A sample of 30 mL blood was drawn from mainly nonfasting participants at baseline and collected in monovettes containing 10% citrate. Samples were aliquoted and stored in tanks of liquid nitrogen (−196°C) or deep freezers (−80°C) (20 (link)). Plasma NT-proBNP, estradiol, testosterone, and sex hormone–binding globulin (SHBG) concentrations were measured using a solid-phase two-site chemiluminescent immunometric assay (IMMULITE 2000 Systems Analyzers; SIEMENS, Eschborn, Germany) at the Institute of Clinical Chemistry, University of Magdeburg, Magdeburg, Germany. The assay has an effective measuring range of 20–35,000 pg/mL. The median within-run coefficient of variation was 5.4% at a concentration of 35.6 pg/mL and 4.1% at a concentration of 29,725 pg/mL. The overall coefficients of variation throughout the analyses were 6.4% and 4.7% at the same sample concentrations (18 (link)). Plasma adiponectin was measured with a sandwich ELISA (LINCO Research, St. Charles, MO); HDL and total cholesterol, triglycerides, hemoglobin A1c (HbA1c), and hs-CRP were measured using an automatic ADVIA 1650 analyzer (Siemens Medical Solutions) at the University of Tübingen, Tübingen, Germany. All plasma biomarker measurements were corrected for the dilution introduced by citrate volume to improve comparability with concentrations measured in EDTA plasma (22 (link)).