Interest in contemporary vitamin D research has been sparked in recent years, stemming from the identification of vitamin D receptors in virtually all cells as well as the enzymatic machinery necessary to produce its active form. Both epidemiological and in-vitro studies have linked vitamin D deficiency to enigmatic diseases including cardiovascular disease; however, a clear mechanistic link remains missing. This review highlights conclusions of observational studies, in-vitro experiments and randomized-controlled trials that aimed to link deficiency of the sunshine vitamin to one of the leading causes of death in the world, cardiovascular disease. Furthermore, putative mechanisms viewed from a novel perspective are also discussed.
Ascorbic acid (AA) functions as an electron donor and scavenges reactive oxygen species such as superoxide, singlet oxygen, and hydroxyl radicals in vitro. However, little is known about the effect of an AA deficiency on protein and lipid oxidation levels in the liver. Therefore, we measured the levels of protein carbonyl and thiobarbituric acid reactive substances (TBARS) in livers from senescence marker protein-30 (SMP30)/gluconolactonase (GNL) knockout (KO) mice. These mice are deficient in AA, because they lack the SMP30/GNL gene, which is essential for the biosynthesis of AA in vivo. To track the effect of an AA deficiency, at 30 d of age, mice were divided into the following four groups: AA (−) SMP30/GNL KO, AA (+) SMP30/GNL KO, AA (−) wild type (WT), and AA (+) WT. The AA (+) groups were given water containing 1.5 g/L AA, whereas the AA (−) groups received water without AA for 57 d. All mice were fed an AA-free diet. Subsequently, protein carbonyl levels in livers from AA (−) SMP30/GNL KO mice were significantly higher than those from the other three groups; however, TBARS levels were not significantly different among the four groups. Therefore, AA must act as an anti-oxidant for proteins but might not directly protect lipid oxidation in the liver.
Micronutrient deficiency is a common condition in HIV-infected individuals and may occur in all stages of the disease. The objective of this cross-sectional study was to compare the concentrations of vitamin A and beta-carotene, micronutrients related to immunity and oxidative stress, in 182 adults with HIV/AIDS, under different highly active antiretroviral therapy (HAART) regimens. Patients were divided into 3 groups according to their HAART regimen: combination of nucleoside analog reverse transcriptase inhibitors (NRTIs) and non-NRTIs; combination of NRTIs, protease inhibitors, and ritonavir; combination of NRTIs and other classes. Multiple linear regression analysis determined the effect of the treatment regimen, time of use, and compliance with the regimen, on vitamin A and beta-carotene concentrations, controlling for the following variables: gender, age, educational level, smoking, physical activity, body mass index, time of infection with HIV, presence of comorbidities, CD4+ T lymphocyte count, total cholesterol and fractions, and triglyceride levels. There was no significant difference in vitamin A or beta-carotene concentrations in patients under the different HAART regimens. However, approximately 4% of the patients had deficient/low concentrations of vitamin A (<0.70 μmol/L), and 98% showed concentrations of beta-carotene <1.0 μmol/L. In conclusion, HIV/AIDS patients in this region will not benefit from vitamin A supplementation, independently of the HAART regimen utilized, but beta-carotene may be of importance, considering its antioxidant effect.
We investigated over time whether contemporary Japanese patients with complicated peptic ulcers have any water-soluble vitamin deficiencies soon after the onset of the complicated peptic ulcers. In this prospective cohort study, fasting serum levels of water-soluble vitamins (vitamins B1, B2, B6, B12, C, and folic acid) and homocysteine were measured at 3 time points (at admission, hospital discharge, and 3 mo after hospital discharge). Among the 20 patients who were enrolled in the study, 10 consecutive patients who completed measurements at all 3 time points were analyzed. The proportion of patients in whom any of the serum water-soluble vitamins that we examined were deficient was as high as 80% at admission, and remained at 70% at discharge. The proportion of patients with vitamin B6 deficiency was significantly higher at admission and discharge (50% and 60%, respectively, p<0.05) than at 3 mo after discharge (10%). In conclusion, most patients with complicated peptic ulcers may have a deficiency of one or more water-soluble vitamins in the early phase of the disease after the onset of ulcer complications, even in a contemporary Japanese population.
