Effects of fatty liver induced by niacin-free diet with orotic acid on the metabolism of tryptophan to niacin in rats

The effects of dietary orotic acid on the metabolism of tryptophan to niacin in weaning rats was investigated. The rats were fed with a niacin-free, 20% casein diet containing 0% (control diet) or 1% orotic acid diet (test diet) for 29 d. Retardation of growth, development of fatty liver, and enlargement of liver were observed in the test group in comparison with the control group. The concentrations of NAD and NADP in liver significantly decreased, while these in blood did not decrease compared to the control group. The formation of the upper metabolites of tryptophan to niacin such as anthranilic acid, kynurenic acid, and 3-hydroxyanthranilic acid were not affected, but the quinolinic acid and beyond, such as nicotinamide, N1-methylnicotinamide, N1-methyl-2-pyridone-5-carboxamide, and N1-methyl-4-pyridone-3-carboxamide, were significantly reduced by the administration of orotic acid. Therefore, the conversion ratio of tryptophan to niacin significantly decreased in the test group in comparison with the control group.     

A Novel Sesquiterpene,Tuberonone, from Solanum tuberosum L.

The effects of the ovarian hormones progesterone and estrone on the conversion of tryptophan to nicotinamide in rats were investigated. Female rats were fed for 35 days with a 20% casein diet, or with a 20% casein diet containing 0.1% progesterone, or 0.001% estrone, or 0.1% progestrone and 0.001% estrone. The conversion ratio of tryptophan to nicotinamide on the last day of the experiment was 2% in the groups fed with the 20% casein diet and the diet containing 0.1% progesterone, but around 1.2% in the group fed with 0.001% estrone, and 0.7% in the group fed with 0.1% progesterone and 0.001% estrone. These results demonstrated that administration of ovarian hormones significantly decreased the conversion of tryptophan to nicotinamide.     

A Simple Bioassay for Chemicals Active to the Photosynthetic or Respiratory Systems of Plants

After male rats of the Sprague Dawley strain, 5 weeks old, were fed a 20% casein diet with or without 0.5% nicotinamide for 13 days, 180 mg/kg body weight of alloxan was injected in- traperitoneally into the rats. The rats were kept for 18 days with the same diet. The level of blood glucose was increased 6-fold in the group on a 20% casein diet by the injection of alloxan, while there was only a 2-foid increase in the group on a nicotinamide-containing diet and the decreased body weight was also lower in the group on the nicotinamide diet than the group on the casein diet. The body weight was indirectly related to the concentration of blood glucose. A marked increase was observed in the activities of tryptophan oxygenase, aminocarboxymuconate-semialdehyde decarboxylase, and nicotinamide methyltransferase upon the injection of alloxan with both diets; on the other hand, the activities of kynureninase and NAD⁺ synthetase were decreased by the injection of alloxan. The activity of kynurenine aminotransferase increased in the group on the 20% casein diet by the injection of alloxan, while in the group on the nicotinamide-containing diet its activity was not increased by the injection. These changes in the above enzyme activities mean that the conversion ratio from tryptophan to niacin is lower in the alloxan diabetic rat than normal rat. It was found that the activities of tryptophan oxygenase, aminocarboxymuconate-semialdehyde decarboxylase, and nicotinamide methyltransferase were directly related to the concentration of blood glucose, and that the activities of kynureninase and NAD⁺ synthetase were inversely related. There was no difference in the activities of 3-hydroxyanthranilic acid oxygenase and nicotinamide mononucleotide adenylyltransferase upon the injection of alloxan with both diets.     

Effect of Amino Acid Supplementation to a Rice Diet on Niacin Requirement of Rats

The effect of amino acid supplementation to a rice diet on the niacin requirement of rats was studied in relation to the phenomenon of niacin or tryptophan deficiency caused by the addition of threonine or gelatin to a low casein diet. Supplementation of a mixture of all limiting amino acids other than tryptophan to a 90% rice diet stimulated the growth of rats only temporarily without additional supplementation of niacin. However, the supplementation of the same mixture of limiting amino acids to a diet containing an amino acid mixture simulating rice protein, clearly decreased the growth of rats after a temporary increase. The growth was then remarkably improved by the further addition of niacin or niacin plus tryptophan. This result supports the hypothesis that the addition of all limiting amino acids other than tryptophan, increases the use of tryptophan for protein synthesis and may lead to niacin deficiency.     

