• Reproductive and Developmental Biology
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• v.35 no.3
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• pp.369-375
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• 2011

The aim of this study was to evaluate the effect of ${\alpha}$-tocopherol on blastocysts development and subsequent cryosurvival of the vitrification. The ${\alpha}$-tocopherol(0, 100, 200, 400 ${\mu}M$) was added in to culture medium for the bovine embryos. The blasocysts from the ${\alpha}$-tocopherol and untreated control groups were then frozen-thawed, and their cryosurvival was assessed by in vitro culture for 48 h. There were no differences in the overall cleavage rate($56.14{\pm}4.66$, $58.18{\pm}4.70$, $62.97{\pm}6.86$ and $51.17{\pm}7.28$) among four treatment groups. However, in blastocyst development and total cell number were significantly higher in ${\alpha}$-tocopherol 200 ${\mu}M$ ($38.60{\pm}7.12$; $106.33{\pm}3.50$) to culture medium than other treatment groups($29.30{\pm}5.24$, $31.60{\pm}7.12$ and $26.37{\pm}4.18$; $101.36{\pm}5.12$, $97.27{\pm}2.87$, and $91.23{\pm}7.52$ respectively). Before and after vitrification, the total cell number and blastocyst development of embryo were significantly higher in July to August than September to October. In conclusion, addition of ${\alpha}$-tocopherol 200 ${\mu}M$ to in vitro bovine embryo culture medium was beneficial for improving embryo quality by decreasing the embryo damage blsstocysts cell number and improving the tolerance of the embryos to cryopreservation.

• Clinical and Experimental Pediatrics
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• v.50 no.9
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• pp.875-881
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• 2007

Purpose : Chromosome analysis is important in genetic study and genetic counseling. This study was performed to evaluate the type and incidence of chromosome abnormalities in a single hospital for 25 years. Methods : Chromosome analyses were performed on peripheral blood lymphocytes, obtained from 4,856 patients with suspected chromosomal aberrations, referred to cytogenetic laboratory in Department of Pediatrics, Kyungpook National University Hospital from May 1981 to October 2005. Results : We analyzed 4,567 cases. Children were 3,014 cases (66.0%) and adult were 1,553 cases (34.0 %). The most common purpose of the chromosomal analysis was growth and developmental abnormality in children and infertility in adults. Total chromosomal aberration rate was 16.9% (770/4,567). Among those cases, the numerical abnormalities were 12.2% (558 cases), the structural abnormalities were 4.1% (187 cases), and others were 0.5% (25 cases). The relative frequencies of autosomal abnormalities were 6.4% (294 cases) in Down syndrome; 0.2% (7 cases) in Edwards syndrome; 0.1% (4 cases) in Patau syndrome; 0.2% (10 cases) in other abnormalities, of sex chromosome, 2.9% (131 cases) in Klinefelter syndrome; 2.2% (99 cases) in Turner syndrome; 0.2% (8 cases) in 47, XXX; 0.1% (3 cases) in 47, XYY. Among the structural abnormalities, translocation was 1.8% (84 cases), inversion was 0.8% (37 cases), deletion was 0.4% (17 cases), and insertion was 0.3% (13 cases), in order of frequency. Conclusion : In this study, the type, incidence and distribution of cytogenetic abnormalities by karyotype were reviewed. We hope that our study could be used as a basic information on the diagnosis, treatment and genetic counseling for chromosome abnormalities in Korea.

• Korean Journal of Animal Reproduction
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• v.20 no.2
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• pp.119-126
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• 1996

This study was carried out to determine the optimal condition for successful and efficient c cryopreservation of zygotes, 1-cell embryos, using EFS40 which was 40% (v/v) ethylene glycol diluted in DPBS medium containing 30% Fic-oll (w/v) and 0.3 M sucrose. After mouse zygote produced by IVF was vitrified by two freezing methods, the post-warming survival rates of 1-cell zygotes were assessed as cleavage to the 2-cell stage and development into the hatching blastocysts at 5 day. In the one-step method, when embryos were directly exposed to the vitrification solution at 25$^{\circ}C$ for 1 min., survival and development rates of zygotes were 85.5% and 31.9% In the two-step method, embryos were equilibrated with a dilute 20% EG for 1, 3, 5 min. before 1 min. exposure to EFS40, re-spectively. However, the rates of development (17.7, 3.3, 0%) were lower than that of one-step method. The highest survival rate (95.9%) was obtained by one-step method which exposes embryos in EFS40 for 30 sec. In this condition, 63. 8% of cleaved 2-cell developed into hatching blastocysts. In the cell number of Total and ICM using differential labelling technique, there are no significant differences in the cell number of Total and ICM between blastocysts devel oped in vitrified-thawed embryos (63.2${\pm}$16.9, 1 13.5${\pm}$4.0) and control balstocysts (54.0${\pm}$15.2, 1 12.3${\pm}$4.6). Therefore, these results show that mouse zygotes can be successfully cryopreserved by a simple vitrification method although developmental rates of vitrified embryos were reduced. In conclusion, this proposed vitrifi cation procedures can be useful in the cryopreservation of mouse IVF zygotes.

