• Proceedings of the KSAR Conference
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• 2004.06a
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• pp.230-230
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• 2004

Polyamines, namely putrescine, spermidine, and spermine, are biogenic low-molecular-weight aliphatic amines. Polyamines play important roles in DNA stabilization, RNA and protein synthesis, membrane stabilization, modulation of ion channels, and protection against oxygen radicals and are essential for cell homeostasis, cell growth, and tumorigenesis. (omitted)

• Journal of Nutrition and Health
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• v.31 no.4
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• pp.799-808
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• 1998

To investigate the antitumor activity of mugwort (Artemisia princeps Pampan), petroleum ether extract of mugwork was partially purified by a silica gel chromatography. Among several fractions, the fraction which was obtained under the elution with acetone, showed potent cytotoxicity against mouse leukemia cell line(Ll210), human colon cancer cell line (HCT-48) and human hepatoma cell line (Hep G2) , but was less effective with normal cell line(mouse embryo cell). Acetone fraction appeared to be glycolipid by Benedict test and the major fatty acids of the lipid were C16 ; 0 , C 18: 3by GC/MS analysis.

• Proceedings of the Korean Society of Crop Science Conference
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• 1996.05a
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• pp.30-31
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• 1996

• Korean Journal of Environment and Ecology
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• v.26 no.5
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• pp.675-681
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• 2012

In this experiment, I investigated toxicity evaluation of chemicals using domestic frog embryos, along FETAX (Frog Embryo Teratogenesis Assay-Xenopus) protocol. I investigated $Cu^{2+}$ and Tebuconazole effect on the tree frog, Hyla japonica, embryos by probit analysis. Mortality and malformation rates increased and larval body length decreased depending on the concentrations of $Cu^{2+}$ and Tebuconazole. The teratogenic concentration ($EC_{50}$) of $Cu^{2+}$ and Tebuconazole were 0.05, 5.0mg/${\ell}$, respectively and the embryo lethal concentration ($LC_{50}$) of $Cu^{2+}$ and Tebuconazole were 0.16, 38.5, respectively. The teratogenic index (TI) appeared 3.0 in $Cu^{2+}$ and 7.7 in Tebuconazole, which showed teratogenicity in embryonic development of Hyla japonica. These results reveal that $Cu^{2+}$ and Tebuconazole in this experiment suppressed the development of embryos at relatively low concentration. Much of Hyla japonica embryos can be secured, and easy to incubate. In addition, mortality, malformation ratios, malformation patterns and growth rates are similar to the results from the other assay systems. Therefore, the Hyla japonica embryo teratogenesis assay system could be a useful tool to evaluate toxicity of pollutants in environment.

• Journal of Ginseng Research
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• v.20 no.1
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• pp.83-87
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• 1996

The effects of gibberilin ($GA_3$) on levels of endogenous indole-3-acetic acid (IAA) and zeatin in both fresh and stratified American ginseng (Panax quinquefolium) seeds were investigated. In our first experiment, the fresh seeds were stratified after soaked in 80 ppd $GA_3$ solution for 24 hours. We found that the IAA concentration in embryo increased by 50.7% and 82.1% respectively at the 120th day and the 188th day of stratification, and the zeatin concentration also increased by 3.8% and 51.6% respectively. In our second experiment, we treated the seeds after 134 days stratification with 80 ppm GA3 for 24 hours and then continued to stratify them. We found that the IAA concentration in embryo increased by 32.9% and 17.7% respectively at the 164th day and the 208th day of stratification while zeatin concentration increased by 22.7% and 30.6% respectively In our another experiment, we studied the effects of $GA_3$, abscislc acid (ABA) and GA, plus ABA on germination rate of seeds treated with these plant hormones during stratification. We found that when the stratified seeds whose ratio of embryo had reached 75% were treated with 80 ppm GA3 for 24 hours and then were allowed to be stratified for another 88 days, the weight and length of embryo (p < 0.05), and germination rate (p < 0.01) increased. In contrast, the 25 ppm ABA treated with for 24 hours was found to Inhibit the growth of embryo (p < 0.05) and reduce the germination rate (p < 0.05) . The experiment of combination treatment of $GA_3$ and ABA showed that $GA_3$ could relieve the inhibitory effects of the ABA on the development of the seeds.

