Modulatory Effects of Single and Complex Vitamins on the In Vitro Growth of Murine Ovarian Follicles.

Background: Vitamin is a well-known co-factor for many metabolic processes and its roles in fertility and follicular growth have been studied. Vitamin supplementation is frequently achieved by daily ingestion in the form of a complex capsule. However, the role of single and complex vitamins in in vitro maturation of murine follicles is not fully elucidated. Methods: In this study, we evaluated the effects of two forms of vitamins. Pure L-ascorbic acid, and multi-vitamin (vitamin C + vitamin B complex) was treated at two different concentrations (50 and 100 µg/ml), to pre-puberty murine follicles during in vitro maturation. To determine the specific stage of growth that is affected by treatment with vitamins, the vitamins were treated from day 0, 4, 9, and 13. Growth of each follicle was assessed by measuring diameters of whole expanded area and of the granulosa cells. Expression of follicular and oocyte growth-related genes and the effect of vitamin on the viability of follicles was assessed using senescence associated ß-galactosidase staining. Results: Treatment with vitamins promoted the in vitro growth of murine follicles and the upregulated the expression of granulosa cell- and oocyte-specific genes such as BMP15, Fsh receptor, and GDF9. The proliferation of the granulosa cells was enhanced by the treatment of vitamin. Fifty µg/ml concentration vitamin showed greater effects compared to higher concentration. The viability of in vitro grown follicles was also significantly improved in vitamin-treated follicles. The effects of single L-ascorbic acid and complex vitamin were not significantly different to those of day 4 and day 9 follicles. Vitamins promoted murine follicle development in vitro with different effects on specific growth stage. Conclusion: Supplementation of vitamins during in vitro maturation of murine follicles is an efficient strategy for in vitro expansion of follicular cells. These results could be customized to the sophisticated culture of follicles retrieved from aged or cancer-survived female that contain smaller number of follicles with reduced potential to develop into mature follicles.

Deer Bone Extract Supplementation for Mild-to-Moderate Knee Osteoarthritis Symptoms: A Randomized, Double-Blind, Placebo-Controlled Trial.

In this randomized, double-blind, placebo-controlled study, we evaluated the efficacy of deer bone extract (DBE) in participants with knee osteoarthritis (OA). We enrolled 50 participants aged 50-70 years, having knee OA with a Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC) score ≥5.0. The participants were assigned to the placebo or DBE group (550 mg/day) for 12 weeks. The outcome measures were as follows: pain score on the visual analog scale (VAS); WOMAC score; and blood and urine biomarkers. In the DBE group, VAS scores, WOMAC total scores, and WOMAC subscores (for pain, stiffness, and physical function) improved significantly compared with the baseline values. However, there was no significant difference in outcomes between the DBE and placebo groups. The present findings suggest that DBE may mildly reduce joint pain and stiffness and improve joint function in patients with painful knee OA.

Ginsenosides may enhance the functionality of human embryonic stem cell-derived cardiomyocytes in vitro.

Various chemicals have been reported to induce the differentiation of human embryonic stem cells (hESCs) into cardiomyocytes (CMs), however, their contributions to the functionality of hESC-derived CMs are still limited. In this study, we evaluated the effects of red ginseng extract (RGE), ginsenoside-Rb1 (gRb1, panaxadiol), and ginsenoside-Re (gRe, panaxatriol) on the differentiation of hESCs and the functionality of derived CMs. Undifferentiated hESCs were treated with 0.25 mg/mL RGE, 10 µmol/L gRb1, or 10 µmol/L gRe for 48 hours at the differentiation induction (early stage) or maturation (late stage) period. The expression of mesodermal and cardiac transcription factor genes was upregulated in the ginsenoside-treated groups from early stage. The expression of cardiac sarcomeric genes was significantly upregulated at the late stage. The gRb1- and gRe-treated groups upregulated the expression of potassium voltage-gated channel subfamily E member 1 (KCNE1) and the gRe-treated group showed a longer beating duration compared to the control. Taken together, ginsenosides may enhance the functionality of hESC-derived CMs in vitro.

Yam tuber mucilage as a candidate substance for saliva substitute: in vitro study of its viscosity and influences on lysozyme and peroxidase activities.