Pantothenic acid (PaA) is involved in the metabolism of amino acids as well as fatty acid. We investigated the systemic metabolism of amino acids in PaA-deficient rats. For this purpose, urine samples were collected and 2-oxo acids and l-tryptophan (l-Trp) and its metabolites including nicotinamide were measured. Group 1 was freely fed a conventional chemically-defined complete diet and used as an ad lib-fed control, which group was used for showing reference values. Group 2 was freely fed the complete diet without PaA (PaA-free diet) and used as a PaA-deficient group. Group 3 was fed the complete diet, but the daily food amount was equal to the amount of the PaA-deficient group and used as a pair-fed control group. All rats were orally administered 100 mg of l-Trp/kg body weight at 09:00 on day 34 of the experiment and the following 24-h urine samples were collected. The urinary excretion of the sum of pyruvic acid and oxaloacetic acid was higher in rats fed the PaA-free diets than in the rats fed pair-fed the complete diet. PaA deficiency elicited the increased urinary excretion of anthranilic acid and kynurenic acid, while the urinary excretion of xanthurenic acid decreased. The urinary excretion of l-Trp itself, 3-hydroxyanthranilic acid, and quinolinic acid revealed no differences between the rats fed the PaA-free and pair-fed the complete diets. PaA deficiency elicited the increased excretion of N1-methylnicotinamide, N1-methyl-2-pyridone-5-carboxamide, and N1-methyl-4-pyridone-3-carboxamide. These findings suggest that PaA deficiency disturbs the amino acid catabolism.
We investigated the postprandial thermic effect of chicken and its mechanisms in rats. A chicken diet showed a strong thermic effect after consumption, and the removal of fat induced more rapid and stronger thermogenesis. Although thermogenesis induced by a purified chicken protein diet was also strong, the thermic reaction was not so rapid and a remarkable rise of peripheral temperatures was not observed. Defatted chicken and purified chicken protein activated the thyroid hormone system and up-regulated rate-limiting enzyme genes of glucose metabolism and the tricarboxylic acid (TCA) cycle in the liver. Moreover, chicken protein up-regulated the mRNA expression of a rate-limiting enzyme of hepatic lipid metabolism. It is possible that the mechanisms by which body temperature is raised are different between chicken protein and defatted chicken. On the other hand, it is possible that chicken fat suppressed the expression of energy metabolism-related genes that was induced by the consumption of lean chicken. As a result, a rise of postprandial body temperature might not have been induced after consumption of chicken fat. These results suggest that the consumption of lean chicken activates the thyroid hormone system and hepatic energy metabolism and consequently induces the postprandial thermic effect of chicken.
We previously demonstrated that the circulating concentrations of interleukin (IL)-1β, which is known to induce the development and progression of type 2 diabetes and its complications, were positively associated with γ-glutamyltransferase (γ-GTP) activity in middle-aged apparently healthy Japanese men. It was still unknown if the association between IL-1β concentrations and γ-GTP activity is within the normal range in apparently healthy Japanese women. In this study, we conducted a cross-sectional study of 824 apparently healthy women aged 40-64 y [mean±standard deviation age, 53.1±7.1 y; body mass index (BMI), 22.0±3.1 kg/m2] who participated in health checkups in Japan, and whose γ-GTP activity was within the normal range (<38 U/L). Associations of γ-GTP with IL-1β and other clinical or lifestyle factors were determined using analysis of variance (ANOVA) and one-way analysis of covariance (ANCOVA) followed by Tukey's multiple-range test. Multivariate logistic regression analyses (MLRA) were performed with γ-GTP activity as the dependent variable; independent variables included IL-1β plus clinical and lifestyle factors. ANOVA and ANCOVA indicated that IL-1β concentrations were positively associated with γ-GTP activity. MLRA showed that γ-GTP activity showed trends for higher IL-1β concentrations after adjusting for age, BMI, energy intake, alcohol intake, and smoking status. Together, IL-1β concentrations are positively associated with γ-GTP activity within the normal range in middle-aged apparently healthy Japanese women. Our results suggest that γ-GTP activity would be useful for assessing inflammation from the healthy state in Japanese women.
