mRNA Fold increase over controls with 14 Day Ureaplasma exposure.

<p>* p&lt;0.05 vs saline controls.</p

Pro-inflammatory cytokine and acute phase mRNA responses in the lung.

<p>(A) IL-1β did not increase with V_T (<i>p</i>>0.05). (B, C) IL-6 and IL-8 mRNA increased similarly for the V_T and V_T+SI groups, with no increase with SI relative to PEEP alone. (D, E) The acute phase genes Egr-1 and Cyr61 also increased with V_T and V_T+SI, with no increases with SI relative to PEEP alone. (F) CTGF mRNA did not change. *<i>p</i><0.05 vs PEEP only.</p

Groups, intra-amniotic (IA) exposures, gestational age and ventilation variables.

<p>(a) Gisslen <i>et al</i>. Innate immunity 2013 <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0096087#pone.0096087-Gisslen1" target="_blank">[12]</a>, (b) Collins <i>et al</i>. AJP-Lung 2010 <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0096087#pone.0096087-Collins1" target="_blank">[10]</a>, (c) Polglase et al. Pediatr Res 2010<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0096087#pone.0096087-Polglase1" target="_blank">[11]</a></p

Pancytokeratin expression.

<p>Representative photomicrographs for pancytokeratin (1:2500) immunohistochemical staining of fetal skin. (A) control; (B) LPS at 16 h; (C) LPS and IL-1ra at 16 h exposure. (D) Graph shows the comparison between groups in the percent of positive cells of different intensity stain; intense (arrow), moderate and no stain (magnification bar represents 20 μm). <b>*</b>indicates a significant difference in the percent of positive cell between LPS at 16 hours and untreated control group. ɸ indicates a significant difference in the percent of positive cell between LPS group and LPS plus IL-1ra group at 16 hours.</p

In situ localization of EREG and AREG with LPS exposure and mechanical ventilation in lung.

<p>(<b>A–C</b>) EREG mRNA was not found in (A) unventilated controls or (B) IA LPS exposed lungs, but dramatically increased in peripheral lungs of lambs receiving (C) mechanical ventilation. EREG mRNA is localized to dome-shaped cells in peripheral lung (C, Insert). (<b>D–F</b>) AREG mRNA is found scattered in the peripheral lung of (D) Unventilated controls with a mild increase with (E) IA LPS exposure. (F) Mechanical ventilation increases the mRNA signal in similar cells to EREG. Scale bar = 50 µm</p

Total Protein and Heat Shock Proteins in FLF and BAL.

<p>Heat shock protein (HSP) Fetal Lung Fluid (FLF) Bronchoalveolar lavage fluid (BALF) Values: Mean ± SEM.</p><p>* p<0.05 vs PEEP and SI groups.</p><p>Total Protein and Heat Shock Proteins in FLF and BAL.</p

Cytokine PCR Analysis.

<p>Quantitative PCR analysis of cytokines in fetal rhesus macaque skin following intra-amniotic exposure with LPS, IL-1β or UP and for different times of exposure (Panel a, b and c). Cytokine expression for LPS and LPS plus IL-1ra exposure groups (Panel d, e and f). Graphs represent mean mRNA fold change ± SD. <b>*</b>indicates a significant increase in transcript expression relative to untreated control group normalized to a value for 1.0. ɸ indicates a significant decrease in transcript expression relative to LPS at 16 hours.</p

Summary of PCR sets.

<p>Summary of PCR sets.</p

<i>Treatment groups</i> Intra-amniotic injections of LPS, IL-1β, and <i>Ureaplasma parvum</i> (UP) or saline were performed in multiparous macaques of similar weight and age.

<p>Separate animals were used for controls for each treatment group. Mean fetal weights at tissue collection were not different between groups.</p

Angiotensin converting enzyme (ACE) 1 and 2 mRNA in lung with exposure to LPS, UP and mechanical ventilation.

<p>(<b>A–C</b>) mRNA values for unventilated controls (UVC), IA-LPS, and mechanical ventilation at 7 mL/kg (V_T7). (A) ACE1 was decreased by LPS but unaffected by V_T7. (B) ACE2 was increased by LPS and LPS+V_T7. (C) ACE2/ACE1 mRNA ratio increased with LPS and LPS+V_T7. * p&lt;0.05 vs UVC.</p