the dietary addition level of GSH was increased. The dietary addition of a mixture of glutamic acid, cysteine and glycine, or cysteine alone corresponding to 2.5% GSH resulted in a cholesterol-lowering effect which could not be distinguished from the effect of GSH in rats fed on the 25% casein diet. When 1.5% GSH was added to a low (10%) casein diet, the plasma cholesterol-lowering effect of GSH was also observed and the effect was comparable to that of cysteine. These results indicate that dietary-added GSH has a plasma and liver cholesterol-lowering efficacy and that this effect is largely attributable to the cysteine residue of GSH rather than to the tripeptide itself or the other amino acid residues.     

Increased response of liver microsomal delta 6-desaturase activity to dietary methionine in rats

The effects of dietary casein level (5-40%) on the liver microsomal phospholipid profile, delta 6-desaturase activity and related variables were investigated in rats to examine whether the dietary protein level affected the delta 6-desaturase activity through an alteration of the liver microsomal phospholipid profile. The effects of supplementing a 10% casein diet with certain amino acids were also investigated. The concentration of hepatic S-adenosylmethionine (SAM), the ratio of phosphatidylcholine (PC) to phosphatidylethanolamine (PE) and the delta 6-desaturase activity in liver microsomes, and the ratio of arachidonate to linoleate of microsomal PC increased with increasing dietary casein level. There were significant correlations between the dietary methionine content and hepatic SAM concentration, hepatic SAM concentration and microsomal PE concentration, and microsomal PE concentration and delta 6-desaturase activity. Supplementation of the 10% casein diet with methionine significantly increased the hepatic SAM concentration, PC/PE ratio, delta 6-desaturase activity, and arachidonate/linoleate ratio, whereas cystine supplementation had no or little effect on these variables. These increases induced by methionine were significantly suppressed by additional glycine. The results obtained here, together with those in our previous report, suggest that quantity and type of dietary protein might affect the delta 6-desaturase activity through an alteration of the liver microsomal profile of phospholipids, especially PE, and that the alteration of phospholipid profile might be mediated by a hepatic SAM concentration that reflects the dietary methionine level.     

Studies on the mechanism of induction of hypercholesterolemia in rabbits by high dietary levels of amino acids

Increasing the proportion of an amino acid mixture corresponding to casein in a low-fat, cholesterol-free, semipurified diet fed to rabbits causes a progressive increase in serum total and low density lipoprotein cholesterol, and the effect appears to be due primarily to the essential amino acids in the mixture. Our recent studies have also shown that the variations in serum cholesterol in response to different levels of the amino acid mixture are not associated with any changes in fecal excretion of cholesterol or bile acids. Further attempts to understand the mechanism of action of dietary amino acids on serum cholesterol levels have shown the following: (1) no correlation with levels of plasma amino acids, either in the fasting or postprandial state: (2) no correlation with serum levels of thyroid hormones: (3) no relationship to activity of hepatic or intestinal microsomal hydroxymethylglutaryl coenzyme A reductase: (4) no corresponding effects on the activities of cholesterol esterifying enzymes of intestinal mucosa: and (5) no correlation with the degree of esterification of cholesterol in very low or low density lipoproteins. Further studies are required to identify the specific amino acids responsible for the hypercholesterolemic effects and to determine the mechanisms involved.     

Fish protein hydrolysate elevates plasma bile acids and reduces visceral adipose tissue mass in rats

Conjugation of bile acids (BAs) to the amino acids taurine or glycine increases their solubility and promotes liver BA secretion. Supplementing diets with taurine or glycine modulates BA metabolism and enhances fecal BA excretion in rats. However, it is still unclear whether dietary proteins varying in taurine and glycine contents alter BA metabolism, and thereby modulate the recently discovered systemic effects of BAs. Here we show that rats fed a diet containing saithe fish protein hydrolysate (saithe FPH), rich in taurine and glycine, for 26 days had markedly elevated fasting plasma BA levels relative to rats fed soy protein or casein. Concomitantly, the saithe FPH fed rats had reduced liver lipids and fasting plasma TAG levels. Furthermore, visceral adipose tissue mass was reduced and expression of genes involved in fatty acid oxidation and energy expenditure was induced in perirenal/retroperitoneal adipose tissues of rats fed saithe FPH. Our results provide the first evidence that dietary protein sources with different amino acid compositions can modulate the level of plasma bile acids and our data suggest potential novel mechanisms by which dietary protein sources can affect energy metabolism.     

