The effect of betaine status on folate deficiency-induced hyperhomocysteinemia was investigated to determine whether folate deficiency impairs homocysteine removal not only by the methionine synthase (MS) pathway but also by the betaine-homocysteine S-methyltransferase (BHMT) pathway. For this purpose, we investigated the effect of dietary supplementation with betaine at a high level (1%) in rats fed a folate-deprived 10% casein diet (10C) and 20% casein diet (20C). We also investigated the effect of choline deprivation on folate deficiency-induced hyperhomocysteinemia in rats fed 20C. Supplementation of folate-deprived 10C and 20C with 1% betaine significantly suppressed folate deprivation-induced hyperhomocysteinemia, but the extent of suppression was partial or limited, especially in rats fed 10C, the suppression of plasma homocysteine increment being 48.5% in rats fed 10C and 69.7% in rats fed 20C. Although betaine supplementation greatly increased hepatic betaine concentration and BHMT activity, these increases did not fully explain why the effect of betaine supplementation was partial or limited. Folate deprivation markedly increased the hepatic concentration of N,N-dimethylglycine (DMG), a known inhibitor of BHMT, and there was a significant positive correlation between hepatic DMG concentration and plasma homocysteine concentration, suggesting that folate deficiency increases hepatic DMG concentration and thereby depresses BHMT reaction, leading to interference with the effect of betaine supplementation. Choline deprivation did not increase plasma homocysteine concentration in rats fed 20C, but it markedly enhanced plasma homocysteine concentration when rats were fed folate-deprived 20C. This indicates that choline deprivation reinforced folate deprivation-induced hyperhomocysteinemia. Increased hepatic DMG concentration was also associated with such an effect. These results support the concept that folate deficiency impairs homocysteine metabolism not only by the MS pathway but also by the BHMT pathway.

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The mechanism by which feeding a higher casein diet results in resistance to choline deprivation-induced hyperhomocysteinemia was investigated in rats. Plasma homocysteine concentration was significantly lower in rats fed a 30% casein diet (30C) than in rats fed a 10% casein diet (10C). Choline deprivation did not enhance plasma homocysteine concentration in rats fed 30C, while it significantly enhanced plasma homocysteine concentration in rats fed 10C. The choline deprivation-induced enhancement of plasma homocysteine concentration in rats fed 10C was significantly suppressed by methionine supplementation in a dose-dependent manner in the range of 0.1 to 0.3%, but the suppressive effect of methionine became smaller with an increase in supplementation level in the range of 0.3 to 0.5%. At a 0.5% supplementation level, methionine did not exhibit any suppressive effect on choline deprivation-induced hyperhomocysteinemia. The higher plasma homocysteine concentration in rats fed choline-deprived 10C+0.5% methionine was significantly decreased by concurrent supplementation with 0.32% glycine+0.94% serine to the level of rats fed 10C. Raising dietary total amino acid level by adding 3.61% branched-chain amino acids (BCAA)+4.5% acidic amino acids (AAA) to choline-deprived 10C+0.5% methionine+0.32% glycine+0.94% serine resulted in a further decrease in plasma homocysteine concentration to a level lower than the level in rats fed 10C. Choline deprivation-induced increases in hepatic S-adenosylhomocysteine and homocysteine concentrations were significantly suppressed by supplementation with glycine+serine and further suppressed by BCAA+AAA. Hepatic cystathionine β-synthase activity and its gene expression were significantly increased by BCAA+AAA. Hepatic triglyceride concentration changed in a manner similar to that of plasma homocysteine concentration. The results indicate that there are at least three factors contributing to the resistivity of rats fed a higher casein diet (30C) to choline deprivation-induced hyperhomocysteinemia, i.e., higher intake of methionine, higher intake of glycine and serine, and higher intake of other amino acids such as BCAA and AAA.

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Moderate-intensity running (treadmill velocity of 21 m/min) increased blood lactate and actived transforming growth factor-β (TGF-β) concentration in rat cerebrospinal fluid (CSF). On the other hand, low-intensity running (15 m/min) did not increase blood lactate and caused no change in CSF TGF-β. Intraperitoneal (i.p.) administration of lactate to anesthetized rats caused an increase in blood lactate similar to that observed after a 21 m/min running exercise and increased the level of active TGF-β in CSF. Intraperitoneal administration of lactate at the same dose to awake and unrestricted rats caused a decrease in the respiratory exchange ratio, that is, enhancement of fatty acid oxidation and depression of spontaneous motor activity (SMA). Given that intracisternal administration of TGF-β to rats has been reported to enhance fatty acid metabolism and to depress SMA, we surmise that the observed changes caused by i.p. lactate administration in this study were mediated, at least in part, by TGF-β in the brain.

