Regulation of xanthine oxidase activity and immunologically detectable protein in rats in response to dietary protein and iron

To investigate the mechanism for changes in xanthine oxidase activity in response to dietary protein and iron, we fed rats diets containing 50, 20 or 5% casein with either normal iron (35 mg Fe/kg diet) or low iron (5 mg Fe/kg diet). Xanthine oxidase activity changed in liver and intestinal mucosa in response to protein and iron, but immunologically detectable xanthine oxidase protein did not change. When total liver RNA isolated from these rats was translated by a rabbit reticulocyte lysate, we found no difference in the amount of xanthine oxidase that was translated. These results demonstrated that the changes in xanthine oxidase activity were not accompanied by changes in the amount of protein. Since xanthine oxidase can exist in an inactive desulfo form, we asked if xanthine oxidase activity was changed by the content of sulfur-containing amino acids in the diet. Xanthine oxidase activity in intestinal mucosa of the rats fed the 5% casein + methionine diet was significantly greater than that of the rats fed the 5% casein diet alone. These findings suggest that xanthine oxidase activity may be regulated by interconversion of active and inactive desulfo enzyme.     

Spinal reflexes in normal and sulfite oxidase deficient rats: effect of sulfite exposure

Sulfites, which are commonly used as food preservatives, are continuously formed in the body during metabolism of sulfur-containing amino acids. Sulfite is oxidized to sulfate ion by sulfite oxidase (SOX, EC. 1.8.3.1). Although sulfite treatment has been reported to increase the excitability of some neurons in vitro, the possible effects of sulfite on neuronal excitability in vivo remain unclear. The aim of this study was to investigate the possible effects of sulfite treatment on spinal reflexes in anesthetized SOX competent and deficient rats. For this purpose, male albino rats used in this study were divided into four groups such as control group (C), sulfite group (CS), SOX deficient group (D), and SOX deficient + sulfite group (DS). Rats in SOX deficient groups were made deficient in SOX by the administration of low molybdenum (Mo) diet (AIN 76, Research Dyets Inc., USA) with concurrent addition of 200-ppm tungsten (W) to their drinking water in the form of sodium tungstate (NaWO4). Sulfite in the form of sodium metabisulfite (Na2O5S2, 70 mg/kg) was given orally by adding to drinking water to the S and DS groups. Monosynaptic reflex potentials were recorded from the ipsilateral L5 ventral root. SOX deficient rats had an approximately 15-fold decrease in hepatic SOX activity compared with normal rats. This makes SOX activity of SOXD rats in the range of human SOX activity. The results of this study show that sulfite treatment significantly increases the amplitude of the monosynaptic reflex response in both S and DS groups with respect to their respective control groups (C and D). SOX deficient rats also had enhanced spinal reflexes when compared with control rats. In conclusion, sulfite has increasing effects on the excitability of spinal reflexes and we speculate that this compound may exhibit its effects on nervous system by affecting sodium channels.     

Effect of dietary methionine on tissue selenium and glutathione peroxidase (EC 1.11.1.9) activity in rats given selenomethionine

1. The effect of dietary methionine on the utilization of selenium from dietary selenomethionine [( Se]Met) for tissue Se deposition and for glutathione peroxidase (EC 1.11.1.9; GSH-Px) synthesis was studied in male weanling rats. 2. When rats were given 0.5 mg Se as [Se]Met/kg diet supplemented with 0, 4 or 9 g methionine/kg, Se in plasma, erythrocytes, liver and muscle increased significantly over the 20 d period for all methionine-treatment groups. The increases in erythrocyte and muscle Se, however, were significantly higher in rats fed on the methionine-deficient diet compared with the methionine-supplemented diets. 3. In contrast to the increases in tissue Se, GSH-Px activity in liver, plasma and muscle decreased in methionine-deficient rats given 0.5 mg Se as [Se]Met/kg whereas GSH-Px activity was maintained or increased in rats supplemented with methionine. 4. The percentage of tissue Se associated with GSH-Px was calculated from the measured Se concentration and GSH-Px activity. A significantly lower percentage of Se was associated with GSH-Px in methionine-deficient rats compared with methionine-supplemented rats. 5. These results show that Se from dietary [Se]Met is preferentially incorporated into body proteins rather than used for GSH-Px synthesis when methionine is limiting in the diet. 6. These results further suggest that [Se]Met might not be the optimum Se compound to use for Se supplementation because metabolism of dietary [Se]Met to a biochemically active form, such as GSH-Px, was impaired when [Se]Met was provided in diets low in methionine.     

