• Clinical and Experimental Reproductive Medicine
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• v.31 no.4
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• pp.225-234
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• 2004

Objectives: To evaluate the efficacy of GnRH antagonist cetrorelix in women undergoing controlled ovarian hyperstimulation (COH) for in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI) and to determine changes in serum hormone concentrations during cetrorelix administration. Methods: We performed a clinical trial on 30 patients undergoing COH with highly purified follicular stimulating hormone (HP-FSH) and gonadotropin releasing hormone antagonist (GnRHant), cetrorelix. FSH was administrated from day 2 or 3 of cycle with fixed dose and adjusted according to individual response. 0.25 mg of cetrorelix was injected daily subcutaneously from stimulation day 5 until the day of hCG administration. Daily ultrasound monitoring was performed for growing follicles and serum levels of luteinizing hormone (LH), estradiol ($E_2$) and progesterone were measured daily during cetrorelix administration. Up to 4 embryos were transferred. Results: Mean age of enrolled patients was $32.0{\pm}3.4$ years (mean $\pm$ S.D.). All of 30 patients underwent oocyte pick-up, and embryo transfer was done in 28 patients. The total and mean numbers of received oocytes were 196 and $6.5{\pm}4.7$, the number of fertilized eggs was 111, and the fertilization rate was 56.6%. Total duration of FSH administration was $9.2{\pm}2.2$ days and mean of $24.3{\pm}7.7$ ampules of HP-FSH was administered. Total duration of cetrorelix administration was $5.7{\pm}1.9$ days. Serum LH and progesterone levels were maintained in the range of $1.4{\sim}2.9\;mIU/mL$ and $0.3{\sim}0.6\;ng/mL$, which respectively reflected effective prevention of premature LH surge. Clinical pregnancies were achieved in 9 patients, and overall clinical pregnancy rate was 30.0% per oocyte retrieval, and 32.1% per embryo transfer. Conclusion: GnRH antagonist is safe and convenient for COH for IVF-ET and effective with optimal pregnancy rate.

• Journal of Aquaculture
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• v.7 no.3
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• pp.151-158
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• 1994

To evaluate the reproductive ability of gynogenetic diploid male. Paralichthys olivaceus. histological analysis of testis. cytological analysis of spermatozoa and fertilization test with normal aggs were studied and the results are as follow; The gonads of gynogenetic diploid male were histologically normal. and many spermatozoa were observed in their testis. Number of spermatozoa from the control and gynogenetic diploid male were $2.58\times10^9$ and $2.42\times10^9$ cells per 1 ml of milt. respectively (P> 0.05). Amount of milt per kg body weight from the gynogenetic diploid male was significantly higher (P< 0.01) than that from the control male (8.3ml). Size and morphology from the two experimental groups were not different (P>0.05). More than $80\%$ of fertilization rates and hatching rates were observed when the eggs from the control were fertilized with the gynogenetic diploid male sperms.

• Clinical and Experimental Reproductive Medicine
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• v.21 no.1
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• pp.21-29
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• 1994

Subzonal insemination(SUZI) has been proposed for patients with severe male factor and previous fertilization failure. However, very low fertilization rates still persisted. The aims of this study were firstly, to examine the relationships between the fertilization rate and sperm parmeters, sperm incubation media and time, secondly, to evaluate the outcome of 119 cycles of SUZI applied the modified sperm preparation method. The fertilization rates were influenced more sensitively by sperm preincubation media and time than by sperm parameters. According to preincubation media and time, the fertilization rates were 43.3% in 50% follicular fluid (HFF), 36.6% in 10% fetal cord serum(FCS), and with the time, increased in FCS, but decreased in HFF. In regrd with sperm parameters, the fertilization rates were 42.9% in normal and 37.6% in subnormal group. The best results were obtained from SUZI by the spermatozoa incubated in 50% HFF for 6-8 hours. So we tried 119 cycles of SUZI(normal; 39 cycles, subnormal; 80 cycles) using the preparation method of 6-8 hour incubation in 50% HFF. There were no signigicant differences in the fertilization rates between normal(125/269, 46.4%) and subnormal sperm(264/635, 41.6%). Contrary to the fertilization rates, pregnancy outcomes were different between both groups. Better results obtained from the subnormal group than the normal in the number of transferred embryos, that of good embryos, and developmental rate of the fertilized eggs. The pregnancy rates per transfer were totally 13.3%(13/98),20.0%(13/65) in subnormal group. In the normal group, 2 patients showed ${\beta}$-hCG positive, but resulted in chemical pregnancy. Of 13 clinical pregnancies, two aborted, 6 on-going, and 5 delivered. In conclusion, SUZI is an effective technique to overcome fertilization failure for male factor and unexplained. The fertilization rate is influenced by sperm parameters, sperm incubation media and time. Also the quality of oocytes might be important for pregnancy as same as that of sperm.

