The modulating effect of amino acids on an organotypic culture of lymphoid tissue

The effect of 20 essential and nonessential L-amino acids on the dynamics of development of spleen explants from 1-and 21-day-old rats on an organotypic tissue culture was studied. The hydrophilic amino acids with a higher molecular mass (asparagine, lysine, arginine, and glutamic acid) induced an inhibitory effect on the growth zone of explants of immature tissue from 1-day-old animals and an opposite, stimulating effect on the mature spleen tissue of 21-day-old rats. An immunohistochemical analysis revealed a reciprocal correlation between the expression of the proapoptotic protein p53 and the cell proliferation upon the action of lysine, asparagine, and glutamic acid. The role of polar amino acids in the modulation of cell proliferation and apoptosis in dependence on the period of ontogenesis was determined.     

The modulating effect of amino acids on an organotypic culture of lymphoid tissue

The effect of 20 essential and nonessential L-amino acids on the dynamics of development of spleen explants from 1- and 21-day-old rats on an organotypic tissue culture was studied. The hydrophilic amino acids with a higher molecular mass (asparagine, lysine, arginine, and glutamic acid) induced an inhibitory effect on the growth zone of explants of immature tissue from 1-day-old animals and an opposite, stimulating effect on the mature spleen tissue of 21-day-old rats. An immunohistochemical analysis revealed a reciprocal correlation between the expression of the proapoptotic protein p53 and the cell proliferation upon the action of lysine, asparagine, and glutamic acid. The role of polar amino acids in the modulation of cell proliferation and apoptosis in dependence on the period of ontogenesis was determined. The English version of the paper: Russian Journal of Bioorganic Chemistry, 2006, vol. 32, no. 3; see also http://www.maik.ru.     

The modulating action of amino acids upon development in an organotypic culture

The effect of L-amino acids in concentration 0.05 ng/ml was investigated in organotypic tissue culture of spleen, liver and brain cortex at the age of 1 and 21 days. The high molecular and low hydrophobic amino acids: asparagin, lysine, arginine, and glutamine, are active inducing both a less intensive growth zone in explants of all three tissue types in 1-day old rats and more intensive growth zone in spleen, liver explants in 21-day rats as compared to control. Another group of low molecular and high hydrophobic amino acids is active inducing a more intensive growth zone in brain cortex explant in 21-day rats, as compared to control. The data obtained suggest a modulating role of amino acids in the tissues at different stages of maturation.     

The effect of the coded amino acids on meso- and ectodermal tissue development in organotypic culture

The effect of L-amino acids was investigated in organotypic tissue culture ofmesodermal tissue (spleen, myocardium) and ectodermal tissue (brain cortex) in mature rats. The low hydrophobic amino acids: asparagine, hystidine, serine, lysine, arginine and glutamine acid, induced the proliferation stimulation. The high hydrophobic amino acids had both the apoptose effect (spleen) and no effect at all (myocardium). The proliferation stimulation occurred in the ectodermal tissue under the effect of the high hydrophobic amino acids (asparagines acid, valine, threonine, methionine, leucine, isoleucine), whereas the low hydrophobic amino acids had no effect on the nervous tissue development. The combination of two amino acids one of which stimulated and another one inhibited the explant growth zone (or was not active in myocardium) lead to an increase of the stimulatory effect in meso- and ectodermal tissue. The amino acid modulated properties can be taken in consideration in synthesis of new regulatory peptides.     

General control of amino acid biosynthesis in mutants of Candida spec. EH 15/D

The general control of amino acid biosynthesis was investigated in Candida spec. EH 15/D, using single and double mutant auxotrophic strains and prototrophic revertants starved for their required amino acids. These experiments show that starvation for lysine, histidine, arginine, leucine, threonine, proline, serine, methionine, homoserine, asparagine, glutamic acid or aspartic acid can result in derepression of enzymes. A correlation was found between the degree of derepression, growth of strains, and concentration of required amino acids. The amino acids pool pattern of mutants and revertants is different from that in the wild type strain.     

