Measurements and cytokine analyses
leukemia, tumor necrosis, TNF‐α
BLOOD, TUMOR NECROSIS, LEUKEMIA
Blood pressure, weight, and height were measured, and 12‐hour fasting blood samples were taken before randomization at the screening visit (0 weeks) and at the 4‐, 12‐ and 20‐week visits. EDTA‐plasma was separated from the blood using standard procedures. In addition, a 24‐hour urine collection was performed before the 1‐, 4‐, 12‐ and 20‐week visits. Plasma and urine samples were frozen and stored at –70°C until assayed.Daily urinary sodium excretion was determined using an ion‐selective electrode (Optima analyzer, Thermo Electron Oy, Vantaa, Finland). Based on the measured 24 h urinary sodium (U‐Na) excretion rate we calculated the estimated daily salt (sodium chloride, NaCl) intake using a formula NaCl (g/24h) = U‐Na (mmol/24h)x58.44×0.001.The concentrations of 45 cytokines were measured from previously unthawed plasma samples using antibody‐based magnetic bead kits (Procarta Plex, Thermo) for multiplexed protein quantification according to the manufacturer's instructions. The cytokines analyzed were: interleukins (IL) IL‐1α, IL‐1β, IL1‐RA (receptor antagonist), IL‐2, IL‐4, IL‐5, IL‐6, IL‐7, IL‐8, IL‐9, IL‐10, IL12p70, IL‐13, IL‐15, IL‐17A, IL18, IL‐21, IL‐22, IL‐23, IL‐27, IL‐31, interferons (IFN) IFN‐α, IFN‐γ, tumor necrosis factors TNF‐α, TNF‐β, chemokines CCL2, CCL3, CCL4, CCL5, CCL11, CXCL1, CXCL10, and CXCL12, and growth factors bNGF (nerve growth factor beta), EGF (epidermal growth factor), FGF‐2 (fibroblast growth factor‐2), HGF (hepatocyte growth factor), LIF (leukemia inhibitory factor), SCF (stem cell factor), BDNF (brain‐derived neurothrophic factor), GM‐CSF (granulocyte‐macrophage colony stimulating factor), PDGF‐BB (platelet derived growth factor‐BB), PlGF1 (placental growth factor), VEGFA (vascular endothelial growth factor A), and VEGFD.All measurements and analyses were done blinded to the intervention allocation of the subject. The baseline and follow‐up samples were always analyzed in one analytical run.