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ATP-binding cassette transporters (ABC) A1 and G1 are key molecules in cholesterol efflux from macrophages, which is an initial step of reverse cholesterol transport (RCT), a major anti-atherogenic property of high-density lipoprotein (HDL). Astaxanthin is one of the naturally occurring carotenoids responsible for the pink-red pigmentation in a variety of living organisms. Although astaxanthin is known to be a strong antioxidant, it remains unclear through what mechanism of action it affects cholesterol homeostasis in macrophages. We therefore investigated the effects of astaxanthin on cholesterol efflux and ABCA1/G1 expressions in macrophages. Astaxanthin enhanced both apolipoprotein (apo) A-I- and HDL-mediated cholesterol efflux from RAW264.7 cells. In supporting these enhanced cholesterol efflux mechanisms, astaxanthin promoted ABCA1/G1 expression in various macrophages. In contrast, peroxisome proliferator-activated receptor γ, liver X receptor (LXR) α and LXRβ levels remained unchanged by astaxanthin. An experiment using actinomycin D demonstrated that astaxanthin transcriptionally induced ABCA1/G1 expression, and oxysterol depletion caused by overexpression of cholesterol sulfotransferase further revealed that these inductions in ABCA1/G1 were independent of LXR-mediated pathways. Finally, we performed luciferase assays using human ABCA1/G1 promoter-reporter constructs to reveal that astaxanthin activated both promoters irrespective of the presence or absence of LXR-responsive elements, indicating LXR-independence of these activations. In conclusion, astaxanthin increased ABCA1/G1 expression, thereby enhancing apoA-I/HDL-mediated cholesterol efflux from the macrophages in an LXR-independent manner. In addition to the anti-oxidative properties, the potential cardioprotective properties of astaxanthin might therefore be associated with an enhanced anti-atherogenic function of HDL.

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Numerous preclinical studies have suggested that micronutrient status is associated with the progression of human immunodeficiency virus (HIV) disease, but results from observational studies are still controversial. The objective was to systematically review the efficacy of multiple micronutrient supplementation on mortality and morbidity in HIV-infected adults. A comprehensive search of the PubMed/MEDLINE, EMBASE and Cochrane Library was performed. Six randomized controlled trials assessing the effect of multiple micronutrient supplementation on HIV-infected adults were included. Relative risk was used as an effect measure to compare the intervention and control groups with fixed-effects or random effects models. Sensitivity analyses were applied to further evaluate heterogeneity. Multiple micronutrient supplementation decreased the mortality and morbidity of HIV-infected adults nonstatistically significantly (RR=0.90; 95% CI, 0.80 to 1.02; p=0.09). Sensitivity analyses revealed that multiple micronutrient supplementation decreased the mortality and morbidity of adults infected with HIV alone statistically significantly (RR=0.75; 95% CI, 0.58 to 0.95; p=0.02), but not adults infected with both HIV and pulmonary tuberculosis (RR=0.97; 95% CI, 0.84 to 1.11; p=0.65). Multiple micronutrient consumption was correlated with reduction of the mortality and morbidity of HIV-infected adults, at least those in developing countries and infected with HIV alone, and should be prescribed by local doctors for those in earlier stages especially.Numerous preclinical studies have suggested that micronutrient status is associated with the progression of human immunodeficiency virus (HIV) disease, but results from observational studies are still controversial. The objective was to systematically review the efficacy of multiple micronutrient supplementation on mortality and morbidity in HIV-infected adults. A comprehensive search of the PubMed/MEDLINE, EMBASE and Cochrane Library was performed. Six randomized controlled trials assessing the effect of multiple micronutrient supplementation on HIV-infected adults were included. Relative risk was used as an effect measure to compare the intervention and control groups with fixed-effects or random effects models. Sensitivity analyses were applied to further evaluate heterogeneity. Multiple micronutrient supplementation decreased the mortality and morbidity of HIV-infected adults nonstatistically significantly (RR=0.90; 95% CI, 0.80 to 1.02; p=0.09). Sensitivity analyses revealed that multiple micronutrient supplementation decreased the mortality and morbidity of adults infected with HIV alone statistically significantly (RR=0.75; 95% CI, 0.58 to 0.95; p=0.02), but not adults infected with both HIV and pulmonary tuberculosis (RR=0.97; 95% CI, 0.84 to 1.11; p=0.65). Multiple micronutrient consumption was correlated with reduction of the mortality and morbidity of HIV-infected adults, at least those in developing countries and infected with HIV alone, and should be prescribed by local doctors for those in earlier stages especially.

