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Serum samples were stored upon collection in local freezers at ?20��C and at ?80��C upon arrival to Providence in 1995. Glucose level, determined with an automatic analyzer, Beckman CX4, and insulin level, BMS-777607 concentration determined with a direct radioimmunoassay (RIA) kit from Diagnostic Products Corporation, were measured upon arrival. Leptin was measured in 1997 with an RIA kit from ALPCO. Estradiol (E2) was measured in 1999 at the Reproductive Ecology Laboratory at Harvard University using a time-resolved fluroimmunoassay (FIA) kit from Wallac. At the time the blood sample was drawn, each woman identified on a calendar the first day (Day 1) of her last menses. Day 1 was then used to count forward and identify women in the follicular phase of their menstrual cycle. The ��forward count�� method, although less precise than hormonal profiles, offers a fairly accurate means of identifying the follicular phase for one-time, cross-sectional data such as these. To control for variation in E2 level attributed to cycle day, we identified women whose E2 values fell within the RVX-208 normal reference range for follicular levels prior to the midcycle peak established by the assay manufacturer. The typical E2 trajectory is marked by relatively low levels at the beginning of the cycle followed by a gradual increase and a rapid rise at midcycle. Linear correlation analysis showed no significant relationship between cycle day and E2 level in our study sample (r?=?0.188, P?=?0.252, n?=?34). Participants were 34 regularly cycling, nonlactating, premenopausal women aged between 32 and 49, of whom 26 were overweight (BMI?>?26 km/m2, adjusted for Pacific Islanders). The sample Protein Tyrosine Kinase inhibitor tended to consist of relatively older overweight women, yet all participants showed indicators of normal glucose tolerance, with physiological values in the normal range of variation for a well-nourished population: fasting glucose levels?