Growth-promoting activity of pyrazinoic acid,a putative active compound of antituberculosis drug pyrazinamide,in niacin-deficient rats through the inhibition of ACMSD activity

We have recently reported that the antituberculosis drug, pyrazinamide (PZA), caused a significant increase in the conversion ratio of tryptophan to niacin in rats. In the present work, we investigated whether or not pyrazinoic acid (POA), a putative metabolite of PZA, increased the conversion ratio of tryptophan to niacin. Weaning rats were fed with a niacin-free and tryptophan-limited diet (negative control diet), or with the negative control diet supplemented with 0.003% nicotinic acid (positive control diet) or 1% POA (test diet) for 27 days. The growth rate was almost same between the groups fed on the positive control diet and the test diet. Dietary POA significantly increased the conversion ratio of tryptophan to niacin. Although POA did not directly inhibit the activity of alpha-amino-beta-carboxymuconate-epsilon-semialdehyde decarboxylase (ACMSD), the rate-limiting enzyme in the tryptophan-niacin pathway, liver ACMSD activity was only not detected in the test diet group. These results suggest that a derivative of POA metabolized by rats inhibited the ACMSD activity.     

Comparison of the Activity of the Tryptophan-NAD Pathway between Rats Fed a Fat-free Diet and a Fat Diet

The effect of dietary fat on tryptophan-NAD metabolism was investigated. Weanling male rats of the Sprague Dawley strain were fed a 40% casein diet (nicotinic acid-free) with or without 20% fat for 13 days. Although the food intake in 13 days was significantly higher in the fat-free group than in the fat group, the gains in body weight in the two groups were almost the same, because of the same energy intakes. The urinary excretion of tryptophan metabolites such as quinolinic acid, niacin and N¹-methylnicotinamide was greatly increased in the fat group in comparison with that in the fat-free group. The urinary excretion of xanthurenic acid was almost the same in the two groups. The blood NAD level of the fat group was significantly increased. The activities of liver amino-carboxymuconate-semialdehyde decarboxylase and liver nicotinamide methyltransferase in the fat group were significantly reduced, and that of liver NMN adenylyltransferase was significantly increased. The changes of these three enzymes could be advantageous for the increased formation of NAD from tryptophan. As a result, the feeding of a high fat diet to rats increased the formation of niacin and niacin-related compounds.     

Lunatoic Acid A,a Morphogenic Substance Inducing Chlamydospore-like Cells in Some Fungi

The effect of the addition of 0.26 % free tryptophan (Trp) to a 20 % casein diet containing 6 mg of nicotinic acid per 100 g of diet on the ratio of N¹-methyl-2-pyridone-5-carboxamide (2-py) plus N¹-methyl-4-pyridone-3-carboxamide (4-py) to -methylnicotinamide (MNA) excretion was investigated in rats. The urinary excretion of MNA, 2-py and 4-py, respectively, increased statistically significantly with the feeding of a 0.26% Trp (the same as the content of the 20% casein diet) supplemented 20% casein diet, although it did not increase with the feeding of a 40% casein diet, compared with in the case of the 20 % casein diet [Agric. Biol. Chem., 52, 1765 (1988)]. So, the total urinary excretion of Nam and its metabolites was 1.8 times higher in the group fed the Trp supplemented diet than in the group fed the 20 % casein diet. However, the ratio of 2-py plus 4-py to MNA excretion was much lower in the group fed the Trp supplemented diet than in the group fed the 20 % casein diet (13.16 ± 3.75→5.49 ± 2.25). This decreased ratio was considered to be partially due to a decrease in the 4-py forming MNA oxidase, which decreased significantly with the feeding of the Trp supplemented diet. Furthermore, the metabolic fate of Trp was greatly affected by the form of Trp, free or bound.     

Effects of dietary di(2-ethylhexyl)phthalate on the metabolism of tryptophan to niacin in mice.

