Red Ginseng Extract and γ-Aminobutyric Acid Synergistically Enhance Immunity Against Cancer Cells and Antitumor Metastasis Activity in Mice.

The effects of combined administration of red ginseng (RG) extracts and gamma-aminobutyric acid (GABA) on immunostimulatory activity and tumor metastasis inhibition were investigated in mice. For the immunostimulatory activity, splenocyte proliferation, natural killer (NK) cell activity, including the production of granzyme B (GrB) and interferon gamma (IFN-γ), and serum level of cytokine such as IFN-γ, interleukin (IL)-17, and IL-21 were assessed. Peyer's patch cells obtained from mice administered with RG+GABA were cultured, and the cytokine level in the culture supernatant and bone marrow (BM) cell proliferation activity were examined. The proliferative activity of splenocytes was significantly higher in the RG-GABA treatment group than in RG or GABA alone (P < .05). In the experimental tumor metastasis model, oral administration of RG+GABA showed a higher antitumor metastatic effect compared to that of RG or GABA alone. Oral administration of RG+GABA significantly augmented NK cell-mediated cytotoxicity against YAC-1 tumor cells. In addition, the production of GrB and IFN-γ was stimulated in the culture supernatant of NK cells and YAC-1 cells. Serum concentrations of IFN-γ, IL-17, and IL-21 in mice with RG+GABA were significantly higher compared to the corresponding blood levels in mice administered with RG or GABA alone. The RG+GABA group showed significant BM cell proliferation and increased production of IL-6 and granulocyte-macrophage colony-stimulating factor compared to that in the monotherapy groups. Therefore, RG may have a synergistic effect with GABA for enhancing the host defense system such as BM proliferation and NK cell activity in a tumor metastasis model.

Children's Dental Sealant Use and Caries Prevalence Affected by National Health Insurance Policy Change: Evidence from the Korean National Health and Nutrition Examination Survey (2007-2015).

We evaluated the effect of the National Health Insurance (NHI) policy including dental sealant on changes in the prevalence of sealant and caries, and examined how NHI affected sealant utilization and untreated caries in children from diverse income groups in South Korea. We used a multivariate logistic regression analysis to explore the effects of three stages of dental sealant policy (pre-policy: 2007-2009, first post-policy: 2010-2012, and second post-policy: 2013-2015) on the prevalence of dental sealant and untreated caries. Participant data (N = 8161, aged 6-14 years) were derived from the Korea National Health and Nutrition Examination Survey (2007-2015). We also conducted subgroup analysis to determine the effects of the NHI policy on dental sealant and untreated caries by income level. Implementation of dental insurance coverage was associated with higher likelihood of using dental sealant (odds ratio (OR) = 1.39 (95% confidence interval (CI): 1.18-1.63) for the first period and OR = 1.58 (95% CI: 1.33-1.87) for the second period) and lower odds of having untreated caries (OR = 0.79 (95% CI: 0.64-0.98) for the first period and OR = 0.65 (95% CI: 0.51-0.83) for the second period) after controlling for covariates. Results revealed that there was a greater prevalence of dental sealant and a lower prevalence of untreated caries in both middle- and low-income households compared to high-income households. The higher prevalence of dental sealant and lower untreated caries after the policy implementation. Moreover, we demonstrated children from low-or middle-income households were more associated with increasing dental sealant use and a declining prevalence of caries.

Procyanidins from the stem wood of Machilus japonica and their inhibitory effect on LDL oxidation.

The stem wood of Machilus japonica Siebold & Zucc were extracted with 80 % aqueous MeOH, and the concentrated extract was successively partitioned with ethyl acetate (EtOAc), normal butanol, and water. From the EtOAc fraction, five procyanidins, procyanidin A1 (1), procyanidin A2 (2), procyanidin B7 (3), cinnamtannin B1 (4), and aesculitannin B (5), were isolated. Their chemical structures were identified through spectroscopic data analyses including NMR, MS, and IR. This is the first time any of these compounds have been isolated from this plant. The compounds were evaluated for inhibition activity on LDL oxidation. All of these compounds and the positive control, BHT, showed a very high inhibition effect with IC50 values of 0.94, 2.1, 1.8, 1.1, 1.0, and 1.9 µM, respectively.

