• Journal of Animal Reproduction and Biotechnology
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• v.35 no.1
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• pp.50-57
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• 2020

The objective of this research work was to know ovarian dynamics and pregnancy rate of cyclic Murrah buffalo cows with induced estrus by administration of prostaglandin F2α (PGF2α) and timed artificial insemination (TAI) with frozen thawed semen. A total of 31 female buffaloes were selected for the study. The buffalos having matured CL observed by ultrasonography were given one intra muscular injection of cloprostenol 500 ㎍ and TAI was performed using frozen thawed semen of Indian Murrah buffalo bull. Results showed that 90.32% (significantly, at p < 0.05) cows explore the sign of heat after injection of PG and 67.85% (significantly, at p < 0.05) cows were become pregnant out of 28 inseminated (TAI) cows. In the 28 inseminated (TAI) cows, average number of smaller and larger size of follicles were non-significantly (p > 0.05) higher at day 3 post PG injection, but the medium size of follicles was nonsignificantly (p > 0.05) higher at PG injection. At day 3 post PG injection the diameter of follicles was significantly (p < 0.05) higher, but the diameter of CL was significantly (p < 0.01) lower compared at PG injection. At PG injection the diameter of largest follicle was non-significantly differences (p > 0.05) in between pregnant and non-pregnant cows. But at day 3 post PG injection it was significantly (p < 0.01) higher in pregnant cows compared to non-pregnant cows. The number of small, medium, and large follicles at PG injection and at day 3 post PG injection were non-significantly (p > 0.05) difference in between pregnant and non-pregnant buffalo cows. Finally, it is concluded that the CL was effectively regresses and induced the sign of heat in buffalo cows and after AI the cows were become pregnant with significant rate. The study will help to the veterinarian and researcher to know the efficacy of PG injection and AI for reproductive efficiency in buffalo cows.

• Clinical and Experimental Reproductive Medicine
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• v.32 no.4
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• pp.315-324
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• 2005

Objectives: To compare the efficacy of GnRH antagonist multiple dose protocol (MDP) with that of GnRH agonist long protocol (LP) in controlled ovarian hyperstimulation for in vitro fertilization in patients with high basal FSH (follicle stimulating hormone) level or old age, a retrospective analysis was done. Methods: Two hundred ninety four infertile women (328 cycles) who were older than 41 years of age or had elevated basal FSH level (> 8.5 mIU/mL) were enrolled in this study. The patients had undergone IVF-ET after controlled ovarian hyperstimulation using GnRH antagonist multiple dose protocol (n=108, 118 cycles) or GnRH agonist long protocol (n=186, 210 cycles). The main outcome measurements were cycle cancellation rate, consumption of gonadotropins, the number of follicles recruited and total oocytes retrieved. The number of fertilized oocytes and transferred embryos, the clinical pregnancy rates, and the implantation rates were also reviewed. And enrolled patients were divided into three groups according to their age and basal FSH levels; Group A - those who were older than 41 years of age, Group B - those with elevated basal FSH level (> 8.5 mIU/mL) and Group C - those who were older than 41 years of age and with elevated basal FSH level (> 8.5 mIU/mL). Poor responders were classified as patients who had less than 4 retrieved oocytes, or those with $E_2$ level <500 pg/mL on the day of hCG injection or those who required more than 45 ampules of exogenous gonadotropin for stimulation. Results: The cancellation rate was lower in the GnRH antagonist group than in GnRH agonist group, but not statistically significant (6.8% vs. 9.5%, p=NS). The amount of used gonadotropins was significantly lower in GnRH antagonist group than in agonist group ($34.8{\pm}11.3$ ampules vs. $44.1{\pm}13.4$ ampules, p<0.001). The number of follicles > 14 mm in diameter was significantly higher in agonist group than in antagonist group ($6.7{\pm}4.6$ vs. $5.0{\pm}3.4$, p<0.01). But, there were no significant differences in clinical pregnancy rate (24.5% in antagonist group vs. 27.4% in agonist group, p=NS) and implantation rate (11.4% in antagonist group vs. 12.0% in agonist group, p=NS) between two groups. Mean number of retrieved oocytes was significantly higher in GnRH agonist LP group than in GnRH antagonist MDP group ($5.4{\pm}3.5$ vs. $6.6{\pm}5.0$, p<0.0001). But, the number of mature and fertilized oocytes, and the number of good quality (grade I and II) and transferred embryos were not different between two groups. In each group A, B, and C, the rate of poor response did not differ according to stimulation protocols. Conclusions: In conclusion, for infertile women expected poor ovarian response such as who are old age or has elevated basal FSH level, a protocol including a controlled ovarian hyperstimulation using GnRH antagonist appears at least as effective as that using a GnRH agonist, and may offer the advantage of reducing gonadotropin consumption and treatment period. However, much work remains to be done in optimizing the GnRH antagonist protocols and individualizing these to different cycle characteristics.

