Plasma β-carotene, α-tocopherol and retinol were measured in 15 female and 5 male children with insulin-dependent diabetes mellitus (IDDM), and the correlations with plasma hemoglobin Ale (HbA 1 c) and fructosamine were analyzed. Twelve female and 8 male children served as age-matched controls. The plasma β-carotene and α-tocopherol levels of the IDDM children were significantly higher than those of the control children, but there were no differences in plasma retinol or total lipid levels. The plasma β-carotene level, β-carotene/retinol ratio and β-carotene/total lipids ratio each showed significant correlations with serum HbAlc and fructosamine in all subjects studied. Similarly, the plasma α-tocopherol level and α-tocopherol/total lipids ratio were correlated with these indexes of glycemic control. These findings suggest certain mechanisms may exist to prevent lipid peroxidation and vascular complications in IDDM patients.
This study was carried out to resolve the discrepancy of data for the proportion of ascorbic acid and dehydroascorbic acid in persimmon leaves at the final stage of the season and to clarify their cellular distributions using histochemical and biochemical techniques. Fresh persimmon leaves were collected and used on July 31, September 5 and October 7, 1996. Ascorbic acid and dehydroascorbic acid in sub-cellular fractions were determined by the HPLC method that was found to be the most reliable for separation. The percent of dehydroascorbic acid in the total leaves was found to be almost constant (between 32 and 37%) in all preparations tested. In all preparations, more than 90% of the ascorbic acid and dehydroascorbic acid was found in the soluble fraction. The histochemical detection of ascorbic acid and an electron micrograph of persimmon leaf cells showed that the reactive color, after the reduction of silver nitrate under acidic conditions, in the leaves of all three preparations was mainly found on the face side of columned-type palisade parenchyma cells where chloroplasts were not rich and large vacuoles were seen. On the inner side of the palisade parenchyma cells where chloroplasts were the richest, only weak color development was observed. This study demonstrates that the percent of dehydroascorbic acid in persimmon leaves did not exceed 40% at least until October 7. It also shows that in persimmon leaf cells, ascorbic acid is mainly localized in the cytosol of palisade parenchyma tissue cells where large vacuoles are seen.
Analogs (6-deoxyascorbic acid, erythroascorbic acid, and associated glycosides) of L-ascorbic acid (AA) contained in mushrooms were allowed to react with hydrazine to form osazones, and the conditions for separative determination by HPLC using a Zorbax SIL column were examined. Separation was started using solvent system 1 (ethylacetate/n-hexane/acetone/acetic acid, 50:50:1:1, v/v) as the mobile phase, and switching after 15min to solvent system 2 (ethylacetate/acetone/acetic acid, 100: 1:1, v/v). Detection was performed by absorbance at 500nm. Because these analogs showed different formation rates for osazone, calibration curves were prepared for each substance. The recovery rate in the load test was 93-105%. By this method, AA and the analogs contained in eight species of edible mushrooms have been determined. The results revealed that: (1) the main constituents of all mushrooms are AA analogs rather than AA itself; (2) only one species contained AA in a very small amount (2μmol/kg); (3) the types of AA analogs present differed according to the species of mushrooms, and (4) the total amount of AA analogs was between ca. 100-500μmol/kg (2-9mg per 100g, converted to AA). In addition, a new AA analog was found in Pleurotus ostreatus and identified as 5-O-(α-D-xylopyranosyl)-D-erythroascorbic acid in structural analyses by NMR and other methods.
Rats were fed 20 or 70% casein diets with varying amounts of vitamin B6 (B6), and the B6 content and B6-dependent enzymatic activity in their tissues were examined to determine the minimum re-quirement of B6 for animals subjected to different levels of dietary protein (i.e., 20%: 0, 1.45, 2.90, 5.80mg pyridoxine (PN)/kg diet; 70%: 0, 2.90, 5.80, 8.70mg PN/kg diet). B6 requirements for the rats were almost met in the 1.45mg PN/kg 20% casein diet and the 2.90mg PN/kg 70% casein diet when judged from the hepatic B6 content. However, almost twice the PN was required in both 20 and 70% casein diets when judged from PLP-enzymatic activity. The content of B6 vitamers in plasma appeared to be most sensitive to B6 status, though the satisfactory level is not known. It was confirmed that, in any case tested, a high-protein diet increased the requirement of B6.
