Levels Of Free Glutamine Research Articles

Dietary free glutamine supplementation to increase physiological responses and survival rate of clown loach juvenile, Chromobotia macracanthus Bleeker, 1852

Clown loach Chromobotia macracanthus is one of Indonesian native species and as a key species for ornamental aquaculture. The problem in mass production of this species are the low of growth rate which causes a long period of rearing and low of survival rate. The purpose of the present study was to evaluate free glutamine supplementation at different doses in diet to increase physiological response, growth performance and survival rate of clown loach. Expe-rimental diets contained four different free glutamine levels, viz 0, 1, 2 and 3%. These diets were given to six repli-cate groups of 50 juvenile clown loaches. The fish were reared in each aquarium with dimensions of 40×30×30 cm3 for 60 days. Fish were fed four times a day at satiation. Parameters observed including intestinal glutamine concen-tration, villous and intestinal morphometry, intestinal protease activity, feed efficiency, nutrient retention, growth performance, survival rate, superoxide dismutase (SOD) activity, and malondialdehyde (MDA). The results showed that supplementation of 1% free glutamine significantly affected the morphometry of villi. The length and surface area of villi at a dose of 1% showed the highest values i.e., 320.44 ± 10.39 μm and 27,046.79 ± 250.54 μm2, respect-ively. The 1% dose also had a significant effect on protease activity (13.57 ± 1.92 mg units of protein-1) compared to the 0% dose. The 2% dose showed the highest SOD activity (0.82 ± 0.07 mg protein-1 unit) and the lowest MDA level was found at a dose of 3% (0.25 ± 0.02 nmol mg protein-1). Feed consumption with the supplementation of free glutamine has a significant effect on survival rate with the highest value reached 97.00 ± 1.00%, but no significant effect on intestinal morphometry, feed efficiency, nutrient retention and growth performance. Dietary with the sup-plementation of free glutamine is not able to improve growth performance, but can improve the physiological res-ponse and survival rate.

Glutamine and Glutamate Supplementation Increases the Levels of These Amino Acids in the Milk of Pasture-fed Mares

Background: L-Glutamine (Gln), the most abundant free alpha amino acid in the body, plays a major role in the transport of nitrogen and carbon between tissues, and is an important source of respiratory energy for intestinal and immune system cells. Mares lose lean body mass during lactation, when plasma and milk Gln levels change significantly. However, supplementation with Gln combined with other amino acids may not alter equine plasma Gln levels. The work reported here was designed to test the hypothesis that supplementation with a mixture of glutamine and glutamate (AminoGut) alters blood and milk free glutamine and glutamate levels in pasture-fed lactating mares.Materials, Methods & Results: This study involved 31 multiparous Quarter Horse mares, which were divided into three groups immediately postpartum, as follows: G-CON (n = 19); G-50 g supplemented with 50 g of Gln + Glu plus 200 g of concentrate (n = 6); and G-100 g, supplemented with 100 g of Gln + Glu plus 200 g of concentrate (n = 6). Blood and milk samples were collected on the day of parturition prior to supplementation, and monthly until weaning. The milk samples were used to analyze the Gln, Glu composition and levels, while the blood samples were used for further analysis of blood biomarkers. The results were analyzed by ANOVA and by Tukey’s test and the P value was set at 5%. The G-CON group showed a significant reduction of 11-35% in the mean blood glutamine levels from the first month postpartum and throughout lactation. In contrast, blood glutamine levels in groups G-50 g and G-100 g did not change significantly from parturition through 5 months of lactation. The supplemented groups showed no significant differences in blood variables such as protein, albumin, urea, creatinine, cholesterol, triglycerides and minerals. Free glutamine levels in milk did not change from parturition through the end of lactation in the G-CON group, but groups G-50 g and the G-100 g showed a marked rise in milk glutamine levels throughout the first three months of lactation (~3x), which remained high (~2x) until the foals were weaned (P > 0.05).Discussion: The results of this study indicate that Gln + Glu supplementation successfully increased Gln levels in mare milk in the first three months of lactation, and Glu levels in G-100 g in the first four months, without affecting the levels of these amino acids in the animals’ blood, which remained similar to data obtained at parturition. In fact, the Gln levels in both supplemented groups exceeded 1,000 mmol/mL throughout lactation, unlike those of the control group and of the samples obtained at parturition. Moreover, supplementation did not produce significant changes in blood biomarkers, including those pertaining to protein metabolism (urea, creatinine, uric acid, albumin and total proteins), indicating that the product used for supplementation did not interfere in these biomarkers, which remained within the normal physiological variations for the species. It was concluded that daily dietary supplementation with 50 g of a mixture of glutamine and glutamate produced an effect similar to supplementation with 100 g/day. Both supplementation protocols succeeded in raising glutamine levels in mare milk in the first three months of lactation, without interfering in blood biomarkers or milk composition. In view of the cost of the product, we recommend that the daily diet of mares during lactation be supplemented with 50 g of a mixture of Gln + Glu in order to produce the desired nutritional effects.

