Lean mass is a modifiable risk factor for vertebral fracture in postmenopausal women

ovaries from a 60-day-old mouse. Control and transplant recipients were assessed at 16 months of age. Immune function was assessed with a FACSbased T-cell immunoassay. Metabolic changes were assessed with a Glucose Tolerance Test. RESULTS: The results showed that glucose metabolism and immune function in 16-month-old postreproductive mice with young ovaries was not different from six-month-old mice and was significantly improved compared with 16-month-old, non-transplanted postreproductive mice. CONCLUSIONS: In summary, our novel results indicate that restoration of young ovarian function to postreproductive female mice significantly influenced metabolism and immune function as measured by glucose tolerance test and T-cell immunoassay. These observations are important because they imply that, in our model, the presence of young ovarian cells may mimic the positive health span effects seen with dietary restriction. These results also support a potential link between the spectacular advances in health span due to reproductive manipulation in primitive species and dietary restriction in mammals. Supported by: This research was supported by Utah State University Research Initiation Funds and a Utah State University Research Catalyst Program Grant to JM. P-359 Wednesday, October 19, 2016 RETROSPECTIVE ANALYSIS OF THE EFFICACY OF ART AND REPRODUCTIVE OUTCOMES IN FEMALES WITH FRAGILE X ASSOCIATED PRIMARY OVARIAN INSUFFICIENCY. L. Tan,a L. Krichevsky,b E. Greenblatt,c R. Casper,d C. A. Laskin,e S. Sierra,f T. Hannam,g M. F. Karnis,h C. L. Librach,i P. A. Sharma.j aObstetrics & Gynaecology, University of Toronto, Toronto, ON, Canada; bFaculty of Medicine, University of Toronto, Toronto, ON, Canada; cCFRH Mount Sinai Hospital, Toronto, ON, Canada; dProfessor, University of Toronto, Toronto, ON, Canada; eMedicine Obstetrics & Gynecology, LifeQuest Centre for Reproductive Medicine, Toronto, ON, Canada; fDivision of Reproductive Endocrinology & Infertili, Women’s College Hospital, University of Toronto, Toronto, ON, Canada; gReproductive Endocrinology, Toronto, ON, Canada; hONE Fertilty, Burlington, ON, Canada; iCReATe Fertility Centre, Toronto, ON, Canada; jObstetrics and Gynecology, Create Fertility Centre, Toronto, ON, Canada. OBJECTIVE: Fragile X premutation (55-200 CGG repeats) is the most common genetic cause of 46,XX primary ovarian insufficiency (POI) [1,2]. There is currently no published literature regarding the reproductive outcomes and efficacy of artificial reproductive technology (ART) in females with Fragile X associated primary ovarian insufficiency (FXPOI) making counseling about reproductive treatment a challenge. Much of the data regarding FXPOI patients were extrapolated from studies of women with other causes of POI. Trial and error in these women can be associated with loss of time to conception as well as substantial cost to the patient. This study aimed to determine whether ART treatment options and success rates differ between females with FXPOI and those with POI due to other causes. DESIGN: Retrospective cohort study. MATERIALS AND METHODS: We performed a retrospective chart review of patients with POI from seven fertility centers in Ontario from 2007 to 2015. The study group consisted of FXPOI patients, younger than 40 years old (n¼19). The control group consisted of females younger than 40 years old who were diagnosed with POI or approaching POI, with no Fragile X premutation (n¼76). The FXPOI and control patients were grouped and matched by age (<30, 30-34, and 35-39 years old) and by their ovarian reserve assessment utilizing AMH testing (AMH <2.0 pmol/L and AMH ¼ 2-6 pmol/L). RESULTS: 19 FXPOI patients who underwent a total of 51 treatment cycles and 76 control patients who underwent a total of 172 treatment cycles were identified. When comparing the FXPOI and control groups as a whole (stimulated and natural cycles combined), the pregnancy rate per cycle was nearly the same in both groups (24% versus 21%) respectively. However, the pregnancy rate per cycle was significantly lower in the AMH <2.0 pmol/L FXPOI group (6.3%) compared to the control group (20.8%). Ovulation induction with clomiphene citrate was the most efficacious method in the FXPOI group with a 23% pregnancy rate per cycle versus 5.9% in the control group. CONCLUSIONS: FXPOI females with very low ovarian reserve (AMH <2.0 pmol/L) have significantly lower chance of pregnancy success compared to their age matched counterparts with POI due to other causes. This group will benefit from early diagnosis and targeted treatment with milder ovarian stimulation (e.g. clomiphene citrate). While this study is

