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Establishment of Mouse Embryonic Stem Cell-like Cells from In Vitro Fertilized Embryos (체외수정 생쥐 배아에서의 배아 줄기세포 확립)

  • Shin, Yong-Moon;Park, Yong-Bin;Kim, Hee-Sun;Oh, Sun-Kyung;Chun, Dae-Woo;Suh, Chang-Suk;Choe, Young-Min;Kim, Jung-Gu;Lee, Jin-Yong;Kim, Seok-Hyun
    • Clinical and Experimental Reproductive Medicine
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    • v.29 no.1
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    • pp.1-12
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    • 2002
  • Objective: In order to acquire the technique for the establishment of human embryonic stem cells (ESe) derived from the human frozen-thawed embryos produced in IVF-ET program, this study was performed to establish mouse ESC derived from the in vitro fertilized embryos. Materials and Methods: After Fl hybrid (C57BL female $\times$ CBA mael) female mice were superovulated with PMSG and hCG treatment, their oocytes were retrieved and inseminated, and the fertilized embryos were cultured for 96-120 hours until the expected stages of blastocysts were obtained. To isolate the inner cell mass (ICM), either the blastocysts were treated with immunosurgery, or the whole embryos were cultured for 4 days. Isolated ICMs were then cultured onto STO feeder cell layer, and the resultant ICM colonies were subcultured with trypsin-EDTA treatment. During the subculture process, ESC-like cell colonies were observed with phase contrast microscopy. To identify ESC in the subcultured ESC-like cell colonies, alkaline phosphatase activity and Oct-4 (octamer-binding transcription factor-4) expression were examined by immunohistochemistry and RT-PCR, respectively. To examine the spontaneous differentiation, ESC-like cell colonies were cultured without STO feeder cell layer and leukemia inhibitory factor (LIF). Results: Seven ESC-like cell lines were established from ICMs isolated from the in vitro fertilized embryos. According to the developmental stage, the growth of ICMs isolated from the expanded blastocysts was significantly better than that of ICMs isolated from the hatched blastocysts (80.3% vs. 58.7%, p<0.05). ESC-like cell colonies were only obtained from ICMs of expanded blastocysts. However, the ICMs isolated from the embryos treated with immunosurgery were poorly grown and frequently differentiated during the culture process. The established ESC-like cell colonies were positively stained with alkaline phosphatase and expressed Oct-4, and their morphology resembled that observed in the previously reported mouse ESC. In addition, following the extended in vitro culture process, they maintained their expression of cell surface markers characteristic of the pluripotent stem cells such as alkaline phosphatase and Oct-4. When cultured without STO feeder cell layer and LIF, they were spontaneously differentiated into the various types of cells. Conclusion: The findings of this study suggest that the establishment of mouse ESC can be successfully derived from the in vitro fertilized embryos. The established ESC-like cells expressed the cell surface markers characteristic of the pluripotent stem cells and spontaneously differentiated into the various types of cells.

Effects of Cultivated Wild Panax ginseng Extract on the Proliferation, Differentiation and Mineralization of Osteoblastic MC3T3-E1 Cells (산양삼(cultivated wild Panax ginseng) 추출물이 조골세포 활성에 미치는 영향)

  • Seo, Hyun-Ju;Eo, Hyun Ji;Kim, Hyun Jun;Jeon, Kwon Seok;Park, Gwang Hun;Hong, Se Chul;Jeong, Jin Boo
    • Korean Journal of Plant Resources
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    • v.33 no.4
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    • pp.227-236
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    • 2020
  • Panax ginseng C.A. Meyer (P. ginseng) is known to exert a wide range of pharmacological effects both in vitro and in vivo. Although studies on ginsenoside, antioxidant activity, and anticancer effect of the cultivated wild Panax ginseng (CWP) have been conducted, there is little research on the effect of CWP extract on bone metabolism. In this study, we investigated the potential anti-osteoporotic properties of CWP on the growth and differentiation of MC3T3-E1 cells. CWP significantly increased the viability and proliferation of MC3T3-E1 cells. CWP activated intracellular alkaline phosphatase (ALP) activity in MC3T3-E1 cells. In addition, CWP increased the mineralized nodules in MC3T3-E1 cells. Furthermore, CWP increased the expression of genes such as Runx2, ALP, OPN and OCN associated with osteoblast growth and differentiation in a dose-dependent manner.