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While sugar intake is an important factor for obesity, diabetes and dental caries, sugars are also important energy sources, especially for rapidly growing children. Children like sugar-rich sweet foods. However, intake for Japanese children is not known due to a lack of studies and sugar composition data. This study was designed to determine sugar intakes from snacks and beverages in Japanese school children. A nutrition survey was conducted for 3 weekdays for 283 Japanese school children (7, 10 and 13 y old) in 8 prefectures from different areas of Japan. The methods for the survey were the weighing method for school lunches and the 24-h recall method for other foods. To estimate sugar intakes, the sugar composition table that was recently compiled by us for 135 beverages, cakes and other sweets was used. Height and weight were measured. They were similar to Japanese averages. Energy intakes were also similar to the results of the Japanese National Health and Nutrition Surveys. Sugar eaten outside meals was 24.7±15.5 g/d. From the National Health and Nutrition Surveys conducted in 2009, the mean sucrose intake from meals including some home-made cookies for 7-14-y-old children was 5.5 g/d, suggesting the mean total sugar intake of these children was about 30 g/d. This was within the range of FAO/WHO recommendation (less than 10% of energy intake, 49 g for these children. Mean intakes among age groups were not significantly different (p>0.05), but the intake for girls was lower than for boys in the oldest age group (p<0.05). Contributions of each sugar to total intake were sucrose 64%, fructose 14%, glucose 13% and lactose 9%. Fructose and glucose were mainly from isomerized sugar. Contributions of food groups to total intake were beverages 25%, baked goods 19% and ice cream 17%, respectively, covering 61% of all. In conclusion, we revealed that the average sugar intake of Japanese children was within the range of the FAO/WHO recommendation, though the effects of the kind of sugars on health remain to be clarified.

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The present study sought to determine estimated equilibrated dietary intakes (EEDIs) for nine essential minerals: sodium (Na), potassium (K), calcium (Ca), magnesium (Mg), phosphorus (P), iron (Fe), zinc (Zn), copper (Cu), and manganese (Mn), using data from 17 human mineral balance studies conducted from 1986 to 2007 (subjects=178). Among these studies, two used male subjects, two subjected some or all subjects to sodium restriction, and one study utilized a low protein diet; these subjects were not included in the present analysis. Consequently, data from 13 studies of young female subjects (n=131) consuming a standard diet were selected. Balance distribution medians for six of the minerals (Na, K, Mg, Fe, Zn and Cu) were positive, so the data were adjusted to set the medians of the balances to zero. Medians for the other minerals (Ca, P and Mn) were close to zero and were not adjusted. Intake and balance for each mineral were divided by body weight (BW), lean body mass (LBM), and standard body weight (SBW), which was calculated using height and standard body mass index (BMI=22), and EEDIs were calculated as the intercept of a simple regression equation. When relationships between intake and balance of a mineral were not significant in the regression equation, a significant regression equation comparing intake and balance of another mineral was used to calculate the intercept. Significant simple regression equations were not obtained from any of the three parameters of Na or Zn, or for two of the parameters of P; thus, K, Fe and Ca balances were used to determine the intercepts for Na, Zn and P, respectively. EEDIs for the minerals were: Na (67.9, 89.0, 62.5), K (39.5, 53.5, 37.4), Ca (11.0, 14.4, 10.1), Mg (4.18, 5.51, 3.86), P (18.7, 24.6, 17.3) (mg/kg BW/d, mg/kg LBM/d, mg/kg SBW/d), Fe (180, 237, 165), Zn (168, 241, 166), Cu (30.9, 42.6, 29.7), Mn (55.1, 72.1, 50.7) (μg/kg BW/d, μg/kg LBM/d, μg/kg SBW/d), respectively. These values are nearly identical to the mean dietary intakes.

