• Asian-Australasian Journal of Animal Sciences
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• v.21 no.8
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• pp.1127-1133
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• 2008

This study was designed to develop a method of liquid storage of boar sperm at $4^{\circ}C$ by using the modified Beltsville F5 (BF5) diluent with bovine serum albumin (BSA) and N-acetyl-D-glucosamine. Boar sperm were stored in lactose-egg yolk and N-acetyl-D-glucosamine (LEN), BF5 and Golden-Pig liquid 4 (GPL4) diluents at $4^{\circ}C$ for 5 days and were examined for sperm viability, adenosine triphosphate (ATP) and in vitro fertilization (IVF). The percentage of sperm viability in GPL4 diluent was higher than in LEN and BF5 diluent from 1 to 5 days of storage at $4^{\circ}C$. The percentage of sperm viability steadily declined from 1 to 5 days of storage in the three different diluents. Sperm ATP in GPL4 diluent was higher than in LEN and BF5 diluents from 1 to 5 days of storage. Sperm ATP rapidly declined after 5 days of storage in the three different diluents. Porcine oocytes matured in vitro were inseminated with different sperm concentrations of liquid semen stored for 3 days in GPL4 diluent. The percentage of monospermic oocytes did not show any differences from 2.5 to $20{\times}10^5$ sperm/ml. However, the percentage of polyspermic oocytes in the sperm concentration of $2.5{\times}10^5$ sperm/ml was lower than in concentrations of 5, 10 and $20{\times}10^5$5 sperm/ml. The percentage of blastocysts from the cleaved oocytes at $2.5{\times}10^5/ml$ sperm concentration was significantly lower than at 5, 10 and $20{\times}10^5sperm/ml$ concentrations. In conclusion, GPL4 diluent can be stored at $4^{\circ}C$ for 5 days and showed higher sperm viability and sperm ATP concentration compared with LEN and BF5 diluents. Also, we found that GPL4 diluent can be used for IVF of porcine oocytes.

• Clinical and Experimental Reproductive Medicine
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• v.42 no.4
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• pp.163-168
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• 2015

Objective: To investigate the impact of fibroids on the blood flow of the uterine and subendometrial arteries and in vitro fertilization (IVF) outcomes. Methods: In this study, we analyzed 86 IVF/intracytoplasmic sperm injection (ICSI) cycles in which a gonadotropin-releasing hormone antagonist protocol was used for controlled ovarian stimulation between January 2008 and March 2009. The subjects comprised 86 infertile women with (fibroid group, n=43) or without (control group, n=43) uterine fibroids. Results: Patient characteristics were similar between the fibroid and control groups. The IVF/ICSI outcomes in patients with fibroids were similar to those of patients in the control group. The resistance index (RI) and pulsatile index (PI) of the uterine and subendometrial arteries on the day of embryo transfer were also comparable between the two groups. IVF outcomes and uterine hemodynamics in patients with multiple (${\geq}2$) fibroids were similar to those of patients with a single fibroid. However, clinical pregnancy and implantation rates were significantly lower in patients with fibroids who experienced uterine cavity distortion than in patients with fibroids who had a normal uterine cavity (both p<0.05). The RI and PI of the subendometrial artery were significantly higher on the day of embryo transfer in patients with fibroids who experienced uterine cavity distortion than in patients with fibroids who had a normal uterine cavity (both p<0.05). Conclusion: Fibroids which distorting the uterine cavity might impair the subendometrial artery blood flow clinical pregnancy rate and embryo implantation rate in infertile patients undergoing IVF. Otherwise, IVF outcomes were not influenced by the presence of uterine fibroids.

• Clinical and Experimental Reproductive Medicine
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• v.44 no.2
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• pp.63-72
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• 2017

Objective: Hyperstimulation methods are broadly used for in vitro fertilization (IVF) in patients with infertility; however, the side effects associated with these therapies, such as ovarian hyperstimulation syndrome (OHSS), have not been well studied. N-glycoproteomes are subproteomes used for the remote sensing of ovarian stimulation in follicular growth. Glycoproteomic variation in human follicular fluid (hFF) has not been evaluated. In this study, we aimed to identify and quantify the glycoproteomes and N-glycoproteins (N-GPs) in natural and stimulated hFF using label-free nano-liquid chromatography/electrospray ionization-quad time-of-flight mass spectrometry. Methods: For profiling of the total proteome and glycoproteome, pooled protein samples from natural and stimulated hFF samples were selectively isolated using hydrazide chemistry to obtain the total proteomes and glycoproteomes. N-GPs were validated by the consensus sequence N-X-S/T (92.2% specificity for the N-glycomotif at p<0.05). All data were compared between natural versus hyperstimulated hFF samples. Results: We detected 41 and 44 N-GPs in the natural and stimulated hFF samples, respectively. Importantly, we identified 11 N-GPs with greater than two-fold upregulation in stimulated hFF samples compared to natural hFF samples. We also validated the novel N-GPs thyroxine-binding globulin, vitamin D-binding protein, and complement proteins C3 and C9. Conclusion: We identified and classified N-GPs in hFF to improve our understanding of follicular physiology in patients requiring assisted reproduction. Our results provided important insights into the prevention of hyperstimulation side effects, such as OHSS.