Glucose is usually chosen as the energy source for total parenteral nutrition. However, the optimal glucose : fat ratio for peripheral parenteral nutrition has not been examined sufficiently. We compared glucose : fat ratios in hypocaloric nutrition. Male SD rats were given hypocaloric parenteral nutrition (approx. 190 kcal/kg/d) for 5 d after laparotomy. The hypocaloric solutions used contained 0, 33, 50, 67 or 100% of the non-protein energy in the form of fat. Body weight change, nitrogen balance, organ weights, and hepatic, splenic and plasma biochemistries were assessed. Body weight increase in the 67 and 100% fat groups was significantly greater than that in the 0 % fat group. Nitrogen balance was the same in all groups. Hepatic glycogen content was significantly lower in the 100% fat group than that in the 0 % fat group. The weight of epididymal fat deposits was significantly lower in the 0 % fat group than in the 50 and 67% fat groups. On the other hand, tissue triglyceride content and plasma lipid levels in the 100% fat group were significantly higher than in the 0 % fat group, and were also higher than in the control group. It is suggested that combinations of glucose and fat have sparing effects on body fat and hepatic glycogen. Combinations of glucose and fat as non-protein energy sources were superior to glucose or fat alone for hypocaloric parenteral nutrition.
Previous studies have demonstrated that dietary lipid manipulation may modify immune response by affecting lymphocyte proliferation, phagocytosis, cytokine production, etc. In this paper, we investigated the effect of olive oil (00) on, the phagocytic activity and cytokine production by murine peritoneal cells. These results were compared with those obtained from mice fed diets containing sunflower oil (SO) or hydrogenated coconut oil (HCO). Balb/c mice were divided into three groups and fed diets containing 15% by weight of either 00, SO or HCO for 5, 15, 30, 60 or 90 d. Phagocytic activity and interleukin-1 (IL-1) production were increased in 00-fed mice as compared to the other groups. On the contrary, io significant differences were observed in the levels of tumor necrosis factor (TNF) production, although the levels of this cytokine were slightly increased in mice fed the 00 diet. These observations suggest that 00 is able to modify the immune response and therefore, it may be used as an immunomodulatory agent.
The existence of disaccharidases and an enzyme that hy-drolyzes maltitol were investigated in the large intestine of rats. In ad-dition, the properties of disaccharidases were studied in the cecum and colon with hyperplasia induced by the ingestion of nondigestible car-bohydrates such as maltitol and glucomannan. Maltase activity was de-tected in the cecal and colonic mucosa of rats fed a regular diet, although it was a very low level as compared with that in the small intestinal mucosa. Maltitol hydrolysis was notably lower in the cecum and colon than in the small intestine. The Km of maltose was 5.56mM in the small intestine and 5.59mM in the cecum, while that in the colon was 2.56mM. The Vmax of maltose was at very low levels in the cecum (0.38μmol/mg protein/h) and colon (0.37μmol/mg protein/h) in comparison with that in the small in-testine (30.3μmol/mg protein/h). With regard to the maltitol hydrolyzing enzyme, Km and Vmax were 2.00 mM and 2.51μmol/mg protein/h in the small intestine, respectively. Km and Vmax in the cecum and colon could not be measured because the level was too low. The tissue weights of the cecum and colon increased significantly in both the maltitol (p<0.01, p<0.05) and glucomannan (p<0.01, p<0.05) groups in comparison with that of the control group. The specific activity of maltase decreased significantly in the small intestine of the maltitol (p<0.05) and glu-comannan (p<0.01) groups. However, maltase activity in the cecum and colon was not lowered by maltitol ingestion, although it decreased significantly in the cecum of the glucomannan group (p<0.01). Sucrase activity in the small intestine and cecum was decreased significantly by maltitol (p<0.05, p<0.01) or glucomannan (p<0.01, p<0.01) ingestion, whereas it was not decreased in the colon. Maltitol hydrolyzing activity did not decrease significantly in the small intestine of the maltitol group, although that in the cecum and colon was not measured exactly by the methods used here. These results demonstrate that disaccharidases exist in the cecal and colonic mucosa of rat, and that they are not induced even in the tissue with hyperplasia, which is caused by maltitol ingestion.