• Korean Journal of Plant Tissue Culture
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• v.21 no.1
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• pp.7-14
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• 1994

Immature zygotic embryos of five Korean soybean cultivars cultured on Murashige and Skoog's (MS) medium supplemented with various concentrations of 2,4-Dichlorophenoxyacetic acid (2,4-D) produced somatic embryos without forming an intervening callus. The highest frequency (up to 90%) of somatic embryo formation was obtained when zygotic embryos were cultlued on medium containing 1 to 2 mga 2, 0-D in four cultivars. However the frequency was highly variable to the cultivars. Transversely sliced primary somatic embryo halves were also capable of forming secondary embryos at frequencies of up to 70% when cultured on medium containing 0.1 to 1 mg/L 2,4-D. Somatic embryos formed on zygotic embryos cultured on medium containing 0.1 to 0.2 mg/L 2,4-D had two cotyledons more frequently than one horn-type cotyledon and those on medium containing 0.5 to 4mg/L 2,4-LD had a horm-type cotyledon at a prominently higher freequency. However somatic embryos on medium containing 10mg/L or higher concentrations of 2,4-D were usually shunted at the globular stage even after transfer to medium containing lower concentrations of 2,4-D or other growth regulators. non somatic embryos with one or two cotyledons or a hem-type cotyledon were transferred to medium containing $GA_3$, those with two cotyledons converted to plantlets at a higher frequency (25%) than the others.

• Journal of Embryo Transfer
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• v.15 no.1
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• pp.67-75
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• 2000

This study was conducted to establish the optimal conditions for in vitro embryo production using oocytes derived from follicles of slaughter-house ovaries. The ovaries of Hanwoo were obtained from a local slaughter-house. The oocytes were aspirated from visible follicles of 2~7mm in diameter. The recovered oocytes which were completely surrounded by at least 2 layers of cumulus cells and a homogeneous cytoplasmic pigmentation were used. The selected oocytes were washed 3 or 4 times with D-PBS containing 10% bovine calf serum (BCS) and matured in vitor (IVM) in Ham's F-10 supplemented with 10% BCS or 0.01 $\mu\textrm{g}$/ml epidermal growth factor(EGF) at 39$^{\circ}C$ under 5% CO2 in air for 24 hours. They were fertilizqed in vitro (IVF) with fresh sperm separated by Percoll density gradient or swim-up in TALP media. The zygotes were cultrued with or without bovine oviductal epitherial cells(BOEC) in media(HECM-6 supplemented with 11 amino acid and / or TCM-199 supplemented with 10% BCS) for 7 to 10 days. The results obtained were as follow: The cleavage rate and developmental rate to blastocyst after maturation and IVF were not significantly different between Ham's F-10 with EGF(76.0% vs. 44.0%) and BCS(75.9% vs. 43.6%)(P<0.05). The cleavage rate and development rate to blastocyst after fertilizing by swim-up or Percoll method were not signifciantly(P<0.05) different between swim-up (80.2% vs. 29.2%) and Percoll(81.9% vs. 26.5%) (P<0.05). The cleavage rate in TCM 199(80.5) was signficiantly higher than that in HECM-6 (72.0%) (P<0.05). However, developmental rate to blastocyst using TCM 199 following HECM-6 for 3 or 4 days (42.2%) was significantly higher than that in TCM-199 alone(26.7%)(P<0.05). The cleavage rate and development rate of embryos produced in vitro by exchange timing for HECM-6 media were not significantly different between in day 3(78.6% vs. 45.5%) and day 4(75.0% vs. 43.2%)(P<0.05). The cleavage rate and developmental rate to blastocysts according to co-culture system were not significantly different between with (74.2% vs. 41.4%) and without BOEC(73.95 vs. 43.5%) (P<0.05). The number of blastomere in blastocyst stage after co-culture with or without BOEC was not significantly different (106.7$\pm$5.1 and 96.6$\pm$4.0). In conclusion, the most transferable IVP embryos could be produced from Ham's F-10 medium for IVM, Percoll density gradient method for IVF sperm separation and in vitro culture in HECM-6 until day 3 or day 4, and then transferred into TCM-199 until day 9 within adequate embryo density in culture droplets after insemination.

• Journal of Embryo Transfer
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• v.11 no.2
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• pp.125-135
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• 1996