We have reported the effect of di(2-ethylhexyl)phthalate (DEHP) on the tryptophan (Trp)-niacin pathway in rats. To clarify the universal effect of DEHP on rodents, we studied whether DEHP also has an effect on Trp metabolism in mice. Mice were fed a niacin-free, 20% casein diet supplemented with DEHP for 21 days. Feeding with DEHP decreased the body weight gain and increased the liver weight in correlation with the dose level of DEHP. The administration of DEHP significantly increased the formation of quinolinic acid and the lower metabolites of the Trp-niacin pathway. The flux of niacin in the lower part of the Trp-niacin pathway in mice was enhanced by feeding with DEHP.     

Soy protein diet ameliorates renal nitrotyrosine formation and chronic nephropathy induced by puromycin aminonucleoside

It has been shown that reactive oxygen species are involved in chronic puromycin aminonucleoside (PAN) induced nephrotic syndrome (NS) and that a 20% soy protein diet reduces renal damage in this experimental model. The purpose of the present work was to investigate if a 20% soy protein diet is able to modulate kidney nitrotyrosine formation and the activity of renal antioxidant enzymes (catalase, glutathione peroxidase, Cu,Zn- or Mn-superoxide dismutase) which could explain, at least in part, the protective effect of the soy protein diet in rats with chronic NS induced by PAN. Four groups of rats were studied: (1) Control rats fed 20% casein diet, (2) Nephrotic rats fed 20% casein diet, (3) Control rats fed 20% soy protein diet, and (4) Nephrotic rats fed 20% soy protein diet. Chronic NS was induced by repeated injections of PAN and rats were sacrificed at week nine. The soy protein diet ameliorated proteinuria, hypercholesterolemia, and the increase in serum creatinine and blood urea nitrogen observed in nephrotic rats fed 20% casein diet. Kidney nitrotyrosine formation increased in nephrotic rats fed 20% casein diet and this increase was ameliorated in nephrotic rats fed 20% soy protein diet. However, the soy protein diet was unable to modulate the antioxidant enzymes activities in control and nephrotic rats fed 20% soy protein diet. Food intake was similar in the two diet groups. The protective effect of a 20% soy protein diet on renal damage in chronic nephropathy induced by PAN was associated with the amelioration in the renal nitrotyrosine formation but not with the modulation of antioxidant enzymes.     

Induction of pancreatic trypsin by dietary amino acids in rats: four trypsinogen isozymes and cholecystokinin messenger RNA

We previously demonstrated that feeding a diet containing a high level of amino acid mixture simulating casein (AA) induced an increase in pancreatic protease activities in rats. In the present study, this effect of dietary AA was further characterized with three separate experiments. These experiments (1) examined periodic changes in pancreatic and small intestinal trypsin activities after switching from a 20% (a normal nitrogen level) AA diet to a 60% AA (a high nitrogen level) diet; (2) measured the abundance of mRNA for four trypsinogen isozymes and for intestinal cholecystokinin (CCK) and secretin in rats fed 20% and 60% AA diets for 10 days compared with rats fed 20% and 60% casein diets; and (3) measured the abundance of mRNA for four trypsinogen isozymes after chronic administration of CCK. Trypsin activities were gradually increased in both the pancreas and the small intestinal lumen and reached maximum at 5 days after the switch to the 60% AA diet (Exp. 1). This result is evidence that the increase in the protease activity in the pancreas is due to enhancement of pancreatic trypsin production. In experiment 2, pancreatic trypsinogen isozymes I, II, III, and IV mRNA abundance were evaluated by the Northern blotting method using cDNA probes specific for each isozyme mRNA. Abundance of trypsinogen mRNA without trypsinogen I tended to increase in the rats fed the 60% casein diet but tended to decrease in the rats fed the 60% AA diet compared with the respective normal nitrogen level diet groups without significant difference. CCK mRNA abundance in the jejunal mucosa increased as a result of feeding the 60% casein diet, but not the 60% AA diet. Subcutaneous CCK injections (3.5 nmole/kg body weight/day, twice daily, at 8:30 am and 7:30 pm) for 10 days resulted in increased pancreatic trypsin activity, whereas the changes in mRNA of the four trypsinogen isozymes was similar between the 20% and 60% casein groups but differed between the 20% and 60% AA groups (Exp. 3). These results suggest that CCK is not involved in the induction of pancreatic trypsin that occurs with feeding of a high AA diet and that the mechanism of protease induction by dietary AA is different from that in the case of dietary protein.     