Longitudinal Variation of Amino Acid Levels in Human Milk and Their Associations with Infant Gender.

It is discussed that specific amino acids (AAs) have functional roles in early life. Understanding the AA composition in human milk (HM) during lactation assists in specifying these roles. To this end we assessed the levels of free AAs (FAAs), total AAs (free and bound, TAAs) and protein levels in HM in the first 6 months of lactation, and evaluated possible associations with infant gender. HM samples of 25 healthy Dutch mothers participating in the PreventCD study were collected monthly during the first 6 months of lactation. Of the participating mothers, 12 gave birth to a boy and 13 gave birth to a girl. Analyses of the HM samples revealed that levels of free glutamate, glutamine, aspartate, glycine, and serine significantly increased during months 1–3 of lactation, both in absolute sense and relative to TAA levels. Evaluation of gender differences by mixed model analyses revealed an association between female infant gender and higher protein content (p = 0.0465) and TAA content (p = 0.0362) in HM during the first 3 months of lactation. Furthermore, there was a tendency for an association of male infant gender with higher levels of free glutamine (p = 0.0948) in HM during the first 3 months of lactation. These results show that FAA, TAA and protein levels in HM display a time-specific occurrence during lactation. Moreover, although confirmation is necessary in view of the small sample size, this study indicates that the AA composition in HM shows differential effects of the infant’s sex.

Formation of High Levels of Acrylamide during the Processing of Flour Derived from Sulfate-Deprived Wheat

When wheat was grown under conditions of severe sulfate depletion, dramatic increases in the concentration of free asparagine were found in the grain of up to 30 times as compared to samples receiving the normal levels of sulfate fertilizer. The effect was observed both in plants grown in pots, where the levels of nutrients were carefully controlled, and in plants grown in field trials on soil with poor levels of natural nutrients where sulfate fertilizer was applied at levels from 0 to 40 kg sulfur/Ha. Many of the other free amino acids were present at higher levels in the sulfate-deprived wheat, but the levels of free glutamine showed increases similar to those observed for asparagine. In baked cereal products, asparagine is the precursor of the suspect carcinogen acrylamide, and when flours from the sulfate-deprived wheat were heated at 160 degrees C for 20 min, levels of acrylamide between 2600 and 5200 microg/kg were found as compared to 600-900 microg/kg in wheat grown with normal levels of sulfate fertilization.

Characterization of transgenic poplar with ectopic expression of pine cytosolic glutamine synthetase under conditions of varying nitrogen availability

The present study addresses the hypothesis that enhanced expression of glutamine synthetase (GS) in transgenic poplar, characterized by the ectopic expression of pine cytosolic GS, results in an enhanced efficiency of nitrogen (N) assimilation and enhanced growth. Transgenic and control poplar were supplied with low and high N levels and the role of ectopic expression of the pine GS in growth and N assimilation was assessed by using amino acid analysis, (15)N enrichment, biochemical analyses, and growth measurements. While leaves of transgenic poplar contained 85% less (P < 0.01) free ammonium than leaves of nontransgenic control plants, leaves of transgenics showed increases in the levels of free glutamine and total free amino acids. Transgenic poplar lines also displayed significant increases in growth parameters when compared with controls grown under both low (0.3 mm) and high (10 mm) nitrate conditions. Furthermore, (15)N-enrichment experiments showed that 27% more (P < 0.05) (15)N was incorporated into structural compounds in transgenic lines than in nontransgenic controls. Using the methods described here, we present direct evidence for increased N assimilation efficiency and growth in GS transgenic lines.

