The two main sources of vitamin D3 are de novo synthesis induced by exposure to ultraviolet (UV) light from the sun, and diet. Vitamin D3 deficiency causes rickets or osteoporosis. Oak mushrooms (Lentinula edodes) that are exposed to UV radiation contain enhanced vitamin D2 and have much higher calcium content than unmodified (non-irradiated) mushrooms. Such modified edible mushrooms have been proposed as a natural alternative source of dietary vitamin D. In the current study, we have examined whether modified oak mushrooms could improve or prevent osteoporosis-like symptoms in mice fed with low calcium and vitamin D3-deficient diet. Four-week-old male mice were fed low calcium, vitamin D3-deficient diets supplemented with 5, 10, or 20% unmodified, calcium-enhanced, or calcium plus vitamin D2-enhanced oak mushrooms for 4 weeks. To assess the effects of the supplemented diets, we evaluated femur density and length, bone histology, the expression of active calcium transport genes, and serum calcium levels. Mice fed with low calcium and vitamin D3-deficient diet developed osteoporosis-like symptoms within 4 weeks. Femur density and tibia thickness were significantly higher in mice fed calcium plus vitamin D2-enhanced mushrooms, and the expression of duodenal and renal calcium transport genes was significantly induced. These results indicate that in mice, vitamin D2 and/or calcium derived from irradiated oak mushrooms may improve bone mineralization through a direct effect on the bone, and by inducing the expression of calcium-absorbing genes in the duodenum and kidney.

The sap of Acer mono has been called 'bone-benefit-water' in Korea because of its mineral and sugar content. In particular, the calcium concentration of the sap of A. mono is 37·5 times higher than commercial spring water. In the current study, we examined whether A. mono sap could improve or prevent osteoporosis-like symptoms in a mouse model. Male mice (3 weeks old) were fed a low-calcium diet supplemented with 25, 50 or 100 % A. mono sap, commercial spring water or a high calcium-containing solution as a beverage for 7 weeks. There were no differences in weekly weight gain and food intake among all the groups. Mice that were given a low-calcium diet supplemented with commercial spring water developed osteoporosis-like symptoms. To assess the effect of sap on osteoporosis-like symptoms, we examined serum calcium concentration, and femur density and length, and carried out a histological examination. Serum calcium levels were significantly lower in mice that received a lowcalcium diet supplemented with commercial spring water (the negative control group), and in the 25 % sap group compared to mice fed a normal diet, but were normal in the 50 and 100 % sap and high-calcium solution groups. Femur density and length were significantly reduced in the negative control and 25 % sap groups. These results indicate that a 50 % sap solution can mitigate osteoporosis-like symptoms induced by a lowcalcium diet. We also examined the regulation of expression of calcium-processing genes in the duodenum and kidney. Duodenal TRPV6 and renal calbindin-D9k were up-regulated dose-dependently by sap, and the levels of these factors were higher than those attained in the spring water-treated control. The results demonstrate that the sap of A. mono ameliorates the low bone density induced by a low-calcium diet, most likely by increasing calcium ion absorption.

The uterine expressions of epidermal growth factor(EGF) family are examined to elucidate their exact role(s) in rat pregnancy. EGF and its receptors' (EGF-R) mRNA levels increased significantly at implantation after which their expression gradually decreased. Heparin-binding EGF (HB-EGF) and transforming growth factor-alpha (TGF-alpha) showed a modest expression at gestation day (GD), GD4 and GD3, respectively, but were much strongly expressed at mid-pregnancy.Amphiregulin (Areg) was strongly expressed around implantation (GD4) and at mid-pregnancy (GD12).Treatment of pregnant rats with RU486 at GD5 or GD8 blocked the expression of all the genes, and administration of immature rats with progesterone (P4) induced the uterine expression of all the genes except HB-EGF. In addition, HB-EGF,TGF-alpha, and Areg proteins in the uterine and glandular epitheliums may participate in mid-pregnancy. Taken together, all of these activities are likely to be controlled by P4 in the uterus of pregnant rats.

The implantation of the developing blastocyst into the uterine wall is regulated by a precisely timed interplay of the ovarian hormones estrogen and progesterone, which control a set of regulatory factors that make the uterus receptive to implantation. These factors include EGF receptor (Egfr) and members of the epidermal growth factor (Egf) family, namely, EGF, heparin-binding EGF (Hbegf), transforming growth factor-alpha (Tgfa), and amphiregulin (Areg). However, the exact role(s) these factors play in pregnancy remain unclear. To address this, a group of three rats was euthanized every day from gestation day (GD) 0 through to GD21. The uterus, attached uterus (these tissues are mostly composed of stromal cells), and placenta were rapidly excised and used directly for total RNA. We used real-time PCR with the TaqMan system (Applied Biosystems, Foster City, CA, ISA) to examine the uterine expression patterns of these factors in rats during the entire pregnancy. Data were analyzed by nonparametric one-way analysis of variance using the Kruskal-Wallis test, followed by Dunnett's test for multiple comparisons. Egf and Egfr mRNA levels increased significantly at implantation, especially on GD3 and GD6, after which their expression gradually decreased. Hbegf and Tgfa showed a modest spike of transcription around the implantation period (GD4 and GD3, respectively) but were much more strongly expressed at mid-pregnancy, which is when progesterone is secreted at high levels. Areg expression peaked strongly around implantation (GD4) and at mid-pregnancy (GD12). Treatment of pregnant rats on GD5 or GD8 with the progesterone receptor antagonist RU486 (2.5 mg per rat) blocked the expression of all of the genes on the days of treatment. Moreover, injection of immature rats with progesterone induced the uterine expression of all of the genes except Hbegf, while injection with estrogen or estrogen plus progesterone had no effect. Taken together, all genes tested may be assumed to regulate the implantation process. Moreover, Hbegf, Tgfa, and Areg may participate during mid-pregnancy. In addition, all of these activities are likely to be controlled by progesterone in the uterus of rats during pregnancy.