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Biomedical subjects

S A Adibi

Publications and source records attributed to S A Adibi.

At least 19 recordsLinked to original sources
PubMed1992

Mechanism of increased conversion of branched chain keto acid dehydrogenase from inactive to active form by a medium chain fatty acid (octanoate) in skeletal muscle.

We and others have previously shown that octanoate increases the oxidation of branched chain amino acids (BCAA) in skeletal muscle. The present study was designed to investigate the mechanism of this increased oxidation. Studies were performed with rat hind limbs perfused with 0.50 mM L-[1-14C]leucine with or without octanoate. The flux through branched chain keto acid (BCKA) dehydrogenase was measured, and the basal and total activity of BCKA dehydrogenase in skeletal muscle was determined. The rate of flux through BCKA dehydrogenase increased by 37, 119, and 297% with 0.5, 1.0, and 2.0 mM octanoate, respectively. This increase in flux was not due to a change in BCAA aminotransferase activity but was due to an increase in the basal activity of BCKA dehydrogenase. There was a strong correlation (r = 0.96) between increases in flux through BCKA dehydrogenase and increases in the basal activities of BCKA dehydrogenase. Preincubation of BCKA dehydrogenase with Mg2+ caused full activation of this enzyme, but preincubation with octanoate did not activate this enzyme. On the other hand, octanoate completely prevented the ATP-dependent inactivation of fully activated BCKA dehydrogenase. We conclude that octanoate increases the oxidation of leucine in skeletal muscle by increasing the activation of BCKA dehydrogenase. The mechanism of this activation is the inhibition of BCKA dehydrogenase kinase rather than the stimulation of a specific or nonspecific protein phosphatase.

3-Methyl-2-Oxobutanoate Dehydrogenase (Lipoamide)↗
PubMed1992

Determinants of substrate affinity for the oligopeptide/H+ symporter in the renal brush border membrane.

We and others have shown previously the existence of high and low affinity systems for oligopeptide transport in kidney brush border membrane vesicles (BBMV). In the present study we investigated the relationship between the structure of substrates and their affinity for interaction with the high-affinity oligopeptide/H+ transporter in kidney BBMV. Based on competition experiments using [3H]Gly-Gln as a probe we determined the Ki values for more than 60 selected peptides. For a high-affinity interaction with the carrier site the following structural features of substrates are required: (a) both a free amino and carboxyl terminus; (b) the amino group and peptide bond nitrogen located in the alpha-position; (c) a trans peptide bond rather than the cis configuration; (d) L-alpha-amino acid isomers in both COOH and NH2 termini, although D-isomers of hydrophobic amino acids are acceptable in the NH2 terminus; and (e) a backbone of less than 3 amino acid residues. A striking finding of the present study is that, for peptides satisfying these minimal structural requirements, the primary determinant of affinity is hydrophobicity. The fact that there is a highly significant (p less than 0.001) correlation between Ki and hydrophobicity allows the prediction of the affinity for any di- or tripeptide composed of alpha-amino acids in the L-form.

Amino Acid Sequence↗
PubMed1992

Carnitine biosynthesis in hepatic peroxisomes. Demonstration of gamma-butyrobetaine hydroxylase activity.

We have investigated whether hepatic peroxisomes are capable of synthesizing carnitine. When purified peroxisomes were incubated with gamma-butyrobetaine, a precursor of carnitine, formation of carnitine was observed. These results indicate that peroxisomes contain gamma-butyrobetaine hydroxylase, the enzyme which catalyzes the final step in the biosynthesis of carnitine. This enzyme was previously believed to be present only in the cytosol. gamma-Butyrobetaine hydroxylase activity in peroxisomes was not due to cytosolic contamination as evaluated by marker enzyme analysis. When proliferation of peroxisomes was induced by clofibrate treatment, gamma-butyrobetaine hydroxylase/mass liver increased by 7.6-fold and the specific activity by 2.5-fold. We conclude that hepatic peroxisomes synthesize carnitine and this synthesis becomes substantial under conditions of peroxisomal proliferation.

Animals↗
PubMed1992

Characteristics and mechanism of glutamine-dipeptide absorption in human intestine.

