The objective of this study was to examine maturation, fertilization and developmental rate of the in vitro-grown mouse oocytes, and to compare these results with those of oocytes grown and matured in vivo. The preantral follicles isolated from 12-day-old mice were cultured on Transwell-COL membrane inserts. After in vitro growth and maturation, 72.5 % of oocytes grown in vitro produced polar body which can be comparable to in vivo growth (70.5 %). However, the mean oocyte diameter of the in vitro group (69.6$\pm$2.1$\mu$m) was smaller than that of the in vivo group (73.3$\pm$3.0$\mu$m). The fertilization rate was significantly lower (p<0.05) in the in vitro group (76.5%) than in the in vivo group (90.2%), however, there was no difference in the percentage of monospermic and polyspermic oocytes between two groups. The capacities of in vitro grown ova to cleave and develop to blastocyst were (57.8 and 14.4%, respectively) significantly lower (p<0.001) than those of the in vivo counterpart (84.4 and 56.6%, respectively). Moreover, the mean number of cells per blastocyst was significantly lower (p<0.05) in the in vitro group (39.0$\pm$10.8) than in the in vivo group (60.5$\pm$12.5). Live young were produced from transferred 2-cell embryos derived from in vitro-grown and matured oocytes. In conclusion, the results show that in vitro-grown oocytes did not achieve the developmental capacity of in vitro-grown oocytes.
Postharvest decay of onion bulbs was examined by inspecting the commercial packages in the market or in storage. Bulb rot incidence was unexpectedly high, and onion bulbs with 1st quality grade were rotten most severely by 51%, followed by 32% for 2nd and 21% for 3rd grades. This indicates that larger bulbs had higher incidences of bulb rots. Major pathogens associated with basal and neck rots were Fusarium oxysporum and Aspergillus sp. or Botrytis allii, respectively, of which basal rot was most prevalent and damaging during storage. Among the epiphytic microorgani는 from onion plants, several Bacillus and Paenibacillus spp. and previously selected Pseudomonas putida and Trichoderma harzianum had inhibitory efficacy against bulb rot pathogens. Among these B. amyloliquefaciens BL-3, Paenibacillus polymyxa BL-4, and P. putida Cha 94 were highly inhibitory to conidial germination of F. oxysporum and B. allii. P. putida Cha 94, B. amyloliquefaciens BL-3, P. polymyxa BL-4, and T. harzianum TM were applied in the rhizoplane of onion at transplanting. Initially antagonist populations decreased rapidly during the first one month. However, among these antagonists, rhizoplane population densities of BL-3, Cha 94, and TM were consistently high thereafter, maintaining about 10$^4$-10$^{5}$ cells or spores per gram of onion root up to harvest time. The other bacterial antagonist BL-4 survived only for two months. TM was the most effective biocontrol agent against basal rot, with the number of rotten bulbs recorded at 4%, while that of the control was 16%. Cha 94 was effective for the first 20 days, but basal rot increased thereafter and had about the same control efficacy as that of BL-3 and BL-4. When the antagonists were applied to the topping areas of onion bulbs at harvest, TM was the most effective in protecting the stored onion bulbs from neck rotting. The second effective antagonist was BL-3. TM and BL-3 completely suppressed the neck rot in another test, suggesting that biocontrol of postharvest decay of onion using these microorganisms either at the time of transplanting or at harvesting may be promising.
The aim of this study is to investigate the seasonal changes of phytoplankton communities based on the environmental changes in a dense oyster farming area (Hansan-Geoje Island) from June to December 2016. The water temperature varied from $14^{\circ}C$ to $28.8^{\circ}C$ and its salinity ranged from 29.4 to 34.2 psu. Nitrate+nitrite was kept at c.a. $3.0{\mu}M$ on the surface layer from June to July, below the concentration limit in August and early September, and then gradually increased from late September. Ammonia was high on July 20 and August 10, and its seasonal characteristics were not clear. Phosphate ranged from 0.01 to $0.7{\mu}M$ on the surface layer, and its seasonal changes were similar to those of nitrate+nitrite. Mean silicate concentrations were $10.7{\mu}M$ on the surface and $15.7{\mu}M$ in the bottom layer, and it was not acted as a limiting factor for the growth of phytoplankton. Among the phytoplankton community, Bacillariophyceae, Dinophyceae and Cryptophyceae was 61.2%, 22.5%, and 13.6%, respectively. In late June, dinoflagellate Prorocentrum donghaiense was dominant in the outer waters(St. T1), later on, Cryptomonas spp. and Chaetoceros spp. were dominant, respectively. From late September to October, diatoms Pseudo-nitzschia spp. and Chaetoceros spp. were stimulated under non-stratified condition after the typhoon. In December, A. sanguinea was found to be $1.7{\times}10^5cells\;L^{-1}$. Seasonally, relative high phytoplankton biomass may be favorable to maintain high production of filter feeder oyster in the dense oyster farming areas of Hansan and Geoje Island.