Effect of dietary methionine and polychlorinated biphenyls on cholesterol metabolism in rats fed a diet containing soy protein isolate

The effects of supplementation of methionine to a 20% soy protein isolate diet on serum level of cholesterol. 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase activity, cholesterol 7 alpha-hydroxylase activity, and biliary and fecal steroids in rats with or without receiving polychlorinated biphenyls (PCB) were investigated. Supplementation of methionine and PCB did not affect the growth. Serum level of cholesterol was higher in rats fed PCB than in controls. In rats fed PCB, addition of methionine elevated serum level of cholesterol synergistically. The activity of HMG-CoA reductase was higher in rats fed the methionine-supplemented diet than in those fed the unsupplemented diet when PCB was included in the diets. Cholesterol 7 alpha-hydroxylase activity (nmol/h.100 g body weight) was higher in rats fed PCB than in controls. Biliary secretion of bile acids was higher in rats fed PCB than in controls. On the other hand, fecal excretion of bile acids decreased in PCB-treated rats, but total steroids were not affected by PCB. In rats fed PCB, the addition of methionine did not alter cholesterol 7 alpha-hydroxylase activity and biliary and fecal steroid output. The data suggest that the increase in serum level of cholesterol due to dietary addition of methionine together with PCB would be mediated through the stimulation of hepatic synthesis of cholesterol.     

Effect of dietary glycine on methionine metabolism in rats fed a high-methionine diet

The alleviation mechanism of methionine toxicity by dietary glycine was investigated in weanling rats fed a high-methionine diet. When rats were fed a 10% casein diet containing 2% methionine, the activities of methionine adenosyltransferase, cystathionine beta-synthase, and cystathionine gamma-lyase, which participate in the methionine metabolism in the transsulfuration pathway, were significantly enhanced. But the addition of 2% glycine to the high methionine diet did not cause further increase in these enzyme activities; the activities of methionine adenosyltransferase and cystathionine beta-synthase were rather decreased while cystathionine gamma-lyase activity was not altered. Methionine transaminase activity was essentially insensitive to the dietary addition of methionine and glycine. In rats fed a high methionine diet, the hepatic methionine level was significantly increased with a concomitant decrease in the levels of glycine, serine, and threonine. The addition of glycine to the high methionine diet effectively suppressed the enhancement of the hepatic methionine level and almost completely restored the glycine level, but it only partially restored the serine level and further decreased the threonine level. From these results, it is suggested that the alleviating effect of dietary glycine on methionine toxicity is primarily elicited by the restoration of the hepatic glycine level rather than by an increase in hepatic enzyme activity.     

Effect of adding dietary methionine to a low soy protein diet on the brain protein synthesis rate in ovariectomized female rats

A deficiency of sex hormones affects brain function in mammals, including the decrease of protein synthesis. Recently, we have shown that the protein synthesis in the brain depended on the quality of dietary protein in ovariectomized female rats. The methionine is the first limiting amino acid for the recommended dietary allowance of amino acids in soy protein. The purpose of this study was to determine whether the addition of dietary methionine affected the rate of brain protein synthesis in ovariectomized female rats fed on the soy protein diet. Experiments were conducted on two groups of ovariectomized female rats (24 week) given the diets containing 5% soy protein or 5% soy protein + 0.2% methionine for 10 d. The fractional rates of protein synthesis in cerebral cortex and cerebellum significantly increased with an addition of dietary methionine. In the brain, the RNA activity [g protein synthesized/((g RNA) x d)] was significantly correlated with the fractional rate of protein synthesis. The RNA concentration (mg of RNA/g of protein) was not related to the fractional rate of protein synthesis in any organ. The results suggest that the addition of limiting amino acid for the low soy protein elevates the rate of protein synthesis in the brain of ovariectomized female rats, and that RNA activity is at least partly related to the fractional rate of brain protein synthesis.     

Influence of dietary methionine on dieldrin metabolism in rats

Bulletin of Environmental Contamination and Toxicology -     

Effect of dietary protein and energy on protein synthesis during lactation in rats

A series of experiments was carried out in which effects of dietary protein and energy on protein synthesis during lactation were evaluated. Purified diets were fed to female rats from conception until d 15 of lactation. Protein supplied by wheat gluten, gluten + lysine + threonine or casein + methionine was fed at 20 or 25% in the diet. Protein synthesis in mammary gland and liver was measured with a large dose of L-[4-3H]phenylalanine. As lactation proceeded, protein absolute synthesis rates (ASRs) in casein-fed dams increased from 1056 to 2246 mg/d in mammary gland but remained unchanged in liver. In contrast, ASRs in wheat gluten-fed dams remained unchanged in both tissues. Feeding a protein-free diet or starving dams for 3 d reduced ASR in mammary gland 59 and 72%, respectively, when measured on d 15 of lactation. The corresponding reductions in liver ASR were 41 and 46%. A 20% restriction of dietary energy at a constant protein intake reduced protein synthesis in mammary glands 34, 55 and 28% in dams fed gluten, gluten + lysine + threonine or casein, respectively. In summary, protein synthesis during lactation was reduced less in liver than in mammary gland in response to dietary restriction of protein or energy.     

Effect of dietary protein on pyrimidine-metabolizing enzymes in rats.

