Levels of platelet-activating factor (PAF) in blood from patients with ischemic stroke were determined by radioimmunoassay (RIA). Using 2 ml of blood as a starting material, PAF was detected in 11 out of 17 stroke patients and 3 of 25 age-matched healthy controls. This implies that blood level of PAF is higher in stroke patients than in controls. Plasma levels of PAF-like lipid(s) (PAF-LL) were also estimated in the same subjects by a bioassay based on aggregation of human polymorphonuclear neutrophils. PAF-LL was detected in plasma samples of all subjects and the average values in patients and controls were 294 +/- 211 pg/ml and 140 +/- 122 pg/ml, respectively. There was a statistically significant difference between these two values (p less than 0.01). Separation of plasma lipids by HPLC gave a single peak in bioassay, which had the same elution volume as authentic PAF. When each fraction was subjected to RIA, the fractions corresponded to phosphatidylcholine (PC) or lysoPC also showed the immunoreactivity, however, the purification procedure using an octadecylsilica gel cartridge eliminated such cross-reacting compounds. We conclude that blood PAF is higher in patients with ischemic stroke than in healthy subjects. Besides, there may be bioactive phospholipid molecules other than PAF, which level in plasma is also higher in stroke patients.