Effect of Slow-release Fertilizer Levels of Rice in Different Cultural Methods (벼 재배양식에 따른 유효성 비료의 시용량이 생육과 수량에 미치는 영향)

  • 정진일;최원영;최민규;이선용
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.40 no.6
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    • pp.747-756
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    • 1995
  • To find out the optimum application method of slow-releasing fertilizer(SRF) and conventional fertilizer(CF) with different fertilization rate under two culture methods[l0-day old seedling machine transplanting(MT) and direct-sowing on dry paddy(DS)] in the south-western region(clay loam soil) of Korea, used were Chosun slow-releasing fertilizer(silicate latex coated fertilizer: N-P$_2$O$_{5}$-K$_2$O =18-12-13) and conventional fertilizer. Plant height and number of tillers with different two culture methods were higher at MT than DS in early growth. The ratio of dry weight in heading stage was higher at CF than SRF in MT than DS and especially, SRF 80% + CF 20% than SRF 100% or CF 100%. Leaf area index (LAI) in heading stage was higher at CF in MT but higher at SRF in DS than their counterparts. Chlorophyll content was higher at SRF than in CF expect for heading stage(HS), especially in DS. It was highest at HS in CF without its difference during maximum tillering stage(MTS) and panicle formation stage(PFS), while highest at PFS in SRF with tendency of gradual increase and decrease before and after PFS, respectively. Heading was delayed 2~3 days at SRF in two cultrue methods and 4~5 days at SRF in DS in comparison with CF in MT with delay of 2 days at DS compared with MT. Culm length was longer at CF in MT and at SRF in DS than their counterparts. Panicle number per m was more at SRF and in DS. Filled grain ratio was higher at CF and in MT. Yield was obtained 101 and 100% at 100% and 80% level of SRF in DS respectively, and 96% at 80% level of CF in MT, compared with conventional application method (516kg /l0a), and increased 2~4% at DS and 0~3% at MT in SRF. Yield was high in order of 100%(SRF) =80%(SRF) + 20%(CF) > 100%(SRF) + 20%(CF) > 80%(SRF) at MT and 80%(SRF) + 20%(CF) =100%(SRF) > 80%(SRF) =100%(SRF) + 20%(CF) at DS.

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Effects of Different Water Depths on Early Growth of Rice and Barnyard grass(Echinochloa crus-galli) (담수심차이가 벼 품종과 피의 초기생육에 미치는 영향)

  • 박성태;장안철;이수관
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.38 no.5
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    • pp.405-412
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    • 1993
  • This experiment was conducted to investigate the effects of water depths on seedling stand and early growth of califonia rice varieties, S201, M202, A301, Italico livorno and Korean variety, Hwaseongbyeo, and barnyardgrass (Echinochloa crus-galli) The coleoptile length of rice was longer with deep water depth while for the radicle length shorten. As water depth was increased, the percentage of seedling stand were decreased slightly in rice, while sharply increased in barnyardgrass. Plant height of rice with increasing water depth were longer, whereas that of barnyardgrass reduced significantly with weaker. Tiller number of rice and barnyardgrass were significantly reduced as water depth increased. Dry matter weight and healthy score of rice seedling at 35DAS were highest in 7.5cm water depth followed saturated moisture, 15, and 22.5cm water depth, while for barnyardgrass those were especially negatively affected by deep water depth. These results showed that the seedling stand and early growth of barnyardgrass was highly suppressed by deeper water levels compared with rice. Rice cultivars which are showes growth characteristics in deeper water levels at early growth stage were Italico livorno and S201 in Japonica / Indica.

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Varietal Differences in Agronomic Characters of Rice Grown on Salty Water Irrigation (벼의 생육시기별 염수처리에 따른 주요 특성의 품종간 차이)

  • 정진일;김보경;박형만;이선용
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.40 no.4
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    • pp.494-503
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    • 1995
  • The selection of salt tolerant rice variety needs an effective method in its testing. Salinity of irrigated water, 0.5% at seedling stage, 0.6% at tillering stage, and 0.9% at panicle formation stage were treated to test salt tolerance of rice using 45 cultivars. At tillering stage, salty water irrigation reduced plant height to 22.6% in early maturing rices(EMR), 30.5% in medium maturing rices(MMR), and 20.9% in medium-late maturing rices(MLMR), and also reduced number of tillers to 11.2% in EMR, 36.2% in MMR, and 36.0% in MLMR compared to rices grown in non-salty water irrigation. At panicle formation stage of rice, salty water irrigation affected plant height and tiller numbers that showed varietal differences. As salt tolerant rice cultivars, Daegwangbyeo, Namweonbyeo, Sinseonchalbyeo, Gyehwabyeo, and Daeyabyeo were selected. Jinbubyeo, Donghaebyeo, and Tamjinbyeo were weak in salty water irrigation.