High-Casein Diet Suppresses Guanidinoacetic Acid-Induced Hyperhomocysteinemia and Potentiates the Hypohomocysteinemic Effect of Serine in Rats

To determine the effect of dietary protein level on experimental hyperhomocysteinemia, rats were fed 10% casein (10C) and 40% casein (40C) diets with or without 0.5% guanidinoacetic acid (GAA) for 14 d. In addition, rats were fed 10C + 0.75% methionine (10CM) and 40C + 0.75% methionine (40CM) diets with or without 2.5% serine for 14 d to determine the relationship between the dietary protein level and intensity of the hypohomocysteinemic effect of serine. GAA supplementation markedly increased the plasma homocysteine concentration in rats fed with the 10C diet, whereas it did not increase the plasma homocysteine concentration in rats fed with the 40C diet. Although serine supplementation significantly suppressed the methionine-induced enhancement of plasma homocysteine concentration, the decreased plasma homocysteine concentration was significantly lower in rats fed with the 40CM diet than in rats fed with the 10CM diet. The hepatic cystathionine β-synthase and betaine-homocysteine S-methyltransferase activities were significantly higher in rats fed with the 40C or 40CM diet than in rats fed with the 10C or 10CM diet, irrespective of supplementation with GAA and serine. These results indicate that the high-casein diet was effective for both suppressing GAA-induced hyperhomocysteinemia and potentiating the hypohomocysteinemic effect of serine, probably through the enhanced activity of homocysteine-metabolizing enzymes.     

Stimulative effect of dietary protein on the phosphorylation of p70 S6 kinase in the skeletal muscle and liver of food-deprived rats

The effect of dietary protein on p70S6k phosphorylation was examined in rats starved for 18 h and then fed either a 20% casein diet (20C) or a protein-free diet (0C). Refeeding the 20C diet, but not the 0C diet, increased p70S6k phosphorylation in both the skeletal muscle and liver. The plasma insulin concentrations were the same after refeeding the 20C or 0C diet, suggesting that a combination of dietary protein and insulin may be required to stimulate p70S6k phosphorylation.     

Influence of Feeding Paradigm in Rats on Temporal Pattern of: I. Macronutrient Intake and Arterio-Venous Differences in Plasma Glucose,Insulin and Tryptophan

Relationships among feeding paradigm (single diet vs food selection) and arterio-venous differences (δAV) of glucose, insulin and tryptophan were studied by measuring the temporal patterns of food intake and plasma parameters during 8 hr feeding cycles in rats. Adult male Sprague-Dawley rats were offered a single diet of fixed composition (20% casein) or a choice between two isocaloric diets (0% and 60% casein) for 2 weeks under 8-hr daily feeding conditions, food being offered during the dark cycle. Groups of animals were then killed at the beginning and at 2-hourly intervals throughout the feeding period. With both feeding paradigms, rats showed temporal patterns of energy, carbohydrate and protein intakes with a peak at the beginning and a trough at the end of the feeding period. However, in rats offered a dietary choice the intake of carbohydrate was significantly lower, and the intakes of energy and protein significantly higher than those found in rats offered a single diet. Throughout the feeding period, these differences between single and choice diets became less accentuated in the case of carbohydrate intake, but more accentuated for energy and protein intakes. Paradoxically, rats fed a choice of diets had a significantly lower weight gain than rats fed a single diet. The temporal variation of insulin secretion and tryptophan absorption varied inversely with the two diet paradigms. Moreover, in rats offered a choice of diets, macronutrient intake was significantly correlated with insulin secretion and venous glucose concentration. The opposed physiologic and metabolic responses to the feeding paradigms suggest the need for future studies to examine the possibility that such can function as synchronizers of biological rhythms.     