FERTILITY & STERILITYÒ

limited by its small sample size, the data seems to suggest that clomiphene citrate may potentially play an important first line ovulation induction therapy for women with FXPOI. References: 1. Conway GS, Payne NN, Webb J, et al. Fragile X premutation screening in women with premature ovarian failure. Hum Reprod 1999;14:573-5. 2. Wittenberger MD, Hagerman RJ, Sherman S, et al. The FMR1 premutation and reproduction. Fertil Steril 2007;3:456-65. Supported by: Supported by CReATe Program Inc. and CREMS University of Toronto summer studentship. P-360 Wednesday, October 19, 2016 PROTECTIVE EFFECT OF EVODIAE FRUCTUS EXTRACT AGAINST 4-VINYLCYCLOHEXENE DIEPOXIDE- INDUCED OVOTOXICITY IN CHO-K1 CELLS. S. Kim, J. Lee, S. KIM, D. Kim. Dongguk university, Gyeonggi-do, Korea, Republic of. OBJECTIVE: This study investigated the cytoprotective effects and molecular mechanism of Evodiae Fructus (EF) extracts against 4-vinylcyclohexene diepoxide (VCD)-induced ovotoxicity in CHO-K1 ovary cells. DESIGN: We established an in vitro screening tool in which to discover promising new drug candidates from herbal medicines for prevention and treatment of premature ovarian failure. In this model, VCD was used as ovotoxicant due to its ability to destroy ovarian follicles via apoptosis and interference with c-kit/kit ligand signaling pathways. MATERIALS AND METHODS: EF extract was prepared by boiling in water and verified its quality by high performance liquid chromatography (HPLC) analysis. CHO-K1 cells were plated, pretreated with EF for 2 h and continuously treated with 1.5 mM VCD for 24h. Cell viability was measured by MTT assay. Protein levels were assessed by western blot. RESULTS: VCD significantly suppressed cell viability and induced apoptosis at 1.5 mM in CHO-K1 cells. EF dose-dependently blocked the ovotoxicity induced by treatment with VCD. Furthermore, EF significantly activated Akt and its downstream effectors such as mTOR and GSK-3b. The ability of EF to prevent cytotoxicity by VCD was antagonized by pretreatment of LY294002, a PI3K/Akt inhibitor. CONCLUSIONS: We demonstrated that EF has the ability to protect ovary cells against VCD-induced ovotoxicity, which may be associated with Akt activation. These results suggest that the beneficial effects of EF may be used to improve premature ovarian failure or unexplained infertility caused by environmental factor. References: 1. Kappeler CJ, Hoyer PB. 4-vinylcyclohexene diepoxide: a model chemical for ovotoxicity. Syst Biol Reprod Med 2012;58(1):57-62. 2. Fernandez SM1, Keating AF, Christian PJ, Sen N, Hoying JB, Brooks HL, Hoyer PB. Involvement of the KIT/KITL signaling pathway in 4-vinylcyclohexene diepoxide-induced ovarian follicle loss in rats. Biol Reprod 2008;79(2):318-27. 3. Takai Y, Canning J, Perez GI, et al. Bax, caspase-2, and caspase-3 are required for ovarian follicle loss caused by 4-vinylcyclohexene diepoxide exposure of female mice in vivo. Endocrinology. 2003;144(1):69-74. Supported by: This research was supported by a grant of the Korea Health Technology R&D Project through the Korea Health Industry Development Institute (KHIDI), funded by the Ministry of Health & Welfare, Republic of Korea (grant number : HI15C0198). P-361 Wednesday, October 19, 2016 LEAN MASS IS A MODIFIABLE RISK FACTOR FOR VERTEBRAL FRACTURE IN POSTMENOPAUSAL WOMEN. A. W. Tiegs,a R. J. Meislin,b N. M. Sachdev,c M. J. Nachtigall,d L. E. Nachtigall,e a OB/GYN, NYU Langone School of Medicine, New York, NY; bNYU School of Medicine, New York, NY; cObstetrics and Gynecology, New York University Fertility Center, New York, NY; dOb/Gyn, NYU Medical Center, New York, NY; eObstetrics and Gynecology, NYU School of Medicine, New York, NY OBJECTIVE: Vertebral fracture is the most common clinical manifestation of osteoporosis and is significantly associated with an increased risk of future fractures.1 Bone mineral density has traditionally been the best predictor of fragility fractures, however, lean mass may have a greater contribution to the risk of fracture than previously understood. Dual-energy X-ray