Using in vivo and in vitro techniques, the mechanism by which intestinal mucosa obtains glutamine from luminal oligopeptides was investigated in humans. The rate of hydrolysis by mucosal brush border membrane was more than threefold greater for alanylglutamine than for glycylglutamine. Despite this difference, rates of dipeptide and amino acid disappearance during intestinal perfusion were greater from test solutions containing glycylglutamine than alanylglutamine. Furthermore, rates of intraluminal appearance of products of hydrolysis during the infusion of two dipeptides were similar and less than 5% of the disappearance rate of the parent dipeptide. In contrast to free glutamine, uptake of peptide-bound glutamine by brush border membrane vesicles was not inhibited by deletion of sodium or addition of free amino acids to the incubation medium but was inhibited by other oligopeptides and stimulated by a proton gradient. Inhibition constants for the saturable uptake of glycylglutamine and alanylglutamine by vesicles were not significantly different, suggesting similar affinities for the peptide transporter. It is concluded that in human intestine the predominant mechanism for assimilation of glutamine-dipeptides is absorption as intact dipeptide rather than hydrolysis.

Absorption↗
PubMed1992

Liver triglyceride concentration and body protein metabolism in ethanol-treated rats: effect of energy and nutrient supplementation.

The objective of this study was to compare the metabolic effects of long-term ethanol consumption with oral (Lieber-DeCarli) and enteral feeding techniques. Enteral feeding allowed administration of greater amounts of energy and nutrients. After 21 days of treatment using the Lieber-DeCarli technique, the ethanol-treated rats had the following significant (P less than 0.05) differences from pair-fed controls: lower cumulative nitrogen balance (days 5-21; 2.8 +/- 0.1 g N vs. 3.5 +/- 0.1 g N), lower protein content of gastrocnemius muscle (289 +/- 17 mg vs. 358 +/- 11 mg) and intestinal mucosa (461 +/- 19 mg vs. 577 +/- 40 mg), higher plasma leucine concentration (147 +/- 8 mumol/L vs. 102 +/- 8 mumol/L), higher liver protein content (2222 +/- 122 mg vs. 1679 +/- 58 mg), and higher liver triglyceride concentration (38.4 +/- 2.8 mg/g vs. 8.7 +/- 1.0 mg/g). When rats received the same amount of nitrogen (1.5 g.kg-1.day-1) and ethanol (13 g.kg-1.day-1) but 16.3% more energy and nutrients by a surgically implanted gastric cannula (enterally fed), the effects of ethanol on nitrogen balance, tissue protein content, plasma leucine concentration, and liver triglyceride concentration were similar to those observed in the rats fed orally. It is concluded that the metabolic effects observed using the Lieber-DeCarli feeding technique are due to ethanol per se and not the synergism of ethanol and undernutrition as recently suggested.

Animal Nutritional Physiological Phenomena↗
PubMed1992

Investigation of the presence of branched-chain alpha-keto acid dehydrogenase in mammalian hepatic peroxisomes.

1. Rat liver was fractionated into peroxisomes and mitochondria and branched-chain keto acid (BCKA) dehydrogenase activity was measured. 2. All BCKA dehydrogenase activity was associated with the mitochondrial fraction and none with the peroxisomal fraction. 3. BCKA dehydrogenase was also not detected in hepatic peroxisomes of rats treated with clofibrate which induces several peroxisomal enzymes. 4. Hepatic peroxisomes from rabbit, hamster and dog also did not show any BCKA dehydrogenase activity. 5. We conclude that mammalian hepatic peroxisomes do not contain BCKA dehydrogenase.

3-Methyl-2-Oxobutanoate Dehydrogenase (Lipoamide)↗
PubMed1992

Protein sparing during treatment of obesity: ketogenic versus nonketogenic very low calorie diet.

Although it is generally agreed that both ketogenic and nonketogenic very low calorie diets promote weight reduction, there is no consensus on a preference of one diet over the other in regard to protein sparing. In the present study, we compared the effects of isocaloric (600 kcal/d) and isonitrogenous (8 g nitrogen/d) ketogenic (low carbohydrate) and nonketogenic diets on parameters of protein and amino acid metabolism, in 16 morbidly obese women maintained on these diets for 4 weeks while confined to a metabolic ward. Cumulative urinary nitrogen excretion (g/4 wk) was significantly (P less than .01) greater (248 +/- 6 v 207 +/- 12, mean +/- SEM, n = 8), and cumulative nitrogen balance significantly (P less than .02) more negative (-50.4 +/- 4.4 v -18.8 +/- 5.7), during treatment with the ketogenic than with the nonketogenic diet. Plasma leucine concentration (mumol/L) was significantly higher (P less than .05) during treatment with the ketogenic than with the nonketogenic diet at day 14 (210 +/- 17 v 150 +/- 8), but not at day 28 (174 +/- 9 v 148 +/- 8). Whole-body rates of leucine oxidation (mmol/h) were significantly higher (P less than .05) during treatment with the ketogenic than with the nonketogenic diet at day 14 (1.29 +/- 0.20 v 0.92 +/- 0.10) and at day 28 (1.00 +/- 0.16 v 0.75 +/- 0.10). Conversely, proteolysis, as measured by leucine turnover rate and urinary excretion of 3-methylhistidine, was not significantly different between the diets.(ABSTRACT TRUNCATED AT 250 WORDS)

Amino Acids↗
PubMed1992

Uptake and hydrolysis of glycylglutamine at the blood-brain barrier.

