Fatty acids containing stearic acid, which are found in hydrogenated fat, may have a detrimental effect on the cholesterol and triacylglycerol (TAG) content of plasma lipoproteins, and on the absorption of fatty acids and fat-soluble vitamins. The aim of our study was to examine the tissue concentration of lipids and vitamins A and E after feeding a hydrogenated soybean oil (HSO) diet to rats. Twenty male Sprague-Dawley rats were randomly divided into two groups, fed on coconut oil (control) and HSO, respectively in amounts corresponding to 15% of the total feed. Plasma total cholesterol, VLDL- and LDL-cholesterol, lipid peroxidation and daily excretion of the TAG and cholesterol in feces were higher in the HSO than in the control group. TAG values in plasma and liver, and HDL-cholesterol levels in plasma were lower in the HSO than in the control group. The same was true for phospholipids in plasma and for saturated fatty acids, mono- and polyunsaturated fatty acids levels in the liver and vitamin E in plasma, LDL and adipose tissue. The results of this study provide new evidence concerning the effect of dietary hydrogenated fat on lipid, TAG and vitamin E status, which are important for maintenance of good health. Consumption of dietary HSO may be associated with cardiovascular disease.
We constructed an expression vector for rice dehydroascorbate reductase (DHAR) (EC 18.104.22.168) with a polyhistidine tag at the amino terminus and introduced the vector into several strains of Escherichia coli. On conventional induction treatment with isopropylthiol-β-D-galactoside, E. coli harboring rice DHAR cDNA produced doublet polypeptides of about 27 kDa. Induction duration or growth temperature did not affect the ratio of these polypeptides. Only the larger polypeptide, corresponding to full-length recombinant DHAR, was produced in E. coli supplemented with tRNAs for several minor codons. Most of the enzymatic characteristics of the recombinant DHAR were similar to those of the native one, although the recombinant protein showed increased heat susceptibility. Using recombinant DHAR, we developed a method for simple and precise determination of dehydroascorbate concentrations in tissue extracts by spectrophotometry, and we successfully applied the method to several fruit juices and vegetables.
The effects of dietary corn bran hemicellulose (CBH) and neomycin (Neo) on hepatic caspase-3 activity and glycoprotein concentration were investigated to explore the possible mechanism of the alleviative action of dietary CBH and Neo on the development of D-galactosamine (GalN)-hepatitis. Rats were fed a diet containing 5% CBH with or without neomycin (Neo) for 7 or 14 d. On the last day of feeding, the rats were treated with GalN (400 mg/kg body weight, i.p.), and their plasma transaminase activities, hepatic glycoprotein concentrations and hepatic caspase-3 activities were determined 6 or 24 h later. Although the elevations of plasma transaminase activities were suppressed by CBH or Neo 24 h after GalN-treatment, the activities were not affected by CBH or Neo at an early stage (6 h) of GalN action. At 6 h, hepatic caspase-3 activity was elevated by CBH diet alone as high as that of the GalN-injected control-diet group, and the activity was not elevated further by GalN. At the same time, both GalN-treatment and CBH feeding reduced the hepatic glycoprotein (Mw. 64,000-74,000) concentration, but Neo did not affect the caspase activity or the glycoprotein concentration. These results suggest that dietary CBH elevates hepatic caspase-3 activity and reduces hepatic glycoprotein concentration, and may imply that CBH would suppress GalN-hepatitis not at the early- or middle-step of apoptosis but at the late-step of apoptosis or necrosis, although the relation between these phenomena and the alleviative effects of CBH and Neo on GalN-induced hepatitis is yet to be clarified.
Hepatocyte nuclear factor (HNF)-1α and HNF-1β are concerned in sucrase-isomaltase (SI) gene expression, and directly bind two sites (SIF2, SIF3) of the promoter of the SI gene. However, it is not completely clear that HNF-1α and HNF-1β play a role in regulation of SI gene expression. To clarify mechanisms of SI gene expression regulated by HNF-1α and HNF-1β, we established four stable cell lines based on enterocyte-like cell line Caco-2, in which wild HNF-1α or wild HNF-1β, or else mutant HNF-1αT539fsdelC or mutant HNF-1βR177X was overexpressed. In the HNF-1αT539fsdelC cells and HNF-1βR177X cells, but not in the wild HNF-1α cells and wild HNF-1β cells, SI gene expression and enzyme activity were significantly diminished compared with that in Caco-2 cells. Moreover, to clarify whether or not stable cell differentiation was influenced by overexpression of these transgenes, alkaline phosphatase (ALP) gene expression and enzyme activity were measured. There were no changes in ALP gene expression or enzyme activity in these cells. These observations suggest that mutant HNF-1αT539fsdelC and mutant HNF-1βR177X inhibits SI gene at the transcriptional level, resulting in decreased SI enzyme activity in Caco-2 cells. We propose that both HNF-1α and HNF-1β would contribute to constitutive expression of the SI gene in the differentiated state in Caco-2 cells.