Flavonoids from the grains of C1/R-S transgenic rice, the transgenic Oryza sativa spp. japonica, and their radical scavenging activities.

The transgenic rice cultivar of Oryza sativa spp. japonica cv. Hwa-Young, C1/R-S transgenic rice (C1/R-S rice), is a flavonoid-rich cultivar of rice. The grains of C1/R-S rice were extracted with aqueous MeOH, and the concentrated extract was partitioned with EtOAc, n-BuOH, and H2O, successively. Repeated silica gel, octadecyl silica gel (ODS), and Sephadex LH-20 column chromatographies for the EtOAc and n-BuOH fractions afforded four new flavonoids (compounds 2, 3, 7, and 8) along with four known flavonoids: (+)-3'-O-methyltaxifolin (1), brassicin (4), isorhamnetin-4'-O-ß-D-glucosyranoside (5), and 3'-O-methyltaxifolin-5-O-ß-D-glucopyranoside (6). The new flavonoids were identified as 3'-O-methyltaxifolin-7-O-ß-D-glucopyranoside (2), 3'-O-methyltaxifolin-4'-O-ß-D-glucopyranoside (3), isorhamnetin-7-O-ß-D-cellobioside (brassicin-4″-O-ß-D-glucopyranoside) (7), and brassicin-4'-O-ß-D-glucosyranoside (8) from the result of spectroscopic data including nuclear magnetic resonance spectrometry (NMR), mass spectrometry (MS), and infrared spectroscopy (IR). Also, quantitative analysis of major flavonoids (compounds 2, 3, and 8) in C1/R-S rice, O. sativa spp. japonica cv. Hwa-Young (HY), and a hybrid of two cultivar (C1/R-S rice/HY) extracts was performed using HPLC experiment. The isolated flavonoids were evaluated for their radical-scavenging effect on DPPH and ABTS radicals.

Longitudinal study of optic cup progression in children.

PURPOSE: To determine the normative rate of cup-to-disc-ratio (C:D) progression in children and the effect of prematurity and low birth weight on this rate. METHOD: In a single pediatric ophthalmology practice, a single examiner evaluated optic cup size by serial ophthalmoscopy over a minimum of 5 years in 92 patients (184 eyes) without intraocular surgery or optic nerve disease. A cross-sectional analysis of C:D was performed per year of age from 0 to 10 years and linear regression was used to compare C:D progression between preterm and term children and between low versus normal birth weight children. RESULTS: Children exhibited progressive optic cupping. In term children, mean C:D increased by 0.0075 per year. Rate of mean C:D progression was double in children born preterm: 0.0160 (P = .049, comparison to term) per age-year. A similar, nonsignificant trend is observed when comparing low birth weight to normal children (P = .131). CONCLUSION: Prematurity and low birth weight are associated with increased rate of cupping in children. Clinicians should recognize that C:D progression is not a specific sign of glaucoma in children.

Lignans from the flowers of Osmanthus fragrans var. aurantiacus and their inhibition effect on NO production.

A new lignan, (7R,7'R,8R,8'R)-8-hydroxypinoresinol 8-O-ß-D-glucopyranoside 4'-methyl ether (7), was isolated from the flowers of Osmanthus fragrans var. aurantiacus along with six known lignans: (+)-phillygenin (1), phillyrin (2), (-)-phillygenin (3), (-)-epipinoresinol-ß-D-glucoside (4), taxiresinol (5), and (-)-olivil (6). The structure of the new compound was elucidated on the basis of 1D- and 2D-NMR spectroscopic analysis and specific rotation data. The compounds isolated from the flowers of O. fragrans var. aurantiacus were evaluated for inhibitory activities on nitric oxide production in lipopolysaccharide-stimulated macrophage RAW 264.7 cells. (+)-Phillygenin (1), phillyrin (2), and (-)-phillygenin (3) exerted the strongest inhibitory activities on NO production with IC(50) values of 25.5, 18.9, and 25.5 µM, respectively. These compounds may prove beneficial in the development of natural agents for prevention and treatment of inflammatory diseases.