The utilization of nitrogen for growth in mice fed blends of purified proteins

Reasons for differences in growth of weanling mice fed different blends of purified proteins in diets apparently adequate in indispensable amino acids were investigated. Dietary nitrogen provided at increasing levels from a mixture of casein, gelatin, and zein, or from these plus gliadin and lactalbumin resulted in similar weight gains and efficiency of feed utilization in weanling mice. These responses were always lower (P less than 0.05) than for mice fed the control diet. When diets were based on true digestible nitrogen and amino acids, weight gains and feed efficiency decreased (P less than 0.05) as the dietary nitrogen level increased. High plasma glycine concentrations and high phenylalanine to tyrosine ratios were associated with high dietary glycine levels provided by gelatin. When a mixture of constant proportions of protein sources, predominantly casein, supplied most of the dietary nitrogen, small increases in weight gains and feed efficiency were observed as the nitrogen level in the diet increased. Varying the proportions of sources, which increased the percentage of gelatin, resulted in decreased gains in response to more dietary nitrogen. Replacing the amino acids found in gelatin by crystalline L-amino acids and replacing glycine by glutamic acid improved gains and feed efficiency (P less than 0.05) compared with gelatin-containing diets. These studies demonstrated that glycine at dietary concentrations of approximately 1-2%, either from gelatin or as the free amino acid, inhibited growth and feed utilization of growing mice.     

Intrauterine Malnutrition and the Brain: Effects on Enzymes and Free Amino Acids Related to Glutamate Metabolism

Abstract: The effect of feeding pregnant rats with wheat and Bengal gram (black chick pea) diets during the later part of pregnancy on brain growth, enzymes, and free amino acids of glutamate metabolism in 1-day-old rats was investigated. These diets did not induce growth dissociation, and the body and brain weights were equally affected. The concentrations of DNA, RNA, protein, and free α-amino nitrogen in brain decreased significantly and the activities of glutamine synthetase, glutamine transferase, glutaminase 1, glutaminase 11, and glutamate decarboxylase and the concentrations of free amino acids, glutamic acid, glutamine, alanine, and GABA were also decreased. The concentration of aspartic acid, however, was increased. Wheat and Bengal gram diets fortified with lysine and with methionine, cystine, and tryptophan respectively showed various beneficial effects on the changes observed in the brain. A 20% casein diet induced higher body and brain weights and better brain protein and free α-amino nitrogen concentrations than those observed on a 10% casein diet.     

Acetyl Groups in Native Glucomannan from Easter Lily Bulbs

The in vitro digestibility of rice glutelin and wheat glutenin was investigated with a view to assessing their nutritional qualities, using casein and bovine serum albumin (BSA) as references. The following hydrolytic processes were adopted: pepsin-pancreation digestion (a model system before intestinal absorption) and aminopeptidase-prolidase hydrolysis [a model system for the intestinal mucosa (membrane digestion) and after intestinal absorption (intracellular hydrolysis)]. The pepsin-pancreatin digests were first examined. The degree of amino acid released from the proteins was 30% (glutelin), 23% (glutenin), 24% (casein) and 30% (BSA). A similar release pattern of individual amino acids was observed for all the proteins. The amounts of large peptide fractions increased in the order: glutelin < glutenin < casein < BSA. Glutelin was highly digestible. Apart from containing high amounts of glutamic acid (glutamine), cystine and proline, the large peptide fractions of glutelin were also rich in threonine, glycine and isoleucine while those of glutenin were only rich in glycine. The aminopeptidase-prolidase digests were examined next. Glutelin was almost completely hydrolyzed to amino acid, except for a low release of cystine, suggesting that the amino acid residues constituting glutelin could be easily utilized as nutrients in the living tissues. The degree of amino acid released from the proteins was 97% (glutelin), 93% (glutenin), 90% (casein) and 79% (BSA).

The convenient application of these model systems for the assessment of the in vitro digestibility of food proteins have been discussed.     

Intake of Hydrolyzed Casein is Associated with Reduced Body Fat Accretion and Enhanced Phase II Metabolism in Obesity Prone C57BL/6J Mice

The amount and form of dietary casein have been shown to affect energy metabolism and lipid accumulation in mice, but the underlying mechanisms are not fully understood. We investigated 48 hrs urinary metabolome, hepatic lipid composition and gene expression in male C57BL/6J mice fed Western diets with 16 or 32 energy% protein in the form of extensively hydrolyzed or intact casein. LC-MS based metabolomics revealed a very strong impact of casein form on the urinary metabolome. Evaluation of the discriminatory metabolites using tandem mass spectrometry indicated that intake of extensively hydrolyzed casein modulated Phase II metabolism associated with an elevated urinary excretion of glucuronic acid- and sulphate conjugated molecules, whereas glycine conjugated molecules were more abundant in urine from mice fed the intact casein diets. Despite the differences in the urinary metabolome, we observed no differences in hepatic expression of genes involved in Phase II metabolism, but it was observed that expression of Abcc3 encoding ATP binding cassette c3 (transporter of glucuronic acid conjugates) was increased in livers of mice fed hydrolyzed casein. As glucuronic acid is derived from glucose and sulphate is derived from cysteine, our metabolomic data provided evidence for changes in carbohydrate and amino acid metabolism and we propose that this modulation of metabolism was associated with the reduced glucose and lipid levels observed in mice fed the extensively hydrolyzed casein diets.     