This study sought to determine whether oral tolerance to ovalbumin (OVA), responsible for food allergy, is affected by different amounts of protein intake. For this, 6-wk-old BALB/c mice were fed with low protein (5%, LP), normal protein (20%, NP) and high protein (40%, HP) diets, orally given either OVA (OVA-fed) or water (Water-fed) for 4 d, and then immunized intraperitoneally twice at a 3-wk interval with alum-precipitated OVA. After the last immunization, sera were collected to measure total and OVA-specific IgE by enzyme assay (ELISA). Splenocytes were cultured and stimulated with concanavalin A (Con A), lipopolysaccharide (LPS) or OVA and assayed for 3H-thymidine incorporation. The culture supernatants from their splenocytes stimulated with OVA were analyzed for interleukin (IL)-4, interferon (IFN)-γ, and IL-12. Total IgE was significantly higher in OVA-fed HP groups as compared to NP and LP groups (p<0.05). The highest and the lowest OVA-specific IgE were observed in HP and LP diet groups, respectively (p<0.05). OVA-fed mice receiving the LP diet demonstrated significantly lower IL-4 as compared to the other two groups (p<0.05), while IFN-γ was significantly higher in the LP compared to the HP group (p<0.05). Levels of IL-12 did not differ among the OVA-fed groups. Splenocytes of OVA-fed mice kept on the LP and HP diet showed significant impairment of proliferation to OVA as compared to the NP group (p<0.01). Proliferation against Con A was impaired in the LP group compared to the NP group (p<0.05) but not in Water-fed groups. However, it was higher against LPS in the HP than the LP group (p<0.05) both in Water-fed and OVA-fed animals. All these findings indicate that established oral tolerance to OVA is clearly affected by the amount of protein diet. They support the suggestion that dietary protein plays an important role(s) in IgE-mediated food allergies.
Background: The origin of moisture in diarrhea feces is unknown but may represent the unabsorbed part of intestinal contents or alternatively, body fluid excreted into the digestive canal. If the latter mechanism contributes to moisture in the feces, active transport of water (H2O) associated with ion exchange channels may be involved. Objective: To investigate this possibility we measured the content of moisture and minerals (sodium [Na], potassium [K], calcium [Ca], magnesium [Mg], phosphorus [P], zinc [Zn], iron [Fe], copper [Cu] and manganese [Mn]) in feces collected during a 12-d metabolic study on 11 young Japanese female students. Design: The study was carried out as part of a human mineral balance study. The same quantity of food was supplied to each of the subjects throughout the study without consideration of body weight. Fecal specimens were collected throughout the study and were separated into those originating from the diet during the balance period based on the appearance of the ingested colored marker in the feces. Results: The moisture content of the feces ranged between 53 and 92%. Na content in the feces was low and stable when the moisture content was below 80%, whereas it increased up to serum levels when the moisture content increased above 80%. On the other hand, K content increased when compared to dry matter base. However, when comparing concentration/g moisture, K content increased when moisture was below 70%, but decreased when this rose above 70%.
The effect of dietary 5-campestenone (campest-5-en-3-one), a chemical modification product of a naturally-occurring plant sterol, campesterol, on lipid metabolism was examined using a rat liver perfusion system. Male Sprague-Dawley rats weighing about 140 g were fed a diet supplemented with or without 0.2% 5-campestenone for 14 d. 5-Campestenone feeding resulted in a marked reduction in the concentrations of serum lipids, such as triacylglycerol (TG), cholesterol, phospholipid, and free fatty acid, without influencing food intake or growth. Then, isolated livers from both groups were perfused for 4 h in the presence of an exogenous linoelaidic acid substrate. Dietary 5-campestenone markedly elevated hepatic ketone body production, while cumulative secretions of TG, cholesterol, and phospholipid by the livers of rats fed 5-campestenone were all significantly lowered as compared to those fed without the compound; the extent of the reduction was more prominent in the secretion of TG than other lipid components. In addition, the reduction of TG secretion was concomitantly accompanied by the reduced incorporation of both exogenous and endogenous fatty acids into this lipid molecule. These results suggest that dietary 5-campestenone exerts its hypotriglyceridemic effect, at least, in part through an enhanced metabolism of endogenous and exogenous fatty acids to oxidation at the expense of esterification in rat liver.
