Amino Acid Concentrations In Liver Research Articles

Dietary supplementation with cholesterol and docosahexaenoic acid affects concentrations of amino acids in tissues of young pigs

Cholesterol and docosahexaenoic acid (DHA) are important nutrients for neural development of infants. However, little is known about the effect of cholesterol or DHA on concentrations of amino acids (AA) in neonatal tissues. This study was conducted with the piglet (an established model for studying human infant nutrition) to test the hypothesis that dietary supplementation with the lipids may modulate AA availability in tissues. Sixteen newborn pigs were nursed by sows for 24 h and then assigned to one of four treatment groups, representing supplementation with 0.0% (control), 0.2% cholesterol, 0.2% DHA, or cholesterol plus DHA to the basal milk-formula. All piglets were euthanized at 49 days of age. In brain, cholesterol supplementation reduced (P < 0.05) concentrations of glutamate, serine, glutamine, threonine, beta-alanine, alanine, methionine, isoleucine, leucine, and gamma-aminobutyrate but increased (P < 0.05) concentrations of glycine and lysine, whereas DHA supplementation similarly affected (P < 0.05) concentrations of the same AA (except for isoleucine and lysine) and taurine. In addition, concentrations of most AA in liver, muscle and plasma were substantially altered by dietary supplementation of cholesterol and DHA in a tissue-dependent manner. Further, DHA reduced concentrations of carnosine in skeletal muscle, as well as ammonia in both plasma and brain. The results reveal that cholesterol and DHA can regulate AA metabolism and availability in various tissues of piglets. These novel findings have important implications for designing the next generation of infant formula to optimize neonatal growth and development.

Organ and Plasma Amino Acid Concentrations Are Profoundly Different in Piglets Fed Identical Diets via Gastric, Central Venous or Portal Venous Routes

In a previous study in piglets fed identical diets intravenously, intraportally or intragastrically, we determined that small intestinal atrophy affects nitrogen metabolism to a greater extent than liver by-pass. Because whole-body amino acid homeostasis is dependent on interorgan metabolism, we also measured the free amino acid concentrations in liver, small intestinal mucosa and kidney in order to study alterations in amino acid metabolism within these organs. Piglets (n = 15; 2-4 d old) were fed identical diets continuously for 8 d via gastric (IG), portal (IP) or central venous (IV) catheters. Concentrations of all measured amino acids were affected by route of feeding in one or more organs. In particular, urea cycle amino acid concentrations were altered in plasma and mucosa of IV and IP pigs, suggesting that arginine synthesis by an atrophied gut may have been limited. Furthermore, most indispensable amino acid concentrations were lower in IP pigs for all organs vs. IG pigs; however, except for phenylalanine, plasma concentrations of these amino acids were not different, demonstrating the liver's "smoothing" capability. Gut atrophy in both IV and IP pigs resulted in significantly lower concentrations of all indispensable amino acids compared with IG pigs. Alterations of all amino acids in various organs due to route of feeding suggest that more detailed analyses of regulatory mechanisms and amino acid interactions on interorgan amino acid metabolism are necessary for all amino acids.

Changes in hepatic nitrogen balance in plasma concentrations of amino acids and hormones and in cell volume after overnight fasting in perinatal and adult rat.

Important regulatory factors of intrahepatic protein synthesis and proteolysis are amino acids, glucagon, insulin, and cell volume. We have investigated the changes in these factors with development and after an overnight fast and evaluated their contribution to changes in the hepatic nitrogen balance in vivo. In the fed state, glucagon levels were highest in suckling animals and gradually declined in older rats, whereas the concentration of insulin increased during development. The amino acid concentrations in liver and plasma declined during the suckling period to levels that in vitro are highly permissive for induction of autophagic proteolysis. In all age groups investigated, fasting was associated with a drop in hepatic protein content, together with a marked decrease in hepatocellular volume and insulin concentrations. On the other hand, glucagon concentrations and the concentration of many amino acids in plasma and liver responded to fasting with a pronounced decrease in perinatal and suckling animals, but this response had become blunted at weaning and had disappeared in adult animals. These findings suggest that insulin and/or hepatocellular volume are more likely candidates as short-term physiologic regulators of the hepatic nitrogen balance than are glucagon or amino acids. In glucose-supplemented fetuses, high levels of insulin could not compensate for a decreased hepatocellular volume in averting a catabolic state, suggesting that cell volume is the more important factor. Although our study cannot discriminate between the effects of fasting on protein synthesis and degradation, our findings show unequivocally that, for a rapid growth of the liver, suckling animals have to be fed around-the-clock.