Effect of thymus polypeptide fractions on the development of rat thymus and spleen in organ culture

Peptides of the thymus--vilon, thymogen and thymalin, alone or in combination with concanavalin A, were used to investigate their effect on organotypic culture of thymus and spleen explants from 1- and 21-day old rats. Vilon, thymogen and thymalin in concentrations of 2 and 10 ng/ml and 5 ng/ml, resp., exerted stimulating effects in thymus and spleen tissue cultures from 21-day old rats as compared to the control explants. Vilon and thymogen showed inhibiting effect in the thymus tissue cultures from 1-day old rats as compared to the control explants. However, the peptides together with concanavalin A in concentration of 10 mkg/ml resulted in decreasing the action of concanavalin A alone. The polypeptide fractions of thymus and their synthetic analogs play different roles in the regulation of thymus and spleen development in rats of different age.     

Influence of Nitrogen and Sucrose in the Medium and of Irradiance of the Stock Plants on Root Formation in Pelargonium Petioles Grown in vitro

In vitro root formation was studied in petiole explants from Pelargonium×hortorom Bailey cv. Radio exposed to different irradiance (2.5. 11.6 or 23.0 W/m²). Optimal root formation was found when stock plants were subjected to 2.5 W/m² and explants were grown on a medium with 20 g/I of sucrose and 5.0 mM of nitrogen. The number of roots per explant decreased with increasing nitrogen or sucrose concentrations in the growth medium and with increasing irradiance applied to the stock plants. Variation in temperature (17, 21 or 25°C) did not affect root formation. The number of roots per explant was higher in explants from young petioles than in explants from older petioles. The initial nitrogen content of the explants was highest at the lowest irradiance. The content of reducing sugars and sucrose increased with increasing irradiance. The endogenous levels of lysine, arginine and ornithine were highest at 2.5 W/m² and the levels of proline. alanine, γ-aminobutyric acid, glutamic acid, glutaminc, aspartic acid and asparagine were highest at 23.0 W/m². With rising nitrogen concentration in the medium, the final endogenous content of nitrogen in the explants increased, whereas the content of reducing sugars decreased. When the sucrose concentration in the medium increased, the endogenous content of reducing sugars rose, and the nitrogen level was lowered. Variations in the nitrogen concentration in the growth medium influenced the final level of free amino acids in the explants.     

INFLUENCE OF AMMONIUM AND NITRATE ON THE PROTEIN- AND FREE AMINO ACIDS IN SHOOTS OF WHEAT SEEDLINGS

Weissman , Gerard S. (Rutgers U., Camden, N. J.) Influence of ammonium and nitrate on the protein- and amino acids in shoots of wheat seedlings. Amer. Jour. Bot. 46(5): 339–346. 1959.—Total and protein nitrogen per shoot of wheat seedlings grown with endosperm attached increased at a steady rate during a 96-hr. growth period, and protein nitrogen, as a percentage of total nitrogen, remained constant at about 53%. Total and protein nitrogen concentration was greatest for 24-hr. shoots and declined as the shoots became older. Total and protein nitrogen were determined in 96-hr. shoots of seedlings grown with endosperm attached but also supplied with ammonium, nitrate, or both in the culture solution. Total nitrogen was greatest in shoots supplied with ammonium, but only 38% was in the form of protein. Maximum protein synthesis occurred in shoots grown in both ammonium and nitrate and protein nitrogen as a percentage of total nitrogen approximated that achieved in shoots lacking nitrogen in the culture solution. The protein amino acid composition of 48-, 72-, and 96-hr. shoots was very similar but differed from 24-hr. shoots which contained higher percentages of arginine and lysine and lower percentages of alanine and threonine. This may be correlated with the higher proportion of meristematic cells in 24-hr. shoots. The protein amino acids in shoots grown with ammonium resembled that of shoots lacking nitrogen in the culture solution, but nitrate shoot protein contained a higher percentage of arginine and a lower percentage of lysine. Nitrate may stimulate the formation of enzymes, possibly of a nitrate-reducing system, with high arginine- low lysine content. Free asparagine and glutamine were both at a maximum in ammonium shoots and at a minimum in nitrate shoots, but asparagine predominated in shoots supplied with ammonium while glutamine was greatest in nitrate shoots. Aspartic acid, asparagine, and glutamine appeared to have ammonia-storage functions, but glutamic acid appeared to be primarily concerned with protein synthesis. Amino acid accumulation was greatest in shoots supplied with both ammonium and nitrate. Protein synthesis in these appeared to be limited by inadequate concentrations of glutamic acid and proline. A hypothesis is proposed in explanation of the high glutamic acid concentration in shoots provided with ammonium and nitrate.     

