We developed a novel simple method to determine serum inorganic pyrophosphatase (PPase; EC 188.8.131.52) activity using purine nucleoside phosphorylase, xanthine oxidase and peroxidase, which method is applicable to automated analysis. Good reproducibility was demonstrated, with a coefficient of variation of 2.5 to 4.0% depending on the PPase activity. Good stability after color development was also observed. The presence of ascorbic acid, bilirubin, hemoglobin or intra-lipid did not influence the results. We also investigated the relationship between the activities of alkaline phosphatase (ALP; EC 184.108.40.206) and PPase. A strong positive correlation between ALP from bone and placenta and PPase was demonstrated with a correlation coefficient of 0.932 and 0.909, respectively. In contrast, ALP from liver showed a relatively weak correlation, with a correlation coefficient of 0.839. Further, placenta showed a higher ratio of PPase activity to ALP activity than did other tissues. PPase and ALP could not be distinguished by gel filtration or by ion-exchange chromatography.
The effect of diets containing olive oil, sunflower oil or hydrogenated coconut oil on immune cell function was investigated. Lipid administration in assays in vitro as well as in the diet has been demonstrated that affect to lymphocyte proliferation, cytokine synthesis, natural killer (NK) cell activity and antigen presentation. Therefore, unsaturated fatty acids may be used in the treatment of inflammatory diseases due to their immunomodulatory properties. Three groups of Balb/c mice were fed three diets at various times for 5, 15, 30, 60, or 90 days. Diets were different for each group, as each contained 15wt% olive oil, sunflower oil or hydrogenated coconut oil. The number of splenic cells showed significant differences with respect to the type of dietary lipid and the time of dietary administration. At the end of dietary lipid supplementation mitogen-stimulated lymphocyte proliferation from the spleens of mice fed diets that contained olive oil or hydrogenated coconut oil was significantly higher than that of mice fed diet that contained sunflower oil in which mitogen-stimulated proliferation of lymphocytes decreased lightly. Interleukin-2 (IL-2) production was significantly higher in mice fed the olive oil-or sunflower oil-enriched diets for 90 days when compared with that of mice fed the diet containing hydrogenated coconut oil. Dietary lipid manipulation also modified NK cell activity, since the results obtained showed a significant difference between the NK cell activity of mice fed the diet that contained olive oil and that observed in the mice fed the diet containing hydrogenated coconut oil. The results demonstrate that the addition to the diet of unsaturated fatty acids for different times may play an important role as a modulatory agent of immune cell function.
Folate absorption was studied in rats with streptozotocin-induced diabetes. Absorption, as determined by the rise in serum folate levels after ingestion of either synthetic simple or dietary polyglutamylfolates, was observed to be 1.6-1.8-fold higher in diabetic rats than in normal ones. Folate transport in everted intestinal segments was also 60% higher in the diabetic animals. The enzyme folyl conjugase, which breaks down dietary polyglutamylfolates to simple absorbable forms, was induced 2-fold in pancreas 30min following folate ingestion in both control and diabetic animals. Luminal folyl conjugase, which showed very little activity at the onset, was elevated 7-9-fold after dietary folate ingestion in the control as well as in the diabetic group. These results indicate that the induction and release into the lumen of folyl conjugase is not adversely affected in diabetes and that the enhanced folate absorption and transport observed in diabetes might be physiological adaptation due to increased requirements.
Emblica officinalis (E.O) extract was found to dose dependently lower the incidence of liver tumors induced by N-nitrosodiethylamine (NDEA). Control group rats (NDEA alone) had 100% tumor incidence, whereas in animals treated with E.O at a dose of 250mg/kg body weight, the incidence was 0%. The liver weight of the control group was 7.99±1.51g/100g body weight, and that of the E.O-treated group showed a dose-dependent decrease (4.90-3.50g/100g body weight). NDEA treatment also increased serum and liver γ-glutamyl transpeptidase (γ-GTP) activity to 135.5±27.9U/liter and 2.90±0.08nmol/min/mg protein, respectively; and E.O treatment reduced these levels in a dose-dependent manner (99.8-44.8U/liter and 2.25-0.38nmol/min/mg protein). The level of liver aniline hydroxylase, a P-450 enzyme, was increased by the NDEA treatment, and this increase was reduced by the administration of the E.O extract. Liver glutathione-S-transferase (GST) activity, which was increased to 1, 414±137nmol/min/mg protein by NDEA treatment, was found to be reduced in a dose-dependent manner by E.O in vivo. E.O extract was also found to inhibit aminopyrene-N-demethylase activity in vitro, with 301μg/ml as the concentration needed for 50% inhibition. The extract was also found to inhibit DNA adduct formation induced by benzo(a)pyrene [B(a)P] and aflatoxin B1 (AFB1). The concentration of E.O extract required for 50% inhibition of microsome-catalyzed B(a)P adduct formation was 280μg/ml; and that for AFB1 adduct formation, 120μg/ml. The results of the present study indicate that E.O extract can interfere with the metabolism of NDEA and inhibit liver tumor development by this carcinogen, suggesting its use in the prevention of hepatocarcinogenesis.
Muthu Marunthu, a herbal formulation, was used to study its anticancer effect on 3-methylcholanthrene-induced fibrosarcoma in rats. The effects of Muthu Marunthu on the tumor weight, on the activities of glutathione peroxidase (GSH-Px), superoxide dismutase (SOD), and catalase (CAT), and on the levels of lipid peroxide and glutathione (GSH) of liver and kidney were studied in the fibrosarcoma-bearing rats. The tumor weight was found to be reduced by the Muthu Marunthu treatment. In liver and kidney, the level of GSH and the activities of antioxidant enzymes such as GSH-Px, SOD, and CAT were decreased significantly, whereas the lipid peroxide content was increased significantly in fibrosarcoma rats. After Muthu Marunthu treatment, the GSH content and the activities of antioxidant enzymes were increased significantly with a concomitant decrease in the level of lipid peroxide. These observations clearly suggest that Muthu Marunthu can effectively normalize the fibrosarcoma-induced alterations to the antioxidant defense systems.
The effects of a low-calorie diet (LCD) therapy with and without dumbbell exercise (aerobic-resistance exercise) on body fat reduction were studied in twelve mildly obese women aged 19-20 years. The subjects were randomly assigned to one of the following two groups: LCD with dumbbell exercise (LCD+DEx group), and LCD only (LCD group). The subjects received a liquid-formula diet based on Micro Diet for 2 meals of their 3 meals per day. Thus, they were restricted to 960±80kcal (mean±SE) of energy intake per day for 12 weeks. In the LCD+DEx group, subjects also performed dumbbell exercises with a pair of 2-kg dumbbells every day after supper for about 20min. Subjects underwent several measurements (body composition, oral glucose tolerance test, blood biochemical test) before starting the experiment and after 12 weeks while still dieting. During the 12-week experimental period, body weight and body fat decreased significantly in both LCD+DEx and LCD groups without a reduction in fat free mass (FFM). The decreases in body weight and body fat were larger in the LCD+DEx group, but the differences were not significant. These results suggest that LCD used with Micro Diet decreases body weight and body fat without reducing FFM. Dumbbell exercise may strengthen the effect of LCD on body fat reduction.