Utilization of tryptophan, nicotinamide and nicotinic acid as precursors for nicotinamide nucleotide synthesis in isolated rat liver cells

1. Incubation of isolated rat hepatocytes with nicotinamide or nicotinic acid showed that while both vitamers were taken up from the incubation medium, neither was utilized to any significant extent as a precursor of the nicotinamide nucleotide coenzymes, NAD and NADP, and neither was capable of preventing the loss of nucleotides that occurs on incubating the cells. 2. Incubation of hepatocytes with tryptophan showed that de novo synthesis from tryptophan permitted replacement of the nucleotides lost during incubation; at high concentrations of tryptophan there was an increase above the initial intracellular concentration of NAD(P). Incubation of hepatocytes with tryptophan also resulted in the formation and release from the cells of a considerable amount of niacin, as well as the two principal metabolities of NAD(P), N1-methyl nicotinamide and methyl pyridone carboxamide. 3. It is suggested that, in the liver, performed niacin is not utilized for nucleotide synthesis, and indeed the function of the liver appears to be synthesis of niacin from tryptophan, and its release for use by extrahepatic tissues that lack the pathway for de novo synthesis of nicotinamide nucleotides from tryptophan.     

Fate of excess nicotinamide and nicotinic acid differs in rats

Rats administered excess nicotinamide and nicotinic acid were studied to determine the metabolic fate of pharmacological levels of these compounds. When a large amount of nicotinamide (500 mg/kg body wt) was intraperitoneally injected into rats, 32% of the dose was excreted as nicotinamide, 11% as N1-methylnicotinamide (MNA), 10% as nicotinuric acid, 5% as nicotinic acid, 3% as N1-methyl-4-pyridone-3-carboxamide (4-pyr) and 2% as N1-methyl-2-pyridone-5-carboxamide (2-pyr) during d 1 after the injection. Urinary excretion of these compounds gradually decreased with time and returned to normal by d 3. Urinary excretion of nicotinic acid and nicotinuric acid was observed only on d 1. When a large amount of nicotinic acid (500 mg/kg body wt) was intraperitoneally injected into rats, 55% of the dose was excreted as nicotinic acid and 15% as nicotinuric acid during d 1, and no excretion of these compounds was observed thereafter. The increase in excretion of nicotinamide, MNA, 2-pyr and 4-pyr was slight even on d 1. Excretion of nicotinic acid, nicotinuric acid, nicotinamide, MNA, 2-pyr and 4-pyr returned to normal levels on d 2. From these results, the different fates of excess nicotinamide and nicotinic acid are discussed.     

Effects of chronic ethanol feeding on the metabolism of tryptophan and nicotinamide in rats

Oxidation of tryptophan and urinary excretion of N1-methylnicotinamide were studied in rats fed a modified DeCarli-Lieber liquid diet of low (1.1 mg/liter or 5 mg/kg dry diet) and adequate (5.5 mg/liter or 25 mg/kg dry diet) nicotinamide content with and without ethanol replacing carbohydrate to 36% of total calories. Protein was restricted to 12% of calories so that the tryptophan content was about 0.16% of dry weight. Rats were fed for 30 or 40 days and feeding was restricted to 7 or 12 hours per day. N1-Methylnicotinamide was measured in urine collected during the last 2 days of feeding. At the end of the experimental period, fasted rats were injected intraperitoneally with 0.8 muCi of L-[ring-2-14C]tryptophan (0.12 or 0.11 muCi/mg tryptophan) or 0.7 muCi of [14C]formate (0.44 muCi/mg formate) per 100 g body weight. Expired CO2 was collected every 30 minutes or hourly for 5 or 6 hours. N1-Methylnicotinamide excretion was increased in ethanol-fed rats (P less than 0.05). The percent of injected [14C]tryptophan recovered as 14CO2 was increased in rats fed ethanol in a low niacin diet as compared to controls (P less than 0.01). Recovery of 14CO2 from [14C]formate oxidation was not affected by ethanol feeding. These findings suggest that the initial oxidation of tryptophan by tryptophan 2,3-dioxygenase is enhanced following chronic ethanol feeding.     

Effect of nicotinamide on methionine metabolism in rat liver

To test the response to increased utilization of methyl groups, we administered large dosages of nicotinamide to rats fed an adequate diet that contained limited amounts of methionine and choline. During the 4 d after the injection, we observed several significant effects on the hepatic concentrations of the enzymes and metabolites of methionine metabolism. Methionine and S-adenosylmethionine remained at control levels; the concentrations of S-adenosylhomocysteine exceeded the control values from 4 to 16 h; and the levels of serine and betaine were lower after 16 h. Treatment with nicotinamide resulted in higher hepatic levels of methionine adenosyltransferase (after 4 h) and cystathionine synthase (after 16 h). These data indicate that increases in both homocysteine methylation and S-adenosylmethionine synthesis may be components of the response to excessive methyl group consumption. An increased synthesis of cystathionine would provide for the removal of S-adenosylhomocysteine (and homocysteine) derived from the adenosylmethionine-dependent methylation of nicotinamide.     