• Korean Journal of Animal Reproduction
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• v.20 no.1
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• pp.27-34
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• 1996

The objective of this study was to examine the cell number of Total, ICM and TE cells of bovine blastocysts according to development progression cultured in CR1 medium, which was reported as successfully supporting medium for preimplantaion bovine embryo development to the blastocyst stage, by differential labelling of the nuclei with immunosurgery and polynucleot-ide-specific fluorochromes. Blastocysts were obtained at day 8 after in vitro fertilization and classified to early, middle, expanded stage according to the developmental morphology; blastocoel expansion and zona thickness. Also, bias tocysts in the same category were divided into two parts to check the Total cell number by using bisbenzimide only and ICM, TE and Total cell number by using immunosurgery and two polynucleotide-specific fluorochromes. 1) The development rate of blastocysts at day 8 after in vitro fertilization was 29.3% and classified bIas tocysts to early, middle, expanded and hatching stage were 8.7, 9.9, 7.6 and 3.1%, respectively. 2) The numbers of total blastomere using bisbenzimide in the classified blastocysts to early, middie and expanded were 46.9${\pm}$8.6, 66.2${\pm}$12.5 and 122.8 ${\pm}$ 14.4, respectively. This indicated that CR1 is a appropriate culture medium for bovine embryo development. 3) The count of ICM and TE cell number by using differential labelling with immunosurgery and polynucleotide-specific fluorochromes in the classified blastocysts to early, middle and expanded; ICM cell numbers of were 12.8${\pm}$5.9, 26.3${\pm}$8.4 and 35.5${\pm}$15.0, respectively and TE cell numbers were 30.5${\pm}$5.0, 4 41.3${\pm}$8.2 and 81.1${\pm}$13.4, respectively. These results presented that the increase of ICM and TE cell numbers averaged two and three doublings between early and expanded blastocyst stage and also total cell number counted from ICM nuclei and TE nuclei by using differential label-ling showed the increase pattern with development advance level and the results were similar to total cell number obtained from bisbenzimide treatment only. Therefore, the differential labelling of ICM and TE nuclei in situ is a very useful technique to evaluate embryo qualities and can be used as an indicator on study of preim-plantation embryo development.

• Clinical and Experimental Pediatrics
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• v.45 no.2
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• pp.232-239
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• 2002

Purpose : Renal cystic diseases comprise a mixed group of heritable, developmental and acquired disorders. Recently the use of imaging modalities such as ultrasonography and radionuclide scanning has increased the detection rate of renal cystic diseases. We studied to review the clinical features and treatment of renal cystic diseases in children. Methods : This study was performed in 95 children with renal cystic diseases in the Department of Pediatrics, Asan Medical Center from October 1989 to June 2001. Results : In 95 patients, there were 55 cases(58.0%) with multicystic dysplastic kidney(MCDK), 19 cases(20.0%) with simple renal cysts, 13 cases(13.7%) with hereditary polycystic kidney diseases( 7 with autosomal recessive type, 5 with autosomal dominant type, 1 with undetermined), 6 cases(6.3%) with renal cysts in tuberous sclerosis and 1 case(1.0%) with medullary cystic disease. All MCDK patients had no renal dysfunction and hypertension during the follow-up period. Three out of 13 with polycystic kidney diseases had progressed to end-stage renal disease during the follow-up period. One case with a simple cyst underwent laparoscopic malsupialization for decompression. Conclusion : Renal cystic diseases have diverse clinicopathologic features and variable prognosis. We emphasize that routine follow-up should be performed to prevent and to detect early treatable complication in renal cystic diseases. Therefore, their natural history and treatment need further investigation and long term follow-up is required.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.25 no.5
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• pp.341-358
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• 1992