We examined the inhibitional and nutritional effects of total parenteral nutrition (TPN) containing D-amino acids (D-phenylalanine, D-Phe; D-valine, D-Val; D-leucine, D-Leu; D-methionine, D-Met) on tumor growth in AH 109A hepatoma-bearing rats. Five experimental groups were examined: a control amino acid solution group (control group), D-Phe group, D-Val group, D-Leu group and D-Met group. The analysis of tumor volume and weight revealed significant tumor growth inhibition in the D-Val group as compared with the control group. In the D-Val group, decreases of DNA and protein contents in the tumor tissues were also observed. The D-Leu and D-Met groups showed a tendency toward tumor growth inhibition. The protein content in the liver tissues of these two groups was significantly higher as compared with the control group. The DNA content in the liver tissue was also significantly higher in the D-Met group. The body weight including the tumor (on the final day of TPN) was significantly lower in the D-Val group as compared with the control group, but there was no significant difference in the groups for body weights not including tumors (carcass body weight). The hematocrit and hemoglobin values, indicators of anemia, were significantly higher in the D-Val group as compared with the control group. From these results, regarding tumor growth inhibition, the D-Val solution had the strongest inhibitory effect with no negative influence on the host, and improvement of nutritional status was also suggested in the rats that received the D-Leu or D-Met solutions.
The diet and nutritional status dominate a tolerance to en-vironmental xenobiotics. In this study, the cytotoxic action of carbon tetrachloride (CCl4) and 3-amino-1, 4-dimethyl-5H-pyrido[4, 3-b]indole (Trp-P-1), one of the dietary carcinogens, was investigated using primary cultured hepatocytes from rats fed a high-fat (23% corn oil) or high-pro-tein (50% casein) diet for three weeks. Both chemicals showed strong cytotoxicity to hepatocytes, which was judged by measurement with the MTT-test and lactate dehydrogenase leakage test. A dietary effect on cytotoxicity was observed; hepatocytes from rats fed the high-protein diet were more susceptible to cytotoxicity than the cells from rats fed a standard diet. On the other hand, ureogenesis, as a cellular function of hepato-cytes, was markedly decreased in the cells from rats fed the high-fat diet. These activities were affected in the CCl4-treated cells but not in the Trp-P-l-treated cells. The same trend of both diet and chemical effects was observed in gluconeogenesis from fructose. We conclude that the hepatocytes from rats fed a high-protein diet have high susceptibility to the cytotoxicity of CCl4 and Trp-P-1, but cytotoxicity was not related to the reduction of cellular functions.
The effects of water-soluble and -insoluble indigestible saccharides (IDS) on immune responses of the intestinal tract were studied. Male 4-week-old Sprague Dawley rats were fed for three weeks on diets containing several kinds of IDS at 5%. The results revealed that the proportion of is-light chain and IgA-presenting lymphocytes in small intestinal and cecal mucosa differed in increased number depending on the type of IDS. The response of colonic mucosa was not pronounced. The amounts of short-chain fatty acid (SOFA) and lactic acid in the cecal contents of the other test groups except the celfur group tended to be higher than those in the cellulose group, particularly in the lactulose group where many acids showed significant increases. The correlation between the proportion of k-light chain and IgA-presenting lymphocytes in the cecal mucosa and lactic acid in the cecal contents was significant, but that between the proportion of both lymphocytes and SCFA was not. Based on the above, we concluded that the oral administration of IDS induces the proliferation of k-light chain and IgA-producing B lymphocytes in small intestinal and cecal mucosa, but the degree of response differs depending on the type of IDS. It is thus suggested that IDS are involved in the intestinal immune system of rats.