Kynurenine metabolism and xanthurenic acid formation in vitamin B6-deficient rat after tryptophan injection

Normal and vitamin B6-deficient rats received an intraperitoneal injection of 30 mg/100g of body wt, and the contents of metabolites in kidney or plasma and the related enzyme activities in kidney were determined. The contents of kynurenine and 3-hydroxykynurenine in B6-deficient rat plasma and kidney were much higher than those in normal rat. The changes of those contents in plasma were parallel to those in kidney, but not in liver. The contents of kynurenic acid and xanthurenic acid in B6-deficient liver, plasma, and kidney were also much higher than those in normal rats. However, the changes of those contents in plasma were parallel to those in liver, but not in kidney. Xanthurenic acid and kynurenic acid accumulated to a much greater extent in kidney than in plasma and liver. Kidney kynureninase activity was very low, but kynurenine aminotransferase activities were very high. These observations indicated that the production of xanthurenic acid after tryptophan injection was favorable in B6-deficient kidney with respect to enzyme activities and substrate concentrations, and suggested that kidney took up kynurenine or 3-hydroxykynurenine from blood and after conversion of them it excreted xanthurenic or kynurenic acid into urine.     

The effect of high intakes of casein and casein phosphopeptide on calcium absorption in the rat

The effect of the level or source of dietary protein or protein-derived peptides on Ca absorption is not well understood. We determined, therefore, the influence of habitual dietary casein level, meal casein and meal casein phosphopeptide (CPP) on Ca absorption in the rat. True fractional Ca absorption was investigated in male 7-week-old rats, Wistar strain, in three separate studies using a faecal 47Sc: 47Ca ratio method. In studies A and C, rats (n 8 per group) were fed on a purified diet containing 200 g casein/kg for 2 weeks. Rats were then given a 47Ca-labelled meal (10 g) containing (per kg) either 0, 100, 200, or 300 g casein (study A) or 0, 100, 200, 350 or 500 g CPP (study C). In study B, rats (n 24 per group) were fed on a purified diet containing (per kg) either 200, 350 or 500 g casein for 2 weeks. Each group was then further randomized into three groups (n 8 per group) and given a 47Ca-labelled meal (10 g of the same diet) containing (per kg) either 200, 350 or 500 g casein. Ca absorption from a meal was unaffected by increasing meal casein concentration from 0 to 300 g/kg (study A), but was increased with a meal casein content of 500 g/kg (study B). Fractional Ca absorption decreased with increasing usual dietary casein intake in the range 200-500 g/kg (study B), suggesting intestinal adaptation. Ca absorption was unaffected by inclusion of 100 g CPP/kg in a single meal but was significantly (P < 0.001) reduced by 200, 350 and 500 g CPP/kg meal, with no evident dose-relationship. Thus, while Ca absorption was enhanced by high-casein meals, the mechanism remains unclear.     

Absorption,distribution and metabolism of epoxystearic acid in the rat

Conclusion Epoxystearic acid was rapidly absorbed after oral administration and well distributed in all tissues, mainly in the fat and liver of rats. Since this compound was metabolized to CO₂ and was removed in the expired air, little or no bioaccumulation could be expected.     

The effect of moderately and severely restricted dietary magnesium intakes on bone composition and bone metabolism in the rat

Forty 3-week-old male rats, Wistar strain, average weight 59 g, were randomized by weight into five groups of eight rats each. Three groups were fed ad libitum on a semi-purified diet containing (per kg) 400 (adequate), 200 (moderately Mg-restricted) or 20 (severely Mg-restricted) mg Mg for 3 weeks while two groups were pair-fed with the Mg-adequate diet in the same quantities as those consumed by the two Mg-restricted groups respectively. While weight gains and food conversion efficiency values for the Mg-restricted groups were similar to those of the corresponding pair-fed control groups, serum and kidney Mg, and femoral dry weight were reduced by 70, 7 and 9% respectively in the severely Mg-restricted group and were unaffected in the moderately Mg-restricted group. Significant reductions were observed in urinary pyridinoline (Pyr) (by 44 and 34%) and deoxypyridinoline (Dpyr) levels (by 40 and 33%) (markers of bone resorption), serum osteocalcin levels (by 46 and 28%) (marker of bone formation), femoral Mg levels (by 52 and 14%) and osteocalcin mRNA levels (by 46 and 22%) compared with the corresponding pair-fed controls, in the severely and moderately Mg-restricted groups respectively, and these reductions, except for those in urinary Pyr and Dpyr, were more marked in the severely Mg-restricted group. Femoral Ca and P concentrations were unaffected by dietary Mg restriction. These results show that not only severe but also moderate dietary restriction of Mg over 21 d results in qualitative changes in bone (i.e. reduced Mg concentration) as well as in aberrant bone turnover in young growing rats (i.e. severely depressed rates of bone formation and bone resorption), which may impair bone development and bone strength.