OBJECTIVE: To investigate the viscosity of yam tuber mucilage (YTM) and its effects on lysozyme and peroxidase activities in solution phase and on surface phase. METHODS: Two kinds of YTM were extracted, one containing both protein and carbohydrate and the other containing mainly carbohydrate. Hen egg-white lysozyme and bovine lactoperoxidase were used as lysozyme and peroxidase sources, respectively. Viscosity was measured with a cone-and-plate digital viscometer. Lysozyme activity was determined using the turbidimetric method, and peroxidase activity was determined using the NbsSCN assay. Hydroxyapatite beads were used as a solid phase. RESULTS: The viscosity values of YTM followed a pattern of a non-Newtonian fluid. The carbohydrate concentration affected the viscosity values at all shear rates, while the protein concentration affected the viscosity values at low shear rates. It could be suggested that YTM composed of 1.0 mg/ml protein and 1.0 mg/ml carbohydrate has viscosity values similar to those of unstimulated whole saliva at shear rates present at routine oral functions. Hydroxyapatite-adsorbed YTM significantly increased the adsorption and subsequent enzymatic activities of lysozyme, but not those of peroxidase. CONCLUSIONS: Yam tuber mucilage has viscoelastic properties similar to those of human saliva and enhances the enzymatic activity of lysozyme on hydroxyapatite surfaces.

Acupuncture suppresses kainic acid-induced neuronal death and inflammatory events in mouse hippocampus.

The administration of kainic acid (KA) causes seizures and produces neurodegeneration in hippocampal CA3 pyramidal cells. The present study investigated a possible role of acupuncture in reducing hippocampal cell death and inflammatory events, using a mouse model of kainic acid-induced epilepsy. Male C57BL/6 mice received acupuncture treatments at acupoint HT8 or in the tail area bilaterally once a day for 2 days and again immediately after an intraperitoneal injection of KA (30 mg/kg). HT8 is located on the palmar surface of the forelimbs, between the fourth and fifth metacarpal bones. Twenty-four hours after the KA injection, neuronal cell survival, the activations of microglia and astrocytes, and mRNA expression of two proinflammatory cytokines, interleukin-1ß (IL-1ß) and tumor necrosis factor-α (TNF-α), were measured in the hippocampus. Acupuncture stimulation at HT8, but not in the tail area, significantly reduced the KA-induced seizure, neuron death, microglial and astrocyte activations, and IL-1ß mRNA expression in the hippocampus. The acupuncture stimulation also decreased the mRNA expression of TNF-α, but it was not significant. These results indicate that acupuncture at HT8 can inhibit hippocampal cell death and suppress KA-induced inflammatory events, suggesting a possible role for acupuncture in the treatment of epilepsy.

Red ginseng extract facilitates the early differentiation of human embryonic stem cells into mesendoderm lineage.

Human embryonic stem cells (hESCs) have capacities to self-renew and differentiate into all cell types in vitro. Red ginseng (RG) is known to have a wide range of pharmacological effects in vivo; however, the reports on its effects on hESCs are few. In this paper, we tried to demonstrate the effects of RG on the proliferation and differentiation of hESCs. Undifferentiated hESCs, embryoid bodies (EBs), and hESC-derived cardiac progenitors (CPs) were treated with RG extract at 0.125, 0.25, and 0.5 mg/mL. After treatment of undifferentiated hESCs from day 2 to day 6 of culture, BrdU labeling showed that RG treatment increased the proliferation of hESCs, and the expression of Oct4 and Nanog was increased in RG-treated group. To find out the effects of RG on early differentiation stage cells, EBs were treated with RG extract for 10 days and attached for further differentiation. Immunostaining for three germ layer markers showed that RG treatment increased the expressions of Brachyury and HNF3ß on EBs. Also, RG treatment increased the expression of Brachyury in early-stage and of Nkx2.5 in late-stage hESC-derived CPs. These results demonstrate facilitating effects of RG extract on the proliferation and early differentiation of hESC.

Enhanced ginsenoside productivity by combination of ethephon and methyl jasmoante in ginseng (Panax ginseng C.A. Meyer) adventitious root cultures.

Ethephon at 50 microM enhanced both root growth and ginsenoside accumulation in ginseng (Panax ginseng C.A. Meyer) adventitious root cultures, but at 100 microM it inhibited only ginsenoside accumulation. Ginsenoside productivity with 50 microM ethephon was the highest at 1.7 mg l(-1) d(-1) after 8 days of elicitation. However, elicitation with 50 microM ethephon and 100 microM methyl jasmonate (MJ) improved productivity (6.3 mg l(-1) d(-1)) whereas elicitation with 100 microM MJ alone gave only 2.9 mg l(-1) d(-1).