Carnitine Excretion Research Articles

Choline Supplementation Alters Carnitine Homeostasis in Humans and Guinea Pigs

We have previously shown that supplementary choline causes significant decreases in urinary excretion of carnitine in humans. The objectives of the present work were to study this interaction in a different human population and on other body pools of carnitine in an animal model. In young adult women, daily choline supplementation (20 mg/kg body wt) resulted in a 75% lower urinary carnitine excretion than in controls, without significantly altering plasma carnitine concentrations. Supplementary choline was added to the ground diet of Sprague-Dawley rats (2.5 g/kg diet) and guinea pigs (3 g/kg diet). Choline supplementation had no effect on plasma concentrations or urinary excretion of carnitine in the rats. In guinea pigs, however, choline supplementation resulted in a significantly lower urinary excretion and higher plasma total carnitine concentrations. The skeletal muscle carnitine concentration was higher in the choline-supplemented guinea pigs, but not significantly higher in other tissues. These studies demonstrated that choline supplementation results in decreased urinary excretion of carnitine in young adult women, that guinea pigs are a suitable animal model for studying the effect of choline supplementation on carnitine status in humans, and that choline results in a conservation of carnitine in guinea pigs and perhaps in humans.

The ability of guinea pigs to synthesize carnitine at a normal rate from ϵ- N-trimethyllysine or γ-butyrobetaine in vivo is not compromised by experimental vitamin C deficiency

Experimental vitamin C deficiency in guinea pigs is associated with low carnitine concentrations in blood and some tissues. Ascorbic acid is a cofactor for two enzymes in the pathway of carnitine biosynthesis. The effect of experimental vitamin C deficiency on the ability of guinea pigs to synthesize carnitine was investigated in animals fed a vitamin C-deficient diet for 28 days. On days 19 to 28, supplements (0.5 mmol · kg body weight −1 · d −) of the carnitine precursors ϵ- N-trimethyllysine or γ-butyrobetaine were administered orally. Ascorbate-supplemented, ascorbate-deficient, and pair-fed (to ascorbate-deficient) animals showed an increase in the rate of carnitine biosynthesis (as estimated from measured rates of carnitine excretion) of 32 to 40 μmol · kg body weight −1 · d −1 following supplementation with ϵ- N-trimethyllysine. Likewise, animals in each experimental group showed an increase in the rate of carnitine biosynthesis of 41 to 50 μmol · kg body weight −1 · d −1 after supplementation with γ-butyrobetaine. These results indicate that scorbutic guinea pigs are able to synthesize carnitine at a normal or above-normal rate. For guinea pigs not given a carnitine precursor supplement, rates of free and total carnitine excretion for ascorbate-deficient (but not pair-fed) animals were threefold higher than for ascorbate-supplemented animals during days 19 to 28 of the feeding regimen. Thus, carnitine depletion in vitamin C deficiency likely is due to excessive urinary excretion of carnitine and not to a decreased rate of carnitine biosynthesis.

Rhabdomyolysis and acute encephalopathy in late onset medium chain acyl-CoA dehydrogenase deficiency.

A previously asymptomatic 30 year old man presented with rhabdomyolysis, muscle weakness, and acute encephalopathy after strenuous exertion in the cold without adequate food intake. Serum and muscle carnitine concentrations were decreased. Urinary excretion of carnitine and glycine esters and biochemical examination of skeletal muscle and fibroblasts led to the diagnosis of medium chain acyl-CoA dehydrogenase (MCAD) deficiency. A point mutation at nucleotide position 985 of the coding region of the MCAD gene was found. The MCAD protein was synthesised in the patient's fibroblasts at a normal rate, but was unstable. In general, patients in whom the 985 point mutation has been established show much more severe clinical symptoms and other symptoms than those seen in this patient. The relation of the 985 point mutation and the residual MACD activity to the symptoms is not as straightforward as previously thought.

Neural dysfunction and metabolic imbalances in diabetic rats. Prevention by acetyl-L-carnitine.