• Journal of Embryo Transfer
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• v.22 no.1
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• pp.53-61
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• 2007

This study was investigated factors affecting the pregnancy rates after transfer of pronuclear microinjected embryos for the production of transgenic Korean black goats. Embryo transfer was carried out in 343 recipient Korean black goats from September 1999 to June 2000. Estrus was induced by the insertion of intravaginal progesterone devices $CIDR^(R)$ for 2 weeks. A single injection of 400 IU equine chorionic gonadotropin was administered at 48h before $CIDR^(R)$ removal to increase the proportion of does cycling and ovulation rate. Good quality embryos were prepared by microinjection of DNA into the pronuclei of fertilized goat oocyte and cultured in vitro. Pronuclear microinjected $1{\sim}8$ cell stage embryos were surgically transferred into the oviducts of the recipient at day 4 or 5 following $CIDR^(R)$ removal, and morula to blastocyst stage embryos were surgically transferred into uterus at day 9. Pregnancy was diagnosed by transrectal ultrasound scanning at $20{\sim}30d$ and 8 weeks following embryo transfer. The pregnancy rate was affected by several factors, such as estrus induction, the number of previous transfer, transfer site, stage of CL (corpus luteum), the number of recipient CL, stage of embryos and the number of transferred embryo. The pregnancy rate was significantly higher in recipients that came into estrus naturally than recipients that induced to come into estrus with $CIDR^(R)$(59.1% vs. 36.8%; P<0.05). The pregnancy rate was higher when the embryos were transferred into the left oviduct than transferred into the right oviduct (42.9% vs. 35.3%; P<0.05). The pregnancy rate of recipients with $CH_1$ (early) stage corpus hemorrhagicum in ovary was hi틴or than recipient with $CH_3$ (late) stage hemorrhagicum (47.5% vs. 17.9%; P<0.01). Higher pregnancy rates were obtained by transfer of 1-cell stage embryos into oviduct while late blastocysts (51.6% vs. 66.7%; P<0.01) into uterus. The pregnancy rates when 3 embryos were transferred to recipients were significantly higher than when 2 embryos we.e transferred (47.6% vs. 27.0%; P<0.05). Although there were no significant difference among the group, adhesion of reproductive organs, uterine size, ovulation rate of recipients, presence of large follicle and difficulty of transfer affected pregnancy rate of recipient. Higher pregnancy rates were obtained in the recipients with $8{\sim}15m$ diameter uterine horn as compared to the recipients with <5m diameter or >20mm diameter uterine hem (38.9%, 20% vs. 18.2%), in the recipients with large follicle in the ovulated ovary ipsilaterally (53.6% vs. 37.1%) and in the transfer which was carried out easily (39.2% vs. 27.8%, 0%). In conclusion, the high rate of pregnancy was achieved following transfer of pronuclear microinjected embryos when three or four 1-cell stage embryos were transferred into oviduct with $CH_1$ stage corpus hemorrhagicum in the ovary of recipient which came into estrus naturally.

• Journal of Embryo Transfer
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• v.19 no.2
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• pp.113-119
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• 2004