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Oral administration of Terminalia catappa extract (TCE; 1,000 mg/kg) for 5 wk suppressed bone weight loss and trabecular bone loss in ovariectomized mice. An in vitro experiment showed that TCE (1.3-20 μg/mL) did not increase alkaline phosphatase activity, which would indicate osteoclast formation, in osteoblast-like 3T3-L1 cells. On the other hand, TCE (12.5 μg/mL) markedly decreased the number of tartrate resistant acid phosphatase (TRAP)-positive multinucleated cells, which would indicate osteoclast formation, in a co-culture system (bone marrow cells/osteoblastic UAMS-32 cells). A detailed analysis of the stages of osteoclast differentiation revealed that TCE mainly suppressed the differentiation of bone marrow mononuclear cells into osteoclast progenitor cells in the presence of M-CSF and TGF-β. An additional experiment using fractionated TCE revealed that the water-soluble fraction suppressed the bone weight loss in OVX-mice and osteoclast differentiation in vitro. Therefore, the suppressive effects of TCE on bone weight loss in mice might be due to the suppressive effects of highly polar components on the early stage of osteoclast differentiation.

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Mangiferin (3) and genkwanin 5-O-β-primeveroside (5) are the two major bioactive polyphenols with laxative property present in the extracts of agarwood (Aquilaria sinensis) leaves (AL). Here we developed an HPLC method to determine these bioactive components and four other major polyphenols in AL extracts and evaluated the pharmacological equivalence of organic and water extracts. Using mobile phase gradient conditions combined with UV detection at 330 nm, all six compounds were separated and we determined the relative extraction ratios of the six compounds present in A. sinensis extracts that were prepared under different conditions and compared the contents of the two laxative polyphenols present in the 60% ethanol extracts of A. sinensis and A. crassna. The polyphenols present in water extracts of 13 commercially cultivated A. crassna plants have also been analyzed. The laxative properties of 60% ethanol and four water extracts of A. crassna were evaluated by the frequency and weight of stools in loperamide-induced constipation model mice. The pharmacological equivalence of 60% ethanol extract and hot water (95°C) extract was identified in mice.

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A decrease in core temperature during general anesthesia is attenuated by infusion of an intravenous amino acid mixture. The purpose of the present study was to investigate the influence of physical and endocrine changes caused by ovariectomy on the inhibitory effect of amino acid infusion on anesthesia-induced hypothermia. Sprague-Dawley female rats were divided into a sham-operated (Sham) group and an ovariectomized (OVX) group. Saline solution or an amino acid mixture solution was infused for 180 min during sevoflurane anesthesia, and the rectal temperature was measured (4 groups). Intraperitoneal white adipose tissue mass, bilateral gastrocnemius weight and plasma insulin levels were measured. In the Sham rats, no inhibitory effect of the amino acid mixture on anesthesia-induced hypothermia was found, while in the OVX rats, hypothermia was significantly decreased. The intraperitoneal fat weight/body weight ratio was significantly higher in the OVX rats than in the Sham rats, but the gastrocnemius weight/body weight ratio was not significantly different. After amino acid infusion, the plasma insulin level was significantly higher in the OVX rats than in the Sham rats. In conclusion, our findings suggest that, in rats, ovarian function or female hormone affects protein turnover mediated by increase in insulin secretion and, thus, decreases the inhibitory effect of an infusion of amino acid mixture on anesthesia-induced hypothermia.

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The purpose of this study was to improve the efficiency of the microbiological assays for vitamin B₆ and inositol by using lyophilized cells from Saccharomyces cerevisiae ATCC 9080. The use of lyophilized cells as an inoculum was assessed to avoid time-consuming processes like cell precultivation and washing. The authors also examined the effects of various protectants such as skim milk, lactose, maltose, and trehalose on the assay. The viable cell counts of the lyophilized cells were approximately equal. The standard curves for vitamin B₆ and inositol concentrations obtained using lyophilized cells with maltose and intact cells gave similar linear ranges. Furthermore, the measured vitamin concentrations of the standard reference material 1849 were in the range of the established values. Therefore, lyophilized cells with maltose are potential alternative inocula for the turbidimetric method. This will increase the overall convenience associated with microbiological assays.

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