Influence of Feeding Paradigm in Rats on Temporal Pattern of: I. Macronutrient Intake and Arterio-Venous Differences in Plasma Glucose, Insulin and Tryptophan

Relationships among feeding paradigm (single diet vs food selection) and arterio-venous differences (δAV) of glucose, insulin and tryptophan were studied by measuring the temporal patterns of food intake and plasma parameters during 8 hr feeding cycles in rats. Adult male Sprague-Dawley rats were offered a single diet of fixed composition (20% casein) or a choice between two isocaloric diets (0% and 60% casein) for 2 weeks under 8-hr daily feeding conditions, food being offered during the dark cycle. Groups of animals were then killed at the beginning and at 2-hourly intervals throughout the feeding period. With both feeding paradigms, rats showed temporal patterns of energy, carbohydrate and protein intakes with a peak at the beginning and a trough at the end of the feeding period. However, in rats offered a dietary choice the intake of carbohydrate was significantly lower, and the intakes of energy and protein significantly higher than those found in rats offered a single diet. Throughout the feeding period, these differences between single and choice diets became less accentuated in the case of carbohydrate intake, but more accentuated for energy and protein intakes. Paradoxically, rats fed a choice of diets had a significantly lower weight gain than rats fed a single diet. The temporal variation of insulin secretion and tryptophan absorption varied inversely with the two diet paradigms. Moreover, in rats offered a choice of diets, macronutrient intake was significantly correlated with insulin secretion and venous glucose concentration. The opposed physiologic and metabolic responses to the feeding paradigms suggest the need for future studies to examine the possibility that such can function as synchronizers of biological rhythms.     

Short-term, increasing dietary protein and fat moderately affect energy expenditure, substrate oxidation and uncoupling protein gene expression in rats

Macronutrient composition of diets can influence body-weight development and energy balance. We studied the short-term effects of high-protein (HP) and/or high-fat (HF) diets on energy expenditure (EE) and uncoupling protein (UCP1-3) gene expression. Adult male rats were fed ad libitum with diets containing different protein-fat ratios: adequate protein-normal fat (AP-NF): 20% casein, 5% fat; adequate protein-high fat (AP-HF): 20% casein, 17% fat; high protein-normal fat (HP-NF): 60% casein, 5% fat; high protein-high fat (HP-HF): 60% casein, 17% fat. Wheat starch was used for adjustment of energy content. After 4 days, overnight EE and oxygen consumption, as measured by indirect calorimetry, were higher and body-weight gain was lower in rats fed with HP diets as compared with rats fed diets with adequate protein content (P<.05). Exchanging carbohydrates by protein increased fat oxidation in HF diet fed groups. The UCP1 mRNA expression in brown adipose tissue was not significantly different in HP diet fed groups as compared with AP diet fed groups. Expression of different homologues of UCPs positively correlated with nighttime oxygen consumption and EE. Moreover, dietary protein and fat distinctly influenced liver UCP2 and skeletal muscle UCP3 mRNA expressions. These findings demonstrated that a 4-day ad libitum high dietary protein exposure influences energy balance in rats. A function of UCPs in energy balance and dissipating food energy was suggested. Future experiments are focused on the regulation of UCP gene expression by dietary protein, which could be important for body-weight management.     

Dietary niacin requirement of fingerling Channa punctatus (Bloch)

A 16‐week experiment was accomplished to determine the dietary niacin requirement of fingerling Channa punctatus (6.8 ± 0.92 cm; 4.65 ± 0.46 g) by feeding seven casein‐gelatin based isonitrogenous (450 g/kg CP) and isoenergetic (18.39 kJ/g GE) diets with graded levels of niacin (0, 10, 20, 30, 40, 50 and 60 mg/kg diet) twice a day to apparent satiation to triplicate groups of fish. Significantly best absolute weight gain (AWG; 38.19 g/fish), feed conversion ratio (FCR; 1.42) and protein retention efficiency (PRE; 26.47%) were registered in fish fed 40 mg niacin/kg diet. Also, fish fed above diet exhibited maximum carcass protein. Hemoglobin (Hb), RBCs counts and hematocrit (Hct) were improved with the incremental levels of dietary niacin up to 40 mg/kg. However, liver niacin content showed the positive correlation up to 50 mg/kg niacin and then leveled off. Fingerling C. punctatus fed niacin‐free diet showed retarded growth, poor feed utilization, high mortality, difficult motion and skin haemorrhage. Broken‐line regression analysis of AWG, FCR and PRE indicated that fingerling C. punctatus require niacin in the range of 37.1–42.1, whereas that of liver niacin concentration indicated the niacin requirement at 52.3 mg/kg dry diet.     

