We examined the effects of dietary vitamin D deficiency on markers of mitochondrial biogenesis and dynamics in rat soleus muscle. Male Wistar rats were fed a chow with no vitamin D (No-D; 0 IU/kg) or a moderate dose (Mod-D; 2,000 IU/kg) of vitamin D chow for 8 wk. Compared to the Mod-D group, at 8 wk the No-D group showed significantly lower serum 25(OH)D levels. Although vitamin D deficiency had no effect on body composition, the No-D rats showed significantly decreased levels of PGC-1α, a marker of skeletal muscle mitochondrial biogenesis, and DRP1, a marker of skeletal muscle mitochondrial fission. The change in the PGC-1α protein expression and the serum 25(OH)D concentrations were significantly correlated. The change in DRP1 protein expression and the serum 25(OH)D concentrations tended to be correlated. There was no significant between-group difference in markers of mitochondrial fusion (MFN2 and OPA1) and mitophagy (PARKIN) in soleus muscle, and no relationship with serum 25(OH)D concentrations. Collectively our findings suggest that dietary vitamin D deficiency decreased PGC-1α and DRP1 protein expression in rat soleus muscle.

This study investigated the effects of dietary high-dose biotin intake on fat oxidation in rats using respiratory gas analysis, and evaluated fatty-acid oxidation-related enzyme activities and gene expressions in the liver. Five-week-old male Sprague–Dawley rats were fed a control diet and three biotin-supplemented diets (additive biotin concentration: 0.05%, 0.10%, and 0.20% of diet) for 3 wk. In 2 wk, fat oxidation in the 0.20% biotin-supplemented diet group was higher than that in the 0.05% biotin-supplemented diet group; however, the energy expenditure and carbohydrate oxidation were unchanged between the dietary groups. At the end of 3 wk, body weight and epididymal white adipose tissue weight reduced in the 0.20% biotin diet group, and hepatic triglyceride levels tended to decrease. Additionally, increased plasma adiponectin concentration and hepatic mitochondrial carnitine palmitoyltransferase activity as well as decreased hepatic acetyl-CoA carboxylase 2 gene expression were observed in the 0.20% biotin-supplemented diet group compared with those in the control group. These results provide strong evidence that dietary high-dose biotin intake activated fat oxidation due to the increase in hepatic β-oxidation, which may contribute to the decrease in hepatic triglyceride concentration and white adipose tissue weight.

Pregnancy is usually associated with dietary imbalances. We aim to assess the changes in Japanese women’s diet and nutritional adequacy before and during pregnancy. In the Japan Environment and Children’s Study (JECS), we recruited a large cohort of pregnant women (>100,000) from 15 Japanese communities. We assessed their diet before and during pregnancy by a food frequency questionnaire. The before and during pregnancy intakes of energy, macronutrients, and micronutrients were lower than the recommended level for Japanese women by 1% to 49%. The dietary intakes of pregnant women were lower than their intakes before pregnancy, except for milk, dairy products, and calcium. The daily during pregnancy intake of energy, carbohydrate, fat, protein, folic acid, and iron decreased by 85 kcal, 11.6 g, 2.3 g, 2.8 g, 20 μg, and 0.5 mg than the before pregnancy intake. Pregnant women’s declined food and nutrients intakes were moderately correlated to their dietary intakes before pregnancy; the correlation coefficient ranged between 0.47 and 0.67. The inter-individual variability was greater than the intra-individual variability of the dietary intakes. Almost half of the pregnant women remained in the same intake quartile for all nutrients and food groups as before pregnancy. Yet, approximately 10% of pregnant women changed their pregnancy intake by ≥50% (≥2 quartiles up or down) compared to before pregnancy. In conclusion, the study identified low periconceptional dietary intakes among Japanese pregnant women. The low dietary intake of essential nutrients, such as folate, requires revising the Japanese national periconceptional dietary and supplementation guidelines.

The element magnesium (Mg) is involved in various metabolic reactions within the human body, and its deficiency is considered a risk factor for several diseases. In this study, we investigated the relationship between serum Mg levels and mortality in a community-based population. We prospectively assessed the association between serum Mg levels at enrollment and all-cause mortality in 1,314 participants who underwent a community health examination. The mean serum Mg level was 2.4 (±0.2) mg/dL. Patients with serum Mg levels ≤2.3 mg/dL constituted the low Mg group, while those with serum Mg ≥2.4 mg/dL constituted the high Mg group. Ninety-three (7.1%) patients died during the 10-y follow-up period. Kaplan-Meier analysis revealed that all-cause mortality was significantly higher in the low Mg group (log-rank p<0.05). Cox proportional hazards analysis revealed a significant association in the unadjusted model (hazard ratio [HR] 1.72, 95% confidence intervals [CI] 1.14–2.58, p<0.01) and in the fully adjusted model (HR 1.73, 95% CI 1.09–2.76, p<0.05). This association was particularly strong in males (HR 2.08, 95% CI 1.19–3.63, p<0.05). Low serum Mg levels were significantly associated with the risk of all-cause mortality among males in a community-based Japanese population.

