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Comparison of 14C-amino acid mixture and [35S]methionine labeling of cellular proteins from mouse fibroblast C3H10T1/2 cells by two-dimensional gel electrophoresis.

Total cellular proteins from mouse C3H10T1/2 fibroblasts were compared by two-dimensional (2-D) gel electrophoresis after radiolabeling with [35S]methionine (35S-Met) or 14C-amino acids (14C-AA). 35S-Met labeling of protein was three to four times greater than 14C-AA incorporation over a 24 h period. Automated comparative analysis of replicate fluorographs after 6, 12, and 24 h of labeling showed considerable homology between radiolabeling methods. More than 88% percent of 35S-Met and 14C-AA-labeled proteins were common at each time point. However, the total number of 35S-Met-labeled proteins dropped from 6 to 24 h while the number of 14C-AA-labeled proteins increased. Additionally, twenty-one proteins were uniquely labeled by 14C-AA that were not detectable by 35S-Met over the labeling period. Densitometric analysis showed that several 35S-Met and 14C-AA-labeled proteins exhibited time-related differences in radiolabel incorporation while most proteins remained relatively constant. Protein patterns of silver-stained gels from 6 to 24 h were highly registered and showed few qualitative differences. Proteins detected in radiolabeled gels were generally, but not always, found in silver-stained gels. Thus, 35S-Met appears better suited for short-term radiolabeling of cellular protein while more comprehensive labeling of protein occurs with 14C-AA during prolonged incubation of cell cultures under present experimental conditions.

Amino Acids↗

[Enzyme preparations for research on protein hydrolysates and amino acid mixtures not containing peptides].

Some properties and the hydrolysing ability of two novel enzyme preparations, a proteolytic preparation "C" from Acremonium chrysogenum (Cephalosporium acremonium) and a peptidase preparation (the producer from the family Pseudomanadaceae), are described. The preparations can be used for obtaining protein hydrolysates with different ratios of free amino acids and peptides. The protein hydrolysis with the preparation "C" enables one to obtain hydrolysates containing 13-18% of free amino acids. The further treatment of the hydrolysates with the peptidase preparation results either in complete hydrolysis of the remained peptide fractions or in obtainment of solutions containing from 60 to 85% of free amino acids and low-molecular weight peptides.

Amino Acids↗

Subchronic treatment with amino acid mixture of L-lysine and L-arginine modifies neuroendocrine activation during psychosocial stress in subjects with high trait anxiety.

The aim of the present study was to evaluate possible modulatory effect of the treatment with L-lysine and L-arginine on neuroendocrine activation during psychosocial stress in healthy subjects with relatively high trait anxiety in a randomized, double blind placebo controlled trial. In 29 healthy subjects at the upper limit of the normal range of a trait anxiety scale, a mixture of L-lysine and L-arginine (3 g each/day) was administered for 10 days followed by exposure to a psychosocial stress procedure based on public speech. Hormone levels, cardiovascular activation and skin conductance were measured. Amino acid treatment resulted in enhanced adrenocorticotropic hormone, cortisol, adrenaline and noradrenaline levels and galvanic skin responses during stress compared to those in placebo-treated group. Increases in the heart rate and blood pressure in response to public speaking task were not influenced by amino acid treatment. Results of the present study support the hypothesis that L-lysine in combination with L-arginine, which may induce anxiolytic effects, modify hormonal responses during psychosocial stress in humans. Such action may represent a normalization of hormone levels to the pattern observed previously in subjects with low trait anxiety.

Adrenocorticotropic Hormone↗

The use of a crystalline amino acid mixture for parenteral nutrition in low-birth-weight infants.

Thirty-six preterm, sick, low-birth-weight neonates were given either total or partial parenteral nutrition. The patients were divided into three groups according to their birth weights: group A -less than 1,000 gm, 12 patients: group B--between 1,000 and 1,500 gm, 15 patients: group C--more than 1,500 gm, 9 patients. The solution for total parenteral nutrition contained 20% glucose and 2.6% crystalline amino acids plus appropriate amounts of vitamins and minerals. The volume of infusate given was usually 125 ml/kg/day, but varied depending on the clinical condition of the patient; occasionally it was as high as 150 to 175 ml/kg/day. Infusate of one-half strength was administered initially; its concentrations of glucose and amino acids were increased to three quarters and full strength gradually, if tolerated. The solution for total parenteral nutrition was infused into the superior vena cava via a central venous catheter; that for partial parenteral nutrition was given into a peripheral vein to supplement inadequate oral feedings. The period of parenteral nutrition lasted from 5 to 49 days, with an average of 13.2 days. The intake of 500 mg of nitrogen as crystalline amino acids and 100 kcal as glucose was capable of achieving body weight gain. Positive nitrogen balance of various degrees was also observed. Hyperglycemia of a slight to moderate degree was observed in nine patients; only three required insulin therapy. Two patients had thrombotic occlusion of the central venous catheter. The conclusion was reached that total parenteral nutrition or partial parenteral nutrition, when properly managed, is a safe procedure in small, premature infants. The amino acid solution given as a nitrogen source along with adequate calories was effective in promoting weight gain and nitrogen balance; it was apparently well tolerated by low-birth-weight neonates.

