Posters * Reproductive Endocrinology (i.e. PCOS, Menarche, Menopause etc.)

Abstract

Introduction: Ovarian function is controlled not only by endocrine system, but also by autocrine/paracrine regulation in the ovarian cells, which may be mediated by a network of cytokines. The aim of this study was to elucidate the involvement of a multifunctional cytokine, interleukin-6 (IL-6), in human ovarian function especially in human luteolysis. Material and Methods: All subjects were women with normal menstrual cycles (28-35 days), which gave their informed consent to participate in this study. The mean age of the subjects was 39.0 {+/-} 8.7 years (range: 22-53 years, n = 37). Ovarian samples were collected at the time of gynecological operation, and follicular or luteal tissue was isolated macroscopically from each sample. Blood serum was also collected at the same time. All subjects were divided into 3 phases of ovarian cycle (follicular, early-mid luteal, and late luteal phase), according to the date of menstrual cycle and serum hormonal values (classified into follicular phase, if progesterone (P) < 2.0 ng/ml, classified into early-mid luteal phase, if progesterone (P) > 2.0 ng/ml, and classified into late luteal phase, if E2 < 80 pg/ml) at the point of operation. The relative expression levels of IL-6 (n = 37) and its receptors (gp130 and IL-6R{alpha}, n = 21) mRNA in granulosa, theca and luteal cells were analyzed with quantitative RT-PCR using TaqMan technology. The relative values of IL-6 protein in the ovarian tissue were also analyzed with immunoblotting (n = 21). The localization of IL-6 and its receptors, gp130 and IL-6R{alpha}, in the ovarian tissues was examined by immunohistochemical staining (n = 8). All data are presented as the mean {+/-} S.D. Inter-group differences were confirmed with Kruskal-Wallis test, and post-hoc test (Scheffe's F) was used for detecting the significant differences between the groups. Results: The relative level of IL-6 mRNA in late luteal phase (4.46 {+/-} 2.05, n = 8) was significantly higher than those of follicular (1.55 {+/-} 0.94, n = 18: p < 0.00005) and early-mid luteal phase (2.13 {+/-} 0.92, n = 11: p < 0.005). The relative level of mRNA for gp130 in late luteal phase (8.35 {+/-} 5.46, n = 7) was significantly higher than those of follicular phase (1.67 {+/-} 1.93, n = 8: p < 0.05), and the level of IL-6R{alpha} mRNA in late luteal phase (11.66 {+/-} 9.71, n = 7) was significantly higher than those of follicular phase (1.33 {+/-} 2.32, n = 8: p < 0.05) and early-mid luteal phase (2.72 {+/-} 2.40, n = 6: p < 0.05). The relative value of IL-6 protein in the late luteal phase (0.935 {+/-} 0.070, n = 7) was also significantly higher than those of follicular phase (0.751 {+/-} 0.049, n = 8: p < 0.001) and early-mid luteal phase (0.762 {+/-} 0.104, n = 6: p < 0.005). The immunohistochemistry for IL-6 showed positive staining mainly in luteal cells and follicular granulosa cells, and partially in theca cells. The positive staining for gp130 was identified in luteal and granulosa cells as well as the endothelial cells of the capillary vessels. IL-6R{alpha} immunoreactivity was observed in luteal, granulosa and theca cells. Conclusions: Analysis for periodic changes of mRNA and protein revealed that IL-6 was highly expressed in the human ovary of late luteal phase during natural ovarian cycle. The expression of both IL-6 receptors mRNA were also enhanced in the human ovary of late luteal phase during natural ovarian cycle. Immunohistochemical staining showed that IL-6 and its receptors, gp130 and IL-6R{alpha}, were localized in human luteal cells of regressing phase during natural ovarian cycle. These results suggest that IL-6 may act on the corpus luteum of regressing phase and play a significant role to regulate the human ovarian function, possibly as a promoter of human luteolysis. Introduction: It is well established that granulosa cells of larger follicles (>10mm in diameter) express LH receptors, and thus become sensitive to LH stimulation. Previous studies indicate that even when FSH is withdrawn and ovarian stimulation is performed by LH activity alone, the outcomes are comparable to recombinant FSH administration throughout the stimulation cycle. It has also been demonstrated that, after recombinant FSH priming, a low dose of hCG is able to stimulate the development of preovulatory follicles with low or no FSH doses. However, whether the follicular development, stimulated by low doses of hCG, depends on patients age, still needs to be elucidated. Therefore, we designed this investigation to compare the outcomes of controlled ovarian stimulation (COS) in two different age groups of patients (< 35 y-old and [≥] 35 y-old) undergoing intracytoplasmic sperm injection (ICSI) cycles with exclusively recombinant FSH (r-FSH) or in association with recombinant hCG (r-hCG) microdose. Material and Methods: We evaluated the outcomes of COS in 320 patients < 35 y-old and 68 patients [≥] 35 y-old