β-Hydroxy-β-methylbutyrate (HMB), a metabolite of leucine, is known to increase muscle mass and strength. However, the effect of perioperative HMB supplementation in liver surgery is unclear. Moreover, the impact of HMB on the skeletal muscle fiber type also remains unclear. We investigated the impact of HMB on the body composition and skeletal muscle fiber type in sarcopenic rats undergoing major hepatectomy. Nine-week-old male F344/NSlc rats were maintained in hindlimb suspension (HLS) and were forcedly supplemented with HMB calcium salt (HMB-Ca, 0.58 g/kg×2 times) or distilled water in addition to free feeding. After 2 wk of HLS, the rats underwent 70% hepatectomy and were sacrificed 3 d after surgery. Body composition factors and the proportion of slow-twitch fibers in hindlimb muscles were evaluated. HMB maintained the body composition and hindlimb force and acted against their deterioration in sarcopenic rats, exerting a particular effect on lean mass weight, which was significant. In the histological study, HMB significantly increased the proportion of slow-twitch fibers in the soleus (p=0.044) and plantaris (p=0.001) of sarcopenic rats. HMB ameliorated deterioration of the body composition and increased the proportion of slow-twitch fibers in sarcopenic rats undergoing major hepatectomy.

Intestinal-type alkaline phosphatase (IAP) is expressed at a high concentration in the brush border membrane of intestinal epithelial cells and is known to be a gut mucosal defense factor. In humans, a single gene (ALPI) for IAP has been isolated, and its transcription produces two kinds of alternatively spliced mRNAs (aAug10 and bAug10). Recently, we discovered that vitamin D up-regulated the expression of both types of human IAP alternative splicing variants in Caco-2 cells. However, the functional difference of protein encoded by the mRNA variants has remained elusive. In the present study, we aimed to provide further insight into the characterization and structure of IAP isoforms. To analyze the protein translated from the ALPI gene, we constructed two kinds of cDNA expression plasmids (aAug10 and bAug10), and the transfected cells were homogenized and assayed for alkaline phosphatase (ALP) activity. We also designed the homology-modeled 3D structures of the protein encoded by the mRNA variants (ALPI-aAug10 and ALPI-bAug10). The levels of ALP activity of COS-1 cells transfected with the aAug10 plasmid were increased significantly, while cells transfected with the bAug10 plasmid had undetectable ALP activity. The homology-modeled 3D structures revealed that the variant bAug10 lacks the central N-terminal α-helix and residue corresponding to Asp-42 of ALPI-aAug10 near the active site. This is the first report on the characterization and structure of alternatively spliced transcript variants of the human ALPI gene. Further studies on the regulation of aAug10 and/or bAug10 mRNA expression may identify novel physiological functions of IAP.

Morus alba leaf extract (MLE), a strong inhibitor of sucrase, suppresses blood glucose elevation following sucrose ingestion. To investigate that sucrose inhibited from digestion using MLE is utilized through gut microbiota, [U-¹⁴C]-sucrose solutions with or without MLE were administered orally to conventional and antibiotic-treated rats, and the excretion of ¹⁴CO₂ and H₂ produced by gut microbiota was measured for 24 h. After an administration of [U-¹⁴C]-sucrose to conventional rats, the unit excreted ¹⁴CO₂ peaked at 4 h, and the cumulative ¹⁴CO₂ excreted over 24 h was approximately 60% of the radioactivity administered. No H₂ was excreted. Following an administration of [U-¹⁴C]-sucrose and MLE in conventional rats, the unit excreted ¹⁴CO₂ peaked later, at 8 h, and was significantly lower (p<0.05). The cumulative ¹⁴CO₂ excreted over 24 h was equal in both groups, although there was a time lag of 2–3 h in rats given [U-¹⁴C]-sucrose and MLE. The amount of H₂ excreted by these rats peaked 8 h after administration. Following the administration of [U-¹⁴C]-sucrose and MLE to antibiotic-treated rats, the unit excreted ¹⁴CO₂ peaked lower, and the cumulative ¹⁴CO₂ excretion over 24 h was approximately 40%. In this group, H₂ was minimally excreted. H₂ and ¹⁴CO₂ produced by gut microbiota were excreted simultaneously. In conclusion, sucrose inhibited from digestion using MLE was fermented spontaneously by gut microbiota and was not excreted into feces. In addition, it confirmed that H₂ excretion could be used directly to indicate the degree of fermentation of nondigestible carbohydrates.

