• Journal of Embryo Transfer
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• v.30 no.3
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• pp.219-224
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• 2015

The purpose of this study is to develop transgenic cell line expressing targeted human granulocyte colony stimulating factor (hGCSF) and green fluorescence protein (GFP) genes as well as production of Somatic Cell Nuclear Transfer (SCNT) embryos derived from co-expressed transgenic donor cells. Constructed pPiggy-mWAP-hGCSF-EF1-GFP vector was chemically transfected into bovine fetus cells and then, only GFP expressed cells were selected as donor cells for SCNT. Cleavage and blastocyst rates of parthenogenetic, SCNT embryos using non-TG cell and hGCSF-GFP dual expressed SCNT embryos were examined (cleavage rate: $78.0{\pm}2.8$ vs. $73.1{\pm}3.2$ vs. $70.4{\pm}4.3%$, developmental rate: $27.2{\pm}3.2$ vs. $21.9{\pm}3.1$ vs. $17.0{\pm}2.9%$). Result indicated that cleavage and blastocyst rates of TG embryos were significantly lower (P<0.05) than those of parthenogenetic and non-TG embryos, respectively. In this study, we successfully produced hGCSF-GFP dual expressed SCNT embryos and cryopreserved to produce transgenic cattle for bioreactor system purpose. Further process of our research will transfer of transgenic embryos to recipients and production of hGCSF secreting cattle.

• Development and Reproduction
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• v.10 no.4
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• pp.239-245
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• 2006

Experiments were conducted to determine the effects of beta-mercaptoethanol(${\beta}-ME$) supplements to the maturation medium on in vitro fertilization(IVF) and intracellular glutathione(GSH) concentration. Bovine cumulus-intact oocytes were matured in TCM-199 medium containing FBS, hormonal supplements, and ${\beta}-ME$(0, 25 and $50\;{\mu}M$) for 12h and 24 h. After culture, cumulus-free matured oocytes were co-incubated with frozen-thawed spermatozoa for 24h. Maturation rate increased(p<0.05) in ${\beta}-ME$ treatment group, but no significant differences among treatment groups. Also, increases(p<0.05) in intracellular GSH concentration before and after fertilization were observed in $50\;{\mu}M\;{\beta}-ME$ supplements to the maturation medium. Male pronuclear formations after IVF was increased(p<0.05) in ${\beta}-ME$ treatment group, but no significant difference among treatment groups. In conclusion, supplementing ${\beta}-ME$ into the maturation medium increased maturation rates, fertilization rates, and intracellular GSH concentrations.

• Korean Journal of Animal Reproduction
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• v.15 no.1
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• pp.49-57
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• 1991

This study was carried out to investigate the effect of equilibration time, precooling and straw loading method on the post-thaw survival rates of mouse embryos cryopreserved by vitrification method. The effect of the vitrification procedure on the damage of the embryos was also examined by the straining of nuclei using Hoechst 33342. The results obtained were summarized as follows; 1. The equilibration in Medium-1 for 10 minutes was considered to be the optimum equilibration time. Embryos equilibrated in Medium-1 for 10 minutes(81.0%) showed significantly(P<0.05) higher survival rates than those equilibrated for 5 minutes(40.0%) or 15 minutes(74.1%). 2. The survival rate of embryos cryopreserved using the double Medium-2 column(81.0%) was significantly(P<0.01) higher than that using the single Medium-2 column, whish was considered to be due to the double Medium-2 column method being more reliable for preventing contamination of diluent solution of 1M sucrose. 3. The survival rate of compacted morula stage embryos cryopreserved with the precooled and non-precooled Medium-2 was not significantly(P<0.05) different. 4. The number of blastomeres of embryos at late blastocyst stage after culture of mouse morulae for 24 to 28hours was significantly(P<0.05) decreased by freezing embryos using vitrification(53.3${\pm}$1.6 vs 41.4${\pm}$1.5), which was considered to be due to the damage of embryos during vitrification and the delay of embryo development by handling in vitro. 5. The vitrification procedure is considered to be a very simple and efficient method for cryopreservation of embryos at early developmental stage.