• Clinical and Experimental Reproductive Medicine
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• v.42 no.4
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• pp.143-148
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• 2015

Objective: The aim of the current study was to determine the predictive value of anti-$M{\ddot{u}}llerian$ hormone (AMH) levels for pregnancy outcomes in patients over 40 years of age who underwent in vitro fertilization or intracytoplasmic sperm injection-embryo transfer (IVF/ICSI-ET) cycles. Methods: We retrospectively analyzed the medical records of 188 women aged 40 to 44 years who underwent IVF/ICSI-fresh ET cycles due to unexplained infertility in the fertility center of CHA Gangnam Medical Center. Patients were divided into group A, with AMH levels <1.0 ng/mL (n=97), and group B, with AMH levels ${\geq}1.0ng/mL$ (n=91). We compared the clinical pregnancy rate (CPR) in the two groups and performed logistic regression analysis to identify factors that had a significant effect on the CPR. Results: The CPR was significantly lower in group A than group B (7.2% vs. 24.2%, p<0.001). In multivariate logistic regression analysis, AMH levels were the only factor that had a significant impact on the CPR (odds ratio, 1.510; 95% confidence interval, 1.172-1.947). The area under the receiver operating characteristic curve for AMH levels as a predictor of the CPR was 0.721. When the cut-off level of AMH was set at 1.90 ng/ mL, the CPR was 6.731-fold higher in the group with AMH levels ${\geq}1.90ng/mL$ than in the group with AMH levels <1.90 ng/mL (p<0.001). Conclusion: Our study showed that AMH levels were predictive of clinical pregnancy in infertility patients over 40 years of age. Further prospective studies should be conducted to validate the predictive capability of AMH levels for the outcome of clinical pregnancy.

• Reproductive and Developmental Biology
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• v.38 no.3
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• pp.115-121
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• 2014

Humulus japonicus is an ornamental plant in the Cannabaceae family. Although the mode of action of Humulus japonicus is not fully understood, a strong relationship was observed between anti-inflammatory and anticancer in some types of cells. Recent studies also have shown that Humulus japonicus possesses anti-inflammatory activities and may significantly improve antioxidant potential in Raw 264.7 macrophage cells. Thus, the aim of this study was evaluated the effect of Humulus japonicus extract on sperm motility and subsequent preimplantation developmental competence of the bovine embryos. After in vitro maturation, the oocytes with sperms were exposed in in vitro fertilization (IVF) medium supplemented with Humulus japonicus extract (0.01, 0.05, $0.1{\mu}g/mL$, respectively) for 1 day. In our results, exposure of IVF medium to Humulus japonicus extract did not affect sperm motility and percentage of penetrated oocytes but ROS intensity was significantly decreased by $0.01{\mu}g/mL$ compared with other groups (p< 0.05). Moreover, treatment with $0.01{\mu}g/mL$ of Humulus japonicus extract was higher the frequency of blastocyst formation than the any other groups (p<0.05). Otherwise, treatment with $0.01{\mu}g/mL$ of Humulus japonicus extract not increased the total cell number but reduced apoptotic-positive nuclei number. In conclusion, our results indicate that supplementation of Humulus japonicus extract in IVF medium may have important implications for improving early embryonic development in bovine embryos.

• Clinical and Experimental Reproductive Medicine
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• v.26 no.3
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• pp.339-344
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• 1999

Objective: To evaluate incidence of microbial growth from the tip of the embryo transfer catheter after embryo transfer in relation to clinical pregnancy rate following in-vitro fertilization and embryo transfer. Method: This study was performed prospectively at the time of transcervical embryo transfer following conventional in-vitro fertilization and intracytoplasmic sperm injection procedures. Sixty three patients were enrolled in this study. Microbiological cultures were performed on endocervical swabs and embryo transfer catheter tips. Results: Positive microbial growths were observed from endocervical swabs in 45 (71.4%) women and from catheter tips in 30 (47.6%) women. There was no statistically significant difference seen in the mean number of oocytes fertilized or number and grade of embryos transferred between the group of patients without growth and the group of patients with positive microbial growth from catheter tips. The clinical pregnancy rate were 30.3% in the group of patients without growth and 13.3% in the group with positive microbial growth from catheter tips. This difference in clinical pregnancy rates was statistically significant. Conclusion: Our finding is that microbial contamination at embryo transfer may influence implantation rates. The major questions arising from our finding are whether eradication of endocervical micro-organisms is possible and whether their eradication will improve implantation rates.