Utilization and requirements of amino acids by a cell line derived from the butterfly Papilio xuthus

The media, in which a butterfly cell line (Px 58), derived from pharate adult ovaries of Papilio xuthus cultured for 8 days, were analysed to examine the changes in free amino acids in the medium during cultivation. Beta-alanine, arginine, glycine, histidine, lysine, phenylalanine, proline, serine, and tryptophan did not change markedly. Asparagine, aspartic acid, cystine, glutamine, isoleucine, leucine, methionine, threonine, tyrosine, and valine decreased to some extent with culturing. Alpha-alanine increased markedly, and glutamic acid did so to a lesser extent. Requirements of amino acids by the cell line were examined by deleting amino acids one at a time. Deletion of alpha-alanine, beta-alanine, asparagine, glutamic acid, glycine, and phenylalanine did not cause deterioration of the cell. These amino acids were thought to be non-essential or required only a little. Deletion of other amino acids impaired the cell growth severely. These amino acids would appear to be essential for growth of the Px 58 cell line.     

Amino Acids as Nitrogen Sources for the Growth of Euglena gracilis and Astasia longa

SYNOPSIS. Euglena gracilis (bacillaris variety, strain SM-L1, streptomycin-bleached) used the following amino adds (10⁻³ M) as sole nitrogen source for growth on a defined medium: glycine, alanine, valine, leucine, isoleucine, serine, threonine, and glutamic acid. Aspartic acid was used at 10⁻² M. Glutamine and asparagine were used at 10⁻³ M and were better N sources than their parent dicarboxylic amino acids. Not used as sole N source for growth were phenylalanine, tyrosine, tryptophan, cysteine, cystine, methionine, proline, hydroxyproline, histidine, arginine, lysine, and taurine. Astasia longa (Jahn strain) was more restricted than Euglena and used only asparagine and glutamine as N sources for growth.     

Transport of Glutamate and Glutamine in the Light Leaf Spot Pathogen Pyrenopeziza brassicae

The kinetics of glutamic acid and glutamine uptake in the light leaf spot fungus, Pyrenopeziza brassicae , are biphasic. At low and high concentrations, glutamic acid and glutamine share a high-affinity and a low-affinity carrier, respectively, with K_ms of 4.0 and 4.4 μ m for uptake of glutamic acid and glutamine, respectively, by the high-affinity system, and K_m s of 580 and 560 μ m for uptake of glutamic acid and glutamine by the low-affinity system. The data suggest that glutamic acid and glutamine are taken up by a different system to that responsible for the uptake of ornithine, arginine, lysine and asparagine, and may represent system IV described in Neurospora crassa for the uptake of acidic amino acids.     

Plasma amino acid concentrations in pregnant rats and in 21-day foetuses.

Plasma amino acid concentrations were determined in virgin female rats, in pregnant rats (12 and 21 days after impregnation) and in 21-day foetuses. The total amino acid concentration in plasma decreases significantly with pregnancy, being lower at 12 than at 21 days. Alanine, glutamine+glutamate and other 'gluconeogenic' amino acids decrease dramatically by mid-term, but regain their original concentrations at the end of the pregnancy. With most other amino acids, mainly the essential ones, the trend is towards lower concentrations which are maintained throughout pregnancy. These data agree with known nitrogen-conservation schemes in pregnancy and with the important demands on amino acids provoked by foetal growth. In the 21-day foetuses, concentrations of individual amino acids are considerably higher than in their mothers, with high plasma foetal/maternal concentration ratios, especially for lysine, phenylalanine and hydroxy-proline, suggesting active protein biosynthesis and turnover. All other amino acids also have high concentration ratios, presumably owing to their requirement by the foetuses for growth. Alanine, glutamine+glutamate, asparagine+aspartate, glycine, serine and threonine form a lower proportion of the total amino acids in foetuses than in the virgin controls or pregnant rats, probably owing to their role primarily in energy metabolism in the adults. The results indicate that at this phase of foetal growth, the placental amino acid uptake is considerable and seems to be higher than immediately before birth.     