• Reproductive and Developmental Biology
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• v.28 no.2
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• pp.113-117
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• 2004

The present study was carried out to examine the effects of $O_2$ concentrations and culture media (North Carolina State University (NCSU)-23, porcine zygote medium(PZM)-3 or PZM-4) on in vitro development of porcine IVM/IVF embryos. Porcine oocyte-cumulus complexes were cultured in BSA-free NCSU-23 medium containing porcine follicular fluid (10%), cysteine (0.9 mM), $\beta$-mercaptoethanol (25 $\mu\textrm{g}$/$m\ell$), epidermal growth factor (10 ng/$m\ell$) and hormonal supplements (PMSG and hCG: 10 IU/$m\ell$) for 20∼22 h. They were then cultured in the same medium but without hormonal supplements for an additional 20∼22 h. After culture, cumulus-free oocyte were coincubated with liquid boar spermatozoa for 5∼6h. Putative zygotes were transferred to NCSU-23, PZM-3 and PZM-4 medium under the condition of 5% $O_2$ or 20% $O_2$ concentrations. At 48 h, no mean differences were found in cleavage rates. However, the rates of blastocyst formation at day 7 after in vitro fertilization were significantly higher in PZM-3 medium under the condition of 5% $O_2$ concentration than other treatments (19.9$\pm$2.4 vs. 11.1$\pm$2.0 to 16.0$\pm$2.5%, P<0.05). The total cell numbers of blastocysts were significantly higher in 5% $O_2$ than in 20% O2 (P<0.05). However, no differences was found among the culture media within each $O_2$ concentrations. In conclusion, the use of PZM-3 medium in 5% $O_2$ concentration was effective on in vitro development of porcine IVM/IVF embryos.

• Journal of the Korea Academia-Industrial cooperation Society
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• v.13 no.8
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• pp.3785-3791
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• 2012

Dissolved oxygen is one of the most important factors controlling growth in aquatic organisms. Hypoxia is generally defined as dissolved oxygen less than 2.8 mg $O_2/L$ (equivalent to 2 mL $O_2/L$ or 91.4 mM). Therefore, hypoxia zone can cause a serious problem in marine ecosystem. In this study, to investigate embryotoxic (fertilization and embryo development rates) effects of hypoxia on sea urchin Strongylocentrotus nudus were exposed to dissolved oxygen levels of 7.6 mg $O_2/L$ (normoxia) and 1.8 mg $O_2/L$ (hypoxia) for 2 days at $15^{\circ}C$ and 33 ‰. Also, Expression levels of stress related gene (HSP70) and antioxidant related gene (glutathione reductase) in the sea urchins exposed to hypoxia were confirmed by Immunoblotting and RT-PCR analysis. In results, we showed that developmental rates were dramatically reduced in hypoxia condition. Molecular analysis demonstrated that higher HSP70 (5.5 fold) and glutathione reductase gene (2.79 fold) were present in the sea urchin exposed to hypoxia. Our results suggested that hypoxia can cause the abnormal development and elicits a stress and antioxidant response on sea urchin.

• Journal of Life Science
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• v.29 no.2
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• pp.202-208
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• 2019

Brown planthopper (BPH), Nilaparvata lugens Stål (Hemiptera: Delphacidae), is one of the most important migratory pests damaging rice in Korea. It invades annually from tropical and subtropical areas via continental air streams. It is necessary to determine the resistance levels of rice varieties in order to control efficiency. The honeydew excretion, development, and reproduction of the migratory BPH were studied by region in a laboratory at $25{\pm}2^{\circ}C$ and $65{\pm}5%\;RH$ and a 16L: 8D photoperiodism conducted on three BPH resistant genes: Bph1, Bph2, and Bph18. The information obtained was reported using the jackknife method, and we created life table statistics accordingly. The feeding amount of Bph1 resistant gene was lower than that of resistant genes. The developmental periods of immature stages ranged from $13.7{\pm}0.10d$ on Bph2 (Namhae, 2015) to $18.5{\pm}1.06d$ on Bph2 (Sacheon, 2016). Reproductive period and female longevity were longest on the non-resistant genes, Bph2 and Bph18 (except 1980s), and the highest fecundity of N. lugens was observed on the two BPH resistant genes. Highest net reproductive rates ($R_0$) were calculated on Bph2 by region. Intrinsic rates of population increase ($r_m$) showed a difference in resistant genes by region. These population parameters showed that migratory regions and biological characteristics of N. lugens vary annually.

• Proceedings of the Korean Society of Embryo Transfer Conference
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• 2002.11a
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• pp.80-80
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• 2002

This study was carried out that to investigate the effects of amino acids supplemented with culture medium on development of porcine embryos cultured in vitro. Cumulus oocyte complexes (COCs) were cultured in the maturation medium containing hormones (0.5$\mu\textrm{g}$/$m\ell$ LH, 0.5$\mu\textrm{g}$/$m\ell$ FSH and 1$\mu\textrm{g}$/$m\ell$ estradiol-17${\beta}$) for 20-22 h at 39$^{\circ}C$ in an atmosphere of 5% CO$_2$in air. Subsequently, COCs were cultured in hormone-free maturation medium for 20-22 h. After maturation for 40-44h, oocytes were removed cumulus cells by pipetting and cultured with epididymal sperm for 5 h in the mTBM. Embryos obtained were divided in 4 groups (1) cultured in NCSU 23 containing 0.4% BSA to blastocyst stage(Control), (2) essential amino acids (EA), (3) non-essential amino acids (NA), (4) mixture of essential and non essential amino acid (EA+NA). All treated groups(2-4) were used a glucose free NCSU 23 medium supplemented with pyruvate (0.33 mM), lactate (4.5 mM) to morula stage. From morula to blastocyst stage embryos of all treated groups were cultured in NCSU 23 containing 0.4% BSA. The rates of cleaved oocytes at 48 h after IVF were from 82% to 88% in the groups of control, EA, NA and EA+NA, respectively. The in vitro developmental rates into blastocysts in the groups of EA and EA+NA were significantly (P<0.05) higher than those of group of control (35.1, 35.4 vs. 19.4%, respectively), however, no significant (P<0.05) between control and NA. In conclusion, supplemented with essential amino acid or mixture of essential and non essential amino acid in the culture medium at morula stage increased the rate of development to blastocyst on in vitro produced porcine embryos.

