Carnitine content of liquid formulas and special diets.

Radioisotopic analyses for carnitine content were done on infant formula, formulas for tube feeding, food supplements, and chemically defined diets. The carnitine content of the diets depend on the protein source. Products whose main protein source is soy protein isolate, casein, or egg white solids contain 4 nmole carnitine per milliliter or less, with most of them containing undetectable amounts of carnitine. Products based on milk or beef contain 50 to 656 nmole carnitine per milliliter. The daily requirement of the body for carnitine is unknown. Evidence is discussed that indicates that the possible use of carnitine as a supplement to formula diets intended for long-term care needs to be considered.

Carnitine status in Thai adults.

Plasma carnitine and urinary carnitine levels were measured in Thai adults living in Bangkok city and Ubol villages. The mean plasma carnitine and urinary carnitine levels expressed in micromoles per liter in Bangkok adults were higher than those in Ubol adults. Their mean plasma carnitine levels were 56.6 +/- 1.8 and 50.3 +/- 1.7 whereas urinary carnitine levels were 161 +/- 19 and 127 +/- 18 micromole/liter, respectively. The nutritional status in Ubol adults was inadequate. This was evidenced by the significant decrease in urinary creatinine excretion, serum albumin, and hematocrit levels. The dietary assessment agreed with the biochemical findings. Since rice, limiting in carnitine, was the main protein and energy source consumed by Ubol adults their inadequate carnitine status could be due to the low carnitine intake. Sex affects plasma carnitine levels in Bangkok adults and urinary carnitine excretion in both groups. This could be related to the lean body mass in which most of the body carnitine resides. This is supported by the higher urinary creatinine excretion in males and the significant positive correlation between carnitine excretion and creatinine-height index.

Buthionine sulfoximine, an experimental tool to induce glutathione deficiency: elucidation of glutathione and ascorbate in their role as antioxidants.

The development of buthionine sulfoximine, a selective inhibitor of glutathione biosynthesis, is an important new tool to elucidate the in vivo role of glutathione. Recent investigations have shown that ascorbic acid can serve as an essential antioxidant in the presence of severe glutathione deficiency.

Muscle carnitine levels in neuromuscular disease.

The production of energy in muscle from long-chain fatty acid oxidation is dependent upon the presence of carnitine. An abnormally low level of muscle carnitine, as seen in patients with the carnitine deficiency syndrome, results in marked muscle weakness. Muscle from 83 consecutive patients undergoing diagnostic muscle biopsy was assayed for carnitine. Carnitine levels (mean +/- SEM, expressed as nmoles carnitine per mg noncollagen protein) in muscle from patients with Duchenne dystrophy (8.1 +/- 1.7) and possible Becker dystrophy (10.6 +/- 3.0) were significantly (P less than 0.001) different from histologically normal muscle (24.0 +/- 1.4). Carnitine levels in patients with limb-girdle dystrophy (16.1 +/- 3.1) and polymyositis/dermatomyositis (16.6 +/- 3.2) were also low, although not as low as in Duchenne dystrophy. Carnitine levels from patients with denervation atrophy (22.1 +/- 3.6), nonspecific fiber atrophy (21.3 +/- 1.3), and a group of miscellaneous neuromuscular diseases (20.4 +/- 1.4) were not significantly different from histologically normal muscle. The low values of carnitine seen in Duchenne dystrophy and a group of possible Becker dystrophy patients may be a nonspecific effect, related to severe muscle damage.

Effects of slaframine on ruminant digestive function: resting salivary flow and composition in cattle.

Four esophageal- and ruminal-cannulated Angus steers (avg weight, 308 kg) were used to investigate how salivation is affected by the administration of purified slaframine (SF; 1-acetoxy-6-aminooctahydroindolizine), a cholinergic secretagogue isolated from Rhizoctonia leguminicola. Steers were fed a concentrate diet at twice the net energy requirement for maintenance in hourly increments. In trial 1, a single injection of SF was administered to four steers intramuscularly at 0, 6, 12 and 24-micrograms/kg body weight (BW) in a 4 X 4 Latin-square design. Saliva was collected via esophageal cannula at 15-min intervals 30 min after each feeding, weighted, sampled and reinfused via ruminal cannula over a 10-h period. At 12- and 24-micrograms SF/kg BW, salivary flow was 31 to 43% greater (P less than .01) than at 0- or 6-micrograms SF/kg. Response peaked within the first 3 h and returned to baseline levels at 8 h. Buffering capacity and pH of saliva were not different (P greater than .10); however, osmolality and Na concentration increased and K concentration decreased (P less than .10) as salivation rates increased. Feed intake of steers did not appear affected at any level of SF administration. In trial 2, 0-, 12- and 24-micrograms SF/kg BW were repeatedly administered intramuscularly to three steers in a 3 X 3 Latin-square design at 8-h intervals for 24 h. Salivary flow was measured and sampled over the entire 24-h period, as in trial 1. Flow rates were increased 50 to 70% (P less than .01) by SF treatment.(ABSTRACT TRUNCATED AT 250 WORDS)

Effects of slaframine on ruminant digestive function: ruminal motility in sheep and cattle.