Using an organ balance technique in dogs, we recently found that liver, skeletal muscle, kidney, and intestine participate in clearance of glycylglutamine from plasma. The purpose of the present study was to investigate whether brain does the same. The study of arteriovenous differences of glycylglutamine across brain, during continuous infusion of this dipeptide (12 mumol.min-1.kg-1) in dogs, showed an arteriovenous difference that was never significantly different from zero. To establish a basis for this lack of clearance, we investigated uptake and hydrolysis of glycylglutamine at the blood-brain barrier. The study of brain uptake index of glycylglutamine in rats showed that it was not significantly different from that of sucrose, an impermeable marker (3.2 +/- 0.4 v 3.5 +/- 0.4, n = 4 to 5). When isolated brain capillaries were incubated with glycylglutamine, uptake was only modestly above background activity and appeared to be due to nonspecific binding. Finally, the plasma membrane of brain capillaries lacked hydrolase activity against glycylglutamine. In conclusion, brain appears to be unique among organs in lacking any mechanism for clearance of glycylglutamine from plasma.

Animals↗
PubMed1992

Glycylglutamine: metabolism and effects on organ balances of amino acids in postabsorptive and starved subjects.

The present study was designed to investigate the metabolism of glycylglutamine and its effects on organ balances of amino acids during intravenous infusion of this dipeptide (100 mumol.h-1.kg-1) in postabsorptive and briefly starved (84-86 h) human subjects. Arterial concentrations of glycylglutamine were not significantly different in postabsorptive (265 +/- 18 microM) and starved (241 +/- 13 microM) subjects. Among the organs examined, kidney predominated in clearance of glycylglutamine from plasma. Moreover, renal clearance of glycylglutamine was reduced by starvation (87 +/- 7 vs. 52 +/- 5 mumol/min, P less than 0.01), whereas neither splanchnic nor muscle clearance was significantly affected. Infusion of glycylglutamine raised plasma concentrations of glycine and glutamine by increasing renal release of these amino acids. In postabsorptive subjects the infusion significantly increased splanchnic balances of glycine and glutamine with little or no effect on the muscle balances; the opposite was found in starved subjects. As far as other amino acids are concerned, the infusion decreased the muscle release of alanine and increased renal release of serine. We conclude that the amino acid residues of glycylglutamine are largely metabolized by the splanchnic organs in postabsorptive subjects and by peripheral organs in starved subjects. The latter results in selective inhibition of muscle release of amino acids.

Adult↗
PubMed1992

Oligopeptides: mechanism of renal clearance depends on molecular structure.

We have investigated the relative contribution of hydrolysis, intact transport and urinary excretion to the renal clearance of Gly-Sar, Gly-Sar-Sar, and Gly-Gly-Sar in fed and starved rats. The results obtained from isolated kidney perfusion studies are summarized as follows: 1) clearance was fastest for Gly-Gly-Sar and slowest for Gly-Sar-Sar, 2) urinary excretion of Gly-Sar-Sar exceeded that of Gly-Gly-Sar or Gly-Sar, 3) there was accumulation of products of hydrolysis of Gly-Gly-Sar in the perfusate but not of Gly-Sar or Gly-Sar-Sar, 4) isolated brush-border and basolateral membranes of renal tubular cells lacked hydrolytic activity against Gly-Sar and Gly-Sar-Sar but possessed hydrolytic activity against Gly-Gly-Sar, 5) an excess amount of Gly-Sar-Sar reduced the rate of clearance of Gly-Gly-Sar by approximately 40% and significantly increased urinary excretion of this peptide, 6) the nonfiltering kidney cleared Gly-Gly-Sar at a rate which was 50% of that of the filtering kidney but did not clear Gly-Sar, and 7) starvation for 96 h was without a significant effect on the renal clearance of either Gly-Sar or Gly-Sar-Sar but significantly reduced the renal clearance of Gly-Gly-Sar and the brush-border membrane hydrolase activity against this peptide. We conclude that the molecular structure determines the affinity of oligopeptides for the membrane transport and hydrolytic systems, which, in turn, determines their efficiency for clearance by the kidney.