Amino Acids↗

[Changes in rat liver chromatin after administration of a mixture of amino acids].

It was shown that injections of an amino acid mixture essentially increase the number of nuclease-sensitive regions of chromatin and its active fraction (Mg2+-soluble fraction), while hydrocortisone increases the amount of the latter, which is less sensitive to the effect of DNAase II. Genome activation during nonhormonal induction after administration of the amino acid mixture or during hydrocortisone injection reflects in different ways on the parameters of melting of chromatin active fractions and on the relative content of its protein fractions.

Amino Acids↗

Eutectic formation analysis of amino acid mixtures using molecular dynamics simulations.

The mechanism of eutectic formation was investigated via computer-aided molecular dynamics techniques based on experimental results. The CBZ group mixtures CBZ-l-Asp/d-AlaNH2 x HCl/methanol, CBZ-l-Asp/l-PheOMe x HCl/methanol, and CBZ-l-Tyr/l-ArgNH2 x 2HCl/methanol formed transparent eutectic melts. The non-CBZ group mixtures l-Asp/d-AlaNH2 x HCl/methanol, l-Asp/l-PheOMe x HCl/methanol, and l-Tyr/l-ArgNH2 x 2HCl/methanol did not form eutectic melts. According to molecular dynamics simulation results, increase in the kinetic energy values of eutectic forming mixtures was much larger than the increase in potential energy over a temperature shift from 298 to 333 K. However, the results for non-eutectic forming mixtures were reversed. The Coulomb interaction energies of eutectic forming mixtures significantly decreased, because eutectic melting can increase the mobility of molecules in the mixtures. The enhancement of molecular mobility was confirmed by increased self-diffusion constant values, and the change of solid-to-liquid phase was detected by radial distribution function results. The periodic boundary conditions for calculation of molecular dynamics were found to be reliable.

Alanine↗

Comparison of an amino acid mixture and protein hydrolysates in treatment of infants with phenylketonuria.

This study compares three feeding regimens for infants with phenylketonuria diagnosed by neonatal screening. Group 1 (five children) received Minafen (Cow & Gate) until they weighed twice their birthweights; Aminogran (Allen & Hanbury) was then added to the feeds in increasing amounts and replaced Minafen at between 8 and 10 months of age. Group 2 (five children) received Aminogran from the neonatal period. Group 3 (five children) received Minafen until they weighed twice their birthweights; Cymogran (Allen & Hanbury) was then added in increasing amounts and replaced Minafen at between 8 and 10 months of age. In all three groups growth was normal and control of phenylalanine levels satisfactory. During the first few months of life the Aminogran regimen proved more complicated and caused more practical difficulties than the regimens starting with Minafen. Later in the first year, when mixed feeding was introduced, and particularly when the bottle was exchanged for the cup, Aminogran had advantages over Cymogran because of its low calorie content, small bulk, and less unpleasant taste. At this age feeding problems were fewer and easier to manage with Aminogran than with Cymogran. A method of using Aminogran in the management of such problems is described. For these reasons, the regimen fed group 1, in which Minafen is used initially and then replaced by Aminogran, is preferred to the other two.

Alkaline Phosphatase↗

[Comparative studies on the enzymatic absorption of protein hydrolysates in the small intestines of the rat. 3. The absorption of trypsin, thermitatic and trypsin thermitatic wheat gluten hydrolysates in comparison with equimolar mixtures of free amino acids].