The modulating role of essential and non-essential amino acids in organotypic tissue culture in rats of different ages

The effect of amino acids L-lysin, L-asparagin, L-arginin, L-glutamate was investigated in organotypic tissue culture of spleen, liver and brain cortex of rats at different age. The amino acids in concentration 0.05 ng/ml are active inducing a less intensive growth zone, as compared to control, in 1-day and in older rats--an intensive growth zone, as compared to control in 21-day rats. The data obtained suggest a modulating role of amino acids in the tissues at different stages of maturation.     

Response of a wild type and a non-nitrogen-fixing mutant of Anabaena doliolum towards different amino acids

The effects of various amino acids on growth and heterocyst differentiation have been studied on wild type and a heterocystous, non-nitrogen-fixing (het+ nif-) mutant of Anabaena doliolum. Glutamine, arginine and asparagine showed maximum stimulation of growth. Serine, proline and alanine elicited slight stimulation of growth of wild type but failed to show any stimulatory effect on mutant strain. Valine, glutamic acid, iso-leucine and leucine at a concentration of as low as 0.1 mM were inhibitory to growth of parent type. Methionine, aspartic acid, threonine, cysteine, and tryptophan did not affect growth at concentrations lower than 0.5 mM. But at 1 mM, these amino acids were inhibitory. In addition to the stimulatory effects of glutamine, arginine and asparagine, the heterocyst frequency was also repressed by these amino acids. Glutamine and arginine at 2 mM completely repressed heterocyst differentiation in the mutant strain; however, other amino acids failed to repress the differentiation of heterocysts. Our results suggest that glutamine and arginine are utilized as nitrogen sources. This is strongly supported from the data of growth and heterocyst differentiation of mutant strain, where at least with glutamine there is good growth without heterocyst formation. Studies with glutamine and arginine on other N2-fixing blue-green algae may reveal the regulation of the heterocyst-nitrogenase sub-system.     

Stimulation of Lysine Decarboxylase Production in Escherichia coli by Amino Acids and Peptides

A commercial hydrolysate of casein stimulated production of lysine decarboxylase (EC 4.1.1.18) by Escherichia coli B. Cellulose and gel chromatography of this hydrolysate yielded peptides which were variably effective in this stimulation. Replacement of individual, stimulatory peptides by equivalent amino acids duplicated the enzyme levels attained with those peptides. There was no indication of specific stimulation by any peptide. The peptides were probably taken up by the oligopeptide transport system of E. coli and hydrolyzed intracellularly by peptidases to their constituent amino acids for use in enzyme synthesis. Single omission of amino acids from mixtures was used to screen them for their relative lysine decarboxylase stimulating abilities. Over 100 different mixtures were evaluated in establishing the total amino acid requirements for maximal synthesis of lysine decarboxylase by E. coli B. A mixture containing all of the common amino acids except glutamic acid, aspartic acid, and alanine increased lysine decarboxylase threefold over an equivalent weight of casein hydrolysate. The nine most stimulatory amino acids were methionine, arginine, cystine, leucine, isoleucine, glutamine, threonine, tyrosine, and asparagine. Methionine and arginine quantitatively were the most important. A mixture of these nine was 87% as effective as the complete mixture. Several amino acids were inhibitory at moderate concentrations, and alanine (2.53 mM) was the most effective. Added pyridoxine increased lysine decarboxylase activity 30%, whereas other B vitamins and cyclic adenosine 5′-monophosphate had no effect.     

