Hexokinase method

Plasma triglyceride and insulin levels were measured using commercially available kits (Roche Molecular Biochemicals, Indianapolis, IN, and Crystal Chem Inc., Downers Grove, IL, USA) and glucose levels were determined according to the hexokinase method (Instruchemie). Plasma levels of multiple cytokines were evaluated simultaneously by using pre-coated multisport plates in an ELISA-based electrochemiluminescence assay (Meso Scale Discovery, Gaithersburg, MD).
Coagulation factor levels were measured as previously described [11 (link)] and pooled mouse plasma was used to generate standard curves. Global coagulability of the plasma was determined by measuring the prothrombin time (PT) and activated partial thromboplastin time (aPTT) on the STart 4 analyzer (Diagnostica Stago, Leiden, The Netherlands) using the STA Neoplastine Plus (Diagnostica Stago) and the TriniCLOT Automated APTT reagent (TCoag, Ireland), respectively.
The in vivo clearance rates of human coagulation factors VII, VIII and IX were analyzed with home-made ELISAs specific for human proteins which did not cross-react with mouse plasma proteins. Standard curves were generated by adding Cofact or Aafact to pooled mouse plasma (final concentration 20%) to calculate human antigen levels, and the level measured directly after injection (1 minute) was set as a reference (100%).

For patients included in the study, the diagnosis of CMD (type 2 DM, IGT, or IFG) was checked in the aforementioned 5-year period. For each selected patient, all data from electronic medical records were downloaded.
Table 1 shows the examination methods.
Laboratory parameters were measured by using standard laboratory methods. Glucose was measured in venous plasma using hexokinase method (Roche, Germany).
Statistical patterns influencing the development of CMDs and death within 5 years after AMI were identified.

Blood glucose concentrations were measured using hexokinase-method (Roche, Basel, Switzerland). Insulin was assessed by human insulin ELISA, Merck Millipore, Billerica, USA; sensitivity of the assay 1 μU/mL. Liver enzymes were assessed by routine diagnostics by standardized International Federation of Clinical Chemistry protocol using pyridoxale-5phosphate.