Table 4 Blood sampling procedure

EDTA tube (4 × 6.0 mL)

Myeloperoxidase, neopterin, soluble intercellular adhesion molecule, soluble vascular cell adhesion molecule-1, soluble E selectin, soluble P selectin, CD40 ligand, interleukin-6, tumor necrosis factor alpha, Homocysteine

Transported on ice to the Biobank Maastricht UMC+ where the blood is preprocessed before storing. EDTA plasma is centrifuged and stored at −80 °C until analysis.

Natriumfluoride/EDTA tube (1x 4.0 mL) Coagulation tube (1 × 5.0 mL)

Glucose, total cholesterol high-density lipoprotein-cholesterol, low-density lipoprotein-cholesterol, triglyceride, folate, vitamine B12, high sensitivity C-reactive protein

Direct analyses of these markers are performed at the laboratory on the day of sample collection after centrifuging.

Buffycoats from the 4 EDTA tubes for DNA extraction

Apoliprotein E, clusterin, phosphatidylinositol binding clathrin assembly protein, encoding complement [3b/4b] receptor 1, angiotensin-converting enzyme,methylenetetrahydrofolate reductase

Buffycoats are taken from four EDTA tubes and separately stored at the Biobank until analysis.

Epigenetics 1 PAXgene tube (2.5 mL)

RNA

Blood is collected in PAXgene tubes to ensure long-term stability of ribonucleic acid and the tubes are stored at the Biobank. RNA extraction and subsequent analysis of expression of inflammatory and stroke-related genes at the mRNA level will be done.