Effects of melatonin on in vitro maturation of porcine oocyte and expression of melatonin receptor RNA in cumulus and granulosa cells.

Melatonin is a multifunctional molecule that mediates several circadian and seasonal processes in animal reproduction. Melatonin and its metabolites are antioxidants and free radical scavengers. We investigated the effects of melatonin on porcine oocyte maturation and embryo development. We then investigated the local expression of the melatonin receptor 1 (MT1) gene in cumulus cells, granulosa cells, and the oocytes with the reverse transcription-polymerase chain reaction (RT-PCR) method. We further evaluated the antioxidant effects [reactive oxygen species (ROS) levels in cumulus-oocytes complexes] of melatonin supplementation during in vitro maturation (IVM). Compared with control, melatonin supplementation (10 ng/mL) during IVM resulted in a greater proportion of oocytes extruding the polar body (75.6% versus 84.6%). Significantly greater proportion of parthenogenetically activated oocytes developed to blastocysts when the in vitro medium was supplemented with melatonin; however, cleavage frequency and blastocyst cell number were not affected by the treatment. RT-PCR analysis revealed the expression of MT1 gene in cumulus and granulosa cells but not in oocytes. Melatonin-treated oocytes had significantly lower levels of ROS than did control (untreated) oocytes. We conclude that exogenous melatonin has beneficial effects on nuclear and cytoplasmic maturation during porcine IVM. Some of the observed effects may be mediated by receptor binding and while others may have been receptor independent, e.g., direct free radical scavenging.

A randomized double-blind, placebo-controlled study to establish the effects of spirulina in elderly Koreans.

AIMS: This study was conducted to determine the antioxidant capacity, immunomodulatory and lipid-lowering effects of spirulina in healthy elderly subjects and to document the effectiveness of spirulina as a functional food for the elderly. METHODS: A randomized double-blind, placebo-controlled study was performed. The subjects were 78 individuals aged 60-87 years and were randomly assigned in a blinded fashion to receive either spirulina or placebo. The elderly were instructed to consume the spirulina or placebo at home, 8 g/day, for 16 consecutive weeks. RESULTS: In male subjects, a significant plasma cholesterol-lowering effect was observed after the spirulina intervention (p < 0.05). Spirulina supplementation resulted in a significant rise in plasma interleukin (IL)-2 concentration, and a significant reduction in IL-6 concentration. A significant time-by-treatment intervention for total antioxidant status was observed between spirulina and placebo groups (p < 0.05). In female subjects, significant increases in IL-2 level and superoxide dismutase activity were observed (p < 0.05) after spirulina supplementation. There were significant reductions in total cholesterol in female subjects. CONCLUSIONS: The results demonstrate that spirulina has favorable effects on lipid profiles, immune variables, and antioxidant capacity in healthy, elderly male and female subjects and is suitable as a functional food.

Improved cryopreservation of bovine preimplantation embryos cultured in chemically defined medium.

