O.         METSIM study

1.         Population (M. Civelek)
2.         Phenotypes (M. Civelek)
3.         Adipose biopsies (M. Civelek)
4.         Adipose expression arrays (M. Civelek)
5.         Adipose RNA-Seq (P. Pajukanta)

 

1.         Population (M. Civelek)

Metabolic Syndrome in Men (METSIM) is an ongoing population-based cross-sectional study conducted in Kuopio, Finland. Subjects, aged from 45 to 70 years, were randomly selected from the population register of the town of Kuopio in eastern Finland (population 95,000).

 


2.         Phenotypes (M. Civelek)

Every participant had a 1-day outpatient visit to the Clinical Research Unit at the University of Kuopio, including an interview on the history of previous diseases and current drug treatment and an evaluation of glucose tolerance and cardiovascular risk factors. Fasting blood samples were drawn after 12 h of fasting followed by an oral glucose tolerance test (OGTT). Plasma glucose was measured by enzymatic hexokinase photometric assay (Konelab Systems reagents; Thermo Fischer Scientific, Vantaa, Finland). Insulin was determined by immunoassay (ADVIA Centaur Insulin IRI no. 02230141; Siemens Medical Solutions Diagnostics, Tarrytown, NY). Height and weight were measured to the nearest 0.5 cm and 0.1 kg, respectively. BMI was calculated as weight (kg) divided by height (m) squared. Waist (at the midpoint between the lateral iliac crest and lowest rib) and hip circumference (at the level of the trochanter major) were measured to the nearest 0.5 cm. Body composition was determined by bioelectrical impedance (RJL Systems) in subjects in the supine position after a 12-h fast. Lipid and amino acid measurements were done with NMR spectroscopy. Glucose tolerance was evaluated according to the ADA criteria.

 


3.         Adipose biopsies (M. Civelek)

The abdominal subcutaneous adipose tissue was surgically extracted. The protocol for this experiment can be found in the following reference:

Stančáková A, Paananen J, Soininen P, Kangas AJ, Bonnycastle LL, Morken MA, Collins FS, Jackson AU, Boehnke ML, Kuusisto J, Ala-Korpela M, Laakso M. Effects of 34 risk loci for type 2 diabetes or hyperglycemia on lipoprotein subclasses and their composition in 6,580 nondiabetic Finnish men. Diabetes. 2011 May;60(5):1608-16. Epub 2011 Mar 18. PubMed PMID: 21421807; PubMed Central PMCID: PMC3292337.

 


4.         Adipose expression arrays (M. Civelek)

Total RNA was isolated from 200 METSIM subjects that underwent subcutaneous fat biopsies, using Qiagen miRNeasy kit according to manufacturer’s instructions. RNA Integrity Number (RIN) values were assessed with the Agilent Bioanalyzer 2100 instrument. Samples with RIN values greater than 7.0 were used for transcriptional profiling. High quality RNA samples were hybridized to Illumina Human HT-12 v3 Expression BeadChips. Genome Studio software (2010.v3) was used for obtaining fluorescent intensities. HT-12 BeadChip contains 48,804 expression and 786 control probes. 19,306 probes were removed due to (i) lack of mapping to any genomic or transcriptomic location, (ii) mapping to multiple genomic or transcriptomic locations, and (iii) presence of single nucleotide polymorphisms in the probe sequence that may affect hybridization efficiency using the methodology developed by Barbosa-Morais et al. (1). The remaining 29,497 probes were processed using non-parametric background correction followed by quantile normalization with both control and expression probes utilizing the neqc function in the limma package (R v2.13.0) (2). 16,223 probes with detection p-values <0.01 in any of the 200 samples were used for further analysis.

 


5.         Adipose RNA-Seq (P. Pajukanta)

In development.