Tryptic, thermitatic, and tryptic-thermitatic wheat-gluten hydrolyzates as well as their equimolar amino-acid mixture were perfused through proximal and distal parts of the intestine (10 cm length) of non-narcotized rats. The total amino-acid concentration of the perfused solution was 50 mM. The tryptic hydrolyzate showed a significantly lower absorption of nitrogen and total amino acids than the amino-acid mixture. Both the supplied forms were very different as to their absorption pattern of the amino acids. The high variability of the percental absorption of the individual amino acids of the tryptic hydrolyzate results in a high coefficient of variation. The absorption of nitrogen and total amino acids from thermitatic and tryptic-thermitatic hydrolyzates is equal to that from the amino-acid mixture. In a peptidic form glutamic acid is more rapidly absorbed from the two hydrolyzates, and methionine from the tryptic-thermitatic hydrolyzate in both the intestinal parts. As to alanine and glycine this concerns only the distal intestinal part for both the hydrolyzates. There are no differences between the absorption patterns of the two hydrolyzates but in comparison with the amino-acid mixture and the tryptic hydrolyzate differences were evident. The coefficients of variation of both the hydrolyzates are significantly lower as compared to those of the tryptic hydrolyzate and the amino-acid mixture. All forms of supply are more rapidly absorbed in the distal than in the proximal part of the intestine.

Amino Acids↗

Chemical specificity of the effects of branched chain amino acids on glucagon and insulin release in the suckling rat.

The work presented here aims at investigating the in vitro relationship between amino acids and pancreatic hormonal release in the suckling rat. Among the 20 most usual amino acids, 12 of them were tested on perifused pieces of pancreas in 5.5-day-old rats at a low glucose concentration. Their effects were compared to those induced by a 20 amino acid mixture. A 20 amino acid mixture (10.3 mM) stimulated glucagon and insulin release (154 and 208%, respectively). In the mixture, the 4 pooled amino acids Ala-Gly-Ser-Thr (at 1.4 instead of 4.2 mM) did not induce further modification of the glucagon or insulin release. However, the 3 branched chain amino acids Ile-Leu-Val (at 2.9 instead of 1.4 mM) or the following amino acids Arg-Asn-Phe-Pro-Tyr (at 3.2 instead of 1.6 mM) modulated glucagon or insulin release in the presence of the amino acid mixture. In the absence of Ile-Leu-Val, the amino acid mixture had no effect on the insulin release: 122 compared to 208%. This seems specific to the chemical nature of Ile-Leu-Val since in the absence of Arg-Asn-Phe-Pro-Tyr, the amino acid mixture was still effective on the insulin release (178 compared to 208%). In the absence of Ile-Leu-Val, the amino acid mixture was effective on glucagon release, and revealed the cellular specificity of the branched chain amino acid action.

Amino Acids↗

Inhibitory effect of L-methionine-deprived amino acid imbalance using total parenteral nutrition on growth of ascites hepatoma in rats.

The antitumor effect of an amino acid imbalance characterized by the absence of L-methionine was investigated in AH-109A ascites hepatoma-bearing rats with the technique of total parenteral nutrition (TPN). A central venous catheter was placed at the superior vena cava of the rat and hypertonic glucose solution plus an amino acid mixture, an ordinary amino acid compound but lacking L-methionine and L-cysteine, was administered as protein source for seven days. Increases in the volume of ascites and the number of tumor cells were significantly inhibited in the rats receiving glucose plus L-methionine-free amino acid mixture compared to the control rats given glucose only, glucose plus ordinary amino acid mixture and laboratory rations. Unfavorable side effects of TPN with amino acid compound lacking L-methionine and L-cysteine were loss of body weight and development of hypoproteinemia. No evidence of hepatic injury and bone marrow suppression due to this treatment were obtained from the results of biochemical and hematological studies. In liver cells and ascites tumor cells, morphologically, prominent enlargement of the nucleoli was observed in the rats treated with the amino acid mixture lacking of L-methionine and L-cysteine.

Amino Acids↗

Amino acid-restricted diets in the treatment of mammary adenocarcinoma in mice.

The effect of a diet moderately restricted in tryptophan on the tumor and the nutritional status of C3H mice bearing mammary adenocarcinoma was examined. Five groups of female tumor-bearing mice were fed ad libitum a diet containing 1) 15% casein, 2) 15% amino acid mixture, 3) 15% amino acid mixture restricted in tryptophan 4) 15% zein supplemented with lysine and tryptophan or 5) 15% zein supplemented with lysine and restricted in tryptophan. Another group of tumor-free mice of similar age fed 15% casein served as a control for the tumor-bearing group fed the 15% casein diet. Moderate restriction of tryptophan in the diet appeared to have no effect on the tumor status of the animal. Tumor burden usually induced malnutrition in the animal. However, tryptophan restriction did not further aggravate this malnutrition. The source of protein, whether casein, amino acid mixture or zein, appeared to make a difference in nutritional status of tumor-bearing animals though not in tumor status. Casein-fed animals generally tended to have higher values for all the nutritional markers studied than did animals fed the amino acid mixture or zein.

Adenocarcinoma↗