Previous studies in adults have shown that chronic pulmonary hypertension is associated with decreased endothelial nitric oxide synthase (eNOS) expression in pulmonary arteries. However, the role of decreased eNOS expression in persistent pulmonary hypertension of the newborn (PPHN) is unknown. We investigated the hypothesis that umbilical vein endothelial cells cultured from infants with PPHN will have decreased eNOS expression. Umbilical cords were collected from meconium-stained infants at birth, and endothelial cells were isolated if the infants developed PPHN. Endothelial cells were grown in primary culture, and total RNA was isolated. cDNA was reverse transcribed from mRNA and amplified by PCR. An expected product of approximately 550 bp was found in all control infants but only in two of the six infants with PPHN. Identity of the PCR product was confirmed by Southern hybridization to a separate internal eNOS-specific probe. Amplification of beta-actin cDNA, an internal control, was detected in all controls and in all infants with PPHN, including the four infants without the eNOS band. There was no difference in the course and outcome of patients with presence or absence of the eNOS band. However, there was an acidotic arterial blood pH (7.19-7.29) and intrapartum fetal heart rate decelerations in all four infants without eNOS expression. In conclusion, eNOS mRNA was detected in all normal term infants but was notably absent in the majority of infants with PPHN in this pilot study. The development of PPHN is multifactorial, and a decrease in eNOS gene expression may occur in some infants. Whether the decreased eNOS transcript is a cause of PPHN or a result of intrapartum stress remains to be determined.

OBJECTIVE: Congenital cytomegalovirus (CMV) infection is a leading cause of hearing loss and mental retardation throughout the world. Detection of the CMV DNA by polymerase chain reaction (PCR) offers a sensitive, rapid, and specific means of identification. Meconium, the stool formed in utero, may be an ideal specimen for CMV detection. The objective of this study was to develop a PCR-based methodology for the detection of CMV in the meconium of neonates. METHODS: Meconium was collected from 10 newborn infants (seven with positive viral cultures and three uninfected infants born to CMV-seropositive mothers). For each, DNA was isolated from meconium by organic extraction and attachment to a DNA-binding matrix, and PCR was performed using amplimers specific for the major intermediate early (MIE) and late antigenic (LA) regions of CMV. RESULTS: Gel electrophoresis demonstrated an anticipated PCR product of 250 base pairs (bp) corresponding to the MIE region of CMV in all infected and positive control meconium samples. Furthermore, a single band of 150 bp corresponding to the LA region of CMV was also amplified in several of the infected infants. Conversely, no amplification of these antigenic regions was noted in either uninfected infants born to CMV-seropositive mothers or negative controls. CONCLUSIONS: CMV is present within the meconium of infected neonates and is readily detectable by PCR.