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Avoidance of sunlight and self-restraint due to the COVID-19 pandemic may contribute to reduced vitamin D status. This study provides comparable data on vitamin D status in Japanese young women and assesses the effect of lifestyle, including changes caused by the COVID-19 pandemic, on vitamin D status. In study 1, 39 young healthy Japanese women aged 21-25 y were recruited from May 2016-June 2017. Serum 25-hydroxyvitamin D (25OHD) concentration and diet and lifestyle information were obtained from participants each month (n=124). In study 2, using the same parameters as study 1, young women aged 21-23 y (n=10) were recruited in September 2020. In the results of study 1, we found the frequencies of vitamin D deficiency (25OHD20 ng/mL) in spring, summer, fall, and winter were 90.5%, 62.5%, 81.5%, and 91.3%, respectively. The substantial difference of serum 25OHD concentration was obtained in spring (Δ3.6 ng/mL) and summer (Δ5.1 ng/mL) depending on the frequency of sunscreen use (0-2 d/wk, 3-7 d/wk). In study 2, serum 25OHD concentration in September 2020 was extremely lower than in September 2016 (13.2 ng/mL vs. 21.7 ng/mL). The number of days spent outside in 2020 decreased drastically compared with 2019. In conclusion, vitamin D deficiency was highly common in Japanese women in their early 20s, and frequent sunscreen use contributed to low vitamin D status. Moreover, because the decrease in days outside due to the COVID-19 pandemic obviously resulted in a decline in vitamin D status, both appropriate sunbathing and increased dietary vitamin D intake are recommended to young women.

The effect of dietary phytate-free soybean protein (PFS) on intestinal mineral absorption and retention was examined in growing male rats using a three-day mineral balance technique. The rats were fed diets containing PFS, soybean protein isolate (SPI) or casein at a 20% level for 5 wk. Total calcium (Ca), magnesium (Mg), phosphorus (P), iron (Fe) and zinc (Zn) contents in diets were adjusted to 0.35, 0.05, 0.7, 0.0035 and 0.003%, respectively, by supplementation of the diet with their salts. Mineral absorption and retention ratios in rats fed the PFS diet were significantly higher than those in rats fed either the SPI or casein diet. These results suggest that PFS may be a promising dietary protein source for improving the mineral bioavailability in humans.

Powdered bovine marrow-free bone was completely solubilized with lactic and citric acids under reduced pressure. The resulting solution was lyophilized to obtain a stable powder form (total bone extract, TBE), and the calcium (Ca) absorbability of TBE from intestine was investigated in normal rats. Each animal perorally received 10 mg of Ca in 1 mL of distilled water as extrinsic 45Ca-labeled TBE, intrinsic 45Ca-labeled Ca lactate, or intrinsic 45Ca-labeled Ca carbonate. The amount of radioactivity in plasma was measured periodically up to 34 h after dosing, and pharmacokinetic parameters were calculated from the radioactivity in plasma. The time taken to reach the maximal 45Ca level (Tmax) did not differ among the three groups. The area under the plasma 45Ca level/time curve (AUC infinity) and the radioactivity at Tmax (Cmax) values for the TBE group were significantly higher than those of the Ca carbonate group. Similar results were observed between the Ca lactate and the Ca carbonate groups. No significant difference was observed in the AUC infinity and the Cmax values between the TBE and the Ca lactate groups. Radioactivity in a femur 34 h after dosing was highest in the Ca lactate group and lowest in the Ca carbonate group among the three groups. Both the TBE and the Ca lactate groups showed significant higher whole-body 45Ca retention than the Ca carbonate group did, although no significant difference was found between the TBE and the Ca lactate groups. These findings indicate that the Ca absorbability of TBE is almost comparable with that of Ca lactate and higher than that of Ca carbonate. Therefore TBE would be useful as a Ca supplement with relatively high absorbability from intestine.