The purpose of this study was to examine whether retrograded tapioca starch (RS3-tapioca) prevents ovarian hormone deficiency-induced hypercholesterolemia. Six-month-old Wistar female rats were subjected to sham-operation or ovariectomy, and fed a cholesterol-free purified diet with or without RS3-tapioca (150 g/kg diet) instead of digestible cornstarch for 28 d. Body weight gain and food intake increased in ovariectomized rats (OVX-rats). Plasma total cholesterol concentration was lowered by RS3-tapioca in OVX rats, but not in sham-operated rats. Liver lipids increased in OVX-rats, but liver cholesterol concentration was not affected by ovariectomy and RS3-tapioca. CYP7A1 activity, small intestinal and cecal bile acid content, and fecal bile acid excretion were increased by RS3-tapioca. The ratio of cholic acid groups to chenodeoxycholic acid groups in the bile acid of the small intestine was increased by RS3-tapioca. Thus, the preventive effect of RS3-tapioca on the ovarian hormone deficiency-associated increase in plasma cholesterol concentration appears to be mediated by accelerated fecal excretion of bile acid and an increase in the intestinal pool of bile acid.
Since NADH has been implicated in medication for some symptoms and as a possible supplement for health, we characterized the metabolic fate of NADH orally given to mice by comparing with those of nicotinamide (Nam), NAD+ and NADH intraperitoneally or orally administered. Mice were individually housed in metabolic cages, and divided into two sets of four groups. Within each set, one group was intraperitoneally or orally administered saline and the other three groups received intraperitoneal or oral administration of a pharmacological dose of Nam, NAD+ or NADH (5 μmol/mouse). Twenty-four hour urine samples for the day before and days 1 to 4 after administration were collected and analyzed for Nam and its metabolites. When mice were administered saline alone, urinary excretion of Nam and its metabolites, such as nicotinamide N-oxide (Nam N-oxide), N1-methylnicotinamide (MNA), N1-methyl-2-pyridone-5-carboxamide (2-Py), and N1-methyl-4-pyridone-3-carboxamide (4-Py), was unchanged from day 0 to day 4. Intraperitoneal injection of Nam, NAD+ and NADH produced significant increases in urinary excretion of Nam and its metabolites. Similar results were obtained when Nam and NAD+ were given orally. On the other hand, oral administration of NADH did not bring about an increase in urinary excretion of Nam and its metabolites, suggesting that NADH in digestive organs has been decomposed to a compound(s) that cannot yield Nam. In fact, incubation of NADH at acidic pH to mimic the stomach resulted in rapid conversion of NADH to an unknown compound. Better understanding of the fate of oral NADH is needed for its therapeutic and supplemental use.
The inhibitory activity of six groups of flavonoids against yeast and rat small intestinal α-glucosidases and porcine pancreatic α-amylase was compared, and chemical structures of flavonoids responsible for the inhibitory activity were evaluated. Yeast α-glucosidase was potently inhibited by the anthocyanidin, isoflavone and flavonol groups with the IC50 values less than 15 μM. The following structures enhanced the inhibitory activity: the unsaturated C ring, 3-OH, 4-CO, the linkage of the B ring at the 3 position, and the hydroxyl substitution on the B ring. Rat small intestinal α-glucosidase was weakly inhibited by many flavonoids, and slightly by the anthocyanidin and isoflavone groups. 3-OH and the hydroxyl substitution on the B ring increased the inhibitory activity. In porcine pancreatic α-amylase, luteolin, myricetin and quercetin were potent inhibitors with the IC50 values less than 500 μM. The 2, 3-double bond, 5-OH, the linkage of the B ring at the 3 position, and the hydroxyl substitution on the B ring enhanced the inhibitory activity, while 3-OH reduced it.
Bioelectrical impedance analysis (BIA) is commonly used to measure the percentage of body fat (%BF), but its accuracy is controversial. In addition, the equations are specific to the group for which they were established. As far as we know, there is no study examining the accuracy of BIA in Japanese middle-aged and older people. We compared %BF assessed using BIA with that of dual-energy X-ray absorptiometry (DEXA) in 102 female and 51 male local residents aged 40 to 78 y. Simple correlation coefficients were 0.79 for females and 0.69 for males, which are statistically significant (p<0.001). However, BIA tended to overestimate %BF in the lower BF group and underestimate it in the BF higher group, and only 45.1% for female and 47.1% for male subjects were measured accurately, i.e., within 10% of the measurement by DEXA. This result suggests that this model of BIA is an alternative for estimating %BF in Japanese middle-aged and older people as well as when subjects are within a normal body fat range, but greater accuracy is needed for lean and overweight subjects.
Lemon, lime and sudachi juices were tested for antibacterial activity against seven strains of Vibrio species. All juices were effective in inhibiting the growth of the Vibrio strains. Citric acid, the major organic acid in these juices, was found to be responsible for inhibiting the growth of Vibrio parahaemolyticus. Sauce prepared from sudachi juice showed a strong bactericidal activity against Vibrio parahaemolyticus, whereas the sauce adjusted to higher pH values had no bacterial activity. Diluted sudachi juice or citric acid solution also had antibacterial activity independently. These results suggest that citrus fruit juices are effective in preventing infection with Vibrio species.