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absorptiometry (DXA) allows for highly accurate measurements of bone mass as well as both fat and lean body mass. The primary objective of this study is to determine if there is an association between lean body mass and the incidence of vertebral fragility fractures in postmenopausal women. The presence of an association between number of vertebral fractures and T-score, Z-score, body mass index (BMI), muscle mass index (lean mass (kg)/ height (m2)), and fat mass are secondary outcome measures. DESIGN: Retrospective cohort study. MATERIALS AND METHODS: All women between the ages of 40 and 100 years, who underwent body composition and bone density testing using DXA scan at the NYU Bone Density and Body Composition Unit from May 2011 to November 2014 were identified. All indications were included. Patients with DXA that did not include a lateral vertebral assessment were excluded. Parametric variables were confirmed by Shapiro-Wilk testing and compared by analysis of variance (ANOVA). Chi-square testing was used for nominal variables. RESULTS: A total of 358 women met inclusion criteria. The average age was 70.2 years +10.3 (Range 46 to 93 years), average weight was 139.6lbs + 25.9 (Range 90 to 267 lbs) and average body mass index (BMI) was 25.0 + 4.5 kg/m2 (Range 16.7 to 42.3). A total of 124 vertebral fractures were identified in 85 patients (23.7%), with an average of 0.35 (+ 0.7) vertebral fractures per patient. Both lean body mass and Z-score were noted to have an inverse association with number of vertebral fractures (p¼0.03 and p¼0.02, respectively). Women without vertebral fractures had an average lean mass of 62.4 lbs (+8.5) (average BMI 24.9 +4.5), while women with 3 vertebral fractures had an average lean mass of 59.5 lbs (+8.7) (average BMI 26.0 +4.5). Women with at least one vertebral fracture were more likely to have an average T-score of at least -0.8 (+1.5), but T-score was not found to be significantly associated with number of fractures (p¼0.26). Additionally, fat mass (p¼0.82), BMI (p¼0.19), and muscle mass index (p¼0.36) did not prove to be predictive of vertebral fracture in this population. CONCLUSIONS: Irrespective of BMI, a lean mass of greater than 62.4lbs was associated with lower incidence of vertebral fracture in our population. These results suggest the importance of assessing lean mass in postmenopausal women, as it is a modifiable risk factor for osteoporotic fractures. Reference: 1. Lindsay R, Silverman SL, Cooper C, Hanley DA, Barton I, Broy SB, Licata A, Benhamou L, Geusens P, Flowers K, Stracke H, Seeman E. Risk of new vertebral fracture in the year following a fracture. JAMA 2001;285(3):320.

REPRODUCTIVE ENDOCRINOLOGY - CLINICAL P-362 Wednesday, October 19, 2016 HIGHER 25-HYDROXYVITAMIN D (25(OH)D) IS ASSOCIATED WITH INCREASED FECUNDABILITY. A. Z. Jukic,a C. R. Weinberg,b A. Z. Steiner.c aYale School of Public Health, New Haven, CT; bNational Institute of Environmental Health Sciences, Durham, NC; c University of North Carolina, Chapel Hill, NC. OBJECTIVE: In rodents, vitamin D deficiency has been associated with dramatic reductions in fertility and supplementation with vitamin D improves reproductive success (1). In women undergoing fertility treatment, vitamin D sufficiency has been associated with improved success rates, however, no studies have prospectively examined vitamin D status and spontaneous conception in women with no known fertility problems. DESIGN: We used data from a prospective community-based cohort study of time to pregnancy, Time to Conceive, to examine the association of vitamin D status with fecundability. MATERIALS AND METHODS: Fecundability was measured by the number of menstrual cycles required to conceive a clinical pregnancy. Women between the ages of 30-44, with no known fertility problems, were enrolled in Time to Conceive early in their attempt to become pregnant. Women provided a whole blood sample that was spotted, dried, and stored frozen. Women kept daily diaries that included menstrual bleeding and daily vitamin and supplement use for up to four months. 25(OH)D2 and 25(OH)D3 were measured in duplicate in the baseline blood spots using liquid chromatography tandem mass spectrometry. We developed a predictive model to adjust the measured total 25(OH)D (referred to hereafter as 25(OH)D) for changes in season and supplement use across each menstrual cycle. We analyzed the association between time to pregnancy and cycle-specific 25(OH)D using a