Nicotinamide and serotonin are synthesized from L-tryptophan in mammals. It is important to know the nutritional factors affecting the synthesis of nicotinamide and serotonin. We investigated the effects of amino acid composition. Young adult rats were fed ad libitum for 21 d a low-protein (9% casein) diet( control), or one of the low protein diets supplemented with following amino acids:  glycine, L-threonine, and L-cystine,  L-threonine and L-cystine,  glycine and L-cystine, and  glycine and L-threonine. The amounts of glycine, L-threonine and L-cystine supplementations were 2%, 0.078%, and 0.2%, respectively, and the amino acid contents of all diet were adjusted with supplementation of L-glutamic acid. The body weight gain, food efficiency ratio, and the amino acid nutrition biomarker, which is the urinary excretion ratio of (N1-methyl-2-pyridone-5-carboxamide+N1-methyl-4-pyridone-3-carboxamide)/N1-methylnicotinamide, improved by adding the amino acids glycine, L-threonine and L-cystine to a 9% casein diet. The conversion percentage of L-tryptophan to nicotinamide decreased with the addition of the amino acids glycine, L-threonine and L-cystine to a 9% casein diet, while the concentrations of serotonin in the brain, stomach and small intestine were not affected at all. The effects of each amino acid on body weight gain and the conversion ratios were also investigated. Glycine did not affect these variables. L-Cystine improved the body weight gain, the food efficiency ratio and the urine ratio, and decreased the conversion percentage. L-Threonine did not affect body weight gain or food efficiency ratio; however, it improved the urine ratio and decreased the conversion percentage.
(−)-Epigallocatechin-3-gallate (EGCG), which is largely found in green tea, is known to eliminate reactive oxygen species and associated inflammatory responses in vitro and in cells. However, the in vivo mechanisms underlying the effects of EGCG on the amelioration of metabolic disorders are not fully understood. In this study, we examined whether dietary supplementation with EGCG reduces inflammatory responses in peripheral leukocytes of a non-obese type 2 diabetes animal model, Goto-Kakizaki (GK) rats. GK rats at 9 wk of age were fed a control high-fat diet (46 energy % from lard and corn oil) or a high-fat diet containing 0.1%, 0.2%, or 0.5% EGCG (w/w) for 25 wk. The oxidative stress markers 8-hydroxydeoxyguanosine (OHdG) and total malondialdehyde (MDA) were reduced by supplementation with EGCG at 0.1%, but not at 0.2% or more. Significant reductions in the mRNA levels of genes related to inflammatory responses (TNF-α, IFN-γ, IL-1β, IL-6, IL-18, MCP-1, CD11b, and S100a6), 8-OHdG, and total MDA were induced in peripheral leukocytes of GK rats by EGCG supplementation at 0.1%, but not at 0.2% or more, compared with rats fed the control diet. The present results suggest that supplementation with a low dose of EGCG reduces oxidative stress and the expressions of genes involved in inflammation in peripheral leukocytes of GK rats.