Gilson Khang;Park, Myoung-Kyu;Jong M. Rhee;Lee, Sang-Jin;Lee, Hai-Bang;Yasuhiko Iwasaki;Nobuo Nakabayashi;Kazuhiko Ishihara
Macromolecular Research
/
v.9
no.2
/
pp.107-115
/
2001
Poly(L-lactide-co-glycolide)(PLGA) was blended with poly[$\omega$-methacryloyloxyethyl phospho-rylcholine-co-ethylhexylmethacrylate (PMEH)] (PLGA/PMEH) to endow with new functionality i.e., to improve the cell-, tissue- and blood-compatibility. The characteristics of surface properties were investigated by measurement of contact angle goniometer, Fourier-transform infrared spectroscopy with attenuated total reflectance (FTIR-ATR) and electron spectroscopy for chemical analysis (ESCA). NIH/3T3 fibroblast and bovine aortic endothelial cell were cultured on control and PLGA/PMEH surfaces for the evaluation of ceil attachment and proliferation in terms of surface functionality such as the concentration of phosphoryl-choline. Also, the behavior of platelet adhesion on PLGA/PMEH was observed in terms of the surface functionality. The contact angles on control and PLGA/PMEH surfaces decreased with increasing PMEH content from 75$^{\circ}$ to about 43$^{\circ}$. It was observed from the FTIR-ATR spectra that phosphorylcholine groups are gradually increased with increasing blended amount of MPC. The experimental P percent values from ESCA analysis were more 3.28∼7.4 times than that of the theoretical P percent for each blend films. These results clearly indicated that the MPC units were concentrated on the surface of PLGA/PMEH blend. The control and PLGA/PMEH films with 0.5 to 10.0 wt% concentration of PMEH were used to evaluate cell adhesion and growth in terms of phosphorylcholine functionality and wettability. Cell adhesion and growth on PLGA/PMEH surfaces were less active than those of control and both cell number decreased with increasing PMEH contents without the effect of surface wettability. It can be explained that the fibronectin adsorption decreased with an increase in the surface density of phosphorylcholine functional group. One can conclude the amount of the protein adsorption and the adhesion number of cells can be controlled and nonspecifically reduced by the introduction with phosphorylcholine group. Morphology of the adhered platelets on the PLGA/PMEH surface showed lower activating than control and the number of adhered platelets on the PLGA/PMEH sample decreased with increasing the phosphorylcholine contents. The amount of fibrinogen adsorbed on the PLGA/PMEH surface demonstrated that the phospholipid polar group played an important role in reducing protein adsorption on the surface. In conclusion, this surface modification technique might be effectively used PLGA film and scaffolds for controlling the adhesion and growth of cell and tissue, furthermore, blood compatibility of the PLGA was improved by blending of the MPC polymer for the application of tissue engineering fields.
In the present study, the total content of polyphenols and flavonoids, the antioxidant activities, and cytotoxic effects of the ethanol extracts from different parts of Taraxacum coreanum Nakai were investigated for their use as functional foods. The extract yields of the flower, leaf, and root were $32.15{\pm}3.21%$, $31.63{\pm}0.63%$, and $27.48{\pm}2.47%$, respectively. Total polyphenol and flavonoid content of the flower extract were $61.29{\pm}2.11mg/g$ and $46.11{\pm}1.88mg/g$, respectively, which were much higher than those of any other plant parts. The antioxidant activities of the flower, leaf, and root extracts were $89.99{\pm}2.83%$, $85.29{\pm}2.22%$, and $37.88{\pm}2.34%$, respectively, at a concentration of 1.0 mg/mL. Cell cytotoxicity effects of AGS (human gastric carcinoma), HCT116 (human colon carcinoma), and A549 (human pulmonary carcinoma) cells were the highest in the flower extract, with values of $62.85{\pm}4.63%$, $69.89{\pm}3.44%$, and $85.72{\pm}4.17%$, respectively, at a concentration of 400 mg/kg. Both the antioxidant activities and cytotoxic effects of the ethanol extracts from all the parts of the T. coreanum Nakai increased dose-dependently. These results provide preliminary data for the development of T. coreanum Nakai as an edible functional food material.