EFFECTS OF CONFINEMENT/CROWDING ON UREOGENESIS IN THE GULF TOADFISH OPSANUS BETA

In order to elucidate the cues for, and the biochemical mechanisms of, the transition to ureogenesis in the gulf toadfish Opsanus beta, experiments on the effects of confinement/crowding were carried out. Confinement of toadfish to small volumes of water initiated a switch to nearly complete reliance on ureogenesis for nitrogen excretion within 24&shy;48 h. Further experiments suggested that this switch was probably due to the physical confinement per se, rather than to a measurable build-up of ammonia in the water. However, the possibility of the response being triggered by a very low concentration of a pheromone-like substance was not excluded by our experimental design. The activities of several enzymes of ureogenesis, ornithine&shy;citrulline transcarbamoylase, aspartate amino transferase and glutamine synthetase, increased in the liver of toadfish during confinement. Notably, glutamine synthetase activity increased almost fourfold within 24 h, and this increase preceded increases in urea excretion. A number of plasma and liver amino acid concentrations changed during confinement: there were declines in plasma asparagine, glutamate and glycine levels and an increase in plasma valine, as well as a decline in liver alanine and an increase in liver arginine concentrations. Liver glutamine was not detectable. When the amino acid data are taken together with the enzyme activity changes, it appears that the switch to ureogenesis occurs primarily upstream of the ornithine&shy;urea cycle, at the level of supply of nitrogen to the pathway. The results are discussed in the context of the habitat of toadfish.

Similar effects of ethanol and tert-butanol on amino acid concentrations in rat serum and liver.

Both metabolic and nonmetabolic mechanisms have been proposed to the plasma amino acid decreasing effect of an acute ethanol load. We used tert-butanol, an alcohol that is only minimally metabolized, as a tool to explain the mechanism behind the amino acid decreasing effect of ethanol. Acute administration of tert-butanol was found to exert a decreasing effect on rat serum amino acid concentrations similar to that of ethanol, indicating that the mechanism of the amino acid decreasing effect of ethanol is primarily due to ethanol itself and not to its oxidation. Ethanol and tert-butanol also had similar effects on liver amino acid concentrations, including an increase in the glycine concentration and decrease in the concentrations of glutamate, alanine, leucine, and tyrosine.

Elevation of Retinol Levels and Suppression of Alanine Aminotransferase Activity in the Liver of Taurine-Deficient Kittens

In taurine-deficient cats, the secretion of bile acids is impaired, and this impairment may reduce intestinal uptake of lipophilic vitamins. It was therefore hypothesized that retinol deficiency is involved in the generation of retinal lesions in taurine-deficient kittens. To this end, the concentration of retinol in plasma and liver was determined in taurine-deficient kittens. Further, the effects of taurine deficiency on amino acid concentrations of heart, liver and kidney were investigated. To see whether taurine deficiency adversely affects the liver, hepatic enzymes were measured in plasma and liver of kittens suffering from taurine deficiency. In addition, liver morphology, growth and food intake were studied. Taurine was the only amino acid whose concentration was consistently decreased in plasma of the experimental group. Unexpectedly, retinol level was increased in plasma and liver from taurine-depleted kittens. Several alterations were noted in amino acid concentrations in liver and kidney, but not in heart. Plasma alanine aminotransferase activity was diminished, probably reflecting decreased activity in the liver. Perivenular steatosis was found in both groups. Controls grew linearly, in contrast to deficient animals, which nevertheless consumed more food. The results demonstrate that retinol deficiency is not involved in taurine-deficiency retinopathy. Moreover, taurine is required for linear growth of juvenile cats and for the maintenance of hepatic and renal pools of certain amino acids.

