Doo Seok Choi

The role of autophagy in follicular development and atresia in rat granulosa cells

OBJECTIVE To investigate the involvement of autophagy in folliculogenesis and its correlation with apoptosis. DESIGN Animal model-based study. SETTING University medical center. ANIMAL(S) Twenty-one day-old female Sprague-Dawley rats. INTERVENTION(S) Ovaries obtained from established immature rat models primed with pregnant mare serum gonadotropin (PMSG) were used for the induction of follicular development and atresia. Granulosa cells isolated from developing follicles were cultured in serum-free condition with or without follicle-stimulating hormone. MAIN OUTCOME MEASURE(S) Microtubule-associated light-chain protein 3 (LC3) and autophagic vacuoles were used as autophagic markers, and cleaved caspase-3 was used as an apoptotic marker in ovaries and/or granulosa cells. RESULT(S) The LC3 protein was expressed mainly in granulosa cells during all developmental stages. In granulosa cells isolated from PMSG-primed immature rat ovaries, LC3-II expression showed a similar expression pattern to cleaved caspase-3. In addition, granulosa cells of atretic follicles that showed intense cleaved caspase-3 staining also showed intense LC3 immunoreactivity. An in vitro culture experiment revealed that the levels of LC3-II and cleaved caspase-3 proteins were gonadotropin-dependent. The induction and the gonadotropin dependency of granulosa cell autophagy were confirmed by the observation of autophagic vacuoles under transmission electron microscopy. CONCLUSION(S) These preliminary results suggest that autophagy is induced mainly in granulosa cells during folliculogenesis and shows good correlation with apoptosis.

Effects of Activation Timing on the Fertilization Rate and Early Embryo Development in Porcine ROSI Procedure

AbstractPurpose: This study was undertaken to evaluate the optimal exposure time for nuclear envelope breakdown (NEBD) after injection of a round spermatid and to investigate the effect of oocyte activation timing on the fertilization rate and early embryo development in porcine round spermatid injection procedure. Methods: Injected oocytes were fixed at 0.5, 1, 2, 3, and 4 h before activation, and NEBD state was examined. The three groups of oocytes were activated before and after the injection of spermatid using a single direct current pulse (100 V/mm, 50 μs): group 1) at 2 h before the injection (pre), group 2) within 0.5 h after the injection (immediate), and group 3) at 2 h after the injection (post). Activated oocytes were cultured and pronucleus formation and blastocyst development was evaluated at 15–18 h and 7–8 days after the injection, respectively. Results: The proportion of oocytes with NEBD significantly increased in the groups with over 2 h of exposure time (p<0.05) and oocyte with premature chromosome condensation began to appear 3 h after the injection. Normal fertilization and development rate to the blastocyst stage were significantly higher in the post group than in those of the pre or immediate group (p<0.05). Conclusion: The optimal exposure time for NEBD is 2 h after the injection, and activation in 2 h after round spermatid injection improved the normal fertilization and early embryo development rate.

Clinical utility of newly developed highly purified human menopausal gonadotrophins: a randomized controlled trial

The aim of this study was to evaluate the safety and efficacy of IVF-M HP, a newly developed highly purified human menopausal gonadotrophin preparation, for ovarian stimulation in women with infertility undergoing IVF, intracytoplasmic sperm injection (IVF-ICSI) and embryo transfer using a GnRH antagonist protocol. This was a multicentre, randomized, active-controlled, parallel design, open-label, non-inferiority clinical study. Of the 112 patients randomized for treatment using the GnRH antagonist protocol, 111 were treated. No significant difference was found in the number of oocytes retrieved from the IVF-M HP and Menopur groups (13.1 ± 7.6 versus 10.3 ± 6.7, respectively). The lower limit of the one-sided 97.5% confidence interval for the difference between the groups was -0.25, i.e., greater than the pre-defined non-inferiority margin (-5). Therefore, the IVF-M HP treatment was considered non-inferior to Menopur. Furthermore, no significant difference was observed between the groups in the number of good-quality oocytes, leading follicles, good-quality embryos, or in fertilization, implantation, positive beta-HCG and clinical pregnancy rates. The safety analysis revealed that 40.4% and 35.2% in the IVF-M HP and Menopur groups, respectively, reported adverse events. In conclusion, IVF-M HP had comparable clinical efficacy and safety profiles to Menopur.

Effects of Hormone Therapy on Serum Lipid Levels in Postmenopausal Korean Women

Objectives This study was conducted to examine the effects of hormone therapy on serum lipid levels in postmenopausal Korean women. Methods This retrospective cohort study included 154 healthy postmenopausal Korean women. Seventy-nine women took oral estrogen (conjugated equine estrogen 0.625 mg/day or equivalent), and 75 applied estrogen transdermally using 0.1% 17β-estradiol gel. Micronized progesterone (MP) was added to 40 women of oral group and 49 women in transdermal group. Serum levels of triglyceride, low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol (HDL-C), and lipoprotein (a) were measured before, 3 and 6 month after hormone therapy. Results At baseline, mean body mass index (BMI) were lower (22.76 vs. 23.74 kg/m2) and proportion of family history of cardiovascular disease (CVD) (61 vs. 39%) were higher in oral group. In oral group, LDL-C and lipoprotein(a) levels decreased, and triglyceride and HDL-C levels increased significantly after 3 and 6 months. There was no significant change in lipoprotein levels compared to the baseline in transdermal group. There were also no differences with additional MP. Changing pattern of HDL-C during 6 months was significantly different by the route of estrogen administration. Conclusion Oral estrogen therapy might be more beneficial than transdermal estrogen in terms of lipid in postmenopausal Korean women. The estrogen effects are not influenced by adding MP.