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ASRM Abstracts

discrete time hazard model that adjusted for age, race, exercise, recent estrogen use, and body mass index. RESULTS: There were 474 women in our sample who contributed 1210 menstrual cycles (of which 234 were conception cycles). The mean 25(OH)D was 35 ng/ml and 30% of women had a 25(OH)D measure below 30 ng/ml. A ten-unit increase in the natural log of 25(OH)D was associated with an approximately 20% higher odds of conception (Fecundability odds ratio(FOR)(95% Confidence interval (CI)): 1.17 (1.01, 1.36). 25(OH)D was not associated with anti-Mullerian hormone (Spearman correlation: -0.017, p ¼ 0.70). CONCLUSIONS: In this population of late reproductive-aged women, increasing 25(OH)D was associated with an increased odds of conceiving a pregnancy. Clinicians should assess vitamin D status in women who are attempting to conceive a pregnancy. References: 1.Lerchbaum E, Obermayer-Pietsch B. Vitamin D and fertility: a systematic review. Eur J Endocrinol 2012;166:765-78. Supported by: Research reported in this abstract was supported by the Eunice Kennedy Shriver National Institute of Child Health and Human Development of the National Institutes of Health (NIH) under award number R00HD079659 (PI: Jukic) and under award number R21 HD060229 R01 HD067683 (PI: Steiner). P-363 Wednesday, October 19, 2016 PATIENTS WITH UNEXPLAINED INFERTILITY DO NOT SHOW EVIDENCE OF DECREASED OVARIAN RESERVE COMPARED TO CONTROLS. H. Huddleston,a N. Santoro,b E. Eisenberg,c D. Haisenleder,d M. P. Diamond,e M. Cedars.f,g aUniversity of California at San Francisco School of Medicine, San Francisco, CO; bObstetrics and Gynecology, University of Colorado School of Medicine, Aurora, CO; c NICHD, Bethesda, MD; dUniversity of Virginia, Charlottesville, VA; e Augusta University, Augusta, GA; fObstetrics, Gynecology and Reproductive Sciences, University of California, San Francisco, San Francisco, CA; g for the Reproductive Medicine Network, Bethesda, MD. OBJECTIVE: To test the hypothesis that women with unexplained infertility (UI) will demonstrate evidence of diminished ovarian reserve across standard measures when compared to a population of community controls. DESIGN: Cross-sectional cohort study. MATERIALS AND METHODS: Patients with UI enrolled in a multi-center trial (AMIGOS) had transvaginal ultrasound determinations of antral follicle count (AFC) (n¼872). A randomly selected subset of subjects from the cohort had AMH levels determined by a central laboratory (ELISA immunoassay, Ansh Labs, University of Virginia Core Ligand Laboratory) using banked specimens (n¼ 245). Controls included participants in the Ovarian Aging (OVA) study and were recruited at one of the AMIGOS study sites (UCSF). The study obtained AFC measurements (n¼962) from a community-based cohort of healthy, ovulatory women not seeking treatment for fertility. A random subset of OVA also had AMH measurement (n¼286) from banked specimens determined by the same assay and at the same central laboratory. CV for the Ansh assay was <5%. Considering UI as the predictor, linear regressions were performed for the following outcomes: AMH, AFC and AMH: AFC ratio while controlling for age, BMI, study site. RESULTS: Results of regression analyses are shown in table 1 CONCLUSIONS: This is the largest study to compare predictors of ovarian reserve in women with UI against a well-characterized control population. Contrary to our hypothesis, women with UI did not show evidence of decreased markers of ovarian reserve. Our findings suggest that ovarian reserve in isolation may not serve as a predictor of future fertility. Supported by: National Institutes of Health Eunice Kennedy Shriver National Institute of Child Health and Human Development (Grant R01 HD044876) and the National Institute on Aging (Grant K08 AG03575); and UCSF Junior Investigator Award.

Adjusted Association between Markers of Ovarian Reserve in Controls vs. UI

Outcome AMH AFC AMH:AFC ratio

B-Coeff (95% CI) Control vs. Unexplained Infertility -1.6 (-5.8,2.5) -3.1 (-8.0,1.8) -.02 (-.32,.28)

p-value NS NS NS

Vol. 106, No. 3, Supplement, September 2016