The purpose of the present study was to examine whether collection time affects results of oocyte recovery from superovulated goats. Fiftyty-one mature Korean native goats, maintained in a pen under natural day length and fed hay ad libitum, were pretreated with progestagen impregnated CIDR for 10 days and then the goats were divided into two groups. One group of the goats received a single intramuscular injection of 1,000 IU PMSG on Day 8 of CIDR insertion. The other group of the goats received twice daily intramuscular injections of a total of 70 mg FSH for 3 days from Day 8 of CIDR. All the gonadotropin treated goats were injected with 10 mg $PGF_{2\alpha}$ on Day 8 and 400 IU hCG in the afternoon on Day 10. For oocyte recovery, donor goats were fasted 24 h before operation. Anesthesia was induced by intravenous injection of 2% xylazine(0.2 mg/kg body weight) and ketamin(11 mg/kg body weight). In vivo oocytes were recovered by follicle aspiration or oviduct flushing at 29 to 34, 35 to 40 and 41 to 50 h after hCG injection through mid-ventral incision. There was no significant difference in the mean number of CL and oocytes recovered. Oocyte collection at 29 to 40 h after hCG increased(P<0.05) the recovery rate of ovulated oocytes in oviducts compared to 41 to 50 h. The same results were also observed in the recovery of follicular oocytes. Oocyte grade was not affected by collection time. When oocytes were collected from follicular oocytes at 41 to 50 h after hCG, the recovery rate of Grade II oocytes was the lowest(P<0.05). From these results, it is suggested that oocyte recovery at 35 to 40 h after hCG will be successful for further use.

• Ocean and Polar Research
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• v.37 no.1
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• pp.49-59
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• 2015

Spatial variation in the reproductive effort of Manila clam Ruditapes philippinarum is often closely associated with variation in the seawater temperature and food availability, which determines gonad maturity and the quantity of gamates produced during spawning. Previous studies also have reported that severe infection by the protozoan parasite Perkinsus olseni exerts a negative impact on clam reproduction, retarding gonad maturation or decreasing the reproductive effort. In the present study, we investigated impacts of P. olseni infection on the reproductive condition of Manila clam during a spawning season. Histology revealed that 54% of female clams in Wando off the south coast were in spawning, while only 10% of the female from Gomso and 0% of the female from Seonjaedo in Gyeonggi bay off the west coast were engaged in spawning at the end of May in 2004. Ray's fluid thioglycollate media (RFTM) assay was applied to assess P. olseni infection and indicated that the infection intensity in Wando ($3,608,000{\pm}258,000cells/g$ wet tissue) was significantly higher than the levels in Gomso ($1,305,000{\pm}106,000cells/g$ wet tissue) and Seonjaedo ($1,083,000{\pm}137,000cells/g$ wet tissue, p < 0.001). The size of the ripe female follicle determined from histology was significantly smaller in Wando ($0.032mm^2$) compared to the sizes in Gomso ($0.059mm^2$) and Seonjaedo ($0.052mm^2$, p < 0.05). Accordingly, the number of ripe eggs in the follicle was significantly fewer among clams in Wando (14) compared to the numbers determined in Gomso (23) and Seonjaedo (22). The absolute quantity of egg in ripe clams from Wando (31.01 mg) was also significantly smaller than Seonjaedo (61.79 mg) and Gomso (133.3 mg). Quantity of total protein, carbohydrate, and lipid in the tissue in the Wando samples was significantly smaller than the quantities determined in Gomso and Seonjaedo (p < 0.001). The observed poor reproductive condition and proximate tissue composition of the females in Wando were, in part, explained by the extremely high level of the parasites, sapping the ability to store energy in the host tissues, which is used in tissue growth and the egg production.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.32 no.5
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• pp.629-636
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• 1999

Gonadal development, gametogenesis, reproductive cycle, egg-diameter and composition, condition factor, and the first sexual maturity of the clam, Potamocorbula amurensis were investigated by histological observation. Samples were collected monthly from the tidal flat of Moonpo, Puan-gun, Chollabuk-do, west coast of Korea from November 1996 to October 1997. P. amurensis is dioecious and oviparous. The gonads were composed of a number of gametogenic follicles. The oogonia and fully ripe oocytes were $9\~12\mu$m and $50\~60\mu$m in diameter, respectively. Each of the spermatogenic follicle formed stratified layers composed of spermatogonia, spermatocytes spermatids, and spermatozoa in groups on the follicular wall. The reproductive cycle of P. amurensis could be classified into five successive stages: early active, late active, ripe, partially spawned, and recovery. Spawning occurred twice a year from May to July and from September to October, the main spawning seasons also appeared twice a year between May and June, and in October when the water temperatures reached above $18^{\circ}C$. The monthly changes in the condition factor were closely related with the reproductive cycle. Minimum size for the sexual maturation of female and male were 8.1 mm in shell length. There were two patterns for the gametogenesis: 1. After spawning, the undischarged ripe oocytes and spermatozoa in the follicles were degenerated and absorbed, but in part, the existing follicles were not contracted significantly and then they took part in new gametogenesis within one or two months (especially, in summer). 2. After spawning, each follicle was contracted, thereafter gametogenesis again occurred in newly formed follicles.