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Effect of Media, Synchronization of Fibroblast Cells, Culture Time, $\textrm{O}_2$ Concentration and Activation on Developmental Rate of Nuclear Transferred Porcine Oocytes (배양액, 섬유아세포, 배양시간, 산소 농도 및 활성화 처리가 돼지 핵이식 배의 체외발달에 미치는 영향)

  • Quan J. H.;Rhee M. H.;Kim S. K.
    • Journal of Embryo Transfer
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    • v.19 no.3
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    • pp.191-199
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    • 2004
  • This study was conducted to examine in vitro development of porcine embryos constructed by the microinjection of cultured fetal fibroblast cells into porcine oocytes matured in vitro. Single fetal donor cells were deposited into the perivitelline space of enucleated oocytes, followed by electrical fusion and activation. Activated embryos were cultured in NCSU-23 medium supplemented with 5% FBS, at 38.5$^{\circ}C$ for 6 to 8 days in 5% $CO_2$ and air. In experiment 1, fusion rates of nuclear transfer embryos did not differ for fetal fibroblast cells incubated in 5% FBS + NCSU-23 or 5% FBS + TL Heaps medium, nor did fusion rates of donor cells differ between 1-8 hr incubation durations. Fusion rates for the four treatment subclasses ranged from 72.1% to 78.0%. In experiment 2, Pre-synchronization in medium containing 0.1 $\mu\textrm{g}$/m Hoechst 33342 an increase from 0 and 8 versus 15 h culture an increased percentage of porcine fibroblast cells in G2/M at the end of the synchronization period (12.4%, 17.5% and 47.6%). Neither an increase in the concentration of H 33342 (0.2-1.6 $\mu\textrm{g}$/$m\ell$) nor a longer exposure time (12h, 18h and 24h) increased the proportion of porcine G2/M fibroblasts. In experiment 3, fusion rates did not differ significantly far nuclear transfer embryos constructed using donor cells cultured in 5% FBS + NCSU-23 medium for 1-2, 6-8 or 12-14 days (60.0%, 73.3% and 62.5%), respectively. The cleavage rate for nuclear transplant embryos using fetal fibroblast cells cultured for 1-2 days was 44.0%, significantly less than 56.7% and 50.0%. for 6-8 or 12-14 days duration of culture, respectively. In experiment 4, the proportions of nuclear transfer embryos that developed to the $\geq$2 cell and to the blastocyst stage were not affected significantly by culture medium (5% FBS + NCSU-23 or 5% FBS + TL-Heaps) or by $O_2$ concentration of the culture (5% vs 10%). Rates of development to the $\geq$2 cell stage ranged from 65.9% to 70.1%, and development rates to the blastocyst stage ranged from 9.8% to 12.5% for the four treatment subclasses. Developmental rate was highest for embryos cultured in 5% FBS + NCSU-23 under a gas atmosphere of 5% $O_2$ in air.

Comparison of Passive Diffusional Organic Vapor Samplers with Charcoal Tubes for the Measurement of Mixed Organic Solvents (확산형 포집기와 활성탄관을 이용한 공기중 혼합 유기용제 측정에 관한 연구)