Allele-specific conditional destabilization of glutamine repeat mRNAs.

Several late-onset neurological diseases are caused by the inheritance of an expanded CAG repeat coding for polyglutamine. To date there is no effective means of halting the progression of these diseases, and their underlying molecular mechanisms remain a mystery. Strategies designed to specifically reduce the levels of long repeat mRNA might provide an effective therapy for these diseases. An emphasis on allele specificity is necessary to avoid the potential toxicities associated with reduction of expression. The experiments described here are based on the relationship between translation and mRNA stability and the idea that translation of a repeated codon might be extremely sensitive to reductions in levels of cognate aminoacylated tRNA. Consistent with this hypothesis, we have discovered that reduced glutamine concentration destabilizes mRNAs coding for long glutamine repeats while sparing short repeat versions of the same mRNAs. These results suggest therapy might be attained with existing compounds or environmental conditions known to decrease free glutamine levels.

Glutamine metabolism in uricotelic species: variation in skeletal muscle glutamine synthetase, glutaminase, glutamine levels and rates of protein synthesis

High intracellular glutamine levels have been implicated in promoting net protein synthesis and accretion in mammalian skeletal muscle. Little is known regarding glutamine metabolism in uricotelic species but chicken breast muscle exhibits high rates of protein accretion and would be predicted to maintain high glutamine levels. However, chicken breast muscle expresses high glutaminase activity and here we report that chicken breast muscle also expresses low glutamine synthetase activity (0.07±0.01 U/g) when compared to leg muscle (0.50±0.04 U/g). Free glutamine levels were 1.38±0.09 and 9.69±0.12 nmol/mg wet weight in breast and leg muscles of fed chickens, respectively. Glutamine levels were also lower in dove breast muscle (4.82±0.35 nmol/mg wet weight) when compared to leg muscle (16.2±1.0 nmol/mg wet weight) and much lower (1.80±0.46 nmol/mg wet weight) in lizard leg muscle. In fed chickens, rates of fractional protein synthesis were higher in leg than in breast muscle, and starvation (48 h) resulted in a decrease in both glutamine content and rate of protein synthesis in leg muscle. Thus, although tissue-specific glutamine metabolism in uricotelic species differs markedly from that in ureotelic animals, differences in rates of skeletal muscle protein synthesis are associated with corresponding differences in intramuscular glutamine content.

Skeletal muscle protein mobilization during the progression of lactation

To investigate changes in muscle metabolism during lactation, serial biopsy of the triceps brachii was conducted in first-parity sows subjected to three degrees of selective protein mobilization through restriction of dietary protein intake (see Clowes EJ, Aherne FX, Foxcroft GR, and Baracos VE. J Anim Sci 81: 753-764, 2003). Muscle biopsies were taken 7 days before parturition and at 12 and 23 days of lactation. The following changes occurred after parturition, were progressive, and were significantly magnified in animals under the greatest degree of dietary protein restriction and hence of protein mobilization. Decreased RNA-to-DNA ratio (capacity for protein synthesis) was observed. The presence of increased expression of several elements of the ubiquitin proteasome proteolytic pathway suggested a robust catabolic response. However, as lactation progressed, and especially under conditions of increased dietary protein restriction, protein mobilization increased, muscle RNA-to-DNA ratio fell further, protease gene expression continued to rise, tissue free glutamine levels rose dramatically, and essential amino acid levels, especially branched-chain amino acids and threonine, fell to below prepartum levels.

Herbicidal pyridyl derivatives of aminomethylene-bisphosphonic acid inhibit plant glutamine synthetase.