We aimed to verify antitumor effects of zinc acetate on hepatocellular carcinoma (HCC) in vitro. Five HCC cell lines (HepG2, Hep3B, Huh7, HLE and Alex) were used to evaluate the antitumor effects of zinc acetate. Cell viability was determined by the Cell Counting Kit-8 assay. The cell-cycle alteration was evaluated by a flow cytometric analysis and the detection of cell cycle-related proteins. Apoptosis was determined based on the caspase-cleaved cytokeratin 18 (cCK18) levels. The microRNAs (miRNAs) related to an antitumor effect of zinc acetate were identified using microarrays. Zinc acetate significantly inhibited the proliferation of HCC cells in a dose-dependent manner. The treatment with zinc acetate resulted in significantly increased cCK18 levels in the supernatant and enhanced the expression of heme oxygenase-1 (HO-1) in HCC cells. The flow cytometric analysis revealed an increase of HCC cells in the S and G₂/M phases by the administration of zinc acetate, and the expressions of Cdk2 and cyclin E were increased. The miRNA expression profile of the HCC cells treated with zinc acetate was extremely different from that of the untreated HCC cells. These results suggest that the zinc acetate supplementation induces the apoptosis of HCC cells, but does not affect the cell cycle progression. Upregulation of HO-1 and the alteration of miRNAs’ profile may be involved in antitumor effects of zinc acetate in HCC cells.

Tryptophan is an essential amino acid important as a protein building block, but it also serves as substrate for the generation of several bioactive compounds with important physiological roles. Furthermore, tryptophan has been reported to have a unique role as a nutritional signaling molecule that regulates protein synthesis in mouse and rat liver. In the present study, the acute effects of tryptophan on protein synthesis were confirmed and compared with those of leucine in rats. Eighteen hours fasted rats were orally administered of tryptophan or leucine at a dose of 135 mg/100 g body weight by gavage and then sacrificed 1 h after administration. The effects of tryptophan and leucine on the rate of protein synthesis were evaluated by the surface sensing of translation (SUnSET) method. We also examined the ability of tryptophan to induce activation of the mTOR pathway by measuring phosphorylation of 4E-BP1 and S6K1. Oral administration of tryptophan led to a stimulation of the rate of protein synthesis concomitant with activation of mTOR pathway in the liver, but not in skeletal muscle. We also investigated the sensitivity of liver protein synthesis to tryptophan administration. The half-maximal effective doses (ED50) of tryptophan in stimulating 4E-BP1 and S6K1 phosphorylation were both about 60% of daily intake. The effect of tryptophan on hepatic protein synthesis was similar to that of leucine on muscle protein synthesis, and the sensitivity of liver protein synthesis to tryptophan administration appeared to be almost the same or slightly lower than that of muscle protein synthesis to leucine administration.

Liver can be directly involved in the synthesis and decomposition of fatty acids. Liver lipid deposition is one of the most common chronic liver diseases. Estrogen deficiency can cause lipid deposition and energy metabolism disorders in the liver. Sheep bone collagen peptide (SBCP) has been shown to have estrogen-like effects in previous studies. And SBCP has high bioavailability, safety and non-toxic side effects. This study aimed to investigate the effect of SBCP on liver lipid deposition (LLD) caused by estrogen deficiency. Female Wistar rats were treated as follows (n=10): sham group: underwent peri-ovary fat removal operations, ovariectomized rats (model group), ovariectomized rats receiving SBCP treatments: SBCP high dose group (SBCP-H), SBCP medium dose group (SBCP-M) and SBCP low dose group (SBCP-L). After 8 wk, the model group demonstrated severe LLD and liver pathological changes, with increased malondialdehyde (MDA) and free fatty acid (FFA) levels (p<0.05). Additionally, the total superoxide dismutase (T-SOD) activity (p<0.05), serum albumin-to-globulin (A/G) ratio (p<0.05), amount of butyric acid-producing bacteria and short-chain fatty acids (SCFAs) content decreased. SBCP intervention could inhibit the occurrence of LLD and alleviate the liver histopathological damage induced by estrogen deficiency by relieving oxidative stress, preventing the loss of butyric acid-producing bacteria, and decreasing the abundance of Lactobacillus reuteri in the gut. The results suggested that SBCP could improve the LLD indecued by estrogen deficiency.