  • Ahn, Kyu-Dong;Yeon, You-Yong;Lee, Byung-Kook
    • Journal of Korean Society of Occupational and Environmental Hygiene
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    • v.4 no.2
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    • pp.127-136
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    • 1994
  • Diffusional sampling devices offer many advantages for measuring concentration levels of industrial contaminants than the conventional pump and charcoal tubes because they are lightweight, require no power, pump or tubing. This study designed to evaluate and compare the sampling performance of passive sampler to charcoal tube from mixed organic solvent workplace with 181 organic solvent using workers working in different concentration of organic solvents. All study workers kept both devices in their breathing zone simultaneuosly in the workplaces, and the sampling analytical results were compared with those of charcoal tube. The results obtained are as follows: 1. The concentrations of toluene and xylene measured by passive sampler were slightly higher than those of charcoal tube, but there were no significant statistical differences between two methods. 2. The concentrations of MEK and cyclo-hexanone measured by passive sampler in low exposure workplace (below 0.20 of MEK TLV levels and 0.1 of cyclo-hexanone TLV levels) were about 2 times higher than that of charcoal tube sampling. While, absorption efficiency of passive sampler was reduced according to increasing concentration measurements of MEK and cyclo-hexanone in air. 3. The ratios of concentrations of toluene, xylene, MEK and cyc1o-hexanone measured by passive sampler over those measured by charcoal tube were 1.11, 1.07, 1.63 and 3.65 respectively. 4. The percentages of concentration of passive samplers within 0.75 and 1.25 of charcoal tube value as a reference value of 1.0 were 57% in toluene, 74% in xylene, 34% in MEK and 32% in cyclo-hexanone respectively. 5. The correlation coefficients of toluene, xylene, MEK and cyclo-hexanone between passive sampler and charcoal tube sampler were 0.963, 0.957, 0.943 and 0.562 with statistical significance.

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A Study on the Water Quality of Reservoir Tank in the Building Complex on Jeonnam Area (대형건축물 저수조의 수질실태 및 개선방안에 관한 연구)

  • Lee, J.H.;Lee, H.H.;Kim, H.B.;Ahn, G.W.;Park, K.N.;Kim, Y.K.;Bae, J.S.;Mun, H.;Park, C.U.;Oh, E.H.;Park, S.I.;Seo, Y.G.
    • Journal of environmental and Sanitary engineering
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    • v.15 no.4
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    • pp.59-77
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    • 2000
  • This study was carried out to investigate on several factors, which contaminative the water quality through the water pipe during feeding water, in 42 largescaled apart-ments(total 84 cases) and assayed the Volatile Organic Compounds(VOCs) and concen-tration of heavy metals that inflow and outflow in reservior water in Jeonnam area(Mokpo, Suncheon, Yeosu) from January 1999 to December 1999. The results obtained were summarized as follows ; 1. The quality of the water pipe composition in the order of frequency in the quality of water pipes were Copper(45.2%)> Zinc(38.9%)> Stainless steel(9.5%)> PVC(4.8%)> PM(2.4%) in observing 42 sites. All of the drain pipes were used the cast iron quality. 2. The result of certification curve from 12 items(17kind) of VOCs was $1.0-4.0{\mu{g}}/{\ell}$ range, a coefficient of correlation was shown 0.99 over. A MDL of each substance range was within $0.1-1.0{\mu{g}}/{\ell}$. 3. The result of the assay, 5 kinds(Viny chloride, Dichloromethane, Ethylbenzene, M,P-xylene, Styrene) of the VOCs of 14 kinds was not detected and the other items were detected slightly. The detection rate of one item and over in total VOCs samples, were 25.9% in inflow and 27.9% in outflow. And frequency of detect in inflow/outflow of THM(Chloroform, Bromodichloro-methane, Dibromochloromethane, Bromoform) were shown higher than 94.1%, 97.0% each stages. It comes to the conclusion that all of the samples were reason able for drinking water standards. 4. The coefficient of correlation were reasonable, it shown 0.999 over in $0.1-1.0{\mu{g}}/{\ell}$ of a measuring range conditions of 4kinds in organic substance(Zn, Cu, Fe, Mn). 5. The results were showed suitability in 78 cases(92.9%) and unsuitability in 6 cases (7.1%), in 84 cases of in organic substances. Compare to inflow stage, mean concentrations of heavy metal, were increased slightly in Zn, Cu, Fe except Mn than outflow stage. The result of the mean concentration in organic substance inflow and outflow in the apartment water tank using Pair-compared T-test, in 95% reliance index, were $0.179mg/{\ell}(0.151-0.307mg/{\ell})$ in Zinc, $0.136mg/{\ell}(0.113-0.230mg/{\ell})$ in Copper, $0.052mg/{\ell}(0.048-0.098mg/{\ell})$ in Fe, and there was a bit growing tendency but there was no differece in Mn. 6. The mean concentration of Copper which used Cu pipe as a water supply pipe in apartment were $0.216mg/{\ell}(0.161-0.338mg/{\ell})$ in case of the Zine pipe were $0.286mg/{\ell}(0.204-0.435mg/{\ell})$. It shows that the detection rate was more higher than the other material used in Cu or Zn as the water supply pipe. We supposed to Cu and Zn substance were gushing out water supply pipe.