• Korean Journal of Ichthyology
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• v.26 no.4
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• pp.267-273
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• 2014

This research has carried out to make a data base of taxonomic research. That Data base explains about spawning behavior, egg development, and morphologic development of variable sabretooth blenny. Fertilized egg was demersal egg which is white and opaque. The number of oil glouble were 10~11, and the size of egg was 0.90~1.43 mm (average rate $1.11{\pm}0.23mm$, n=10). Breeding water temperature was $25.5{\sim}28.5^{\circ}C$ (average rate $27.0^{\circ}C$), and salinity was 32.5~33.5‰ (average rate 33.0‰). After 24 hours from 2 cells, the process of egg development was reached to Blastula stage. Moreover, after 330 hours from 2 cells, nostrils and eyes were formed. Egg membrane was pierced by the head, and the hatch began. After the hatch, postlarvae had 2.59~3.02 mm (average rate $2.81{\pm}0.25mm$, n=5) of whole length, and the mouth and anus were opened. Yolk sac and oil glouble were absorbed. After three days from hatch, prelarvae were 3.02~3.07mm(average rate $3.04{\pm}0.04mm$) of whole length, and caudal fin was grown with round shape. After 13 days from hatch, prelarvae had 3.04~3.20 mm (average rate $3.12{\pm}0.11mm$) of whole length, and they could eat food with upper jaw and bottom jaw.

• Korean Journal of Ichthyology
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• v.10 no.2
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• pp.184-190
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• 1998

The early life history of black bullhead, Pseudobagrus koreanus, endemic to Korea was investigated to get biological information needed in artificial production of seedlings and in recovering natural resources. The fertilized eggs showed some characteristics in having heavy sticky material and minute folds which is formed radical pattern on the egg membrane. The shape of egg was spherical and $2.59{\pm}0.08$(2.45~2.70, n=10)mm in diameter. The yolk had not oil globule. The first cleavage was observed 2 hrs after insemination at $21{\sim}23^{\circ}C$, and the progressive cleavage were done about 30 min. interval. The characteristic changing of the yolk surface started at morula stage and continued to the end of gastrula. Hatching was started 72 hrs and completed 90 hrs after fertilization. The size of the larvae were 5.41~5.81mm in total length and 2.76~2.94mm in preanal length, and the number of so mites was 15-16+33~34(48~50). The barbels and swimbladder were completed and all the fins except second dorsal were appeared 1 week after hatching. The larvae attained 9.67~10.52mm in total length and 5.20~5.65mm in preanal length. All the fin sets and color pattern were completed 2 weeks after hatching and body mucus was secreted at that stage. The juvenile attained 14.59~16.02mm in standard length, 3.31~4.16mm in head length and 8.07~9.31mm in prenal length.