Pyridoxal (PL) has been shown to suppress lipopolysaccharide (LPS)-induced gene expression of cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS). Commonly used cell culture media contain extremely high concentrations of pyridoxine (PN) compared to total serum levels of vitamin B6. Therefore, we evaluated how physiological concentrations of PN influence LPS-stimulated gene expression of COX-2 and iNOS. The mouse macrophage cell line, RAW264.7, was cultured in PN-free DMEM supplemented with 10% fetal bovine serum (DMEM(−PN+FBS)) for 7 d. Although the level of pyridoxal 5'-phosphate in these cells was decreased by 65%, no change was observed in cell proliferation rate or aspartate aminotransferase activity for 7 d. LPS-induced expression of COX-2 mRNA was compared between DMEM(+FBS) and DMEM(−PN+FBS). COX-2 expression was enhanced by 2.2 or 1.9 times with a 1 or 3 d treatment, respectively; however, no difference was observed at 7 d. PN (0.032-100 μm) added to the DMEM(−PN+FBS) and RAW264.7 cells was cultured in the medium containing each concentration of PN for 1 d. Enhancement of COX-2 and iNOS gene expression was suppressed by PN addition in a concentration-dependent manner. COX-2 and iNOS mRNA were similarly expressed in cells grown in media containing PN at 4 μm or higher. Overall, induction of COX-2 and iNOS by LPS was transiently enhanced when RAW264.7 cells were cultured in physiological PN concentrations.
The purpose of this study was to determine whether the effects of dietary protein on sexual organ development were different in mice and rats kept under constant darkness. Four-week-old mice (ICR strain) and rats (F344 strain) were kept under constant darkness (D) or normal lighting (N; 12-h light/dark cycle) for 4 wk. The dietary protein level was 9% casein with the addition of 0.135% cystine (9PC) or without it (9P); other components of the diet were based on the AIN-93G diet. The testes and epididymides weights (g/100 g BW) of the rats given the 9P diet in the D-group were lower than those of the rats given the 9P diet in the N-group. In the mice, lighting conditions and diet did not affect testes or epididymides weights. Body weight and food intake in the rats were affected by diet, and these values were lower in the 9P diet group; however, body weight and food intake in the mice was not affected by diet. The serum albumin concentration in the rats was lower in the 9P diet group, while that of the mice was lower in the 9PC diet group. In the rats kept under constant darkness, a diet lacking in cystine accelerated the suppression of sexual organ development and decreased serum albumin concentration, but this diet had no such effects on the mice. The finding that the effects of dietary protein were different in mice and rats suggests that protein requirements of mice are different from those of rats.
Mast cells, multifunctional effector cells of the immune system, are implicated in the pathogenesis of hepatic steatosis and fibrosis. Magnesium (Mg) deficiency was reported to increase triglyceride concentration in the liver, and to exacerbate the collagen deposition induced by carbon tetrachloride in the liver. Although Mg deficiency increases mast cells in the small intestine, the kidney and bone marrow, the effect of Mg deficiency on mast cells has not been clarified in the liver. We examined the emergence of mast cells in the liver of Sprague-Dawley rats given an Mg-deficient diet. Rats were fed a control diet or an Mg-deficient diet for 4 wk. Mg deficiency increased the levels of mRNA known to be expressed by mast cells in the liver; the mRNA of α- and β-chain high-affinity immunoglobulin E receptor (FcεR1α, FcεR1β), and the mRNA of mast cell protease 1 (Mcpt1), and mast cell protease 2 (Mcpt2). Histological observation showed that some mast cells were locally distributed around portal triads in the Mg-deficient group but mast cells were scarcely found in the control group. These results clearly indicate that Mg deficiency induces the emergence of mast cells around portal triads of the liver in Sprague-Dawley rats.