Effect of purified slaframine (SF; 1-acetoxy-6-aminooctahydroindolizine), a parasympathomimetic secretagogue isolated from Rhizoctonia leguminicola, on ruminal motility was investigated in cattle and sheep. In trial 1, four ruminal cannulated wethers, fed a pelleted concentrate and hay diet, were injected intramuscularly with 0, 12, 24 and 48 micrograms SF/kg body weight (BW) in a 4 X 4 Latin-square design. Ruminal motility was recorded 1 h before and 1 to 2 h and 3 to 4 h after SF administration by measuring pressure changes exerted upon a fluid-filled, open-tipped catheter inserted into the dorsal sac of the rumen. The frequencies of both primary and secondary ruminal contractions were decreased as much as 20 to 78% with SF (P less than .05) depending upon the dosage level and time after administration. In trial 2, three ruminal-cannulated steers fed a concentrated diet were injected intramuscularly with 0, 12 and 24 micrograms SF/kg BW in a 3 X 3 Latin-square design. A water-filled balloon inserted into the cranial sac of the rumen was used to measure ruminal pressure changes 1 h before and 1 to 2 h, 3 to 4 h and 7 to 8 after SE administration. Frequency of primary and secondary ruminal contractions decreased with SF as much as 27 to 64% depending on the dosage level and time after administration. The frequency of secondary contractions increased 28% (P less than .05) as compared with control during the 7 to 8 h after administration.(ABSTRACT TRUNCATED AT 250 WORDS)

Effects of slaframine on ruminant digestive function: liquid turnover rate and fermentation patterns in sheep and cattle.

Two trials were initiated to determine if slaframine (SF) can be used to alter fluid digesta flow and fermentation patterns in the rumen. In trial 1, a preliminary experiment, four Dorset X Barbados Black-belly ruminal-cannulated wethers (avg weight 41.6 8.7 kg) given ad libitum access to a pelleted concentrate/hay diet were injected intramuscularly with 0, 12, 24 or 48 micrograms SF/kg body weight (BW) in a 4 X 4 Latin-square design. Ruminal fluid dilution rate was determined using a single intraruminal infusion of polyethylene glycol (7 g), followed by seven hourly ruminal fluid samples. The administration of 48 micrograms SF/kg BW increased (P less than .10) ruminal volume and outflow by 27 and 25%, respectively, compared with controls. In trial 2, two Hereford and two Angus ruminal cannulated steers (avg weight 568 +/- 93 kg) were injected with 0, 6, 12 or 24 micrograms SF/kg BW at 8-h intervals over a 24-h period in a 4 X 4 Latin-square design. Steers were fed a concentrate diet at twice maintenance in 24 equal portions daily. Ruminal fluid dilution was measured using a single intraruminal infusion of cobalt-ethylenediamine tetraacetic acid (20 g) administered 9 h after the initial SF injection. Ruminal fluid was collected each hour during 8 to 24 h after the initial SF injection and analyzed for pH, osmolality and volatile fatty acids (VFA).(ABSTRACT TRUNCATED AT 250 WORDS)

Effects of chronic administration of slaframine on production and digestive function in broiler chicks.

Slaframine (SF), 1-acetoxy-6-aminooctahydroindolizine a parasympathomimetic with a high affinity for the gastrointestinal tract, was administered by oral intubation daily to 240 broiler chicks at either 0, 8.9, or 17.8 micrograms/kg body weight.75 (BW.75) in saline for 21 days. Throughout the experimental period weight, feed intake, and fecal output were measured. On Day 21 birds were killed, eviscerated, and wet organ weights were obtained. Pancreas and small intestine digesta were homogenized with saline and frozen for analyses of trypsin, chymotrypsin, amylase, and lipase activity as well as total protein. Weight, feed intake and utilization, pancreatic weight, liver weight, and small intestine digesta weight were not affected by SF treatment. Protein content of the digesta decreased 16.6% with the 17.8 micrograms SF/kg BW.75 treatment. Digesta lipase activity was 13.3% (P greater than .05) and specific activity 24% less (P less than or equal to .02) in 17.8 micrograms/kg BW.75 treated birds in comparison with those of controls, and activities decreased in a linear fashion across treatment levels (P less than or equal to .04). Digesta trypsin-specific activity decreased linearly with SF treatment (P less than or equal to .05), averaging 5.5 to 16.9% lower than control treated birds. Pancreatic chymotrypsin-specific activity was not significantly different among treatments. These results suggest that relatively small dosages of SF may affect digestive function of broiler chicks.