The rationale for these experiments is that administration of L-carnitine and/or short-chain acylcarnitines attenuates myocardial dysfunction 1) in hearts from diabetic animals (in which L-carnitine levels are decreased); 2) induced by ischemia-reperfusion in hearts from nondiabetic animals; and 3) in nondiabetic humans with ischemic heart disease. The objective of these studies was to investigate whether imbalances in carnitine metabolism play a role in the pathogenesis of diabetic peripheral neuropathy. The major findings in rats with streptozotocin-induced diabetes of 4-6 weeks duration were that 24-h urinary carnitine excretion was increased approximately twofold and L-carnitine levels were decreased in plasma (46%) and sciatic nerve endoneurium (31%). These changes in carnitine levels/excretion were associated with decreased caudal nerve conduction velocity (10-15%) and sciatic nerve changes in Na(+)-K(+)-ATPase activity (decreased 50%), Mg(2+)-ATPase (decreased 65%), 1,2-diacyl-sn-glycerol (DAG) (decreased 40%), vascular albumin permeation (increased 60%), and blood flow (increased 65%). Treatment with acetyl-L-carnitine normalized plasma and endoneurial L-carnitine levels and prevented all of these metabolic and functional changes except the increased blood flow, which was unaffected, and the reduction in DAG, which decreased another 40%. In conclusion, these observations 1) demonstrate a link between imbalances in carnitine metabolism and several metabolic and functional abnormalities associated with diabetic polyneuropathy and 2) indicate that decreased sciatic nerve endoneurial ATPase activity (ouabain-sensitive and insensitive) in this model of diabetes is associated with decreased DAG.

Urinary carnitine excretion in patients with heart failure.

To evaluate the fatty acid metabolism in heart failure, the semiquantitative analysis of urinary free carnitine and acylcarnitine was made by fast atom bombardment mass spectrometry (FABMS) in 22 patients (mean age 67.3 years) with heart failure and 19 age-matched healthy controls (average age 60.4 years). Urinary excretion of free carnitine was 0.20 +/- 0.118 ratio/mg creatinine in the healthy controls and 1.32 +/- 1.170 ratio/mg creatinine in the patients with heart failure. The latter value was significantly higher (p < 0.01). Patients with heart failure were classified into two groups according to the urinary free carnitine concentration. One was the high excretion group (2.19 +/- 0.102 ratio/mg creatinine, 12 cases) and the other was the low excretion group (0.37 +/- 0.212 ratio/mg creatinine, 10 cases). In the high excretion group, urinary acetylcarnitine was also increased, but no significant abnormalities were observed in the urinary organic acid profile. In the high group, 1 patient was classified as NYHA class III and 11 as NYHA class IV. Four patients died in the hospital. In the low excretion group, five patients were classified as NYHA class III and five as NYHA class IV. Only one patient died in the hospital. In the high group, patients with severe and prolonged heart failure tended to maintain higher values of urinary free carnitine. We could not find any abnormalities in fatty acid metabolism in patients with heart failure, but it is suspected that the patients who excrete large amounts of free carnitine into the urine, namely the patients with severe heart failure, have some possibility of carnitine deficiency.

Restricting Food Intake Does Not Exacerbate the Effects of a Choline-Deficient Diet on Tissue Carnitine Concentrations in Rats

Previous reports in animals indicate that choline deficiency alters carnitine metabolism. Recent studies in humans suggest that choline deficiency occurs in individuals during long term total parenteral nutrition. Malnutrition is also a frequent complication in this population. We therefore examined the effect of restricting the intake of a choline-deficient diet on carnitine concentrations in plasma and tissues. Adult male rats were randomly assigned to one of four dietary regimens: control, choline deficient, restricted control (85% of control), or restricted choline deficient for 42-43 d. At the end of the experimental period, restricted animals weighed significantly less than their respective controls (P < 0.01). Liver weight relative to body weight and fat concentration were greater in choline-deficient animals (P < 0.01 and 0.001, respectively). Choline-deficient rats fed free access had elevated plasma carnitine concentration (P < 0.01). Urinary carnitine excretion was elevated in both groups of choline-deficient rats (P < 0.01), while liver, heart and muscle carnitine concentrations were lower than in controls (P < 0.05). Restricting dietary intake reduced plasma carnitine concentration in choline-deficient animals (P < 0.01), but did not alter tissue or urine carnitine concentrations in either group. Restricted, choline-deficient animals did not exhibit a worsening of the sequelae of choline deficiency. We conclude that choline deficiency alters carnitine concentrations in plasma and tissues and that restricting the intake of a choline-deficient diet does not alter this effect in tissues.