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Physico-Chemical Properties of Rainfall Interception Culture and Open Field Culture Soils of Rubus sp. In Gochang-gun, Jeollabuk-do (고창지역 복분자 비가림하우스 토양과 노지토양의 물리·화학적 특성)

  • Chung, Byung-Yeoup;Lee, Kang-Soo;Kim, Myung-Kon;Choi, Young-Hee;Kim, Moo-Key;Cho, Jae-Young
    • Korean Journal of Soil Science and Fertilizer
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    • v.41 no.5
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    • pp.303-309
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    • 2008
  • The physico-chemical properties of the rainfall interception culture (85 sites) and the open field culture soils (85 sites) in the area of Gochang-gun, Jeollabuk-do were surveyed. Soil textural distribution of the rainfall interception culture and the open field culture soils was 74% and 64% for silt loam, 16% and 35% for loam, and 10% and 1% for clay, respectively. The percentage of aggregate rates was higher in the open field culture soils (60.06%) than rainfall interception culture soils (55.84%). Electrical conductivity, exchangeable cations, and anions in the rainfall interception culture soils were higher than those in open field soils. Specially, accumulated amount of anion in rainfall interception culture soils was remarkably higher 2~3 times than open field culture soils. The results from the analyses of rainfall interception culture soils suggested that the most critical problem is the salts accumulation caused by over-fertilization of chemical fertilizer and compost. Therefore, application rates of chemical fertilizer and compost should be controlled in order to conservation of soil and water for sustainable agriculture.

Effects of Dietary Beef Tallow, Soybean Oil and Squid Liver Oil on Growth and Body Composition of the Korean Rockfish Sebastes schlegeli, and Biochemical Changes with Starvation (사료 지질원으로 우지, 대두유 및 오징어 간유 첨가에 따른 조피볼락 Sebastes schlegeli의 성장 및 체성분 변화와 절식시 체내 대사)

  • LEE Sang-Min
    • Journal of Aquaculture
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    • v.7 no.1
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    • pp.63-76
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    • 1994
  • The effects of different dietary lipids on growth, body chemical composition, and nutrient partition of the Korean rockfish (43 g of initial mean weight) were studied. Fish were fed three experimental diets containing 1) $4.5\%$ squid liver oil, 2) $1\%$ squid liver oil and $3.5\%$ beef tallow, and 3) $1\%$ squid liver oil and $3.5\%$ soybean oil. Growth and feed efficiency were detarmined during a 19-week feeding period at water temperature of $15.5^{\circ}C$, and at the end of which body chemical compositions were analyzed. Remained fish were fasted for 45 days, and four fish were taken at 6, 12 and 24 hours, and 3, 15 and 45 days after the fasting to determine tissue compositions and hematological changes. Growth, feed efficiency, protein and lipid retention efficiency, hepatosomatic index(HSI), viscerosomatic index (VSI), and chemical composition of whole body and dorsal muscle were not affected by the different dietary lipids. Liver moisture content of fish fed squid liver oil diet was higher than that of fish fed beef tallow or soybean oil diet. Liver lipid content of fish fed beef tallow diet was higher than that of fish fed the other diets. Fatty acid composition of dorsal muscle and liver were affected by the different dietary lipids ; high levels of 20 : 5n-3 and 22 : 6n-3 from fish fed the diet containing squid liver oil, 18 : 1 from fish fed the diet containing beef tallow, and high 18 : 2n-6 and 18 : 3n-3 from fish fed the diet containing soybean oil were observed. Both HSI and VSI of fish fed three diets decreased with time after the begining of starvation. Liver glycogen did not change during the first 15 days of starvation and decreased thereafter, and that was not affected by the different dietary lipids. Lipid and protein contents in the dorsal muscle of fish decreased up to 15 days of starvation and remained unchanged thereafter, these were not different from each other. Glucose, free fatty acid, triglyceride and phospholipid concentrations in Fish serum were varied for the first 15 days of starvation, after that the concentrations of fish serum remained relatively stable in all the treatment groups with prolonged starvation. The results indicate that Korean rockfish can utilize fish oil, animal fat or vegetable oil equally as energy source when n-3HUF A is adquate.

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