Polypeptide synthetase activity of chromatin from eukaryotic cells]

It is shown that the polypeptide synthetase activity (PS-activity) of chromatin from rat liver is increased 9--21 hrs after partial hepatectomy. Among 9 amino acids studied alanine, methionine, lysine, tyrosine and arginine are not incorporated into the system in question. The highest rate of polymerization is observed in case of glutamic acid. The rate of glutamine, asparagine and glycine incorporation is 7--8 times slower. The PS-activity of chromatin is enhanced by chromatin preincubation with NAD (but not with its analogs). The activation is prevented by thymidine and nicotinamide. Storage of chromatin for 18 hrs at 2--4 degrees C results in a complete loss of PS-activity. Ability of "old" chromatin to incorporate of amino acids may be restored by its preincubation with NAD. Storage of chromatin in the presence of 5 mM cAMP does not decrease the PS-activity. It is assumed that in the system described poly-ADP ribose is an energy source for amino acid activation.     

Regulation of Growth and Gametangial Formation in Riccia gangetica Ahmad by Some Amino Acids

The present work deals with the effect of six amino acids: asparagine,aspartic acid, glutamic acid, glycine, serine, and tryptophan,on growth and gametangial formation in Riccia gangetica. Allthe amino acids tested enhance vegetative growth, and amongthese glutamic acid proves best. The total number of rhizoidsis reduced in response to amino acids. Aspartic acid and glutamicacid favour antheridial production. In contrast, asparagine,serine, and tryptophan enhance archegonial formation, and amongthese asparagine elicits the best response. Glycine proves bestfor antheridial production, and also increases the number ofarchegonia. Key words: Riccia gangetica, Amino acids, Growth, Gametangial formation     

A comparison of amide and ureide nitrogen sources in tissue culture of tree legume Prosopis alba clone B2 V50

The effects of medium nitrogen sources on the recalcitrant nature of Prosopis alba clone B2V50 in tissue culture were compared involving shoot development using axillary bud explants from 2 to 4-year-old greenhouse-grown trees. A significant difference (P<0.05) was found between the amino acids aspartic acid and glutamic acid and their corresponding amide-containing compounds asparagine and glutamine. A comparison between amide and ureide nitrogen sources showed that allantoin, a ureide, was an acceptable replacement for asparagine or glutamine. Allantoin, asparagine, and glutamine could be used as the sole nitrogen sources. Allantoin at a concentration of 20 mM was adopted for use in future research. Although shoots were consistently induced, all explants showed complete shoot-tip necrosis after 12 weeks of in vitro culture.     

Amino adids as nitrogen sources for the growth of rice callus tissue

Availability of amino acids for the growth of rice callus tissuewas examined by supplying various kinds of amino acids to thetissue separately or in combination. When an amino acid wassupplied alone as the sole source of nitrogen, only the followingfive amino acids were found to favour the growth of callus tissue;arginine, alanine, asparagine, glutamine and proline. In combinationwith other acids, both AHCH and CMAA were very effective instimulating the growth of tissue, but EHCH was inhibitory. Whenmethionine or arginine was excluded from the CMAA medium, callusgrowth on the medium was reduced significantly. The effect producedby omitting methionine suggested that some amino acid interactionwas involved in this instance. (Received February 19, 1970; )     

E.s.r. of spin-trapped radicals in aqueous solutions of amino acids. Reactions of the hydroxyl radical.

The radicals produced by reactions of hydroxyl radicals with amino acids in aqueous solutions have been investigated. Hydroxyl radicals were formed by U.V.-photolysis of hydrogen peroxide and the short-lived amino acid radicals were spin-trapped by tert-nitrosobutane and identified by electron spin resonance spectroscopy. Nineteen amino acids were studied, and several radicals were identified which have not been observed previously by other methods. Only side-chain radicals were identified for alanine, threonine, aspartic acid, asparagine, lysine, phenylalanine, tyrosine, proline and hydroxyproline; whereas for glycine the C(2) carbon radical was spin-trapped. Both C(2) carbon radicals and side-chain radicals were assigned to valine, leucine, isoleucine, serine, glutamic acid, glutamine, arginine and methionine.