Primary defect of juvenile visceral steatosis (jvs) mouse with systemic carnitine deficiency is probably in renal carnitine transport system

We investigated the reabsorptional system for carnitine in the kidney to elucidate the mechanism of carnitine deficiency in juvenile visceral steatosis ( jvs) mice. Jvs mice had a higher rate of carnitine excretion at 10 days after birth than the controls, in spite of having no pathological acylcarnitine in the urine. In an experiment to assay the uptake of carnitine using kidney slices, homozygous mutants showed significantly lower rates of Na-dependent carnitine uptake than controls. Heterozygous mice showed values of transport activity intermediate between homozygous mutants and homozygous controls. Scatchard plots (transport activity versus transport activity/carnitine concentration) revealed that the homozygous mutants had a defect in the hihg affinity site ( K m = 58 μM) in the Na-dependent carnitine transport system in the kidney. These results indicate that the primary defect of jvs mice is most probably related to the system for reabsorption of carnitine in the kidney.

Intravenous L-Carnitine and Acetyl-L-Carnitine in Medium-Chain Acyl-Coenzyme A Dehydrogenase Deficiency and Isovaleric Acidemia

The purpose of this study was to determine whether treatment with L-carnitine or acetyl-L-carnitine enhances the turnover of lipid or branched-chain amino acid oxidation in patients with inborn errors of metabolism. Increasing i.v. doses of L-carnitine and acetyl-L-carnitine were given to one patient with medium-chain acyl-CoA dehydrogenase deficiency and to another with isovaleric acidemia. Both patients were in stable condition and receiving oral L-carnitine supplements. The excretion of carnitine and disease-specific metabolites was measured. The incorporation of L-carnitine in the intracellular pool was demonstrated using stable isotopes and mass spectrometry. Increasing doses of either i.v. L-carnitine or acetyl-L-carnitine did not stimulate the excretion of octanoylcarnitine in the patient with medium-chain acyl-CoA dehydrogenase deficiency, nor did it raise the plasma levels of either cis-4-decenoate or octanoylcarnitine. Similarly, increasing doses of either i.v. L-carnitine or acetyl-L-carnitine did not enhance the excretion of isovalerylcarnitine in a patient with isovaleric acidemia. The excretion of isovalerylglycine actually decreased. We conclude that there was no evidence of enhanced fatty acid beta-oxidation or enhanced branched-chain amino acid oxidation in vivo by the administration of high doses of L-carnitine or acetyl-L-carnitine in these two patients. Because only one individual with each disorder was studied, the data are only indicative and may not necessarily be representative of all individuals with these disorders. Definite settlement of this issue will require further studies in additional subjects.

Effect of dietary macronutrient content on carnitine excretion and efficiency of carnitine reabsorption

We examined the effect of macronutrient content on glomerular filtration rate (GFR), and excretion, reabsorption, and filtered load of carnitine. Ten subjects consumed five diets [high protein (HP), low protein (LP), control, high fat (HF), and high carbohydrate (HC)] of equal energy and carnitine content for 6 d each, in a randomized crossover manner. The rate of carnitine excretion was lower after the LP diet than after the HP diet because of lower GFR after the LP diet. The rate of carnitine reabsorption was lower after the LP diet than after the HP diet, also because of the lower GFR after the LP diet. The rate of carnitine reabsorption was not different after the HF and HC diets, nor was GFR. The filtered load of carnitine, however, was greater after the HF diet, resulting in a higher rate of carnitine excretion.