The immunomodulatory effect of fermented non-salty soybean powder (NSBP) was investigated in C3H/HeN mice. The number of splenic CD11b+, CD49b+, and interferon (IFN)-γ+CD4+ cells increased significantly, while that of interleukin (IL)-4+CD4+ and CD19+ cells decreased significantly in cultures containing NSBP. Similarly, in the spleen and Peyer's patches of mice fed a diet containing NSBP, the number of IL-12+CD11b+, CD49b+, and IFN-γ+CD4+ cells increased noticeably, whereas the number of splenic IL-4+CD4+ and CD19b+ cells was lower compared to mice fed an NSBP-free diet. Superoxide production by peritoneal macrophages was significantly higher in mice fed an NSBP-containing diet. Both intestinal total IgA and serum total IgG levels declined in mice fed the NSBP-containing diet. Microarray analysis of mRNAs extracted from Peyer's patch cells of mice fed the NSBP-containing diet indicated an increase in the expression of several genes related to cellular immune responses, while the expression of genes related to immunoglobulin production decreased. These results indicate that NSBP stimulates the cellular immune response, but suppresses the acquired humoral immune response in C3H/HeN mice.
In this study, chemical evidence for the potent xanthine oxidase inhibitory activity of “kakidoushi-cha” (dry leaves of Glechoma hederacea var. grandis), a traditional folk tea consumed in Japan, was clarified on the basis of structure identification of the active constituents. Assay-guided fractionation and purification afforded 15 compounds from the most active chromatographic fraction of an extract of the tea. Two flavonoids, apigenin and luteolin, showed remarkable inhibitory activity against xanthine oxidase (XO). The contribution of these flavonoid constituents to the observed XO inhibitory activity of the methanol and boiling-water extracts of the tea was estimated to be ca. 35% and ca. 18%, respectively.
This study was performed to: (1) assess the prevalence of dietary supplement and fortified food use, (2) examine the differences in vitamin E intake with and without dietary supplementation and/or fortified food use, and (3) determine whether some individuals consume vitamin E above the tolerable upper intake level (UL). Data were obtained from 64,624 individuals (age, ≥1 y; 47.4% males) who completed a 1-d household dietary assessment that was part of the National Health and Nutrition Survey conducted in Japan, 2003-2009. The survey also obtained information on the brand or generic name of each dietary supplement or fortified food reported, including their ingredients, through dietary assessment. The prevalence of a potential risk of excess was estimated by the proportion of persons above the age-/sex-specific ULs provided by the Dietary Reference Intakes for Japanese 2010. Supplement use was reported by 5.8% of men and 7.7% of women, whereas fortified food consumption was reported by only 2.9% of men and 3.6% of women. Use of dietary supplements was most common among older women, whereas use of fortified foods was most common among younger women. Both dietary supplement and fortified food use accounted for maximum vitamin E intake; however, the use of dietary supplements and fortified foods had little effect on the median and 95th percentile intake values. None of the subjects consumed nutrients above the UL. The collected data confirm that the use of both dietary supplements and fortified foods contributes a small amount to nutrient intake in Japanese subjects.
The Adequate Intake (AI) values in the Dietary Reference Intakes for Japanese (DRIs-J) 2010 were mainly determined based on the median intakes from 2 y of pooled data (2005-2006) from the National Health and Nutrition Survey-Japan (NHNS-J). However, it remains unclear whether 2 y of pooled data from the NHNS-J are appropriate for evaluating the intake of the population. To clarify the differences in nutrient intakes determined from 2 and 7 y of pooled data, we analyzed selected nutrient intake levels by sex and age groups using NHNS-J data. Intake data were obtained from 64,624 individuals (age: ≥1 y; 47.4% men) who completed a semi-weighed 1-d household dietary record that was part of the NHNS-J conducted annually in Japan from 2003 to 2009. There were no large differences between the median intakes calculated from 2 or 7 y of pooled data for n-6 or n-3 polyunsaturated fatty acids (PUFAs), vitamin D, pantothenic acid, potassium, or phosphorus. When the AI values and median intakes were compared, there was no large difference in the values for n-6 or n-3 PUFAs, pantothenic acid, or phosphorus. Conversely, the AI values for vitamin D and potassium differed from the median intakes of these nutrients for specific sex and age groups, because values were not based on NHNS-J data. Our results indicate that 2 y of pooled data from the NHNS-J adequately reflect the population's intake, and that the current system for determination of AI values will be applicable for future revisions.