• Korean Journal of Ichthyology
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• v.1 no.1_2
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• pp.9-18
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• 1989

Blenniid fish, Istiblennius stellifer(jordan et Snyder) is distributed in the coastal waters of south-eastern Korea and Japan. Matured adults of blenniid fish were collected from the rocky shore of Namchun-dong, Nam-gu, Pusan, Korea on May 15, 1988. The fertilized eggs were incubated and the larvae were reared in laboratory. The eggs of this species were demersal and adhesive, and their diameters varied from 0.84 to 0.88 mm(mean 0.86 mm, n=30). They have a number of small oil globules. The water temperature throughout incubation ranged from 18.5 to $23.3^{\circ}C$ and salinity was maintained at $28.2-29.5\;^{\circ}/_{\circ\circ}$. The hatching took place in 130 hours after fertilization. The newly hatched larvae were 2.70 mm in total length with 11 (abdominal)+22~25 (caudal)=33-36 myomeres. The larvae absorbed the yolk material and oil globule completely in 10 days after hatching and became postlarvae. Total lengths of the larvae reached 4.65 and 5.75 mm in 10 and 13 days after the hatching, respectively.

• Korean Journal of Ichthyology
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• v.19 no.4
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• pp.350-359
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• 2007

The egg development and morphological change of larvae and juveniles of the starry flounder, Platichthys stellatus were observed in laboratory. Fertilized eggs of the species, 1.09~1.19 mm (mean $1.13{\pm}0.03mm$, n=50) in diameter, were floating, colorless, transparent in shape and lacked in oil globules. The eggs hatched out 121 hours after fertilization at water temperature $8.2{\sim}11.2^{\circ}C$. The size of the hatched larvae were 2.58~2.89 mm (mean $2.67{\pm}0.09mm$) in total length, their mouth and anus were not open yet and myotome number was 14+27=41. Melanophore and xanthophore appeared on the notochord and digestive organ and the margin of membrane fin, on the yolk sac and eyes were lacking in pigment cells. 5 days after hatching the larvae attained 4.30~4.97 mm (mean$4.74{\pm}0.21mm$) in TL, and their mouth and anus were open. 10 days after hatching the larvae transformed to postlarval stage and they were 4.67~5.75 mm in TL (mean $5.30{\pm}0.31mm$), and absorbing the yolk completely. Feeding activity increased as the mouth became larger. At 23 days, the larvae attained 6.69~8.82 mm in TL (mean $7.85{\pm}0.75mm$), and the right eye was started moving to the left side of the head. At 52 days, the juveniles attained 10.99~17.06 mm in TL (mean $13.50{\pm}1.67mm$). The right eye was moved completely onto the left side. All of the fins had completed set of the fin rays (D. 64~67: A. 45~51: P. 11: V. 6: C. 19).

• The Korean Journal of Malacology
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• v.17 no.1
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• pp.45-55
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• 2001

Gametogenes, reproductive cycle, first sexual maturity(biological minimum size), sex ratio and larval development of the marsh clam Corbicula japonica were investigated monthly by histological observations. Samples were collected in brackish water of Gokgang stream, Kyungsangbuk-Do, Korea, from August 1997 to July 1998. Sexuality of Corbicula japonica is dioecious and the species are an oviparous clam. The gonads are irregularly arranged from the sub-region of mid-intestinal gland in visceral cavity to reticular connective tissue of foot. The ovary is composed of a number of ovarian sac which are branched arborescent. Oogonia actively proliferate along the germinal epithelium of ovarian sac, in which young oocytes are growing. The testis is composed of a number of testicular tubules, and the epithelium of the tubule has function of germinal epithelium, along which spermatogonia actively proliferate. A great number of undifferentiated mesenchymal tissue and eosinophilic granular cells are abundantly distributed between developing oocytes and spermatocytes in the early developmental stages. With the further development of the ovary and testis these tissue and cells gradually disappear. Then the undifferentiated mesenchymal tissue and eosinophilic granular cells are considered to be related to the growing of the oocytes and spermatocytes. The spawning period is from July to September, and the main spawning occur between July and August when seawater temperatures reach above 22$^{\circ}C$. The reproductive cycle of this species can be divided into five successive stages; early active (February to April), late active (May to July), ripe (June to September), partially spawned (July to September), degenerative (September to October) and resting stage (October to February). Percentages of first sexual maturity of female and male clams ranging in length from 10 mm to 12 mm are over 50% and 100% for clams over 16.0 mm in shell length. Fertilized eggs or Corbicula japonica were 80-90 ${\mu}{\textrm}{m}$ in diameter. In the early embryonic development of C. japonica, the appearance of polar body, trochophore and D-shaped veliger were observed around 40 min., 27 hours and 4 days after spawning, respectively, at a water temperature of 26.5-28.$0^{\circ}C$. The size of larvae of early umbo stage was about 185-210 ${\mu}{\textrm}{m}$ in shell length, 160-180 ${\mu}{\textrm}{m}$ in shell height around 7 days after fertilization. The correlation of relative growth between the culture day (D) and shell length (SL) was expressed by the following simple formula from D-shaped veliger to metamorphosing stage; SL = 13.300D + 209.36($r^2$= 0.9078).