The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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v.24
no.1
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pp.121-141
/
2011
Objective : This Experimental study was done to investigate the Effect of Taklisodok-um and Hwangryunhaedok-tang(TH) on Atopic Dermatitis. Methods : We assessed effects of TH on the IgE, IL-4, IL-5, IL-6, IgM, IgG1, IFN-${\gamma}$ in vivo, on the IL-4, IL-5, CCR3 in the skin tissues of ear and dorsum with NC/Nga mice. And we assessed effects of TH on the COX-2, IL-$1{\beta}$, TNF-${\alpha}$, IL-6 with RAW 264.7 cell. Results and Conclusion : 1. IgE, IL-4, IL-5, IL-6, IgM, IgGl levels in the serum of TH treated NC/Nga mouse group were decreased compared to the untreated control mice. IFN-r showed a increase in the experimental group compared to the untreated control group. The spleen weight of TH treated NC/Nga mice was decreased compared to the untreated control group. 2. mRNA expression levels of IL-4, IL-5 and CCR3 in the skin tissues of TH treated NC/Nga mice were decreased, and expression levels of IL-6 in the skin tissues of TH treated NC/Nga mice were decreased compared to the untreated control group. IFN-${\gamma}$ mRNA expression levels were increased compared to the untreated control group. 3. Judged from that IL-$1{\beta}$, TNF-${\alpha}$, IL-6 express of gene, effect of inflammatory Cytokines revelation were decreased compared to the untreated control group. 4. Depend on the strength of TH, inflammatory RAW 264.7 in the serum of TH were inhibited compared to the untreated control serum that leaded a COX-2 activity model. 5. Histological observation of the ear and skin tissues showed that the extents of inflammation and infiltrated immune cells in the epidermis and dermis of TH treated NC/Nga mice were highly reduced compared to the untreated control group.
Journal of the Society of Cosmetic Scientists of Korea
/
v.40
no.3
/
pp.279-287
/
2014
Estrogen deficiency results in a reduction of skin quality and function in postmenopausal women. Over the past decade, many studies have supported that estrogen provides anti-aging effects as a result of the ability of estrogen to prevent skin collagen decline, restore skin elasticity, and increase skin hydration in postmenopausal women skin. Due to their structural similarity with estrogen, isoflavones have been called phytoestrogens. Photoprotective effects of isoflavones are well established while their estrogenic-like activities are not fully understood in human skin. In this study, we investigated whether daidzein, an effective isoflavone, has phytoestrogenic activity and induces transcriptional change of extracellular matrix components in dermal fibroblasts. We examined the luciferase activity of daidzein and ${\beta}$-estradiol using transiently transfected NIH3T3-ERE cells. The estrogenic receptor-dependent transcriptional activity was increased in a dose-dependent manner when treated with daidzein, with a maximum of 2.5-fold induction at $10{\mu}g/mL$ of daidzein compared with non-treated control. In addition, daidzein significantly in creased the expressions of collagen type I, collagen type IV, elastin, and fibrillin-1 in human dermal fibroblasts. By comparing with the effects of ${\beta}$-estradiol through out all the experiments, we confirmed that daidzein had estrogenic activity and function in fibroblasts. These results suggest that daidzein-based application, having both photoprotective and phytoestrogenic effects, may be a powerful approach for skin anti-aging of postmenopausal women.