Persimmon is a fruit rich in polyphenols (proanthocyanidins or condensed tannins). Using rats and humans, the effects of Kaki-tannin (Nara-type), persimmon polyphenols prepared using a new method, on postprandial plasma glucose levels were investigated in this study. Kaki-tannin (Nara-type) comprised mainly proanthocyanidins, composed of epicatechin : epicatechin gallate : epigallocatechin : epigallocatechin gallate in a ratio of 1 : 1 : 2 : 2 with a molecular weight of approximately 8,000 Da, with epicatechin gallate as a terminal unit. These polyphenols inhibited amylolytic enzymes, such as α-amylase, maltase, sucrase, and α-glucosidase in vitro, and sodium-dependent glucose transporter 1 in Caco-2 cells. These results suggested that the polyphenols suppressed digestion and absorption in the intestinal tract. The ingestion of 250 mg/kg body weight of the polyphenols significantly suppressed increased blood glucose levels after carbohydrate (2 g/kg body weight of glucose or maltose) loading in rats. In a human trial, 1.88 g of Kaki-tannin (Nara-type) significantly delayed increased plasma glucose levels after carbohydrate (150 kcal of maltooligosaccharides) loading. Thus, Kaki-tannin (Nara-type) holds promise to be developed as a food material that potentially improve blood glucose elevation after meals.

Eurycoma longifolia (Tongkat Ali; TA) is a traditional medicinal herb, commonly known as Malaysian ginseng. The root tea has been traditionally applied to treat fevers, aches, sexual dysfunction and other ailments. We evaluated the effects of TA extract supplementation on diurnal core body temperature (BT) and sleep architecture in model mice. Dietary supplementation with TA extract for 4 wk resulted in significantly and moderately reduced BT during the rest and active phases, respectively. A high dose delayed the onset of BT elevation at the start of the active phase, indicating that the effect was dose-dependent. Electroencephalography findings revealed that dietary supplementation with TA extract changed sleep rhythms and delta power during the inactive phase of NREM sleep, indicating improved sleep quality. Our findings suggested that dietary TA extract could be a promising natural aid that alleviates sleep problems via thermoregulation.

Allitol is a rare sugar alcohol obtained by reducing d-allulose (d-psicose). However, information on the effects of long-term dietary allitol intake is limited. This study aimed to investigate the effect of allitol supplementation, as a sugar substitute, on body fat accumulation in rats compared with sucrose, rare sugar d-allulose, or erythritol. Thirty-two male Wistar rats (3 wk old) were fed experimental diets including 5% sucrose, allitol, erythritol, or d-allulose for 8 wk ad libitum. Weight gain, food intake, and food efficiency did not differ among the groups. The total body fat mass and percentage, and intra-abdominal adipose tissue weights were significantly lower in rats fed with the allitol diet than in those fed with the sucrose diet. These body fat indicators tended to be lower in rats fed with the erythritol and d-allulose diets than in those fed with the sucrose diet, but there was no significant difference. The serum glucose-lowering effect obtained in rats fed with the d-allulose diet did not appear in rats fed with the allitol diet. These results suggest that the anti-obesity effect of allitol may be equal to or greater than that of d-allulose.

Plant-derived polar lipids have been reported to exhibit various beneficial effects on human health. The green alga Chlorella is known to be abundant in nutrients, including lipophilic components, and has varying nutrient content depending on the strain. In this study, to assess the nutritional functions of the strain Chlorella pyrenoidosa, we comprehensively analyzed the composition of fatty acids, polar glycerolipids, and sphingolipids. We found that n-3 polyunsaturated fatty acids (PUFAs) comprised 45.6 mol% of fatty acids in the total lipids and 62.2 mol% of n-3 PUFAs in the total lipids occurred in the glycolipids. Monogalactosyldiacylglycerol was the primary glycolipid class, and n-3 PUFA constituted 73.5 mol% of the fatty acids. Although glucosylceramide was observed in trace amounts, highly polar sphingolipids (HPSs), including glycosyl inositol phosphoryl ceramide, were found in much higher amounts compared to rice bran, which is a common source of sphingolipids. These results suggest that the examined Chlorella strain, which is abundant in glycolipids bearing n-3 PUFAs and HPS, is potentially useful as a dietary supplement for improving human health.