dynorphin (KNDy) neurons by using N-7 cells: immortalized hypothalamic cell line

CONCLUSION: This study showed that lymphangiogenesis is increased in endometrium and myometrium of adenomyosis in comparison to the controls, and that increased LVD in adenomyosis is reduced by use of LNG-IUD. The results suggest that effect of LNG-IUD on lymphangiogenesis could be one possible mechanism of reducing adenoymosis related symptoms. P-622 Wednesday, October 16, 2013 REPRODUCTIVE HISTORY IN RELATION TO PLASMA SEX STEROID HORMONE, PROLACTIN, AND GROWTH FACTOR CONCENTRATIONS IN PREMENOPAUSAL WOMEN. F. Mu,a H. Eliassen,a,b S. Tworoger,a,b S. Hankinson,a,b,c S. Missmer.a,b aDepartment of Epidemiology, Harvard School of Public Health, Boston, MA; b Channing Division of Network Medicine, Department of Medicine, Brigham and Women’s Hospital and Harvard Medical School, Boston, MA; cDivision of Biostatistics and Epidemiology, School of Public Health and Health Sciences University of Massachusetts, Amherst, Amherst, MA. OBJECTIVE: To investigate the relation between reproductive history and plasma sex steroid hormones, prolactin and growth factors levels in premenopausal women. DESIGN: Cross-sectional study. MATERIALS AND METHODS: We assessed the relations of parity, age at first birth, time since last birth and lifetime duration of breastfeeding, with plasma levels of estrogens, androgens, progesterone, prolactin, sex hormone binding globulin (SHBG), growth hormone (GH), insulin-like growth factor1 (IGF-I), and its binding protein (IGFBP-3), among 2745 premenopausal women in the Nurses’ Health Study 2, aged 32-52 years at blood draw. Estrogens and progesterone were measured in samples timed within the menstrual cycle; others were measured in timed and untimed samples. Multivariable general linear models were used to calculate adjusted geometric means. RESULTS: Parity was directly associated with progesterone (percent difference >¼3 vs. 0¼9%, Ptrend¼0.03) and SHBG levels (7%, Ptrend¼0.05). Parity was inversely associated with luteal free estradiol (8%, Ptrend¼0.02) and prolactin (24%, Ptrend<0.01). Age at first birth was directly associated with IGFBP-3(>¼35 vs. <25¼3%, Ptrend¼0.05). Time since last birth was inversely associated with progesterone (>6 vs. %2 years¼11%, Ptrend¼0.05), as well as IGF-1 and IGFBP3 (IGF-1, 7%, Ptrend<0.01; IGFBP3, 4%, Ptrend¼0.02). In contrast, time since last birth was directly associated with total and free testosterone (total, 27%, Ptrend¼0.02; free, 22%, Ptrend¼0.03). Longer history of breastfeeding was associated with higher SHBG (R24 vs. <1 months ¼ 9%, Ptrend¼0.03) and higher IGF-1 (5%, Ptrend¼0.03). These associations remained largely unchanged after exclusion of anovulatory, irregular cycles and cycles with extreme luteal days or additionally adjusting for body mass index (BMI) at age 18, BMI at blood draw, alcohol intake and smoking history. CONCLUSION: These data suggest that reproductive history is moderately associated with hormone levels during the mid-late reproductive years. Supported by: CA67262 and CA050385. P-623 Wednesday, October 16, 2013 MTHFR GENE POLYMORPHISMS AND THE CORRELATION WITH ESTRADIOL AND AMH SERUM LEVELS AND ASSISTED REPRODUCTION OUTCOMES. E. B. Cordts, C. Paiva, B. L. Lechado, D. M. Christofolini, C. P. B. P. Barbosa, B. Bianco. Human Reproduction and Genetics, Faculdade de Medicina do ABC, Santo Andre, Sao Paulo, Brazil. OBJECTIVE: To correlate MTHFR C677T, A1298C and G1793A polymorphisms with estradiol and AMH serum level and assisted reproduction outcomes. DESIGN: Cross-sectional study comprising 186 infertile women that underwent IVF (n¼54 tubeperitoneal factor, n¼82 male infertility and n¼50 idiopathic infertility). All patients were younger than 38 years old, had normal prolactin and TSH levels, ovaries without morphological abnormalities, ovulatory cycle, BMI lower than 30, no history of poor ovulatory response, and no evidence of endocrine disorders or endometriosis. MATERIALS AND METHODS: Genotyping of polymorphisms were performed by real time PCR. The measurement of estradiol and AMH was performed by ELISA. RESULTS: Considering the controlled ovarian hyperstimulation response 67.2% (125/186) showed good response, 24.7% (46/186) poor response; 4.3% (8/186) hyper response, and 3.7% (7/186) developed ovarian hyper-