The aim of this study was to examine the effects of modifications to a standard slow freezing protocol on the viability of in vitro produced bovine embryos. Bovine oocytes were matured, fertilized with frozen-thawed semen, and presumptive zygotes cultured in defined two-step culture media. The standard freezing medium was 1.5M ethylene glycol (EG), 0.1M sucrose, 10% fetal bovine serum (FBS) in Dulbecco's phosphate buffered saline (D-PBS). A preliminary trial showed that in vitro produced embryos cryopreserved in this medium had a survival rate of 74.6% at 24h and 53.5% at 48 h post-thaw. Experiment 1 studied the effects of omitting the sucrose supplement or replacing it with 0.1M xylose. In Experiment 2, the effects of partial (0%, 25% or 50%) or total (100%) replacement of sodium chloride with choline chloride in the cryopreservation medium were examined (the medium with 100% replacement was designated CJ1). The effects of replacing the 10% FBS with 0.4% BSA or 0.4% lipid-rich BSA (Albumax I) in CJ1 was studied in Experiment 3. In Experiment 4, pregnancy/calving rates following the post-thaw transfer of in vitro produced embryos cryopreserved in the standard freezing medium were compared with those of in vitro and in vivo produced embryos cryopreserved in the improved medium (Albumax I in CJ1). Supplementation of the cryopreservation medium with 0.1M sucrose resulted in higher post-thaw survival rates at 24 h (71.3% versus 53.5 and 51.7%; P<0.05), 48 h (51.1% versus 45.3 and 40.2%), and 72 h (34.0% versus 24.4 and 23.0%) than 0.1M xylose or no supplement, respectively, in Experiment 1. Experiment 2 showed that embryos cryopreserved in the standard medium had poorer survival rates at 24 h (72.8% versus 86.5%; P<0.05), 48 h (53.1% versus 66.3%) or 72 h (28.4% versus 44.9%) than those frozen in CJ1. The post-thaw survival rate of embryos frozen in medium supplemented with Albumax I was better than that for the FBS or BSA supplements at 24h (92.0% versus 90.7 and 87.3%), 48 h (87.3% versus 76.9 and 70.9%; P<0.05), and 72 h (70.4% versus 49.1 and 46 4%; P<0.05; Experiment 3). In Experiment 4, in vitro produced embryos cryopreserved in CJ1 medium supplemented with Albumax I resulted in higher pregnancy rates at Day 35 (31.9% versus 22.9%) and Day 60 (24.1% versus 14.3%) of gestation, and calving rates (22.6% versus 10.0%; P<0.05) than similar embryos frozen in the standard medium. However, in vivo produced embryos cryopreserved in Albumax I in CJ1 resulted in higher pregnancy rates at Day 35 (50.7%; P<0.05) and Day 60 (45.1%; P<0.05) of gestation, and calving rate (43.7%; P<0.05). It was concluded that modification of the freezing medium by addition of lipid-rich BSA and replacing sodium chloride with choline chloride improves the post-thaw survival of in vitro produced embryos, and their viability post-transfer.

Improved in vitro bovine embryo development and increased efficiency in producing viable calves using defined media.

In this study, we developed a defined culture medium that supported improved in vitro bovine embryo development and calving rate after embryo transfer (ET). In vitro-matured bovine oocytes from abbatoir-derived ovaries from Korean native, HanWoo cattle were fertilized with frozen-thawed spermatozoa and embryos were cultured in two-step culture media. In Experiment 1, embryos were cultured in media supplemented with 8 mg/mL BSA, or 0.1mg/mL PVA and 8 mg/mL BSA+2.77 mM myo-inositol or 0.1mg/mL PVA+2.77 mM myo-inositol. Although defined culture media containing PVA supported lower developmental competence compared to undefined media (containing BSA; 8% versus 34%, respectively), defined culture media containing 2.77 mM myo-inositol increased rates of blastocyst formation up to 28%. In Experiment 2, the effect of energy substrate (1.5mM glucose or 1.2mM phosphate) in PVA-myo-inositol defined culture medium on in vitro embryo development was investigated. Defined culture media containing PVA, myo-inositol and phosphate supported better embryo development to blastocysts compared to medium supplemented with both glucose and phosphate (43% versus 31%). In Experiment 3, the effect of epidermal growth factor (EGF) in PVA+myo-inositol-phosphate two-step culture medium on in vitro embryo development was investigated. Among 0, 1, 10 and 100 ng/mL EGF concentrations, the maximal effect was observed with 10 ng/mL EGF (52% blastocyst formation). In Experiment 4, total cell number and calving rate were compared between defined PVA-myo-inositol-phosphate-EGF medium and undefined medium containing BSA, glucose and phosphate. No differences in total cell number of blastocysts obtained from the two groups were observed, however, the rate of viable offspring production was increased using the defined culture medium, compared to the undefined culture medium. In Experiment 5, the relative abundance of mRNA transcripts [interferon-tau (If-tau), glucose transporter-1 (glut-1) and insulin like growth factor 2 receptor (Igf2r)] were analyzed in blastocysts derived from undefined or defined culture media. Gene expression of If-tau, glut-1 was significantly increased in defined culture medium compared to undefined medium. In conclusion, chemically defined culture media without BSA or FBS improved developmental competence of in vitro cultured bovine embryos and delivery of viable calves after ET.