Renal adaptation to dietary carnitine in humans

Carnitine homeostasis in humans is maintained by dietary carnitine intake, a modest rate of endogenous carnitine synthesis, and efficient conservation of carnitine by the kidney. To assess the effect of dietary carnitine on the efficiency of carnitine reabsorption in humans, rates of carnitine excretion and reabsorption, indexed to the glomerular filtration rate, were determined over a range of plasma free and total carnitine concentrations in 12 strict vegetarians before and after dietary carnitine supplementation (0.248 mmol/d). This amount of dietary carnitine supplementation did not significantly increase plasma carnitine concentration and did not alter the glomerular filtration rate. At normal physiological plasma carnitine concentrations, the rate of carnitine excretion was increased and the rate of carnitine reabsorption was decreased by carnitine supplementation. We conclude that the kidney adapts to carnitine intake by reducing the efficiency of carnitine reabsorption.

Relationship of Carnitine and Carnitine Precursors Lysine, ɛ-N-Trimethyllysine, and γ-Butyrobetaine in Drug-Induced Carnitine Depletion

Plasma concentrations and rates of urinary excretion of carnitine and some of its precursors were studied in three groups of children receiving drugs known to cause carnitine depletion. Patients in group A received pivampicillin and a molar equivalent of carnitine for 7 d. Patients in group B received pivampicillin with a 5.8-fold molar excess of carnitine for 1 wk. Patients in group C were treated chronically with valproic acid and received a molar equivalent (to valproic acid) of carnitine for 14 d. Patients in group A had markedly increased (16-fold) urinary carnitine ester excretion concomitant with diminished urinary free carnitine and gamma-butyrobetaine output and lower plasma free carnitine concentration. Supplementation with one molar equivalent of carnitine (to pivampicillin) was ineffective in preventing the reduction of plasma carnitine concentration observed with pivampicillin treatment alone. For group B patients, administration of excess carnitine resulted in a further increase (35-fold) of urinary carnitine ester output with no decrease of plasma carnitine concentration, urinary gamma-butyrobetaine, or free carnitine excretion. For patients in group C, the initially low plasma free and total carnitine concentrations and urinary output of carnitine and carnitine esters markedly increased with carnitine supplementation, but urinary excretion of gamma-butyrobetaine remained unchanged. The plasma concentrations and urinary output of L-lysine and epsilon-N-trimethyllysine remained unchanged within each group before and after treatment. A positive linear correlation was found between urinary epsilon-N-trimethyllysine and 3-methylhistidine output, indicating that the rate of epsilon-N-trimethyllysine excretion correlates with the amount of 3-methylhistidine liberated by protein turnover.(ABSTRACT TRUNCATED AT 250 WORDS)

Muscle carnitine repletion by long-term carnitine supplementation in nephropathic cystinosis.

The renal tubular Fanconi syndrome of children with nephropathic cystinosis causes plasma and muscle carnitine depletion. L-Carnitine replacement therapy for up to 18 mo has previously been shown to normalize plasma but not muscle carnitine levels. We treated six cystinosis patients, aged 1 to 4 y, with a mean dosage of 92 mg L-carnitine/kg/d given every 6 h for an average of 62 mo. Despite fractional excretions of free carnitine ranging from 55 to 108%, plasma-free and total carnitine concentrations were maintained at or above normal levels. At the end of the carnitine replacement period, the six children had muscle-free carnitine values ranging from 16.0 to 28.0 nmol/mg noncollagen protein compared with values of 3.0 to 11.4 for cystinosis children not supplemented with carnitine [normal, 22.7 +/- 5.0 (SD) nmol/mg protein]. Total muscle carnitine values were also normalized by L-carnitine replacement. The monthly increase in total body creatinine production, a measure of muscle mass, was higher (p = 0.036) in children with normal plasma free carnitine concentrations (3.4 +/- 0.9 mg/d) than in children with low plasma free carnitine (2.3 +/- 0.7 mg/d). No serious side effects, such as severe diarrhea, were observed. We conclude that oral L-carnitine replacement can normalize muscle carnitine content in children with cystinosis.