of age. In both age groups, patients were divided into a Control-group (n = 160 < 35 y-old and n = 34 [≥] 35 y-old) and a Microdose-group (n = 160 < 35 y-old and n = 34 [≥] 35 y-old). For all patients, down regulation was achieved trough leuprolide acetate. In control-groups follicular growth was stimulated with r-FSH (225 IU daily) and ovulation was triggered with an hCG injection. In Microdose-groups, when at least one follicle [≥] 14 mm was observed, the r-FSH dose was reduced to 75 IU. In this case, r-hCG microdose was administered subcutaneously (7.7 {micro}g) until the day of ovulation triggering. The number of metaphase II (MII) oocytes, percentage of high quality embryos, fertilization, pregnancy, implantation and miscarriage rates were evaluated. Results: In patients < 35 y-old, a higher number of MII oocytes was observed in the Microdose-group (10.2 {+/-} 7.19 vs 8.31 {+/-} 7.01; p = 0.019); however no differences were observed regarding the other evaluated variables. In patients [≥] 35 y-old the Microdose-group showed a higher fertilization rate (75.9% vs 60.7%; p = 0.036), and a higher percentage of high-quality embryos (66.5% vs 51.1%; p = 0.018). Moreover, a trend for a higher implantation rate in Microdose-group was noted (12.3% versus 6.4%; p = 0.085), however, this most likely did not reach statistical significance because of insufficient number of cycles evaluated in this trial. Conclusions: The increased number of MII oocytes, observed in group of patients < 35 y-old treated with the r-hCG microdose, suggests that the association of r-hCG and r-FSH may prevent the decrease on estradiol concentration after GnRH antagonist administration, which in turn, may significantly increase the number of mature retrieved oocytes. For patients [≥] 35y-old the use of hCG microdose was able to improve both fertilization and embryo quality, suggesting an important role of LH during the latter phase of oocyte development. These evidences suggest that the use of hCG in COS protocols beginning at the time of antagonist administration could be recommended. Although its positive role on pregnancy rates remains controversial, it seems clear that hCG supplementation does not play any negative role on estradiol synthesis, providing a more physiological hormonal milieu. In conclusion, our data confirm that the recombinant hCG microdose is an efficient source of LH activity for both younger and older patients, and this strategy can be used to reduce the FSH amounts required in COS protocols. Introduction: To compare the efficacy of Venalafaxine (Serotonin, norepinephrine re-uptake inhibitor, SNRI) and HRT (hormone replacement therapy) in controlling vasomotor symptoms and postmenopausal depression in Indian women Methods: 91 postmenopausal women (age 45-60yrs) were recruited over a period of three years to randomly receive either drug over a period of 12 weeks. Depression was evaluated by Hamilton depression rating scale (HDRS) and vasomotor symptoms by frequency, duration and intensity of hot flushes, presence of sweating and insomnia. Venlafaxine was used in dose of 37.5-70mg /day in 45 cases. HRT as conjugated equine estrogen 0.625 mg and Medroxy progesterone acetate 2.5 mg/day was given in 46 patients.5 patients were lost in follow up and 4 stopped the medications due to not tolerating them. The patients in both groups had matched controls given placebo tablets All patients were evaluated at 8 and 12 weeks after starting the treatment. Results: A clinically significant improvement, up to 60%, occurred in depressive symptoms with Venalafexine while with HRT it was only up to 40%. Improvement in vasomotor symptoms was comparable in both groups at 8 and 12 weeks as 67% and 70% with Venalafaxine and HRT respectively. Conclusion: HRT cannot be considered as an effective antidepressant in postmenopausal women with mild to moderate depression. Venalafaxine is an effective and well-tolerated drug for management.HRT is effective in treating vasomotor symptoms but alternative drugs like Venalafaxine are equally effective, with fewer side effects and better tolerated Introduction: Uterine contractility is important not only for labor, but also for the transport of semen and gametes and successful embryo implantation. Uterine activity is regulated by complex and mutual interactions among sex steroids, neurohypophysial hormones, and neurotransmitters. It also involves the integration of many signal-transducing events, resulting in precise coordination of myometrial contractile activity. There are many uterotonic agents and also many intracellular effectors that are known to be crucial for efficient uterine smooth-muscle contraction -- such as oxytocin, various prostaglandins, and muscarinic receptor agonists. Material and Methods: Each swine uterus was perfused in a perfusion system with Krebs-Ringer solution. After one hour with rhythmical spontaneous contractions present, the substances were administered via the arterial vein catheter or in