Plasma Ammonia, Plasma, Brain and Liver Amino Acids and Urea Cycle Enzyme Activities in Rats Fed Ammonium Acetate

Male Sprague-Dawley rats were trained to eat a 6% casein diet within a 3-h period each day. They were then fed a 6% casein diet for 10-16 d before they were fed either the same 6% casein diet or the 6% casein diet supplemented with 15% ammonium acetate for 1 or 7 d. During the absorptive period, plasma ammonia, plasma amino acids and brain amino acids were measured on d 1 and d 7 after feeding ammonium acetate. Food intake of rats fed 15% ammonium acetate was depressed on d 1 and increased to approximately 75% of the intake of the 6% casein-fed group by d 7. On d 1 plasma ammonia of the rats fed 5% ammonium acetate was 101 microM as compared to 56 microM for the rats fed 6% casein (P less than 0.05). On d 7, plasma ammonia of the rats fed 15% ammonium acetate was 240 microM (P less than 0.05) as compared to 44 microM for the rats fed 6% casein. In rats fed 15% ammonium acetate, after 7 d ornithine transcarbamylase and arginase activities were higher and argininosuccinate synthetase activity was lower (P less than 0.05) while carbamyl phosphate synthetase activity tended to be higher than that of rats fed 6% casein. The results suggest that rats adapt to ingestion of 15% ammonium acetate by some unknown neural mechanism rather than by increases in all urea cycle enzyme activities. Feeding ammonium acetate causes changes in plasma, brain and liver amino acid concentrations.

Effect of lead toxicity on the organ concentration of glutathione and glutathione-related free amino acids in the chick

Total non-protein sulfhydryl (NPSH) and glutathione (GSH) concentrations in liver and kidney and GSH-related free amino acid concentrations in liver, kidney and plasma were examined in the chick following intoxication with 2000 ppm dietary lead (Pb). After 3 weeks, NPSH content was significantly ( P < 0.05) increased by Pb in both liver and kidney while only liver GSH was increased by Pb treatment. The concentrations of glutamate, glycine and methionine were elevated in liver by Pb while glycine, eysteine and cystathionine concentrations were increased in the kidney. In plasma, Pb significantly decreased cysteine, taurine and cystathionine concentrations.

Dietary requirements of rainbow trout for tryptophan, lysine and arginine determined by growth and biochemical measurements

Three separate studies were performed to determine the dietary requirements of rainbow troutSalmo gairdneri for tryptophan (Trp), lysine (Lys) and arginine (Arg) from both growth and biochemical data. The growth studies were carried out over a 12 week period. From graphical plots of % mean weight gain against % amino acid in diet the following requirement values were obtained, Trp 0.25% diet (0.4% dietary crude protein); Lys 1.9% diet (4.3% dietary protein); and Arg 1.6-1.8% diet (3.6-4% dietary protein). Plasma and liver amino acid concentrations measured 20h after feeding did not prove useful for determination of requirement values. Hepatic activities of Trp pyrrolase (TP), Lys α ketoglutarate reductase (LKGR) and arginase were not significantly affected by varying levels of Trp, Lys and Arg respectively in the diet. TP has a cytosolic location and a Km of 0.2 mM for Trp; LKGR is mitochondrial and the Km for Lys is 7.3 mM; arginase is also mitochondrial and has a Km of 4.9 mM for arginine. Measurements of expired(14)CO2, after injection of a tracer dose of(14)C amino acid, did allow estimates of requirement levels to be made. The values obtained from the oxidation studies reinforced the values obtained from the growth data but were not precise enough to justify using this method on its own.