A series of aminomethylene-bisphosphonic acid derivatives, previously synthesized and shown to be endowed with herbicidal properties, were evaluated as potential inhibitors of plant glutamine synthetase. The cytosolic form of the enzyme was partially purified from rice cultured cells and assayed in the presence of millimolar concentrations of the compounds by means of three different assay methods, respectively measuring the hemibiosynthetic, the transferase, and the full biosynthetic reactions. Several compounds were found to exert a remarkable inhibition, with I(50) values similar to those obtained under the same conditions with a well-established inhibitor of glutamine synthetase, the herbicide phosphinothricin. Contrary to the reference compound, enzyme kinetics accounted for a reversible inhibition mechanism. The biological activity of the most active derivatives was further characterized by measuring free glutamine levels in cell suspension rice cultures following treatment with the inhibitors. Results confirmed their ability to interfere in vivo with nitrogen metabolism. A preliminary analysis of structure-activity relationship allowed it to be hypothesized that steric rather than electronic factors are responsible for the inhibitory potential of these compounds.

Utilization of alanyl-L-glutamine and glycyl-L-glutamine during long-term parenteral nutrition in the growing rat

Utilization of intravenously administered alanyl-L-glutamine and glycyl-L-glutamine as a source of glutamine was tested in growing rats receiving total parenteral nutrition for 15 days. In experiment A the two peptides were compared to each other, in experiment B alanyl-L-glutamine was compared to an equimolar mixture of free alanine and glutamine, and in experiment C glycyl-L-glutamine was tested against an equimolar mixture of free glycine and glutamine.There was no difference of statistical significance in weight gain or nitrogen balance between the respective groups participating in the 3 experiments. Plasma levels of free glutamine were similar during infusion with alanyl-L-glutamine or glycyl-L-glutamine. The daily urinary excretion rate recorded for alanyl-L-glutamine was 3.7% and for glycyl-L-glutamine 4.3% of the infused amount. The results show that both peptides are utilized for protein synthesis and growth to approximately the same as the corresponding free amino acids.

Alpha-ketoglutarate given together with TPN improves the free glutamine-levels in glutamine-depleted intensive care patients

Alpha-ketoglutarate given together with TPN improves the free glutamine-levels in glutamine-depleted intensive care patients

The metabolism of 14C-urea by white spruce seedlings in light and darkness

Weak urease activity was detected in 2-week-old white spruce seedlings. The addition of urea to these seedlings increased urease activity in light compared to darkness. Urease was localized in the cytoplasm mainly in the epidermal cells, and to a lesser extent in vascular tissues and at the shoot apex.In light, the early products of 14C-urea indicated the release of 14C-carbon dioxide and ammonium ions. Radioactivity appeared in the anionic and neutral fractions and then in alanine, serine, and glycine, just as in the fixation of 14C-bicarbonate. By 4 h most radioactivity resided in glutamic acid and alanine. This pattern was associated with increased protein synthesis, and with high levels of free glutamine. Serine, glycine, alanine, and carbamyl aspartic acid had high specific activities.In darkness, radioactivity in alanine persisted, but levels in serine, glycine, and protein were low. Most radioactivity eventually resided in asparagine. Radioactive urea and citrulline were kept at higher levels than in light. Glutamic acid, citrulline, and carbamyl aspartic acid had the greatest specific activities. In light and darkness, carbamyl derivatives were formed more readily from 14C-urea than from 14C-bicarbonate.Carbamyl phosphate was implicated as a precursor of citrulline and arginine as well as for carbamyl aspartic acid. In light, radioactivity was recovered from ribosomal (25, 23, 18, 16, and 5 S) and transfer RNA (4 S). This resided mainly in uracil and cytosine. By contrast, in DNA, radioactivity was recovered mainly from thymine, cytosine, and 5-methylcytosine. In rapidly growing seedlings, urea contributed to de novo nucleic acid synthesis mainly through pyrimidine pathways via carbamyl aspartic acid.

Glutamine and glutamine synthetase in fetal, adult and neoplastic rat tissues.

Glutamine synthetase and glutamine concentrations were compared in various rat tissues, in rat liver throughout development and during regeneration, and in transplanted tumors. There was no obvious relationship between the two components in these tissues. The enzyme increased abruptly in liver during the third week of life, but was not significantly induced by hydrocortisone nor ‘repressed’ by treatment with glutamine. Eight tumors with different growth rates all contained glutamine synthetase and glutamine in low but reasonable concentrations. One highly differentiated Morris hepatoma contained more glutamine synthetase than normal liver. There was no depression of glutamine synthetase in liver throughout the growth period of the tumors examined. Levels of free glutamine in plasma and liver were, however, significantly less than normal in the tumor-bearing rats.