Euglycaemic and hyperglycaemic clamp technique combined with tracer technology and indirect calorimetry for the evaluation of insulin action and insulin secretion in vivo and for the estimation of whole-body glucose disposal (insulin sensitivity), endogenous (hepatic) glucose production, glucose and lipid oxidation as well as non-oxidative glucose metabolism (glycogen storage), glucose disappearance and glycolytic flux.
- Frequently-sampled intravenous glucose tolerance test (FSIGT) for the estimation of insulin secretion, insulin sensitivity and glucose effectiveness.
- Muscle biopsies a. m. Bergström for: 1) establishment and studies of cultured skeletal muscle cells (myotubes); 2) isolation of mitochondria for the evaluation of respiration and oxidative enzyme activities; 3) morphometric studies of intramyocellular content of glycogen, lipids, mitochondria and glucose transporters; 4) studies of mRNA expression (real time PCR analysis and Affymetrix gene chips); 5) studies of expression, activity and phosphorylation of muscle enzymes involved in insulin action in muscle; 6) quantitative proteome analysis of muscle based on mass-spectrometry alone and/or using 2-dimensional gel electrophoresis.
- Spectrophotometry, spectroflurometry, luminescence, HPLC, and radiotracer techniques for enzymes kinetic studies and intracellular metabolites concentrations in both lipid and glucose metabolism.
- Hormone profile identification: pulsatile secretion and diurnal variation of hormones. Blood samples collected through heparinised intravenous catheter from Carmeda, Sweden.
- Automatised calculation of abdominal fat deposits after MRI-scanning of the abdomen. Identification of subcutaneous, intra-abdominal anterior and intra-abdominal posterior fat.
- In vitro measurements of insulin receptor numbers, tyrosine-kinase activity, GLUT4, IRS-1, PI-3 kinase, glycogen synthase, morphological measurement of lipid deposition and glucose transporters in skeletal muscle.
- Proteome analysis of skeletal muscle, specifically in relation to phosphorylation of glycogen synthase.
- Ultrasonic estimation of thyroid and parathyroid nodules
- Computer-programmed evaluation of hormone pulsatility and techniques for genetic screening procedures.
- Measurement of bone density and body composition (DXA-scanning).
- Histomorphometric analysis of cortical bone remodelling.
- RNA-extraction from bone tissues.
- Quantitative PCR-analysis of RNA encoding an array of cytokines including TGF-beta, IL-1, IL-6, OPG, and RANK-L.
- Muscular strength in the extremities measured by:
- Jumping height identified by contact mat, New-test, Finland.
- Extension of the extremities by Nottingham Power Rig, England.
- Modified WinGate spurt test on ergometer bike, Monarch, Sweden.
- Isometric strength of biceps brachii by strength coach apparatus, specially constructed unit, Denmark.
- Oxygen uptake carried out on ergometer bike, Monarch, Sweden, with lactate measurement, a direct test, Oxicon pro, Jaeger, Germany.
- Validated questionnaire for health profile/life quality SF36, validated in Denmark 1993 and applied on more than 1500 published studies.
- Dual energy x-ray absorptiometry (DEXA), morphometric x-ray absorptiometry (MXA), bioimpedance measurement, measurement of muscle strength, measurement of broad-band ultrasound attenuation and speed of sound in bone (calcaneus and phalanges). Transiliac bone biopies (Bordier's trepine).
- Cell culture, stem cell cultures, human embryonic stem cells
- RNA, DNA isolation
- Nothern blotting, Southern blotting, Western blotting
- Immunohistochemistry and in situ hybridization
- Confocal microscopy
- Cloning, gene transfer using viral and non-viral vectors
- Quantitative real time PCR
- FACS analysis and cell sorting