• Korean Journal of Animal Reproduction
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• v.24 no.1
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• pp.69-76
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• 2000

This study was carried out to improve of effective culture system on development of IVM/IVF/IVC bovine embryos. The cumulus-oocyte-complexes (COCs) collected from Korean cattle ovaries harvested at a local abattoir were matured in 50 ${mu}ell$ of TCM199 supplemented with 10% fetal bovine serum (FBS) and hormones (35 $\mu\textrm{g}$/$m\ell$ FSH, 10 $\mu\textrm{g}$/$m\ell$ LH, 1$\mu\textrm{g}$/$m\ell$ estradiol 17 $\beta$ under paraffin oil at 39$^{\circ}C$ in a humidified atmosphere of 5% $CO_2$in air. At 24 hrs after culture, matured oocytes were fertilized in vitro for 22~24 hrs with motile semen in which obtained by centrifugation of a frozen thawed semen on Percoll-density gradients (45% vs. 90%) at 500 g for 20 min. The presumptive zygotes were divided into three experimental groups. Single egg (Group 1), 25 (Group 2) or 50 eggs (Group 3) were cultured on cumulus cell in 50 ${mu}ell$ TCM199 supplement with 10% FBS for 6~9 days after fertilization. In vitro developmental rates into the blastocysts in the groups 2 and 3 were significantly (P＜0.05) higher than those of group 1 (37,27 vs. 6%, respectively). Cell number of blastocysts obtained in groups 2 and 3 at day 8 were significantly (P${mu}ell$) resulted in higher developmental competence and cell number of bovine blastocysts produced in vitro than those the culture of single embryos with cumulus cells.

• Korean Journal of Environmental Biology
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• v.23 no.3 s.59
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• pp.293-303
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• 2005

Japanese medaka, Oryzias latipes is widely distributed in the North East Asia including Korea, Japan and east China, and commonly used for freshwater toxicity tests and cytotoxicological studies worldwide. In this study, a series of experiments were conducted to identify the potential of the fish as a standard test species for saltwater toxicity evaluation such as marine receiving waters, ocean-dumped materials and sediment pore waters etc. Hatching, growth and mortality rates of the fish were estimated with the wide ranges of salinity from freshwater to seawater (35 psu). Direct exposure of the fertilized eggs in freshwater to the wide ranges of salinity (from 0 to 35 psu) without pre- acclimation to the saltwater revealed no significant differences in hatching rates by salinities (p =0.24). On the other hand, medaka larvae hatched in freshwater and exposed to saltwater directly showed high mortality at > 25 psu treatment groups (p < 0.0001). However, there was no significant difference in mortality of medaka larvae hatched in 13.8 and 14.2 psu at the wide ranges of salinities ($0\~35$ psu). Growth rates of medaka larvae hatched in the above two salinities showed no differences in body length either from 0 to 35 psu treatment groups (p =0.64 for 13.8 psu group and p=0.32 for 14.2 psu group). The number of gill chloride cell in medaka larvae sharply increased when the larvae were exposed to high salinity. Reference tests with zinc chloride revealed 96h $LC_{50}=8.84(7.19\~10.87)mg\;L^{-1}$ using 7~10 day old medaka larvae. These were comparable or better sensitivity in comparison with the other standard test species such as North American sheepshead minnow Cyprinodon variegatus. Based on the results of these experiments, hatching rates and larvalmortality of medaka must be good toxicity parameters for seawater bioassay and the species seems to be a good standard species for both the freshwater and seawater toxicity test.