The effect of dietary protein on pyrimidine-metabolizing enzymes was studied in the rat. The activities of dihydropyrimidine dehydrogenase and beta-ureidopropionase in the livers of rats fed a protein-free diet were significantly decreased, while the activity of dihydropyrimidinase was unaffected. Protein deficiency (5%) also decreased the activity of beta-ureidopropionase. On the other hand, a high-protein diet (60%) increased the level of beta-ureidopropionase. The activities of beta-alanine-oxoglutarate aminotransferase (aminobutyrate aminotransferase) and D-3-aminoisobutyrate-pyruvate aminotransferase ((R)-3-amino-2-methylpropionate-pyruvate aminotransferase), which are present in mitochondria, depended on the amount of protein in the diet. Ammonium ions supplemented in the diet and given by injection did not affect the activities of rat liver pyrimidine-metabolizing enzymes (dihydropyrimidine dehydrogenase, dihydropyrimidinase, beta-ureidopropionase, beta-alanine-oxoglutarate aminotransferase and D-3-aminoisobutyrate-pyruvate aminotransferase). Dietary uridine resulted in the accumulation of uracil in the liver, but did not affect the activities of pyrimidine-metabolizing enzymes.     

Effect of dietary restriction on liver protein synthesis in rats

At 6 wk of age, male Fischer F344 rats were fed a purified, casein-based diet either ad libitum or in the amount of 60% of the diet consumed by the rats fed ad libitum (restricted diet). Hepatocytes were isolated from the rats between 2.5 and 19 mo of age. The protein content of the hepatocytes isolated from the rats fed the restricted amount of diet was significantly lower than that of hepatocytes isolated from rats fed ad libitum. The DNA and RNA content of the hepatocytes were similar for the rats fed the two dietary regimens. The absolute rate of protein synthesis for hepatocytes isolated from rats fed ad libitum decreased 55% between 2.5 and 19 mo of age. However, the rate of protein synthesis by hepatocytes from rats fed the restricted amount of diet decreased only slightly with increasing age. At 19 mo of age, the rate of protein synthesis by hepatocytes from the rats fed the restricted amount of diet was significantly higher than the rate of protein synthesis for hepatocytes from rats fed ad libitum. Therefore, dietary restriction retards the age-related decline in liver protein synthesis.     

Influence of dietary methionine with or without adequate dietary vitamins on hyperhomocysteinemia in rats

Information on the effects of dietary vitamins involved in homocysteine metabolism on the dietary methionine (Met)-induced hyperhomocysteinemia is limited. Thus, a six-wk study was conducted to investigate the effects of dietary Met with or without adequate vitamins on plasma total homocysteine (tHcy) in rats. Four levels of supplemental L-Met (0, 5, 10 and 20 g/kg) and two levels of vitamins (adequate and deficient in folate plus B-12) were tested in the casein-based diets. The plasma tHcy values in males were higher (p < 0.05) than in females (8.1± 0.6 vs. 6.0±0.6 μmol/L for adequate diet; 66.5± 1.4 vs. 45.5±0.9 μmol/L for folate-B-12 deficient diet). In males, supplementation of the adequate (control) diet with 5, 10 and 20 g/kg Met, increased tHcy to 1.3, 1.9 and 7.9 times control, respectively. In females, the corresponding values were 1.3, 1.7 and 5.6 times control. In rats fed folate-B-12 deficient diets, supplemental Met, however, generally caused reductions in plasma tHcy values in both sexes. These disparate responses to supplementary Met could be partly due to increases in hepatic S-adenosylmethionine (SAM)/S-adenosylhomocysteine (SAH) ratios in folate-B-12 deficient rats.     

Influence of dietary methionine on the metabolism of selenomethionine in rats

To determine the influence of methionine on selenomethionine (SeMet) metabolism, weanling male rats were fed for 8 wk a basal diet marginally deficient in sulfur amino acids, containing 2.0 micrograms selenium (Se)/g as DL-SeMet and supplemented with 0, 0.3, 0.6 or 1.2% DL-methionine. Increased dietary methionine caused decreased selenium deposition in all tissues examined but increased glutathione peroxidase (GSHPx, EC 1.11.1.9) activity in testes, liver and lungs. A positive correlation was found between dietary methionine and the calculated percentage of selenium associated with GSHPx. In a second experiment, 75SeMet was injected into weanling male rats which had been fed the basal diet containing 2.0 micrograms selenium as DL-SeMet with or without the addition of 1.0% methionine. The selenoamino acid content of tissues and the distribution of 75Se in erythrocyte proteins were determined. In comparison to the rats fed the basal diet without added methionine, significantly more 75Se-selenocysteine was found in liver and muscle, more 75Se was found in erythrocyte GSHPx and less 75Se was found in erythrocyte hemoglobin of rats fed 1.0% methionine. These data suggest that methionine diverts SeMet from incorporation into general proteins and enhances its conversion to selenocysteine for specific selenium-requiring proteins, such as GSHPx.