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stimulation syndrome. Women with tubeperitoneal factor have better oocyte maturation rate when compared to patients with idiopathic infertility (p¼0.018), while women with tubeperitoneal factor had higher rate of frozen embryos when compared to male factor (p¼0.032). The assisted reproduction outcomes were not associated both with MTHFR A1298C and C677T polymorphisms. Regarding G1793A polymorphism we observed a statistically tendency in patients with wild allele in relation to the rate of oocyte maturation, p¼0.056. When we compared the ovarian stimulation response with embryo parameters and hormonal measurements we found a statistically significant difference for all parameters and estradiol and AMH serum level. CONCLUSION: Previous studies observed that women underwent assisted reproduction techniques carrying T allele of the MTHFR C677T polymorphism showed reduced ovarian response to recombinant FSH. Furthermore, the patients have significantly lower concentrations of estradiol and showed significantly less oocytes retrieved. In the present study, we just found a tendency of the G1793A polymorphism with oocyte maturation. P-624 Wednesday, October 16, 2013 INVESTIGATION OF THE KISSPEPTIN/NEUROKININ B/DYNORPHIN (KNDY) NEURONS BY USING N-7 CELLS: IMMORTALIZED HYPOTHALAMIC CELL LINE. S. Osuka, A. Iwase, T. Nakahara, M. Kondo, M. Goto, F. Kikkawa. Department of Obstetrics and Gynecology, Nagoya University Graduate School of Medicine, Nagoya, Aichi, Japan. OBJECTIVE: Kisspeptin, a neuropeptide hormone encoded by the Kiss-1 gene, and its receptor have been recently recognized as fundamental activators of the gonadotropic axis with key roles in the control of gonadotropin secretion, especially, in vivo studies. Moreover, recent observations show that neurokinin B (NKB) is coexpressed with kisspeptin and dynorphin in the arcuate nucleus (ARC) in hypothalamus, named the Kisspeptin/NKB/Dynorphin (KNDy) neurons. KNDy neurons are thought to drive pulsatile GnRH secretion and gonadotropin release, and estradiol negatively regulates kisspeptin expression in KNDy neurons. In this study, we investigate the presence of KNDy peptides and its receptors, and the change of Kiss-1 gene expression after NKB receptor agonist and estradiol treatment. DESIGN: In vitro study. MATERIALS AND METHODS: Reverse-transcription polymerase chain reactions (RT-PCR) were performed to investigate the pattern of expression of Kiss-1, Kiss-1 receptor (GPR54), NKB receptor (TCR3; Tachykinin receptor 3), Dynorphin receptor (KOR; Kappa opioid receptor), estrogen receptor a/b (ERa/b) and progesterone receptor (PgR) in N-7 cells. And also we investigated the quantitative real-time RT-PCR (qRT-PCR) of Kiss-1 expression in N-7 cells after TCR3 agonist senktide and estradiol treatment. RESULTS: Expression of the genes encoding TCR3, KOR, estrogen receptor a/b (ERa/b), progesterone receptor (PgR), Kiss1 and its receptor GPR54 were found in N-7 cell. Furthermore, after the treatment of senktide, the expression of Kiss-1 mRNA tended to increase and decrease with additional estradiol treatment. CONCLUSION: Our results suggest that N-7 cells possess similar expression profiles in receptors of KNDy peptides and sex steroids and show the tendency which support the hypothesis about the negative regulation of kisspeptin expression by estradiol in KNDy neuron. P-625 Wednesday, October 16, 2013 THE EVALUATION OF POLYMORPHISMS OF THE GENES ERa AND ERb IN INFERTILE WOMEN AND YOUR CORRELATION WITH THE RESULTS OF ASSISTED REPRODUCTION. V. Cavalcanti, C. M. Trevisan, C. Peluso, M. A. P. Barbosa, E. B. Cordts, B. Bianco. Human Reproduction and Genetics, Faculdade de Medicina do ABC, Santo Andre, Sao Paulo, Brazil. OBJECTIVE: Identify the ER alpha and ER beta polymorphisms in infertility women who underwent assisted reproduction and correlate the polymorphisms with the assisted reproductive outcomes. DESIGN: Case-control study comprising 184 Brazilian infertile women that underwent in vitro fertilization (IVF) [n¼50 Idiopathic infertility, n¼53 tube peritoneal, and n¼81 male factor]. All patients were younger than 38 years, had normal prolactin and TSH serum levels, presence of both ovaries without morphological abnormalities, ovulatory cycle, BMI% 30, no previous history of poor ovulatory response, and no evidence of endocrine disorders or endometriosis.

Vol. 100, No. 3, Supplement, September 2013