• Korean Journal of Ecology and Environment
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• v.41 no.4
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• pp.431-440
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• 2008

Turbid water or suspended sediment is associated with negative effects on aquatic organisms; fish, aquatic invertebrate, and periphyton. Effects of turbid water on fish differ depending on their developmental stage and a level of turbidity. Low turbid water may cause feeding and predation rates, reaction distance, and avoidance in fish, and it could make fish to die under high turbidity and long period. Therefore, it is very important to find out how turbid water or suspended sediment can affect fish in domestic watersheds. The objectives of this study were 1) to introduce international case studies and their standards to deal with suspended sediment, 2) to determine acute toxicity in 4 major freshwater fishes, and 3) to determine in relation to adverse effect of macroinvertebrates and fish. Impacts of turbid water on fish can be categorized into direct and indirect effects, and some factors such as duration and frequency of exposure, toxicity, temperature, life stage of fish, size of particle, time of occurrence, availability of and access to refugia, etc, play important role to decide magnitude of effect. A review of turbidity standard in USA, Canada, and Europe indicated that each standard varied with natural condition, and Alaska allowed liberal increase of turbidity over natural conditions in streams. Even though acute toxicity with four different species did not show any fatal effect, it should be considered to conduct a chronic test (long-term) for more detailed assessment. Compared to the control, dominance index of macroinvertebrates was greater in the turbid site, whereas biotic index, species diversity index, species richness index, and ecological score were smaller in the turbid site. According to histopathological analysis with gills of macroinvertebrate and fishes, morphological and physiological modification of gills due to suspended sediments can cause disturbance of respiration, excretion and secretion. In conclusion, in order to maintain good and healthy aquatic ecosystem, it is the best to minimize or prevent impact by occurrence of turbid water in stream and reservoir. We must make every effort to maintain and manage healthy aquatic ecosystem with additional investigation using various assessment tools and periodic biomonitoring of fish.

• Reproductive and Developmental Biology
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• v.32 no.4
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• pp.217-222
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• 2008

The techniques of IVM, IVF and IVC of canine oocytes may provide useful information for gamete salvage programs and the conservation of endangered canidae. This investigation has been made to determine the efficiency of in vitro maturation (IVM) as a basic experiment to study the development of canine oocytes after in vitro fertilization (IVF). The rate of oocytes developing to the Mil stage was higher in the hormone treated group (10 IU/ml hCG+eCG, 14.7%, p<0.05) than in the control group (0 IU/ml hCG+eCG, 10.0%). The monospermy and pronuclear rates of canine oocytes were investigated after caffeine treatment on IVF. Canine oocytes were fertilized in the Fert-TALP medium supplemented with 0, 10, 20 or 30 mM caffeine (Fert I, Fert II, Fert III or Fert IV, respectively). The highest pronuclear formation rate was obtained in the Fert I for 24 h IVF (6.7%, 6/89). Therefore, it is believed that unlike in other mammals, caffeine in canine IVF does not increase the efficiency of fertilization rate, and is not an important factor.

• Korean Journal of Environmental Biology
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• v.26 no.4
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• pp.349-354
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• 2008

Superoxide dismutase (SOD) activities were investigated from the portions of shoots and roots in accordance with developmental stage and in response to environmental stress and antioxidants using Rhodiola-seedlings. The rates of SOD activities were revealed highly at the portion of roots and its tip of seedlings in the latter stages rather than the initial stages. SOD activities of seedlings in the initial stages treated with sodium chloride and cadmium as environmental stressors showed the decrease by 15 and 30% with respect to the control, respectively. However, in spite of stressor-treatments, the activities in the roots were increased according to the growth period showing a maximum rate of up to 45%. Also, SOD activities of the seedling treated with ascorbic acid as a antioxidant were increased by 46% of control value, but this was similar to the rate revealed in the presence of stressors. These results suggest that SOD activities in Rhodiola-seedlings may be related with the important defence-system against injurious environments.