In order to estimate energy budgets of Palaemon macrodactylus, larvae of the shrimp were reared in the laboratory at constant conditions $(25^{\circ}C: 31-32\%o),$ and then juvenile to adult of the shrimp were reared at $15^{\circ}C\;and\;25^{\circ}C$ in the laboratory. Energy used by the reared shrimps were calculated from estimates of data on feeding, growth, molting, metabolism, nitrogen excretion, and energy content. Juveniles and adults reared in the laboratory, which fed on Artemia nauplii, had an average daily growth rates of 0.079 mm/day at $15^{\circ}C\;and\;of\;0.122mm/day\;at\;25^{\circ}C$. The average growth factor* of P. macrodactylus males and females ranged from $3.2\%$ for adult to $13.2\%$ for juveniles individuals, respectively. Intermolt periods were related to body size of the shrimp and to temperature. Average laboratory growth curves were calculated from data on growth factors and intermolt periods to body size of the shrimp at $15^{\circ}C\;and\;25^{\circ}C$. The calorie contents of the shrimp, their molts, eggs and larvae were determined by biochemical composition and oxygen bomb calorimetry. The average amount of energy used in growth for larvae and juvenile to adult were 4.94 cal and 4.55 cal per dry weight in milligram, respectively. The ammount of oxygen used in metabolism was calculated from size, temperature-specific respiration rate. To convert the ammount of oxygen used in respiration into the equivalent energy lost heat was estimated from the data on chemical composition for the larvae and adult, the values was 4.58 cal/ml $O_2$. The energy content per egg was 0.078 cal. The assimilation efficiency estimated by nitrogen content of food and egested faeces gave $61.5\%$ for the larvae. The efficiencies for juvenile to adult ranged between $79.4\%$ and $90.1\%$ The gross growth efficiencies $(K_1)$ and net growth efficiencies $(K_2)$ of P macrodactylus showed $18.33\%\;and 32.63\%$ for total larval stages, ranged from $21.30\%\;to\;31.04\%\;and\;from\;30.03\%\;to\;39.34\%$ for juvenile to adult, respectively.

• Korean journal of applied entomology
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• v.48 no.2
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• pp.123-158
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• 2009

An attempt was made to stimulate future research by providing exemplary information, which would integrate published knowledge to solve specific pest problem caused by resistance. This review was directed to find a way for delaying resistance development with consideration of chemical(s) nature, of mixture, rotation, or mosaics, and of insecticide(s) compatible with the biological agents in integrated pest management (IPM). The application frequency, related to the resistance development, was influenced by insecticide activity from potentiation, residual period, and the vulnerability to resistance development of chemical, with secondary pest. Chemical affected feeding, locomotion, flight, mating, and predator avoidance. Insecticides with negative cross-resistance by the difference of target sites and mode of action would be adapted to mixture, rotation and mosaic. Mixtures for delaying resistance depend on each component killing very high percentage of the insects, considering allele dominance, cross-resistance, and immigration and fitness disadvantage. Potential disadvantages associated with mixtures include disruption of biological control, resistance in secondary pests, selecting very resistant population, and extending cross-resistance range. The rotation would use insecticides in high and low doses, or with different metabolic mechanisms. Mosaic apply insecticides to the different sectors of a grid for highly mobile insects, spray unrelated insecticides to sedentary aphids in different areas, or mix plots of insecticide-treated and untreated rows. On the evolution of pest resistance, selectivity and resistance of parasitoids and predator decreased the number of generations in which pesticide treatment is required and they could be complementary to refuges from pesticides To enhance the viability of parasitoids, the terms on the insecticides selectivity and factors affecting to the selectivity in field were examined. For establishment of resistant parasitoid, migration, survivorship, refuge, alternative pesticides were considered. To use parasitoids under the pressure of pesticides, resistant or tolerant parasitoids were tested, collected, and/or selected. A parasitoid parasitized more successfully in the susceptible host than the resistant. Factors affecting to selective toxicity of predator are mixing mineral oil, application method, insecticide contaminated prey, trait of individual insecticide, sub-lethal doses, and the developmental stage of predators. To improve the predator/prey ratio in field, application time, method, and formulation of pesticide, reducing dose rate, using mulches and weeds, multicropping and managing of surroundings are suggested. Plant resistance, predator activity, selective insect growth regulator, and alternative prey positively contributed to the increase of the ratio. Using selective insecticides or insecticide resistant predator controlled its phytophagous prey mites, kept them below an economic level, increased yield, and reduced the spray number and fruits damaged.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.40 no.6
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• pp.684-696
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• 1995

A mutant for chalky endosperm and genetic male sterility (GMS) was newly developed in rice. The two characters were found to be controlled by single recessive gene which has pleiotropic effect, indicating that chalky seeds should be GMS seeds in segregating populations. Chalky seeds showed the same shape and size as normal seeds. However, starch composition of central part of endosperm was looser and shape of starch granules was rounder compared with normal endosperm, resulting in significantly lower grain weight, absolute density and grain hardness in chalky grains than in normal ones. Amylose content and gel consistency of chalky grains were much lower and harder, respectively. Male sterile plants showed much shorter plant height, poorer panicle exsertion and lesser panicle number compared with normal plants. Microspore abortion stage in pollen developmental process was observed as before meiosis. Male sterility of the mutant was stable regadless of temperature and day length. A system breeding hybrid rice using this mutant was discussed, comp ring with other systems utilizing cytoplasmic-genic male sterility(CGMS) and environment sensitive GMS(EGMS). Separation of GMS seeds in mixed seed bulks by specific gravity (1.14∼1.16g / cm3) was successful about 85∼90%. But some mixed normal plants were seemed to be easily removed by the apparent difference in growth characters at seedling stage. The highest natural outcrossing rate of this GMS line was as 17.3 % in a plot treated with 2-row pollinator, I-row GMS, and GA3 + cutting of flag leaf + pollen-scattering by rope.

