Inhaled nitric oxide (iNO) improves oxygenation in premature infants, but concern has been raised about its potential oxidative toxicity. We designed this study to assess the oxidative balance in premature infants who were exposed to low dose iNO and the relationship with their clinical outcome on day 28 of life. A total of 274 infants who were Ͻ32 wk gestation were randomized at birth to receive 5 ppm of iNO if they presented with hypoxemic respiratory failure. Nonhypoxemic infants were studied as the reference group. Blood samples were withdrawn 24 h apart, within the first 4 d of life, to assess malondialdehyde (MDA) concentration as oxidative stress marker and total plasmatic glutathione (GSH), intraerythrocyte GSH peroxidase, and GSH reductase activities as antioxidant defenses. After 24 h, the rise in MDA was blunted in the iNO group compared with controls and was close to the reference infants. Conversely, GSH was more stable in the iNO group, when there was no difference for the GSH peroxidase and GSH reductase activities. On day 28, Oxygen dependence was linked with a higher increase in MDA as was the risk for death, whereas intraventricular hemorrhage was associated with a higher initial drop in GSH. Early low-dose iNO in hypoxemic preterm infants improves oxidative balance and seems to be clinically beneficial up to day 28 of life. Since 1992, several randomized trials have shown that inhaled nitric oxide (iNO) significantly improved oxygenation in term or near-term infants, with a significant reduction in the use of extracorporeal membrane oxygenation (1-6). In 1997, Skimming et al. (7) were the first to report a positive response of 23 premature infants with respiratory distress syndrome to low-dose iNO (5 ppm). Since then, a few clinical trials involving a small number of premature infants have shown that the initial improvement in oxygenation seemed to be transient (8 -10) because no improvement in mortality or in morbidity had been demonstrated (11). However, most of these studies were considering iNO therapy in premature infants who had severe hypoxemic respiratory failure or refractory hypoxemia (8,10,11) or were at an advanced stage of respiratory insufficiency (12). Most of them used a high dosage of iNO (10 -20 ppm).Finally, concerns have been raised about specific side effects of this new molecule that could worsen long-term outcome after an initial improvement in this high-risk population (13)(14)(15). Besides the risk for increased brain hemorrhage as a result of a prolonged bleeding time (16), neonatologists remain cautious in the use of this treatment because NO behaves as a "chameleon" molecule for the oxidative balance. Endogenous NO is incriminated in the cascade leading to cerebral lesions in perinatal experimental models of hypoxo-ischemic injuries (17). In addition, iNO may interact with oxygen to form nitrogen dioxide and peroxynitrites, which would enhance pulmonary toxicity. This is a main concern when dealing with Received May 20, 2004; accepted September 27, 2004