• Clinical and Experimental Reproductive Medicine
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• v.34 no.2
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• pp.125-131
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• 2007

Objective: The aim of this study was to compare GnRH antagonist and agonist flare-up treatment in the management of poor responder patients. Methods: One hundred forty-four patients from Jan. 1, 2002 to Aug. 31, 2005 undergoing IVF/ICSI treatment who responded poorly to the previous cycle (No. of oocyte retrieved$\leq$5) and had high early follicular phase follicle stimulating hormone (FSH>12 mIU/ml were selected. Seventy-five patients received agonist flare-up protocol and 71 patients received antagonist protocol. We analyzed the number of oocytes retrieved, number of good embryos (GI, GI-1), total dose of hMG administered, implantation rate, cycle cancellation rate, pregnancy rate, live birth rate. Results: The cancellation rate was high in antagonist protocol (53.5% vs. 30.1%). The number of oocyte retrieved, the number of good embyos were high in agonist flare-up group. There was no statistical difference between GnRH agonist flare up protocol and GnRH antagonist protocol in implantation rate (14.5%, 10.1%), clinical pregnancy rate per transfer (29.4%, 21.2%) and live birth rate per transfer (21.6%, 18.2%). Although the result was not statistically significant, GnRH agonist flare up group showed a nearly doubled pregnancy rate and live birth rate per initial cycle than GnRH antagonist group. Conclusions: The agonist flare-up protocol appears to be slightly more effective than the GnRH antagonist protocol in implantation rate, pregnancy rate, live birth rate but shows statistically no significance. Agonist flare-up protocol improved the ovarian response in poor responders. However, based of the result of the study, we can expect improved ovarian response in poor responders by GnRH agonist flare up protocol.

• Clinical and Experimental Reproductive Medicine
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• v.17 no.1
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• pp.29-44
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• 1990

In 105 patients with the past history of poor response to the previous controlled ovarian hyperstimulation(COH) due to poor follicular growth or premature LH surge, the effectiveness of pituitary suppression with gonadotropin-releasing hormone agonist(GnRH agonist) in IVF/GIFT program was evaluated in 112 cycles of COH using a combination regimen of Leuprolide acetate (Lupron TAP Pharmaceuticals, USA) and FSH/hMG or pure FSH from May to December, 1989 at SNUH. Starting on day 21 of the menstrual cycle(MCD #21, Day 1), Lupron (1.0mg/day, subcutaneous) was administered once a day till next MCD #3(suppression phase). After the confirmation of pituitary suppression, ovarian follicular growth was stimulated with FSH/hMG or pure FSH from MCD #3(Day + 1), and Lupron was continued with hMG or FSH until hCG administration (D 0) (stimulation phase). After suppression phase, serum E2 level decreased from 183.7${\pm}$95.1(Day 1) to 17.4${\pm}$12.3pg/ml (Day +1), and serum progesterone level from 19.17${\pm}$8.67 to 0.12${\pm}$0.05ng/ml. But there was no decresas in serum LH and FSH levels; LH from 12.74${\pm}$6.21 to 15.49${\pm}$4.93mIU/ml,FSH from 7.60${\pm}$3.84 to 8.58${\pm}$3.15 rnlU/ml. There was no occurrence of premature LH surge during COH. Eleven cycles(9.8%) were cancelled due to poor follicular growth during stimulation phase, and 3 cycles (3.0%) failed in the transvaginal oocytes fretrieval. Serum E2 level was 1366.8${\pm}$642.4 on D 0 and 1492.3${\pm}$906.9pg/ml on D+1. 7.00${\pm}$3.32 follicles(FD${\geq}$12mm) were observed on D 0, and 6.11${\pm}$4.15 oocytes were retrieved, with the oocyte retrieval rate per follicle of 95.0%. 3.59${\pm}$2.57 oocytes were fertilized and cleaved with the oocyte cleavage rate of 55.7%. In 83 IVF patients, 4.08${\pm}$2.39 embryos were transferred, and 16 pregnancies were obtained with the pregnancy rate per ET 2.39 mebryos were transferred, and 16 pregnancies were obtained with the pregnancy rate per ET of 19.3%. In 6 GIFT patients, 7.83${\pm}$3.31 oocytes were retrieved and transferred with maximum number of 6, but no pregnancy was obtained. When compared with the previous 108 cycles of COH using FSH/hMG or pure FSH regimen, the cancellation rate during COH was significantly decreased, and all the parameters of the outcome of COH including the pregnancy rate were increased. These data suggest that GnRH agonist therapy for pituitary suppression is an effective adjunct to the current gonadotropin regimens for COH in IVF/GIFT and can increase the probability of oocytes retrieval and pregnancy, especially in the previous poor responders.

