Comparison of Clinical Efficacy between a Single Administration of Long-Acting Gonadotrophin-Releasing Hormone Agonist (GnRHa) and Daily Administrations of Short-Acting GnRHa in In Vitro Fertilization-Embryo Transfer Cycles

This study was aimed to evaluate the efficacy of a single administration of long-acting gonadotrophin-releasing hormone agonist (GnRHa) as compared with daily administrations of short-acting GnRHa in controlled ovarian hyperstimulation (COH) for in vitro fertilization and embryo transfer (IVF-ET) cycles. The mean dosage of recombinant follicle-stimulating hormone (rFSH) required for COH (2,354.5+/-244.2 vs. 2,012.5+/-626.1 IU) and the rFSH dosage per retrieved oocyte (336.7+/-230.4 vs. 292.1+/-540.4 IU) were significantly higher in the long-acting GnRHa group (N= 22) than those in the short-acting GnRHa group (N=28) (p<0.05). However, the mean number of visit to the hospital that was required before ovum pick-up (3.3+/-0.5 vs. 22.2+/-2.0) and the frequency of injecting GnRHa and rFSH (12.8+/-1.2 vs. 33.5+/- 3.5) were significantly decreased in the long-acting GnRHa group (p<0.0001). The clinical pregnancy rate, implantation rate, and early pregnancy loss rate were not significantly different between the 2 groups. So, we suggest that a single administration of long-acting GnRHa is a useful alternative for improving patient's convenience with clinical outcomes comparable to daily administrations of short-acting GnRHa in COH for IVF-ET cycles.

Sperm-specific Cation Channels, CatSper Family; Characteristics, Function, and Clinical Value / 대한남성과학회지

In human reproduction, fertilization is the first step for successful pregnancy. From the perspective of sperm physiology, the progressive motility and capacitation, including hyperactivation and acrosome reaction, are the most important factors in the fertilization of oocytes. Numerous studies have demonstrated the roles of calcium ions, cyclic nucleotides, and bicarbonate in the acquisition of progressive motility and capacitation. Among these factors, calcium ion plays the most important role. Sperm possess several calcium channels, including voltage-gated calcium channel, cyclic nucleotide-gated calcium channel, transient receptor potential channel, and channels of the CatSper family The CatSper family is a newly-identified group of four sperm-specific cation channels. CatSper1 and CatSper2 proteins localize on the sperm tail and play a critical role in sperm motility and fertilization. In contrast, CatSper3 and CatSper4 proteinsare expressed only in the acrosomal region of sperm head, which implies that they may have a role in the acrosome reaction. Taken together, the CatSper family is the most important group of calcium channels for regulating sperm physiology and appear to be an attractive target for non-hormonal male contraceptives.

Expression of Sperm-specific Cation Channel CatSper in Human Spermatozoa / 대한비뇨기과학회지

PURPOSE: We aimed to elucidate the expression and intracellular localization of sperm-specific cation channel CatSper in human spermatozoa. Moreover, the relationship between the expression of CatSper mRNA and the motility of ejaculated human spermatozoa were investigated. MATERIALS AND METHODS: Using cDNAs extracted from the ejaculated sperm of patients (n=39), the expression of CatSper mRNA was observed by RT-PCR. Semi-quantitative analysis of the CatSper mRNA expression was performed by comparing with the expression of GAPDH mRNA. To elucidate the expression and intracellular localization of CatSper protein, double fluorescent immunocytochemistry for CatSper and beta-tubulin was performed. RESULTS: The CatSper mRNA was expressed in all of the sperm samples. Using semi-quantitative analysis for the amount of CatSper mRNA expression, no significant difference was found between the normozoospermia and asthenozoospermia groups (1.5+/-0.6 vs. 1.4+/-0.6, p=0.623). Polyclonal antiserum, generated against a recombinant protein of the N-terminal 160 amino acids of human CatSper, was used. In double fluorescent immunocytochemistry, CatSper protein was found to be expressed in the flagellum of the ejaculated human spermatozoa, and localized in the connecting piece, mid-piece and principal piece, with the exception of the end piece of the flagellum. Moreover, the proportion of CatSper-positive sperm was similar in both the normozoospermia and asthenozoospermia groups. CONCLUSIONS: To the best of our knowledge, this is the first time ejaculated human spermatozoa have been shown to express the mRNA and protein of CatSper. The results of our RT-PCR and immunocytochemistry suggest that CatSper may play a role in the motility of ejaculated human spermatozoa.