Animals↗
PubMed1991

The high and low affinity transport systems for dipeptides in kidney brush border membrane respond differently to alterations in pH gradient and membrane potential.

The principal aim of the present study was to investigate the effects of variation in proton gradient and membrane potential on the transport of glycyl-L-glutamine (Gly-Gln) by renal brush border membrane vesicles. Under our conditions of transport assay, Gly-Gln was taken up by brush border membrane vesicles almost entirely as intact dipeptide. This uptake was mediated by two transporters shared by other dipeptides and characterized as the high affinity (Kt = 44.1 +/- 11.2 microM)/low capacity (Vmax = 0.41 +/- 0.03 nmol/mg protein/5 s) and low affinity (Kt = 2.62 +/- 0.50 mM)/high capacity (Vmax 4.04 +/- 0.80 nmol/mg protein/5 s) transporters. In the absence of a pH gradient, only the low affinity system was operational, but with a reduced transport capacity. Imposing a pH gradient of 1.6 pH units increased the Vmax of both transporters. Kinetic analysis of the rates of Gly-Gln uptake as a function of external pH revealed Hill coefficients of close or equal to 1, indicating that transporters contain only one binding site for the interaction with external H+. The effects of membrane potential on Gly-Gln uptake were investigated with valinomycin-induced K+ diffusion potentials. The velocity of the high affinity system but not of the low affinity system increased linearly with increasing inside-negative K+ diffusion potentials (p less than 0.01). The Kt of neither system was affected by alterations in either pH gradient or membrane potential. We conclude that (a) the high affinity transporter is far more sensitive to changes in proton gradient and membrane potential than the low affinity transporter and (b) in the presence of a pH gradient, transport of each dipeptide molecule requires cotransport of one hydrogen ion to serve as the driving force.

Animals↗
PubMed1990

Uptake and metabolism of dipeptides by human red blood cells.

A function of the abundant cytoplasmic peptidases in red blood cells could be hydrolysis of oligopeptides circulating in plasma. To investigate whether human red blood cells actively transport dipeptides for this purpose, these cells were incubated with 14C-labelled glycylproline, glycylsarcosine, glycine, proline and alanine. There was uptake of each dipeptide, as indicated by their recovery as dipeptides in the cell cytoplasm. However, after a brief time (1-2 min) uptake of dipeptides abruptly ceased, while that of amino acids continued. As a result, after 30 min red blood cell uptake of amino acids was 5-13-fold greater than that of any dipeptide. Investigation of intracellular contents after 1 min of incubation revealed different metabolism for different dipeptides. The composition of intracellular radioactivity was 19-71% as intact dipeptides, 0-20% as free amino acids and 8-77% as neither dipeptides nor constituent amino acids. Investigation of the mechanism of dipeptide uptake by red blood cells showed: (1) a lack of hydrolysis by the plasma membrane, (2) no non-specific binding to the plasma membrane, and (3) a lack of saturation over a wide range of concentrations (0.05-50 mM). The data suggest that the mechanism of uptake of trace amounts of dipeptides by human red blood cells is either by simple diffusion or by a carrier system which has a very weak affinity for dipeptides. Upon entry, depending on the molecular structure, dipeptides are either hydrolysed or transformed into new compounds. The red blood cell uptake, however, does not appear to play any appreciable role in clearance of dipeptides from the plasma in the human.

Alanine↗
PubMed1990

Splanchnic, renal, and muscle clearance of alanylglutamine in man and organ fluxes of alanine and glutamine when infused in free and peptide forms.

The present study was designed to investigate organ metabolism of intravenously (IV) infused (100 mumol.h-1.kg-1) alanylglutamine and its amino acid constituents in a group of healthy subjects. The dipeptide clearance (mumol/min) by kidney (51 +/- 3) was significantly (P less than .01) greater than the clearance by either splanchnic organs (19 +/- 6) or skeletal muscle (21 +/- 8). Infusion of alanylglutamine significantly (P less than .01) increased arterial plasma concentrations of free alanine (260 +/- 31 v 330 +/- 38 mumol/L) and free glutamine (620 +/- 66 v 764 +/- 65 mumol/L) when compared with the baseline period. Concurrently, splanchnic uptake of alanine and glutamine increased and muscle release of alanine ceased. However, muscle release of glutamine remained unaffected. Renal balances of alanine and glutamine changed from neutral to negative (net release) and from positive (net uptake) to neutral, respectively. Infusion of a corresponding mixture of alanine and glutamine had similar effects on arterial plasma concentrations and splanchnic and muscle balances of alanine and glutamine, but had no effect on renal balances of these amino acids. From these studies in man, we conclude that kidney predominates over other organs in clearance of alanylglutamine from plasma and that this may account for the different effect of infusion of alanine and glutamine in free and peptide forms on renal fluxes of these amino acids.