Renal Handling of Carnitine in Secondary Carnitine Deficiency Disorders

Reduced plasma and tissue concentrations of carnitine, a cofactor required for fatty acid oxidation, are present in patients with inherited disorders of mitochondrial acyl-CoA oxidation that are associated with accumulations of acylcarnitines. To determine whether the secondary carnitine deficiency in these patients is due to excessive urinary loss of acylcarnitines, the development of carnitine deficiency was examined in patients with four different acyl-CoA oxidation disorders, including medium-chain and long-chain fatty acyl-CoA dehydrogenase deficiencies, isovaleric acidemia, and propionic acidemia. After a 3-mo period of treatment with oral carnitine to raise plasma total carnitine concentrations to or above normal, patients were started on a carnitine-free diet and the changes in plasma total and free carnitine levels and urinary total and free carnitine excretion were followed for 5 d. Patients with all four disorders showed a return of plasma carnitine levels and urinary carnitine excretion to baseline within 2 to 4 d. The rapidity of these changes could not be explained solely by excessive acylcarnitine wasting. Continued excretion of free carnitine in all patients indicated the additional presence of an impairment in renal transport of free carnitine. Consistent with this interpretation, estimates of renal thresholds for free carnitine gave values that were less than that for a control child in all four disorders and ranged as low as one half those reported in normal individuals. These results suggest that secondary carnitine deficiency in the acyl-CoA oxidation disorders is due to indirect as well as direct effects of accumulated acylcarnitines.(ABSTRACT TRUNCATED AT 250 WORDS)

EFFECT OF MEDIUM CHAIN TRIGLYCERIDE (MCT) ADMINISTRATION ON CARNITINE METABOLISM IN THE ANESTHESIZED PIGLET AND CULTURED KIDNEY EPITHELIAL CELLS

To test the hypothesis that carnitine plays a role in medium chain fatty acid metabolism, blood, urine, liver and bile specimens were collected from anesthesized neonatal piglets prior to, and after 30, 60, 120 and 180 minutes of a continuous intravenous infusion of MCT (1 g/kg-h). Interval fractional tubular reabsorption was calculated throughout the infusion. 3H-L-Carnitine uptake was determined in cultured renal tubular epithelial (Cos-l) cells incubated with increasing amounts of octanoic acid.Results: The ketogenic response to MCT administration was associated with a rise in esterified (15.2-->40.9 μM), particularly acetylcamitine (11.4-->26.2 μM), and a fall in free carnitine plasma concentrations (34.6-->14.4 μM). The appearance in the plasma, bile and liver of butyryl-, hexanoyl-, octanoyl- and decanoylcarniune and the greater than 4-fold increase in ratios of plasma free fatty acids/β-hydroxybutyratc and lactate/pyruvate suggested overloading of both β- and Krebs cycle oxidative pathways. The fractional tubular reabsorption of free carnitine decreased from 96.5% to 48.7% with MCT loading, and was associated with a 6-fold increase of free carnitine excretion. Uptake of 3H-L-Carnitine into Cos-1 cells decreased in a dose-dependent manner with increasing octanoic acid concentration. The results suggest that MCT loading has profound effects on carnitine metabolism affecting intracellular esterifications and renal tubular reabsorption.

Metabolism of S-1108, a new oral cephem antibiotic, and metabolic profiles of its metabolites in humans.

