Liaison insulin

MMTs were performed at baseline and at 18 months after a 10-hour overnight fast. Blood was sampled at 0, 30, 60, 90 and 120 min. Participants were asked to stop the use of metformin, when used, 24 hours before the study visit. Participants consumed a 250ml liquid drink (Fortisip; 18.4 g carbohydrate per 100 ml) over a period of 2–5 min. Plasma glucose (mmol/l) was determined using the enzymatic glucose hexokinase method (Konelab 20 XT Clinical Chemistry analyzer, Thermo Fisher Scientific, Vantaa, Finland). C-peptide (nmol/l) and plasma insulin (pmol/l) were determined using chemiluminometric immunoassay (Liaison Insulin and Liaison C-peptid, DiaSorin S.p.A, Saluggia, Italy). HbA_1c (mmol/mol) was measured by ion-exchange high performance liquid chromatography on a TOSOH G8 analyser (Tosoh Bioscience, Inc, CA, USA). Triglycerides, total and high density lipoprotein (HDL) cholesterol were enzymatically assessed. More details on sample handling and biomarker determination are described elsewhere [6 (link)].

IGF1 and insulin concentrations were measured in serum samples from the cord blood of newborns. IGF1 concentration was measured using a chemiluminescent immunometric assay after pre-treatment with acid using Immulite®2000 (Siemens Healthcare Diagnostics Products Llanberis, UK). Insulin concentration was measured using Liaison®Insulin (DiaSorin, Saluggia, Italy).

Venous blood samples were obtained after an overnight fast of at least 10 hours. Biochemical measurements included liver function tests, fasting blood glucose (FBG) and postprandial blood glucose (PPBG) levels, and fasting lipid profile using standard methods as described earlier [11 (link)]. Serum levels of hsCRP were measured using a commercially available reagent kit based on the principle of solid phase enzyme-linked immune sorbent assay (Biocheck, Inc., Foster City, USA). Fasting insulin was measured using a commercially available reagent kit based on the principle of electrochemiluminescence (Liaison® Insulin, DiaSorin Inc., USA). The inter- and intra-assay coefficient of variation was <5%. The HOMA-IR was calculated by using the following formula: HOMA-IR = fasting insulin (µU/mL) ∗ fasting blood glucose (mmol/L)/22.5.