Effect of dietary proteins on the turnover of rat liver argininosuccinate synthetase.

The rates of synthesis and degradation of arginosuccinate synthetase in rat liver under various dietary conditions were determined. The relative rate of the enzyme synthesis in the livers of rats fed on 70% casein diet was 4.0 times greater than that for rats fed on 5% casein diet. The rate constants of degradation (Kd of argininosuccinate synthetase were estimated to be 0.15 and 0.16 day-1 under 70% and 5% casein feeding, respectively. When the dietary conditions were changed acutely from 70% to 5% casein diet or vice versa, the rates of the enzyme synthesis decreased or increased, respectively, and the rates of enzyme degradation were also affected. The change from 5% to 70% casein diet caused a transient decrease in the rate of degradation. After the enzyme activity had achieved a new steady-state level, the enzyme degradation proceeded at the normal steady rate. On the other hand, the change from 70% to 5% casein diet caused a transient increase in the rate of degradation. Thus, the only factor regulating the amount of enzyme in rat liver is the rate of enzyme synthesis under the steady-state conditions. However, the rates of both enzyme synthesis and degradation are involved in the regulation of the amount of enzyme during dietary transition.     

Analysis of regulatory factors for urea synthesis by isolated perfused rat liver. II. Comparison of urea synthesis in livers of rats subjected to different dietary conditions.

Capacities for urea synthesis and amino acid patterns in the perfused livers isolated from rats fed low and high-protein diets were compared. Urea formation with amjonium chlorode as the nitrogen source in perfused livers isolated from rats fed on a 70% casein diet was rapid and the efficiency of conversion of ammonia to urea was 97.9%. However, that in livers isolated from rats fed on a 5% casein diet was much slower and the efficiency of conversion of ammonia to urea was only 36.1%. The ratios of the rate of urea formation from ammonium chloride to activity of ornithine transcarbamylase [EC 2.1.3.3.] in the perfused livers of rats fed on 5 and 70% casein diets were calculated. The ratio of the former condition was much lower than that of the latter. The ratios reached nearly the same level by the addition of ornithine and N-acetylglutamate, the addition of which to the perfusate caused marked elevation of the ratios in both cases. In the perfused livers from rats fed on a 5% casein diet a considerable portion of the ammonia added to the perfusate was fixed into an amino ro an amide group of amino acids such as alamin, aspartate, and glutamine. On the other hand, in the perfused livers from rats fed on a 70% casein diet most of the ammonia added was converted to urea. The regulation of urea synthesis and the relation between anabolism and catabolism of amino acids in rat livers subjected to different dietary conditions were compared.     

Effects of dietary pyrazinamide, an antituberculosis agent, on the metabolism of tryptophan to niacin and of tryptophan to serotonin in rats

The effects of pyrazinamide on the metabolism of tryptophan to niacin and of tryptophan to serotonin were investigated to elucidate the mechanism for pyrazinamide action against tuberculosis. Weanling rats were fed with a diet with or without 0.25% pyrazinamide for 61 days. Urine samples were periodically collected for measuring the tryptophan metabolites. The administration of pyrazinamide significantly increased the metabolites, 3-hydroxyanthranilic acid and beyond, especially quinolinic acid, nicotinamide, N'-methylnicotinamide, and N1-methyl-4-pyridone-3-carboxamide, and therefore significantly increased the conversion ratio of tryptophan to niacin and the blood NAD level . However, no difference in the upper metabolites of the tryptophan to niacin pathway such as anthranilic acid, kynurenic acid and xanthurenic acid was apparent between the two groups. No difference in the concentrations of trytptophan and serotonin in the blood were apparent either. It is suggested from these results that the action of pyrazinamide against tuberculosis is linked to the increase in turnover of NAD and to the increased content of NAD in the host cells.