• Journal of The Korean Society of Grassland and Forage Science
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• v.38 no.4
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• pp.320-324
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• 2018

In this study, we examined effect of sperm penetration of oocytes after in vitro fertilization (IVF) with cauda epididymal spermatozoa in Hanwoo bull after feeding of timothy hay. One testicle with epididymides was castrated from one Hanwoo bull (14 months of age) and spermatozoa recovered from cauda epididymis and cryopreserved. As control, frozen Hanwoo semen was used. Matured cumulus oocyte complexes were co-incubated with frozen-thawed cauda epididymal spermatozoa for 12 or 18 hours. After IVF, presumptive zygotes were cultured in modified synthetic oviductal fluid. In experiment 1, we examined sperm penetration rate at 12 hours of IVF with epididymal sperm. Total penetration rate among cauda epididymis and control was similar(mean${\pm}$standard error, cauda epididymis and control vs. $49.7{\pm}11.3$ and $54.4{\pm}12.8%$). In experiment 2, cleavage and blastocyst developmental rate were evaluated at day 2 and day 8 after IVF for 18 hours. Cleavage rate among cauda epididymis and control was similar(cauda epididymis and control vs. $81.2{\pm}3.4$ and $82.7{\pm}2.5%$). However, blastocyst developmental rate of cauda epididymis group was significantly higher than that of control group(cauda epididymis and control vs. $24.4{\pm}1.6$ and $12.2{\pm}2.8%$, p<0.05). In conclusion, cauda epididymal spermatozoa in Hanwoo bull has high embryo developmental competence and can be used as an alternative to ejaculated frozen sperm in vitro.

• Clinical and Experimental Reproductive Medicine
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• v.19 no.1
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• pp.31-39
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• 1992

By means of the results of In vitro fertilization(IVF) of supernumerary oocytes, the possibility of predicting a pregnancy outcome following gamete intrafallopian transfer(GIFT) and the prognostic implications for future pregnancy were evaluated in 46 GIFT cycles excluding simultaneous program of GIFT and IVF from February, 1989 to July, 1991. IVF of supernumerary oocytes were identified in 21 cycles, but not in remaining 25 cycles. There was no significant difference in age, duration and etiologic factors of infertility, and serum levels of FSH, LH and $E_2$ on MCD #3 and $E_2$ on the day of hCG injection between fertilized(N=21) and unfertilized group(N=25). The number of oocytes retrieved was similar in both groups. The number of supernumerary oocytes available for IVF after transfer was $5.43{pm}2.95$ ranging from 2 to 12. The prenancy rate in fertilized group, 33.3%(7/21), was higher without statistical significance, compared with 8.0%(2/25) in unfertilized group. Using IVF of supernumerary oocytes as a test of pregnancy following GIFT, sensitivity was 77.8 %; specificity, 61.2%; positive predictive value(PPV), 33.3%; negative predictive value(NPV), 92%. The fertilization rate of supernumerary oocytes in pregnant group, $86.4{\pm}22.8%$ was significantly higher compared with $56.1{\pm}20.2%$ in nonpregnant group. In cases with fertilization rate ${\geq}80%$, pregnancy was expected with PPV of 85.7%. In conclusion, IVF of supernumerary oocytes in GIFT program can be a profitable method as a prognostic indicator of pregnancy following GIFT. More aggressive diagnostic and therapeutic measures should be performed in cases with failure in IVF of supernumerary oocytes.

• Journal of Embryo Transfer
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• v.13 no.2
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• pp.147-157
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• 1998

This study was conducted to investigate the effects of in vitro fertilization, culture and embryo development according to in vitro maturation rate, protectant composition and equilibrium time after frozen /thawing of bovine immature oocytes. This results obtained in studies on the effect of different cryoprotectants on the viability, maturation and development of in vitro bovine oocytes were as follow: 1.The post-thawing of immature oocytes matured to metaphase II during culture time for 0 to 26 h, and those group (62~3%) were low than control group (76.7%). The optimal maturation time of frozen-thawed immature oocytes was at 24 h. 2.The viability of cryopreserved immature oocytes was not affected by sort of cryoprotectants. The developmental competence of frozen4hawed oocytes was not affected by cryoprotectants. These results indicate that an optimal maturation time of frozen /thawed immature oocytes was at 24h. Furthermore the viability of cryopreserved immature oocytes was not affected by sort of cryoprotectants and developmental competence of frozen /thawed oocytes.

• Korean Journal of Animal Reproduction
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• v.24 no.3
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• pp.311-317
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• 2000

This study was carried out to compare the temperature and time of heat shock, and the effect of heat shock on development of embryos after in vitro maturation and fertilization in bovine oocytes. The results obtained were as follows. 1. The optimum temperature and time of heat shock were 41$^{\circ}C$ and 30sec on in vitro development of embryos from 4~8 cell to blastocyst. 2. The rates of cleavage on zygotes produced on in vitro were significantly increased by heat shock after IVM than before IVM(P<0.05). 3. When the oocytes were treated heat shock after IVM and 5 days cultured, developmental rates to blastocyst were increased than other experimental treatments.