To elucidate the effects of soybean protein and casein on postprandial lipemia, oral fat load tests were performed before and 3 weeks after the administration of soy protein isolate (SPI) and casein supplement to normolipidemic men. Eleven normolipidemic male subjects on otherwise identical controlled diets were assigned to either a 20 g/d soy protein isolate (SPI) dietary supplement or a casein dietary supple-ment for three weeks in a crossover design. Fat load tests with 40 g/m2 of bovine milk fat were carried out before and after 3 weeks on the ex-perimental dietary supplements. Fasting plasma concentrations of lipids and apolipoproteins were not significantly different from baseline levels before or after the administration of SPI or casein supplemented diets. Neither SPI nor casein supplement affected the fasting plasma con-centrations of lipids and apolipoproteins. The areas under the incre-mental curve (AUIC) of triglyceride (TG) and remnant-like particles triglyceride (RLP-TG) after both experimental diets were not significantly different from those before the experimental diets. However, the AUIC of remnant-like particles cholesterol (RLP-C) showed a tendency (p=0.07) to decrease after administration of the diet supplemented with SPI than before the diet. The AUIC of RLP-C was significantly (p<0.05) lower after the diet supplemented with SPI than after administration of the diet supplemented with casein. These results suggest that 3 weeks of 20 g/d SPI dietary supplement favorably affects the postprandial remnant lipoprotein response as compared to the casein dietary supplement.
α-Amino-β-carboxymuconate-8-semialdehyde decarboxylase (ACMSD) [EC 184.108.40.206] is a key enzyme of niacin synthesis from tryptophan. In this study, we examined whether dietary linoleic acid alters the protein expression of ACMSD in rat liver. Antibody against rat liver ACMSD was prepared by injecting mice with the purified enzyme. With the use of this polyclonal antibody and analysis by two-dimensional electrophoresis, we studied the mechanism by which the level of liver ACMSD activity was varied in rats fed a linoleic acid diet. In the rats fed a dietary linoleic acid (L), ACMSD protein levels in the liver were strongly suppressed as compared with the rats fed a fat-free diet (FF). These results suggest that the expression level of ACMSD might be modulated by linoleic acid or their metabolites.
Calbindin-D9k expression in intestinal mucosal cells reveals a specific pattern during development in rats. It shows a low basal level in suckling and adult rats, but after weaning at 21 d of age, increases to three times that of the basal level for several days only, around 24 d. We attempted to clarify whether the regulation of developmental change was at the transcriptional or post-transcriptional level. The calbindin-D9k protein and mRNA concentrations during pre- and postweaned de-velopment were determined by Western blot and Northern blot analysis, respectively, and compared with calcium binding activity by 45Ca. For Western blot analysis, a corresponding antibody was raised in rabbit using a bacterially expressed fusion protein, glutathione S-transferase (GST, EC 220.127.116.11), and calbindin-D9k. Calbindin-D9k cDNA was linked to a GST gene within a molecule of vector plasmid and a fusion protein was expressed in Escherichia coll. There were significant (p<0.001) correlations between calbindin-D9k protein, mRNA concentrations and calcium-binding ac-tivity: r=0.90 for protein vs. mRNA, r=0.93 for protein vs. binding activity and r=0.95 for mRNA vs. binding activity. These results indicate that calbindin-D9k expression during postnatal development is regulated at the transcriptional level.
