Patient flow.

<p>Patient flow.</p

Ibuprofen treatment characteristics.

<p>Data are expressed as number (%) or mean (SD: standard deviation). MD: missing data.</p><p>*: p values were obtained with Fisher’s exact test (category 1 versus 2 and 3).</p><p>Ibuprofen treatment characteristics.</p

Risk of failure of the first course of ibuprofen.

<p>GA: gestational age; AGC: antenatal glucocorticoids (defined by at least one complete course of two doses at an interval of 24 hours); PROM: premature rupture of membranes (>24 hours).</p><p>SD: standard deviation; OR: odds ratio; CI: confidence interval.</p><p>Risk of failure of the first course of ibuprofen.</p

Perinatal population characteristics.

<p>Perinatal population characteristics.</p

qRT-PCR analysis of validated mRNA targets of the miRNAs differentially expressed in lungs at P10 and P21 after LPD-induced IUGR.

<p>Five genes were selected from among the target genes according to their described function and quantified by qRT-PCR at P10 and P21. The relative expression was normalized to that of housekeeping genes (GAPDH and MMP16 [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0190445#pone.0190445.ref016" target="_blank">16</a>]). Data are mean ± SEM of 5 animals/group. *, <i>p</i> < 0.05; two-tailed Mann-Whitney test.</p

Flow chart of infants who met inclusion/exclusion criteria for the study.

<p>PNC, postnatal corticosteroids; NICU, neonatal intensive care unit; GA: gestational age.</p

Lung microRNA deregulation associated with impaired alveolarization in rats after intrauterine growth restriction

<div><p>Intrauterine growth restriction (IUGR) was recently described as an independent risk factor of bronchopulmonary dysplasia, the main respiratory sequelae of preterm birth. We previously showed impaired alveolarization in rat pups born with IUGR induced by a low-protein diet (LPD) during gestation. We conducted a genome-wide analysis of gene expression and found the involvement of several pathways such as cell adhesion. Here, we describe our unbiased microRNA (miRNA) profiling by microarray assay and validation by qPCR at postnatal days 10 and 21 (P10 and P21) in lungs of rat pups with LPD-induced lung-alveolarization disorder after IUGR. We identified 13 miRNAs with more than two-fold differential expression between control lungs and LPD-induced IUGR lungs. Validated and predicted target genes of these miRNAs were related to “tissue repair” at P10 and “cellular communication regulation” at P21. We predicted the deregulation of several genes associated with these pathways. Especially, E2F3, a transcription factor involved in cell cycle control, was expressed in developing alveoli, and its mRNA and protein levels were significantly increased at P21 after IUGR. Hence, IUGR affects the expression of selected miRNAs during lung alveolarization. These results provide a basis for deciphering the mechanistic contributions of IUGR to impaired alveolarization.</p></div

Association of NICU practices and PNC use.

<p>Association of NICU practices and PNC use.</p

Primer sequences used for RT-PCR.

<p>Primer sequences used for RT-PCR.</p

Experimentally validated targets (source: Tarbase and Mirtarbase) for the 13 significantly modulated miRNAs, sorted by Fold change in expression, during alveolarization in rats with low-protein diet-induced intrauterine growth restriction.

<p>Experimentally validated targets (source: Tarbase and Mirtarbase) for the 13 significantly modulated miRNAs, sorted by Fold change in expression, during alveolarization in rats with low-protein diet-induced intrauterine growth restriction.</p