Effects of burns on amino acid levels in rat plasma, liver and muscle

In this study the influence of a severe catabolic situation (scalding and nitrogen deprivation) on amino acid (AA) metabolism was investigated in an experimental rat model. Scalding of 25 per cent of the total body surface area (TBSA) and hypocaloric alimentation (5–6 kcal per 100g rat per day, no nitrogen) resulted in mean daily nitrogen losses of −0·27±0·3 g. Compared to anabolic growing rats this nitrogen catabolism significantly reduced the total free AA content of muscle (−47 per cent, P<0·001) and liver (-39 per cent, P<0·001). The total plasma AA concentrations were slightly increased in catabolic rats (+10 per cent). In catabolic rats muscle glycine concentrations dropped significantly (−79 per cent, P<0·001), while glutamine concentrations decreased by 22 per cent, which was not significant. Branched chain AA and phenylalanine were significantly elevated both in muscle and in plasma. Scalding and nitrogen depletion in rats leads to characteristic changes in plasma, muscle and liver AA concentrations, which are comparable to the results obtained in catabolic patients. However, the low muscle glycine concentrations in burned rats differ from the clinical observations where glutamine rather than glycine concentrations in muscle tissue are reduced. The rat model seems to be well suited for studying the influence of various therapeutic approaches such as different forms of parenteral nutrition or hormonal substitution on nitrogen catabolism.

Failure of the normal ureagenic response to amino acids in organic acid-loaded rats. Proposed mechanism for the hyperammonemia of propionic and methylmalonic acidemia.

Propionic and methylmalonic acidemia are both known to be associated with hyperammonemia. Rats injected with 10 or 20 mmol/kg of propionate or 20 mmol/kg of methylmalonate, along with 1.5 g/kg of a mixture of amino acids, developed severe hyperammonemia, whereas rats administered the same dosages of acetate did not. In vitro, neither propionyl nor methylmalonyl CoA affected the activity of carbamyl phosphate synthetase I, ornithine transcarbamylase, nor the activation constant (K(A)) of carbamyl phosphate synthetase I for N-acetyl glutamate. Furthermore, rats injected with propionate showed no alteration of liver amino acid concentrations, which could explain impaired ureagenesis. Animals injected with methylmalonate showed an increase in both citrulline and aspartate, suggesting that argininosuccinic acid synthetase may also have been inhibited. Liver ATP levels were unchanged. Citrullinogenesis, measured in intact mitochondria from livers of injected animals, was reduced 20-25% by 20 mmol/kg of propionate or methylmalonate (compared with acetate). This effect was attributable to an impairment in the normal rise of liver N-acetyl glutamate content after amino acid injection. Thus, carbamyl phosphate synthetase I activation was reduced. Liver levels of acetyl CoA and free CoA were reduced. Levels of unidentified acyl CoA derivatives rose, presumably reflecting the accumulation of propionyl and methylmalonyl CoA. Thus, the principal mechanism for hyperammonemia induced by these acids is depletion of liver N-acetyl glutamate, which is in turn attributable to depletion of acetyl CoA and/or competitive inhibition by propionyl and methylmalonyl CoA of N-acetyl glutamate synthetase. Injection of methylmalonate may also have an additional inhibitory effect on argininosuccinic acid synthetase.

Nutritional aspects of amino acid metabolism. 3. The effects of diabetes on blood and liver amino acid concentrations in the rat.

1. Measurements of the concentrations of amino acids in the plasma of blood from the portal vein and hepatic vein in the livers of rats made diabetic with streptozotocin or alloxan and starved for 1 d were compared with similar measurements from normal rats starved for 1 d.2. The concentrations of many of the amino acids in the blood plasma were lower in the streptozotocin diabetic rats than in the normal animal while the liver concentrations tended to be increased. This suggests that there is enhanced concentration of these amino acids by the liver in diabetics, as is also found in starvation, which is probably due to factors other than the direct absence of insulin.3. The direction of flow of the groups of amino acids into and out of the liver was unchanged in the diabetic compared with the normal rat except that the output of tryptophan was abolished, and that of the branched-chain group was abolished in alloxan diabetes though apparently enhanced in streptozotocin diabetes. The rates of movement of amino acids in both directions appeared to be increased in the streptozotocin diabetic animals.4. The changed amino acid pattern in the alloxan diabetic rats was to some extent similar to that of the streptozotocin diabetic rats but the changes were more difficult to interpret, perhaps because of side-effects of alloxan on tissues including liver.