Modular chemistry analyzer

Weight (nearest 0.1kg), height (nearest 0.1cm), and waist circumference (nearest 0.5cm) were measured without shoes or heavy clothing using standard protocols [21 (link)]. The height and weight measures were used to calculate BMI (kg/m²). Glycated haemoglobin (HbA_1c) was measured from whole blood samples by the high performance liquid chromatography method (ion-exchange with UV detection; Bio-Rad Variant II Turbo, Hemoglobin A_1c Program Reorder Pack, (270–2415)). The method for measuring plasma glucose was enzymatic reference with hexokinase [27 (link)]. HDL-cholesterol and triacylglycerols were measured by an enzymatic colorimetric assay with Roche Modular Chemistry Analyzer (Roche HDL-C Plus Reagent 1 Cat No. 04713311; Roche Triglyceride Reagent Cat No. L1876040).

The urinary KIM-1, NGAL, and H-FABP levels were measured with the duoset enzyme-linked immunosorbent assay (ELISA; R&D systems, Minneapolis, MN, USA). NAG was measured by the modified enzyme assay using p-nitrophenyl-N-acetyl-β-D-glucosaminidase as the substrate. Urinary creatinine was determined on a Roche Modular chemistry analyzer (Roche Diagnostics, Indianapolis, IN, USA). KIM-1, H-FABP, and NAG were measured for the VAPOR-1 study. For this post-hoc analysis, we additionally measured NGAL.