Effects of slaframine on circulating concentrations of growth hormone and glucose.

The ability of slaframine (SF), a parasympathomimetic, to alter blood growth hormone (GH) and glucose concentrations in broiler chicks was investigated. Eighty male broiler chicks (average weight 225 g) were divided into 10 groups and dosed with either saline (control) or 1 mg SF/kg of body weight by oral intubation. Plasma samples were obtained from separate groups of chicks at 1, 2, 4, 8, and 12 hr after SF administration and analyzed for growth hormone and glucose. One hour after SF administration, glucose increased (P less than .05) 21.4% compared with controls. Growth hormone increased (P less than .05) 449 to 948% from 8 to 12 hr after SF administration. Administration of SF at 1 mg/kg of body weight was associated with increased plasma GH.

Memories of microbes and metabolism.

As Jackie Gleason was wont to say: "How sweet it really is!" And--reflecting on the 1940s-1980s, when studies of microbial nutrition revealed exciting structure-function relationships of the B-complex vitamins with relevance to metabolism in humans--it really is. A chemistry degree from Beloit College and a doctorate in biochemistry from the University of Wisconsin set the stage for my life's work at Lederle Laboratories, the University of Illinois, and Vanderbilt University. At Lederle my research contributed to folic acid chemistry: coenzyme forms and function; antimetabolites and cancer chemotherapy. My subsequent university studies centered on lysine biosynthesis and metabolism, e.g. its precursor role in carnitine and in indolizidine alkaloids of physiological interest. There were also many opportunities to reach out and give something back to the system via teaching and diverse service activities, all of which has led to a happy, fulfilling career, one for which I am ever thankful.

Carnitine biosynthesis in Neurospora crassa: enzymatic conversion of lysine to epsilon-N-trimethyllysine.

The enzymatic conversion of L-lysine, epsilon-N-trimethyl-L-lysine the first series of reactions in the biosynthesis of carnitine in Neurospora crassa, proceeds via sequential methylation of free L-lysine, epsilon-N-methyl-L-lysine, and epsilon -N-dimethyl-L-lysine. The latter two compounds have been shown to be intermediates in the biosynthesis of carnitine by radioisotope dilution and incorporation experiments in growing cultures of N. crassa 33933 (lys-) and 38706 (met-). Methionine but not choline, has been recognized as an effective methyl donor in vivo. Inclusion of choline in the growth medium of strain 33933 does, however, enhance incorporation of the methyl groups of L-[methyl-3H]methionine into carnitine in an apparent "sparing" effect on methionine synthesis. Studies in cell-free extracts of the lysine auxotroph strain 33933 of N. crassa have established that lysine and epsilon-N-methyl and epsilon-N-dimethyllysine are enzymatically methylated, with S-adenosyl-L-methionine as the methyl group donor. The enzyme system appears to have no essential cofactors. Lysine does not induce synthesis of the enzyme system in the wild-type strain 262, whereas both carnitine and epsilon-N-trimethyllysine repress its synthesis in strain 33933.

Lysine catabolism in Rhizoctonia leguminicola and related fungi.

The catabolism of lysine was studied in several yeasts and fungi. Results with cell-free extracts of Rhizoctonia leguminicola support a proposed pathway involving (D- and L-) EPSILON-N-acetyllysine, alpha-keto-epsilon-acetamidohexanoic acid, delta-acetamidovaleric acid, and delta-aminovaleric acid in the conversion of L-lysine to shortchain organic acids. Label from radioactive L-lysine was found to accumulate in D- and L-epsilon-N-acetyllysine, delta-acetamidovaleric acid, delta-aminovaleric acid, and glutaric acid in cultures of R. leguminicola, Neurospora crassa, Saccharomyces cerevisiae, and Hansenula saturnus, suggesting that the proposed omega-acetyl pathway of lysine catabolism is generalized among yeasts and fungi. In N. crassa, as is the case in R. leguminicola, the major precursor of L-pipecolic acid was the L-isomer of lysine; 15N experiments were consistent with delta1-piperideine-2-carboxylic acid as an intermediate in the transformation.