Implantation Rate and Clinical Pregnancy Rate According to Dosage and Timing of Progesterone Administration for Secretory Endometrial Preparation in Frozen-Thawed Embryo Transfer Cycles / 대한불임학회지

OBJECTIVE: To evaluate the difference of implantation rate (IR) and clinical pregnancy rate (CPR) between two protocols of endometrial preperation in women undergoing frozen-thawed embryo transfer (FET) cycles. METHODS: This study was performed during the different time periods: A retrospective study from January 2000 to June 2001 (phase I) and a prospective study from July 2001 to March 2002 (phase II). All the patients received estradiol valerate (6 mg p.o. daily) starting from day 1 or 2 of the menstrual cycle without pituitary down regulation. Progesterone was administered around day 14 after sonographic confirmation of endometrial thickness > or = 7 mm and no growing follicle. In Group A (n=88, 99 cycles) of phase I, progesterone was administered i.m. at a dose of 50 mg daily from one day prior to thawing of pronuclear (PN) stage frozen embryo or three days prior to thawing of 6-8 cell stage frozen embryo and then each stage embryos were trasnsferred 2 days or 1 day later after thawing. In Group B (n=246, 299 cycles) of phase I, patients recieved progesterone 100 mg i.m. from one day earlier than group A; two days prior to PN embryo thawing, four days prior to of 6-8 cell embryo thawing. During the phase II, to exclude any differences in embryo transfer procedures, in Group 1 (n=23, 28 cycles) of phase II embryo was transfered by one who have used the progesterone protocol since the phase I. In Group 2 (n=122, 139 cycles) of phase II embryo was transfered by one who use the progesterone protocol from the phase II. RESULTS: When compared across the phase and group, there were no significant differences in the characteristics. During the phase I, there were significant increase in IR (14.4% vs 5.9%, p=0.001) and CPR (28.3% vs 14.5%, p=0.000) in group A. During the phases II, IR (11.8% vs 10.6%) and CPR (27.6% vs 27.3%) show no differences between two groups. CONCLUSIONS: In FET cycles, IR and CPR are increased significantly by the change of dosage and timing of progesterone administraton. And the timing is considered to be more important factor because the dosage of progesterone did not affect implantation window in previous studies. Therefore, we suggest that progesterone administration in FET cycle should begin from one day prior to PN stage embryo thawing and three days prior to 6-8 cell stage embryo thawing.

Recombinant Human Follicle Stimulating Hormone (rFSH) versus Highly Purified Urinary FSH (uFSH): Oocyte and Embryo Quality / 대한산부인과학회잡지

OBJECTIVE: To estimate the efficacy of recombinant human follicle stimulating hormone (rFSH) versus highly purified urinary human FSH (uFSH) in women undergoing controlled ovarian hyperstimulation (COH) for in vitro fertilization and embryo transfer (IVF-ET). METHODS: From 1 January 2001 to 31 August 2001, A total of 254 cycles from 241 patients who attended infertility clinic at Samsung cheil hospital was enrolled in this study. With pituitary down regulation using GnRH agonist by short protocol, rFSH (Puregon(R), Organon, Netherlands) was administered in 131 cycles and uFSH (Metrodin-HP(R), Serono, Switzerland) was administered in 123 cycles. We analyzed ovarian response, pregnancy rate, live birth rate, oocyte quality and embryo quality. RESULTS: The clinical characteristics of two groups were not different. Total FSH dosages (1322.3+/-526.2 IU versus 2124.4+/-881.9 IU, p<0.001) and dosages per retrieved oocyte (90.6+/-36.0 IU versus 138.0+/-57.2 IU, p<0.001) were significantly lower in rFSH group than uFSH group. Clinical pregnancy rate and live birth rate of two groups were not significantly different. The rate of good quality oocyte (Grade I and II) from retrieved oocytes was higher in rFSH group (68.2% versus 64.8%, p=0.024), but after preincubating oocytes for 4 to 6 hours and removing cumulus cells in intracytoplasmic sperm injection (ICSI) cycles, nuclear maturity of oocytes were not significantly different. The quality of transferred embryos were not significantly different too. CONCLUSION: rFSH offered more effective ovarian response in COH and better quality of retrieved oocytes, compared with uFSH.

Molecular Diagnosis of 21-hydroxylase (CYP21) Gene mutations in Congenital Adrenal Hyperplasia / 대한산부인과학회잡지

OBJECTIVES: Congenital adrenal hyperplasia (CAH) is an autosomal recessive disease which is most often caused by a deficiency in steroid 21-hydroxylase (21-OH), a microsomal enzyme encoded by the CYP21 gene. Although several CAH causing mutations have been identified in the CYP21 gene of patients with 21-OH deficiency, genotyping of the 21-OH locus is quite complex because of the high frequency of gene conversion and the presence of multiple mutations on single CAH alleles. This study was aimed to analyze the complete characterization of the CYP21 gene coding region in a Korean CAH patient and to conform the PCR-based single strand conformation polymorphism (SSCP) and heteroduplex analysis as a diagnostic tool. METHODS: We used a highly sensitive, non-radioactive method allowing PCR-based single strand conformation polymorphism (SSCP) analysis. This method was applied to the characterization of all the exons and intron-exon junctions of the CYP21 gene in one patients affected by the salt wasting form and 4 normal controls. RESULTS: In all samples showing SSCP abnormal band patterns, sequence analysis showed the presence of sequence variants. In particular, one mutation (I172N) which is already known to cause the disease and 3 silent mutations were detected. CONCLUSION: PCR-based single strand conformation polymorphism (SSCP) and heteroduplex analysis should be useful for the clinical application as a diagnostic tool for the detection of 21-hydroxylase gene mutations.