In previous reports we demonstrated that ginsenosides, active ingredients of Panax ginseng, affect some subsets of voltage-dependent $Ca^{2+}$ channels in neuronal cells expressed in Xenopus laevis oocytes. However, the major component(s) of ginseng that affect cloned $Ca^{2+}$ channel subtypes such as ${\alpha}_{1C}$(L)-, ${\alpha}_{1B}$(N)-, ${\alpha}_{1A}$(P/Q)-, ${\alpha}_{1E}$(R)- and ${\alpha}_{1G}$(T) have not been identified. Here, we used the two-microelectrode voltage clamp technique to characterize the effects of ginsenosides and ginsenoside metabolites on $Ba^{2+}$ currents ($I_{Ba}$) in Xenopus oocytes expressing five different $Ca^{2+}$ channel subtypes. Exposure to ginseng total saponins (GTS) induced voltage-dependent, dose-dependent and reversible inhibition of the five channel subtypes, with particularly strong inhibition of the ${\alpha}_{1G}$-type. Of the various ginsenosides, $Rb_1$, Rc, Re, Rf, $Rg_1$, $Rg_3$, and $Rh_2$, ginsenoside $Rg_3$ also inhibited all five channel subtypes and ginsenoside $Rh_2$ had most effect on the ${\alpha}_{1C}$- and ${\alpha}_{1E}$-type $Ca^{2+}$ channels. Compound K (CK), a protopanaxadiol ginsenoside metabolite, strongly inhibited only the ${\alpha}_{1G}$-type of $Ca^{2+}$ channel, whereas M4, a protopanaxatriol ginsenoside metabolite, had almost no effect on any of the channels. $Rg_3$, $Rh_2$, and CK shifted the steady-state activation curves but not the inactivation curves in the depolarizing direction in the ${\alpha}_{1B}$- and ${\alpha}_{1A}$-types. These results reveal that $Rg_3$, $Rh_2$ and CK are the major inhibitors of $Ca^{2+}$ channels in Panax ginseng, and that they show some $Ca^{2+}$ channel selectivity.
The present study was conducted to investigate the effects of various co-culture systems and supplemented protein sources on the in vitro development of bovine IVF embryos. Bovine cumulus oocyte complexes (COCs) were matured and fertilized in vitro. Presumptive zygotes with cumulus cells were transferred to TCM-199 or CRlaa containing 10% FBS or 3mg/$m\ell$ BSA, and cultured for 36~40 hr. After primary culture, cleaved embryos were co-cultured with cumulus cells(CC), bovine oviduct epithelial cells(BOEC) or Buffalo rat liver cells (BRLC) in TCM-199 or CRlaa supplemented with FBS or BSA respectively, for further 6 days. Cleavage rate increased with BSA(P<0.01) in the both TCM-199(79%) or CRlaa(74%) When embryos were co-cultured with CC or BOEC in TCM-199, blastocyst development was enhanced with BSA(40% and 43%) compared to FBS (22% and 29%) , whereas in CRlaa no difference observed between BSA(40% and 39%) and FBS (40% and 42%). When embryos were co-cultured with BRLC monolayer, FBS enhanced the blastocyst development (P<0.05) compared to BSA in both TCM-199(41% vs 31%) and CRlaa (44% vs 37%). The result of the present study showed that the cleavage rate of bovine IVF embryos increased with BSA, The result also showed that BSA can enhance the development of IVF embryos in co-culture with CC or BOEC in TCM-199, suggesting the in vitro development is affected by the medium and supplemented protein sources in co-culture with somatic cells.
Campeau, Cindy;Proctor, John T.A.;Murr, Dennis P.;Schooley, Jan
Journal of Ginseng Research
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v.27
no.4
/
pp.188-194
/
2003
Rust-spot on North American ginseng roots (Panax quinquefolius L.) is considered a physiological, not a pathological disorder. Ginseng rust-spot starts as an orange spot on the surface of the root and may spread forming a sunken, round to irregular lesion. 5 mm in diameter. Pieces of root, 7 mm in length and containing a rust-spotted lesion, were embedded in agar and sectioned using a vibratome. These sections and hand sections, cut with a two-sided razor blade, were examined using fluorescence microscopy. The 4-5 cell layers of the periderm were destroyed in the area of the lesion and orange substance:, were deposited in and around the lesion. Sections stained with vanillin-HCI and viewed using bright field microscopy confirmed that the orange substances were phenolic compounds. Scanning electron micros-copy showed that the periderm had pulled away from the root, or was completely destroyed, in the area of the lesion. The smooth surface of the lesion indicates the deposition of phenolic compounds in surrounding cells as a wound response. Roots sprayed or dipped in ethephon (1500 mgㆍL$^{-1}$ ) developed rust-spots, more so at 21$\pm$2$^{\circ}C$ than at 3$\pm$0.2$^{\circ}C$. Roots held at 21$\pm$2$^{\circ}C$ were yellowish and developed white cell proliferations. Comparable control roots also developed rust-spots likely due to the high undecomposed organic matter content of the incubation soilless mix.
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