• Journal of Embryo Transfer
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• v.24 no.3
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• pp.213-220
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• 2009

The objective of this study was to examine the effect of eCG and various concentrations (20, 40, and 80 ${\mu}g/ml$) of porcine FSH on nuclear maturation and intracellular glutathione (GSH) level of oocytes, and embryonic development after parthenogenetic activation (PA) and somatic cell nuclear transfer (SCNT) in pigs. Immature pig oocytes were matured in TCM-199 supplemented with porcine follicular fluid, cysteine, pyruvate, EGF, insulin, and hormones (10 IU/ml hCG and 10 IU/ml eCG or $20{\sim}80{\mu}g/ml$ FSH) for the first 22 h and then further cultured in hormone-tree medium for an additional 22 h. Nuclear maturation of oocytes ($85{\sim}89%$) was not influencem foreCG and various concentrations FSH. Embryonic development to the cleavage stage ($86{\sim}94%$) and mean number of cells in blastocyst ($33{\sim}37$ cells) after PA were not altered but blastocyst formation e-treignificaddlor(p<0.05) improvem forthe supplementation eith 80 ${\mu}g/ml$ FSHr(64%) compared to 47%, io8%, iand 47% in oocytes that were treated with eCG, 20,i and 40 ${\mu}g/ml$ FSH,i numectivelo. In SCNT, fusion ($78{\sim}83%$) of cell-cytoplast couplets and siosequent embryo cleavage ($82{\sim}88%$) were not influencem fordifferent gonadotropins but blastocyst formation tended to increase forthe supplementation eith 80 ${\mu}g/ml$ FSHr(25% vs. $11{\sim}18%$). Our nuults demonstrated that oocyte maturation and embryonic development after PA and SCNT e-frinfluencem fortype of gcem fortype of gits concentration. In this study, supplementation of maturation medium eith 80 ${\mu}g/ml$ FSHrimproved preimplantation development of PA and SCNT pig embryos, probably by increasing intracellular GSH concentration of matured oocytes.

• Journal of Physiology & Pathology in Korean Medicine
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• v.21 no.1
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• pp.136-144
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• 2007

Biotae Orientalis has been widely used for treatment of relaxion of smooth muscle, gastrointestinal hemorrhage and alopecia in Oriental Medicine. This experiment examined the effect of an extracts, obtained from the acetone and MeOH extracts of dried or fresh Biotae Orientalis, on hair growing activity of the C57BL/6N mice after topical application to skin. First, We examined on hair growth activity of extracts of Biotae Orientalis compare to control and 1% minoxidil groups. Second, We investigated on the number of hair follicle and mast cells after topical application of extracts of the Biotae Orientalis to skin for 16 day. Third, We investigated immunoreactive density of vascular endothelial growth factor(VEGF), protein kinase C-${\alpha}$(PKC-${\alpha}$) and stem(mast) cell factor(SCF) in skin of C57BL/6N mice by immunohistochemical methods. I fourth investigated changes of subpopulation of splenocytes and thymocytes in C57BL/6N mice for 16day using laser flow cytometry. The results were as follows : Hair growing effect of acetone and MeOH extracts of dried and fresh Biotae Orientalis was observed in 70%, 90% and 60% in hair removed skin area in 16 day respectively. Immunoreactive density of VEGF and PKC-${\alpha}$ in skin of experimental groups was weakly stained compare to control group in 10 day. Immunoreactive density of stem cell factor in skin of experimental group was heavily stained compare to control group in 10 day. Splenic TH/TC Iymphocytes of lived MeOH extracts group significantly increased compare to control group. TH cells in thymic T lymphocytes were increased compare to control group. These experiment suggest that acetone and MeOH extracts of Biotae Orientalis may be used for topical treatment of alopecia areata.