The metabolism and pharmacokinetics of pivalic acid, a major metabolite of S-1108, were studied with three healthy volunteers. Concentrations of S-1006 (the active compound), pivalic acid, and pivaloylcarnitine in plasma and urine were measured after administration of S-1108. Recoveries in urine at the doses of S-1108 given (100 and 200 mg) were 33 to 41% for S-1006, 93% for total pivalic acid, and 89 to 94% for pivaloylcarnitine in 24 h, and maximum concentrations in plasma were 2 micrograms of S-1006 per ml, 1 micrograms of total pivalic acid per ml, and 2 micrograms of pivaloylcarnitine per ml after a 200-mg oral administration of S-1108. More than 90% of the pivalic acid was excreted as pivaloylcarnitine, and no measurable amount of free pivalic acid was present in urine samples, indicating that the pivalic acid liberated from S-1108 was almost quantitatively conjugated with carnitine in the human body. The level of free carnitine in plasma was unaffected by a single 200-mg administration of S-1108, whereas urinary excretion of free carnitine decreased as levels of acylcarnitine increased. The acylcarnitines were excreted primarily in the form of pivaloylcarnitine. This study clearly showed how the pivalic acid was metabolized and excreted in humans. The importance of monitoring carnitine, an essential cofactor in fatty acid metabolism, was also discussed in terms of its utilization by pivalic acid.

Prevalence of carnitine depletion in critically ill patients with undernutrition

The aim of this study was to evaluate to what extent secondary carnitine deficiency may exist based on the prevalence of subnormal carnitine status in patients with critical illness and abnormal nutritional state. Healthy control patients (n = 12) were investigated and compared with patients with possible secondary carnitine deficiency, ie, patients with overt severe protein-energy malnutrition (PEM, n = 28), postoperative long-term (> 14 days) parenteral glucose feeding (250 g glucose/d, n = 7), severe liver disease (n = 10), renal insufficiency (n = 7), and sustained septicemia with increased metabolic rate (n = 8). Nutritional status, energy expenditure, creatinine excretion, and blood biochemical tests were measured in relationship to free and total carnitine concentrations in plasma and skeletal muscle tissue, as well as urinary excretion of free and total carnitine. The overall mortality rate was 48% within 30 days of the investigation in study patients with the highest mortality in liver disease (90%). The hospitalization range was 14 to 129 days in study patients. Most study patients had lost weight (4% to 19%) and had abnormal body composition. Patients with liver disease, septicemia, renal insufficiency, and those on long-term glucose feeding had significantly higher than predicted metabolic rate (+25% ± 3%), while patients with severe malnutrition had decreased metabolic rate compared with controls. Patients with liver disease had increased plasma concentrations of free (96 ± 16 μmol/L) and total (144 ± 27 μmol/L) carnitine compared with controls (45 ± 3,58 ± 7 μmol/L, respectively). No other differences among the groups were seen in plasma or muscle carnitine concentrations as compared with control patients, although some patient groups (liver disease, sever PEM, long-term glucose feeding) showed indices of increased free and total carnitine excretion. There was no correlation between muscle and plasma carnitine concentrations. The present study demonstrates that the overall prevalence of statistically significant carnitine depletion was less than 5% in a large heterogeneous group of patients suffering from critical illness and abnormal nutritional state.

Quantitative estimation of absorption and degradation of a carnitine supplement by human adults

Results of kinetic and pharmacokinetic studies have suggested that dietary carnitine supplements are not totally absorbed, and are in part degraded in the gastrointestinal tract of humans. To determine the metabolic fate of dietary carnitine supplements in humans, we administered orally a tracer dose of [ methyl- 3H] l-carnitine with a meal to five normal adult males, who had been adapted to a high-carnitine diet plus carnitine supplement (2 g/d) for 14 days. Appearance of [ methyl- 3H] l-carnitine and metabolites in serum, and urinary and fecal excretion of radiolabeled carnitine and metabolites was monitored for 5 to 11 days following administration of the test dose. Maximum concentration of [ methyl- 3H] l-carnitine in serum occurred at 2.0 to 4.5 hours after administration of the tracer, indicating relatively slow absorption from the intestinal lumen. Total radioactive metabolites excreted in urine and feces ranged from 47% to 55% of the ingested tracer. Major metabolites found were [ 3H]trimethylamine N-oxide (8% to 49% of the administered dose; excreted primarily in urine) and [ 3H]γ-butyrobetaine (0.44% to 45% of the administered dose; excreted primarily in feces). Urinary excretion of total carnitine was 16% to 23% of intake. Fecal excretion of total carnitine was negligible (less than 2% of total carnitine excretion).

