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Evaluation of newborns for hypoxic ischemic encephalopathy (HIE) includes laboratory and clinical parameters, as well as amplitude integrated electroencephalogram (aEEG). Based on qualifying criteria, selective head cooling (SHC) is initiated for infants with evidence of moderate to severe HIE. However, some newborns may not qualify for hypothermia therapy based on normal aEEG.To compare levels of serum glial fibrillary acidic protein (GFAP), ubiquitin c-terminal hydrolase-1 (UCHL-1) protein and phosphorylated axonal neurofilament heavy chain (pNF-H), in newborns who met initial screening criteria for HIE but did not qualify for head cooling, to the levels in healthy newborns.Newborns ≥36 weeks of gestational age at risk for HIE, who were evaluated but did not qualify for SHC from July 2013 through June 2014 at NYU Langone Medical Center and Bellevue Hospital center were enrolled. A control group included healthy newborns from the newborn nursery (NBN). Serum samples were collected between 24 and 48 h of life from both groups.There was no significant difference in the serum levels of GFAP, UCHL-1 protein and pNF-H between the two groups of infants.Newborns at risk for HIE who met the initial criteria for head cooling but who were excluded based on normal aEEG did not show significant elevation of biomarkers of brain injury compared to healthy newborns. These findings may help to validate using aEEG as an additional evaluation criteria in cooling.