Adult↗
PubMed1990

Mechanism of clearance of dipeptides by perfused hindquarters: sarcolemmal hydrolysis of peptides.

The objective of the present experiment was to investigate the mechanism of clearance of a load of dipeptides (10 mumols) by perfused hindquarters of rats. The clearance was progressive over 60 min and was significantly (P less than 0.01) greater for glycylleucine than for glycylglycine (99 vs. 58% disappearance from the medium). Insulin had no significant effect on clearance of these dipeptides but stimulated the net uptake of their constituent amino acids. Investigation of the fate of peptides considered resistant to membrane hydrolysis showed a modest (24%) clearance for glycylsarcosine but a substantial one (89%) for glycylproline. Investigation of hydrolysis by sarcolemmal vesicles of skeletal muscle showed hydrolase activity against glycylglycine and glycylleucine but none against glycylsarcosine and glycylproline. Investigation of hydrolysis in the medium previously used to perfuse hindquarters for 60 min showed considerable activity against glycylleucine and glycylproline but none against glycylglycine and glycylsarcosine. These activities were entirely abolished by p-hydroxymercuribenzoate, an inhibitor of cytoplasmic peptide hydrolases. In conclusion, our data show that the mechanism of clearance of dipeptides by the perfused hindquarters is largely by hydrolysis, and the site of this hydrolysis differs for different dipeptides; hydrolysis is mediated either by plasma membrane enzymes, cytoplasmic enzymes released into the medium, or a combination of both.

Animals↗
PubMed1989

Possible sources of glutamine for parenteral nutrition: impact on glutamine metabolism.

Due to its instability, glutamine is not included in solutions for parenteral solution. This problem can be obviated by providing glutamine as acetyl-, glycyl-, or alanylglutamine. Using an organ balance technique in conscious dogs, we investigated metabolism of these three sources of glutamine. Liver, gut, kidney, and muscle participated in clearance of glycyl- and alanylglutamine from plasma, but among these organs only kidney cleared acetylglutamine. Furthermore, there was a large urinary excretion for acetylglutamine (38 +/- 6% of amount infused) but only a trace amount for either dipeptide. The infusion of glutamine-dipeptides resulted in similar increases in blood level of free glutamine. The main source of this increase appeared to be hydrolysis of dipeptides by kidney and release of free glutamine to circulation. During the infusion of both dipeptides, glutamine balance (free and dipeptide forms) was always positive (net uptake) across liver, gut, and kidney but was neutral across muscle. Liver or gut glutamine balances were not significantly different during the infusion of dipeptides, but kidney glutamine balance was twofold greater during the infusion of glycyl- than alanylglutamine. We conclude that among these three sources of glutamine, acetylglutamine is least desirable for use in parenteral nutrition. Glycylglutamine may be preferable over alanylglutamine if the objective is to target glutamine for kidney.

Animals↗
PubMed1989

Nitrogen absorption in pancreatectomized patients: protein versus protein hydrolysate as substrate.

To investigate nitrogen absorption in the absence of the pancreas, six patients with total pancreatectomy, all in stable nutritional and metabolic condition, underwent two periods of enteral nutrition identical in all respects except for the nitrogen source. Nitrogen source was either lactalbumin or its hydrolysate. The quantity and quality of calories infused simulated the patient's usual diet, which was a high-protein diet (2.0 +/- 0.4 gm/kg body weight). Pancreatic enzyme replacement therapy was discontinued during each period of enteral nutrition. All patients had greater nitrogen absorption during the enteral nutrition with lactalbumin hydrolysate than that with lactalbumin (91% +/- 2% vs 61% +/- 6% of nitrogen intake, p less than 0.02). Despite this difference in absorption, nitrogen balances during the two periods of enteral nutrition were not significantly different. This appeared to be caused by a urea production rate that was greater during the enteral nutrition with lactalbumin hydrolysate than the rate during that with lactalbumin (26 +/- 1 gm/24 hr vs 16 +/- 3 gm/24 hr, p less than 0.05). Plasma concentrations of amino acids and proteins did not differ significantly during the two treatments. In conclusion, the data suggest that (1) the intestine plays a significant role in protein digestion and that (2) enteral feeding with a protein hydrolysate could eliminate the need for a high-protein diet in patients with pancreatic insufficiency.

Absorption↗