Dietary carnitine effects on carnitine concentrations in urine and milk in lactating women

The effect of dietary carnitine on urinary excretion of free and total carnitine and on breast-milk secretion of the carnitine fractions in 15 control and 16 lactating women aged 21-40 y was measured. Free and total carnitine excretions, obtained from 24-h urine collections, correlated with carnitine consumed on the collection day (P less than 0.03, P less than 0.01, respectively) but not with the mean intake calculated from 3-d diet records. The immediate responses of the control and lactating groups were not significantly different. Urinary excretion of carnitine (n = 31) was 82 +/- 13 mumol/d for free excretion and 226 +/- 22 mumol/d for total excretion. Milk free, acid-soluble acyl-, acid-insoluble acyl-, or total carnitine did not correlate with dietary carnitine or with the duration of lactation (1-10 mo). Milk total carnitine was 45 +/- 3 mumol/L. With the carnitine content of breast milk remaining stable for greater than or equal to 10 mo, the importance of exogenous carnitine throughout infancy is suggested.

Increased Dietary Branched-Chain Amino Acids Do Not Improve Growth in Developing Rats with Chronic Biliary Obstruction

We studied dietary branched-chain amino acid enrichment in cholestatic weanling rats. Growth was assessed with body weight, muscle weight and nitrogen balance. Systemic metabolic measurements that reflect liver function were evaluated, including plasma ammonia, albumin, amino acids, glucose, triglyceride and branched-chain ketoacids, as well as urinary carnitine excretion. Twenty-two rats underwent bile-duct ligation at 14 d of age. At weaning, 11 rats were fed a control diet and 11 an isoenergetic, isonitrogenous branched-chain amino acid-enriched diet for 3 wk, each with a sham-operated, pair-fed control. Body weights were similar in all four groups. Changes due to bile-duct ligation and not affected by the diet manipulation included lower plasma glucose, nitrogen balance and muscle weight, and higher triglyceride concentration, carnitine excretion and liver weight. Changes due to ligation that were normalized by dietary manipulation included plasma albumin, ammonia and total amino acid concentrations. The ratio of branched-chain to aromatic amino acids was decreased in ligated animals fed both diets; however, branched-chain amino acids were lower in the two groups fed more branched-chain amino acids.

Metabolic effects of pivalate in isolated rat hepatocytes

Pivalate (trimethylacetic acid) administration in humans or rat has been reported to cause metabolic changes and increased urinary carnitine excretion secondary to pivaloylcarnitine generation. As pivaloylcarnitine formation is dependent on intracellular activation of pivalate, the effects of pivalate on cellular coenzyme A and acyl-CoA contents and oxidative metabolism were defined using isolated rat hepatocytes. During incubations with pivalate (1.0 mM), hepatocyte coenzyme A content fell to less than 0.05 nmol/10(6) cells (vs 0.97 nmol/10(6) cells in the absence of pivalate) as pivaloyl-CoA accumulated. Pivalate (5 mM) inhibited 14CO2 generation from 10 mM [1-14C]pyruvate by 34%, but had no effect on 0.8 mM [1-14C]palmitate oxidation. Pivaloyl-CoA was a substrate for hepatocyte carnitine acyltransferase activity, but supported acylcarnitine formation at rates only 10-20% of those observed with equimolar acetyl-CoA or isovaleryl-CoA as substrates. Thus, hepatocytes activate pivalate to pivaloyl-CoA, which can then be used as a substrate for pivaloylcarnitine formation. The sequestration of hepatocyte coenzyme A as pivaloyl-CoA is associated with inhibition of pyruvate oxidation. As with other organic carboxylic acids, activation of pivalate to the coenzyme A thioester is an important aspect in the biochemical toxicology of the compound.