• Asian-Australasian Journal of Animal Sciences
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• v.14 no.9
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• pp.1260-1266
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• 2001

The effect of protein supplementation, $O_2$ concentration and co-culture on the development of embryos produced by nuclear transfer using cultured cumulus cell was investigated. Recipient oocytes and cumulus cells were obtained from the ovaries of the slaughtered Hanwoo cows. Donor cumulus cells were cultured in Dulbecco's modified Eagle medium containing 10% fetal bovine serum at 5% $CO_2$ in air at $38.5^{\circ}C$. The 1 to 6 passages of cumulus cells were isolated and used as donor cells. The in vitro matured oocytes were enucleated and then the isolated donor cells were introduced. One $15{\mu}s$ pulse of 180 volts was applied to induce the fusion between karyoplast and cytoplast. The fused embryos were activated with $10{\mu}M$ calcium ionophore for 5 min and 2 mM 6-dimethylaminopurine for 3 h. To examine the effect of protein supplementation, nuclear transfer (NT) embryos were cultured in one of the following 4 treatments : 1) CR1aa + 3 mg/ml BSA for 7 days ; 2) CR1aa + 10% FBS for 7 days ; 3) CR1aa + 1.5 mg/ml BSA + 5% FBS for 7 days ; and 4) CR1aa + 3 mg/ml BSA for first 3 days and then CR1aa + 1.5 mg/ml BSA + 5% FBS for 4 days. Culture took place at 5% $CO_2$, 5% $O_2$ and 90% $N_2$ at $38.5^{\circ}C$. Although there were no significant differences in cleavage rate among different protein supplements, the rates of blastocyst formation were significantly different. When NT embryos were cultured in the medium supplemented with only BSA, they could develop to only morula not to blastocyst. However, when FBS was supplemented, NT embryos developed to blastocyst stage. In order to investigate the effect of $O_2$ concentration and co-culture, NT embryos were cultured in CR1aa + 1.5 mg/ml BSA + 5% FBS with or without cumulus cell co-culture at an atmosphere of 5% $CO_2$ in air (20% $O_2$) or 5% $CO_2$, 5% $O_2$, 90% $N_2$ (5% $O_2$) at $38.5^{\circ}C$ for 7 days. The percentage of blastocyst development was significantly higher when the NT embryos were cultured at an atmosphere of 5% $O_2$ than that of 20% $O_2$ (p<0.05). However, there was no significant difference between with and without cumulus cell co-culture at an atmosphere of 5% $O_2$ or 20% $O_2$. Fifty embryos were transferred to 25 recipients and 5 recipients were pregnant at 100 days. From 5 pregnant cows, only one cow was delivered of female twin. In conclusion, the embryos reconstructed by enucleation of metaphase II oocytes and introduction of the cycling and quiescent cumulus donor cells in Hanwoo had developmental potential to term after embryo transfer to recipient cows.

• Development and Reproduction
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• v.13 no.4
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• pp.353-362
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• 2009

GnRH and its receptor are known to express locally in the ovary and to regulate the ovarian function by affecting on granulosa and lutein cells. It has been reported that GnRH directly causes apoptosis in the granulosa and lutein cells of the ovary. However, whether the apoptosis of the cells by GnRH is recovered by FSH as an anti-apoptotic factor is not yet known. In this study, we evaluated the apoptosis and the production of progesterone $(P_4)$ and estradiol $(E_2)$ after treatment with 5, 50, and 100 ng/$m\ell$ GnRH and 1 IU/ml FSH in the granulosa-lutein cells that are obtained during oocyte-retrieval for IVF-ET. Results of DNA fragment analysis and TUNEL assay demonstrated that DNA fragmentation and the rate of apoptotic cells were increased in a dose-dependent manner showing a significant increase in the cells treated with 100 ng/$m\ell$ GnRH. In addition, we found that FSH suppresses the apoptosis of the cells induced by GnRH. In the results of chemiluminescence assay for $P_4$ and $E_2$, $P_4$ production was decreased by GnRH treatment, whereas $E_2$ production was not changed. We also demonstrated that FSH inhibits the suppressive effect of GnRH on $P_4$ production as the result of apoptosis. The present results suggest that GnRH agonist using in ovarian hyperstimulation protocol might induce the dysfunction of the ovary, but its function could be recovered by FSH. These results also will be expected to use as the basic data to elucidate the physiological role of GnRH and to develop new ovarian hyperstimulation protocols for IVF-ET.