The present study was carried out to investigate the effects of cryoprotectants, equilibration step, freezing rate, culture condition following in vitro fertilization, and age and development stage of embryo by freezing with conventional slow freezing and vitrification on survival of frozen-thawed Korean native cattle(KNC) blastocysts produced in vitro. The KNC blastocysts produced in vitro were equilibrated in 1.8M ethylene glycol or 1.4M glycerol and cooled from -6$^{\circ}C$ to -35$^{\circ}C$ at -0.3$^{\circ}C$ or -O.6$^{\circ}C$ /minute. When equilibrated in 1.8M ethylene glycol, survival rate of fiozen4hawed blastocysts was sarne in both -0. 3$^{\circ}C$ /min and -0.6$^{\circ}C$ /min cooling rate(71.4%). With the equilibration in 1.4M glycerol, survival rate was higher in -0.3$^{\circ}C$ /min(63.6%) than in -0.6$^{\circ}C$ /min cooling rate(53.8%). For vitrification of the KNC blastocysts produced in vitro, they were equilibrated in 2-step or 3-step exposure to vitrification solution(25% ethylene glycol + 25% glycerol). Survival rate was sirilar in both 2-step(45.0%) and 3-step exposure(47.4%). According to culture condition following in vitro fertilization, higher survival rate was obtained for blastocysts co-cultured with bovine oviductal epithelial cell(BOEC, 77.3%) than for those cultured with epidermal growth factor(EGF, 65.7%) or for those co-cultured with BOEG + EGF (54.8%). According to embryo age and development stage, higher survival rate was obtained for 7-day ernbryos(70.0%) than 8-day(56.8%) or 9-day(20.0%) for blastocyst stage and obtained for 8-day embryos(74.3%) than 7-day(62.5%) or 9-day(42.9%) for exponded blastocyst. In surnmary, higher survival rate of frozen4hawed KNC blastocysts produced in vitro were obtained by using ethylene glycol for cryoprotectant and -0.3$^{\circ}C$ /min for cooling rate. And higher survival rate were obtained with co-culture with BOEC for culture condition following in vitro fertilization and with 7-day blastocyst or 8-day expanded blasto cyst for embryo age and development stage.

• Journal of Embryo Transfer
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• v.22 no.4
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• pp.245-249
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• 2007

The aim of present experiment was to examine hatching rate as in vitro indicator of viability of porcine embryos before early stage embryo transfer such as zygotes or 2-cell stage embryos. Cumulus-oocyte complexes (COCs) collected from ovaries were matured in North Carolina State University 23 (NCSU-23) containing 10% porcine follicular fluid (pFF), 10 ng/ml epidermal growth factor (EGF), $10{\mu}g/ml$ follicle stimulating hormone (FSH), $35{\mu}g/ml$ luteinizing hormone (LH), and 1mg/ml cysteine. After 24 hours, the COCs were transferred to the same medium without hormones. After 65h of maturation, oocytes were exposed to phosphate buffered saline (PBS) with 7% ethanol (v/v) for 7 minutes, and then the oocytes were washed and cultured in tissue culture medium (TCM) 199 containing 5 ug/ml cytochalasin B for 5h at $38.5^{\circ}C$ in an atmosphere of 5% $CO_2$ and 95% air with high humidity. After cytochalasin B treatment, the presumptive parthenotes were cultured in porcine zygote medium (PZM)-5 and cleavage of the parthenotes was assessed at 72h of activation, Normally cleaved parthenotes were cultured for an additional 8 days to evaluate their ability to develop to blastocyst and hatching stages. The fetal bovine serum (FBS) were added at Day 4 or 5 with concentrations of 2.5, 5 or 10%. The blastocyst rates were ranged within $39.1{\sim}70%$ in each treatment. However hatching rate was dramatically decreased in non-addition group. In this experiment, embryo viability in female reproductive tract may be estimated before embryo transfer with in vitro culture adding FBS by hatching ability.

• Korean Journal of Environmental Biology
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• v.39 no.3
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• pp.311-318
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• 2021

In this experiment, we investigated the toxicity of tebuconazole (fungicide) using domestic frog embryos, along the FETAX (Frog Embryo Teratogenesis Assay-Xenopus) protocol. Bufo gargarizans, Hyla japonica, and Pelophylax nigromaculatus embryos were incubated, and investigation of the tebuconazole effect was performed by the probit analysis. As a result, depending on the concentrations of tebuconazole, the mortality and malformation rates were increased and larval body length was decreased. The teratogenic concentrations (EC₅₀) of tebuconazole were 34.4mg L^-1, 10.6mg L^-1, and 14.9mg L^-1, respectively, and the embryo lethal concentrations(LC₅₀) of tebuconazole were 74.7 mg L^-1, 38.5 mg L^-1, and 39.1 mg L^-1, respectively. The teratogenic index (TI) valuesof tebuconazole were 2.19, 3.58, and 2.65; thus, it showed teratogenicity in embryonic development of these three frogs. These results revealed that in this experiment, tebuconazole suppressed the development of embryos at a relatively low concentration. In addition, mortality, malformation ratios, malformation patterns, and growth rates were similar to the results from the other assay systems. Therefore, tebuconazole was thought to have an effect on the embryo development of domestic frogs. In future, it will be necessary to identify species specificity in order to the clarify the causes of differences in mortality, malformation rate, and malformation patterns depending on the species.