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The role of neutrophils on the development of reperfusion injury was evaluated in canine hearts. Reperfusion was performed with a closed reperfusion system. 18 dog hearts were arrested by a single dose of National Institute of Health cardioplegia and stored cold (0.5 degrees C) for 24 hours. Thereafter they were randomly attached to one of the following groups. Group I: reperfusion with whole blood (n = 6), group II: reperfusion with low-leukocyte blood (n = 6), group III: reperfusion with low-leukocyte blood plus phenyl-isopropyl adenosine (PIA) (n = 6). Hearts in group I showed an increase of high energy phosphates (HEP) within the first 30 minutes, and a significant decrease during the subsequent 30 minutes. Concomitantly the initially soft hearts turned into stone hearts. In group II and III a significant increase of HEP was seen during the entire reperfusion. After 60 minutes of reperfusion myocardial ATP levels in group III were significantly higher than in group I. All hearts in group III were soft after reperfusion. It is concluded that reperfusion injury appears to be dependent upon the presence and activation of neutrophils, which can be partially prevented by reperfusion with low-leukocyte blood and more so by the simultaneous administration of PIA.