Several reports indicate an important role for vitamin K in bone health as well as blood coagulation. However, the current Adequate Intakes (AI) might not be sufficient for the maintenance of bone health. To obtain a closer estimate of dietary intake of phylloquinone (PK) and menaquinones (MKs), PK, MK-4 and MK-7 contents in food samples (58 food items) were determined by an improved high-performance liquid chromatography method. Next, we assessed dietary vitamin K intake in young women living in eastern Japan using vitamin K contents measured here and the Standard Tables of Food Composition in Japan. PK was widely distributed in green vegetables and algae, and high amounts were found in spinach and broccoli (raw, 498 and 307 microg/100 g wet weight, respectively). Although MK-4 was widely distributed in animal products, overall MK-4 content was lower than PK. MK-7 was observed characteristically in fermented soybean products such as natto (939 microg/100 g). The mean total vitamin K intake of all subjects (using data from this study and Japanese food composition tables) was about 230 microg/d and 94% of participants met the AI of vitamin K for women aged 18-29 y in Japan, 60 microg/d. The contributions of PK, MK-4 and MK-7 to total vitamin K intake were 67.7, 7.3 and 24.9%, respectively. PK from vegetables and algae and MK-7 from pulses (including fermented soybean foods) were the major contributors to the total vitamin K intake of young women living in eastern Japan.

Vitamin K is a cofactor for gamma-glutamyl carboxylase (GGCX), which is an essential enzyme for the gamma-carboxylation of vitamin K-dependent proteins such as osteocalcin (OC). Associations among dietary vitamin K intake, vitamin K status, and bone metabolism have not been thoroughly investigated. Recently, it has been reported that single nucleotide polymorphisms of GGCX (R325Q, 974GA) were associated with age-related bone loss and the kinetic affinity for the substrate. In the present study, we investigated the associations among dietary vitamin K intake, the level of serum vitamin K, and the ratio of undercarboxylated OC (ucOC) to intact OC. The subjects were 60 healthy young male volunteers (mean age, 22.6 y; standard deviation, 1.6). Dietary nutrient intake was assessed by consecutive individual 3-d food records taken before the day of blood examinations. Serum concentrations of vitamin K (phylloquinone: PK, menaquinone 4: MK-4, and menaquinone 7: MK-7), ucOC, and intact OC were measured. All subjects were genotyped for polymorphism (R325Q) presence. Dietary vitamin K intake from vegetables was significantly correlated with the level of serum PK, and vitamin K intake from fermented beans, natto, was also significantly correlated with the level of serum MK-7. The ratio of ucOC to intact OC showed a negative association with the total vitamin K intake (r=-0.331, p=0.010) and serum MK-7 (r=-0.394, p=0.002). Interestingly, grouped by the GGCX genotype, a significant interaction between the ratio of ucOC to intact OC with serum MK-7 was observed in 325R homozygotes (r=-0.572, p=0.003), but not in heterozygotes, nor in 325Q homozygotes. This is the first report to suggest the effects of the single nucleotide polymorphism R325Q in the GGCX gene on the correlation between the level of serum MK-7 and gamma-carboxylation of serum OC.

We recently identified 1alpha,25-dihydroxy-3-epi-vitamin D3 [1alpha,25(OH)2-3-epi-D3] as a metabolite of 1alpha,25-dihydroxyvitamin D3 [1alpha,25(OH)2D3] produced in rat osteosarcoma cells (UMR 106). We now report the isolation of 24R,25-dihydroxy-3-epi-vitamin D3 [24R,25(OH)2-3-epi-D3] as a metabolite of 24R,25-dihydroxyvitamin D3 [24R,25(OH)2D3] by high-performance liquid chromatography (HPLC) with chiral column and its structure assignment by proton nuclear magnetic resonance (1H-NMR) and liquid chromatography-mass spectrometry (LC-MS) analysis. We also demonstrated the production of 24R,25(OH)2-3-epi-D, in two other cell lines [human colon carcinoma cells (Caco-2) and porcine kidney cells (LLC-PK1)] which were previously shown to convert 1alpha,25(OH)2D3 into 1alpha,25(OH)2-3-epi-D3. It can be seen that the production of 24R,25(OH)2- 3-epi-D3 from 24R,25(OH)2D3 is lower than that of 1alpha,25(OH)2-3-epi-D3 from 1alpha,25(OH)2D3 in all the cells studied. 24R,25(OH)2-3-epi-D3 was found to be inactive in terms of its ability to bind to the vitamin D receptor (VDR), in inhibiting proliferation and in inducing differentiation of human promyelocytic leukemia cells (HL-60). Thus, our study indicates that the C-3 epimerization pathway is common to both 1alpha,25(OH)2D3 and 24R,25(OH)2D3 and may play an important role in modulating the concentration and the biological activity of these two major vitamin D3 metabolites in target tissues.