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Immediately before assay, all samples were brought to room temperature. No sample was thawed more than once. Quality control samples, consisting of four pairs at high, medium, and low concentrations were included in each assay. IGF-I and IGFBP-3 were measured by chemiluminescence technology using reagents from Immunodiagnostics Systems, Boldon Business Park, Boldon, Tyne and Wear, UK. C-peptide and IGF-II were measured using ELISA with reagents from ALPCO Diagnostics, Keewaydin Drive, Salem, NH, USA. Leptin was also measured by ELISA using reagents from R&D Systems, Minneapolis, MN, USA. Selleckchem ABT-263 The manufacturer-stated limits of detection for IGF-I, IGF-II, IGFBP-3, C-peptide, and leptin are 10�C1,200 ng/mL, 0.02�C3,600 ng/mL, 80�C10,000 ng/mL, 0.011�C10.8 ng/mL, and 0.0078�C100 ng/mL, respectively. Statistical analysis of the data was carried out using STATA Version 11 (STATA Corporation, College Station, TX, USA). Differences in mean IGF levels between serum and plasma samples were assessed using paired t-tests. Within-batch coefficients selleck chemical of variation (CVs) were calculated from four pairs of quality control samples in each batch. Correlations were determined using parametric and nonparametric analyses. Data were log-transformed but untransformed data are reported. Lin��s7 concordance correlation coefficient (rho-C) was calculated to test for agreement in concentrations in the two types of specimens. The concordance correlation coefficient combines measures of both precision and accuracy to determine how far the observed data deviate from the line of perfect concordance (ie, the line at 45�� on a square scatterplot).8 Accuracy is depicted by the nearness of the data��s reduced major axis to the line of perfect concordance and precision is reflected by the tightness of the data about its reduced major axis. Results Assay characterization The experimental intraassay CVs, which represent both intrabatch laboratory error and the difference associated with the two types of specimens, ROR1 ranged from 0.4%�C10% (Table 1), with the CVs being highest for IGF-II and C-peptide. Differences were noted between the concentrations of the five analytes measured in serum versus those measured in EDTA plasma, with mean concentrations in serum being consistently higher (Table 1; all P