To characterize the energy metabolism in individuals with mental retardation (MRs), we measured energy cost at several physical activity levels (basal, supine, sitting, standing, and walking at 30, 50 and 70m/min), maximal oxygen consumption (Vo2max), and body composi-tion in 23 male MRs and the same number of volunteer male controls. Both groups were individually matched for age, body height, and body weight. Energy cost was measured by the Douglas bag technique. The recently developed sulfur hexafluoride (SF6) dilution technique was employed for measuring body composition. In addition, 3-dimensional accelerometry was used for evaluating body movements, and plasma indices of macronutrients were also measured. The energy cost of MRs, when sitting, standing, and walking at 30 and 50 m/min, was significantly higher than that of controls (p<0.05), while the basal and resting metabolic rates were similar in both groups. Vo2max was significantly lower (p<0.05) in MRs than controls. Accelerometry demonstrated excessive movement by MRs, which may explain their higher energy cost of exercise. In contrast, no significant difference was observed in percent body fat or lean body mass. Concentrations of plasma total cholesterol, triacylglycerols and albumin were significantly lower in MRs as compared with the controls. Our findings suggest that MRs are burdened with an energy metabolism less economical than non-MRs. Limited physical activity in their daily life may be the cause. These characteristics of MRs' energy metabolism should be considered for planning their proper dietary schedules and physical activity programs.
Ethanol in the presence of disulfiram (N, N, N', N'-tetra-ethylthiuram disulfide, an inhibitor of aldehyde dehydrogenase) inhib-ited liver β-alanine-oxoglutarate aminotransferase (β-AlaAT I) activity yet activated tyrosine aminotransferase (TAT) in weanlirrg rats in vivo. The effect on fi-AIaAT I was followed by the inhibitory expression of β-AIaAT I mRNA. The β-AIaAT I activity was reduced with a pseudo-first-order profile with time, and the half-life was calculated to be 12.3±0.83h with the rate constant (Kd) of 0.056±0.004h-1. The synthesis of β-AIaAT Tin rat liver was estimated to be 1.56×10-10 mol/g of wet tissue per hour at a steady state. A combination of ethanol and disulfiram also reduced β-alanine-pyruvate aminotransferase (β-AIaAT II) activity to 60% of the control after 24h.
Sake lees obtained by brewing from liquefied rice were deprived of water and alcohol by lyophilization, and then examined for nutritional availability with the aid of proximate food analysis, amino acid analysis and animal experiment. Freeze-dried sake lees powder was comprised of 44.6% protein, 37.4% carbohydrate, 2.5% fat, 6.7% fiber, 1.8% ash and 7.2% moisture (alcohol <0.1%), of which the nutritive value (amino acid score) was estimated as 89.6 when compared with the amino acid requirements for preschool children (FAO/WHO/UNU, 1985). Sake lees protein had been, however, appreciably improved in the limiting amino acid “lysine” relative to polished rice protein. As a result of an animal experiment, the rats fed a 50% sake lees powder diet proved to be equal in growth to those fed a 20% casein (control) diet, although the former diet had to be supplemented with vitamins and minerals, which were in shortage as compared to the control diet. On the other hand, the feeding of sake lees powder was effective in lowering the serum triacylglycerol concentration. Accordingly, sake lees powder can be assessed as a favorable candidate for not only protein-rich but also hypolipidemic provisions.
The effects of supplementing Bifidobacterium longum SBT 2928 and Lactobacillus acidophilus SBT 2062 to a high-fat, low-calcium diet on bile acid concentration, fatty acid concentration, cytolytic activity and intestinal alkaline phosphatase (ALP) activity of fecal water in rats injected with and without 1, 2-dimethylhydrazine dihydrochloride (DMH) were examined. Male Wistar rats at 8 weeks of age were fed a diet containing 18% coconut oil, 2% corn oil and 0.1% calcium for 15 d. Lyophilized cultures were supplemented to test diets at a concentration of 1%. The feeding of a high-fat, low-calcium diet elevated the bile acid concentration, cytolytic activity and ALP activity of fecal water as compared to the AIN-76A diet, whereas the fatty acid concentration was not changed. None of the cultures had any effect on these parameters. Furthermore, 8 week-old rats were given a single subcutaneous injection of DMH at 40 mg/kg body weight, and fed the same diets for 15 d. The DMH injection had no effect on the bile acid concentration but increased the fatty acid concentration and cytolytic activity of fecal water. In contrast, ALP activity was lower in the DMH-treated rats than in the non-treated rats. The ingestion of B. longum lowered cytolytic activity but had no effect on the bile acids, fatty acids and ALP activity of fecal water. L. acidophilus had no effect on these parameters.