Vitamin requirements of larvae of Sitophilus oryzae

Improvements were made in the meridic larval diet for Sitophilus oryzae by replacing the minerals and vitamins supplied by dietary brewer's yeast and wheat germ with mineral and vitamin mixtures. The effects of different concentrations of individual vitamins were studied with the improved diet containing 20% casein. In later tests the dietary casein was replaced with a mixture of 15 amino acids. The results indicated that these larvae, which contain an associated bacteria-like micro-organism, required thiamine, nicotinic acid, pyridoxine, folic acid, and biotin in the diet. No requirement for riboflavin or pantothenic acid could be demonstrated on either the 20% casein diet or the amino acid diet possibly due to contamination of the cornstarch with these two vitamins. In addition, the larvae did not require choline or inositol for the growth of one larval generation.The asymbiotic larvae of S. granarius failed to develop on the improved casein diet indirectly implicating the symbiotes in a nutritional rôle; however, the symbiotes present in S. oryzae apparently do not provide B vitamins. Larvae of S. oryzae failed to develop when the concentration of casein was reduced to 10% while growth was maintained with a 10% concentration of the amino acid mixture. Casein is not an optimal source of amino acids for this species.     

Effect of the Supplementation of Sulfur Amino Acids to a Diet Containing Soy Protein Isolate on Lipid Metabolism in Rats

The effect of the supplementation of sulfur amino acids to a low casein or soy protein isolate diet on tissue lipid metabolism was investigated. Supplementation of methionine to a 8% casein diet produced a fatty liver in rats, however, supplementation of methionine to a 8.8% soy protein diet (corresponding to a 8% casein diet as to sulfur amino acids content) did not produce a fatty liver. At the level of 8% or less of soy protein in the diet, the accumulation of liver lipids and serum triglyceride was observed. An amino acid mixture simulating the composition of soy protein isolate caused significant accumulation of liver lipids, but serum triglyceride was not changed. Serum cholesterol in rats fed the soy protein diet was lower than that in rats fed the casein diet, but on feeding the amino acid mixtures simulating these protein diets, there was no difference between the two groups. The small differences between soy protein isolate and casein as to lipid metabolism might be due to the small differences in the contents of sulfur amino acids or the specific nature of the soy protein or casein. The supplemental effect of methionine and cystine was also studied. About 60% of total sulfur amino acids could be substituted by cystine for maximum growth.     

Regional distribution of homocarnosine and other ninhydrin-positive substances in brains of malnourished monkeys

—Eight male monkeys (Macaca nemestrina) aged 6–9 months were divided into two groups and fed either an adequate protein diet (20% casein) or a protein deficient diet (2% casein). After 3- 5 months of receiving the low protein diet, the malnourished monkeys showed extensive fatty metamorphosis of the liver cells, distorted patterns of plasma and hepatic free amino acid pools, and other features consistent with the diagnosis of protein-calorie malnutrition. Examination of the cerebrum, cerebellum and brain stem in the malnourished animals revealed profound accumulation of 3-methylhistidine, histidine and homocarnosine in all three regions. For histidine, the cerebral, cerebellar and brain stem levels in the protein deficient animals increased by 145, 104 and 101 per cent over levels observed in corresponding regions of the brain in well-fed monkeys. Similarly, there were significant elevations in homocarnosine contents of the cerebrum (+ 99 per cent), cerebellum (+ 140 per cent) and brain stem (+ 146 per cent) in comparison to levels in control animals. In contrast, the levels of valine, serine and aspartic acid were markedly reduced in all three brain areas in the malnourished animals. Protein-calorie deficiency also produced reductions in the brain levels of taurine, glutamic acid, isoleucine, leucine and threonine which varied in magnitude in the three major regions of the brain examined. These biochemical alterations which may in part underlie some of the psychomotor changes often observed in protein-calorie malnutrition, were discussed not only in relation to the role of amino acids as precursors for the synthesis of neuroregulatory substances but also with due regard to the possibility that some of these ninhydrin-positive substances such as GABA, homocarnosine, glycine and the dicarboxylic amino acids may possess neuroexcitatory or inhibitory properties in various parts of the central nervous system.