branching in chick lung embryo. There is currently little to no information regarding the role of NF-kB in mammalian lung development. Objectives: To determine the role of NF-kB during mouse lung embryogenesis by (1) characterizing NF-kB expression pattern in wild-type mice and (2) generating hypermorphic transgenic mice for NF-kB to determine the phenotypic impact of NF-kB over- expression. Design/Methods: Pregnant wild-type C57BL/6 mice and newborn pups were sacrificed to obtain fetal lungs at designated gestational ages ranging between E12 and E18 for mRNA and protein analysis of NF-kB. A transgenic mouse line was generated by first subcloning human NF-kB (p65) cDNA into a SpC-promoter-driven expression vector followed by subsequent injection into single- to two-staged mouse embryos and reimplantation into pseudopregnant female mice. The offspring were genotyped to establish expression of the NF-kB transgene and lungs were harvested for mRNA, protein, and histologic analysis. Results: NF-kB mRNA expression in the developing lung is maximal between E12 to E15 and decreases to minimal levels from E18 through adulthood. Transgenic mice overexpressing NF-kB in type II alveolar precursor cells demonstrated increased NF-kB mRNA levels via Northern blot and increased protein expression via immunohistochemical analysis compared with wild-type controls. Importantly, overexpression led to phenotypic changes characterized by a paucity of airspaces. Finally, immunohistochemical analysis revealed an increase in markers for alveolar type II cells. Conclusions: The phenotypic changes due to overexpression of NF-kB in the developing mouse lung suggest that NF-kB may play an important role during lung morphogenesis and subsequently during lung injury and repair. The molecular signals regulating NF-kB expression remain to be elucidated. 128 FETAL LUNG-TO-HEAD RATIO AND LIVER HERNIATION IN CONGENITAL DIAPHRAGMATIC HERNIA: PROGNOSTIC VALUE VARIES BASED ON SITE OF NEONATAL CARE. A. Baraghoush, 1 J. Farrell, 2 R. Bisgaard, 2 R.A. Filly, 2 H. Lee, 2,3 K.K. Nobuhara, 2,3 R.L. Keller, 1,2 1 Department of Pediatrics, 2 Fetal Treatment Center, and 3 Department of Surgery, University of California, San Francisco, CA. Background: Congenital diaphragmatic hernia (CDH) causes fetal and neonatal lung hypoplasia. Severe fetal CDH has been defined by midgestation ultrasonography (US) and characterized by the presence of liver herniated into the thorax with a low lung-to-head ratio (LHR) (# 1.4). LHR is the area of the contralateral lung at the level of the cardiac atria divided by head circumference. We previously performed a single-center randomized study of fetal tracheal occlusion (FTO) versus standard care for severe fetal CDH. The trial was stopped prematurely due to an unexpectedly high survival rate in the control group (compared with historical data). Aim: To determine if survival to hospital discharge (DC) in infants receiving neonatal care at a single center (University of California, San Francisco [UCSF]) differs from survival at other centers (Other), after accounting for fetal liver herniation and LHR. Methods: We identified all UCSF US evaluations with diagnosis of CDH and LHR at 20 to 296/7 weeks’ gestational age (GA, 1991–2006, n 5 237). Patients were excluded if palliative care was elected (n 5 37) or survival was unknown (n 5 20). The primary outcome was survival to DC. If liver was herniated, fetal CDH severity was defined by LHR group (LHR [139} 1.0 [LHR 1.0], 1.0 , LHR # 1.4 [LHR 1–1.4], and LHR . 1.4 [LHR1.4]). Data were analyzed by chi square, Fisher’s exact, or rank sum test; p , .05 was considered statistically significant. Results: Survival for infants with liver herniation and standard care at UCSF versus Other was 26 of 44 (59%) versus 40 of 82 (49%) (p 5 .27). Evaluation by the LHR group revealed significantly better survival at UCSF in the lowest LHR group only (LHR 1.0: 17 of 30 [57%] vs 8 of 37 [22%], p 5 .003). In other LHR groups there were no significant survival differences (LHR 1–1.4: 6 of 8 [75%] vs 22 of 33 [67%], p 5 1.0; LHR 1.4: 3 of 6 [50%] vs 10 of 12 [77%], p 5 .27). Without liver herniation, survival was not different by site of neonatal care (UCSF vs Other: 14 of 14 [100%] vs 12 of 16 [75%], p 5 .10). GA at delivery was comparable by site of care and CDH severity. There was no survival difference among UCSF infants with severe fetal CDH after FTO versus standard care (23 of 37 [62%] vs 23 of 38 [61%], p 5 .88). There were also no differences by LHR group. Conclusions: Survival differences based on fetal CDH severity were significantly different by site of neonatal care only for infants with the most severe fetal CDH (liver herniated, LHR 1.0). This difference may be explained by consistent care at UCSF, a high- volume neonatal referral center. There were no survival differences seen when evaluating only UCSF infants with severe fetal CDH, regardless of whether FTO was performed. 129 HISTONE DEACETYLASE INHIBITORS VALPROIC ACID AND TRICHOSTATIN A PERMIT ALVEOLAR FORMATION IN PRETERM LAMBS MANAGED FOR 72 HOURS BY MECHANICAL VENTILATION. K.H. Albertine, S. Amundsen, L. Dong, M.J. Dahl, R.A. McKnight, D.M. Null, B. Yoder, R.H. Lane, Pediatrics, University of Utah, Salt Lake City, UT. Objective: Chronic lung disease (CLD) of prematurity is characterized by DNA hypermethylation and H3 hypoacetylation. Long-term changes in gene expression and phenotypic changes suggest persistent changes in gene expression by altering determinants of chromatin structure. We hypothesized that disruption of histone deacetylation will permit alveolar formation. Materials/Methods: Preterm lambs (<132 days’ gestation; term <148 days), treated with antenatal steroids and postnatal surfactant, were managed by mechanical ventilation (MV) for 3 days and treated daily with valproic acid (VPA; 25 mg/ kg, IV; n 5 4), trichostatin A (TSA; 0.10 mg/kg, IV; n 5 4), or vehicle (n 5 4). The positive gold standard was management by nasal CPAP (n 5 4). At the end of 3 days, the lungs were analyzed morphometrically. Results: VPA-treated and TSA-treated preterm lambs had more advanced alveolar formation than vehicle controls, as shown by morphometric measurements of radial alveolar count (6 6 2 and 6 6 1 vs 2 6 1, respectively; p , .05), secondary septal volume density (11 6 3 and 10 6 3 vs 5 6 2, respectively; p , .05), and alveolar wall thickness (2.89 6 0.07 and 2.97 6 0.08 vs 3.88 6 0.10, respectively; p , .05). Alveolar formation in the VPA- or TSA-treated lambs was comparable to that in nasal CPAP-managed preterm lambs. Conclusions: Management of preterm lambs with MV leads to alveolar simplification. Treating preterm lambs with valproic acid or trichostatin A, histone deacetylase inhibitors, permitted alveolar formation. We conclude that histone acetylation, which affects patterns of gene expression, is necessary for alveolar formation. HL62875, HL56401, HD41075, CHRC. 130 INVOLVEMENT OF CYCLOOXYGENASE 2 IN LIPOPOLYSACCHARIDE-INDUCED PRENATAL LUNG INFLAMMATION IS INDEPENDENT OF THE PARATHYROID HORMONE–RELATED PROTEIN SIGNALING PATHWAY. L. Cerny, Y. Wang, J. Santos, R. Sakurai, J.S. Torday, V.K. Rehan, Department of Pediatrics, Los Angeles Biomedical Research Institute at Harbor-UCLA Medical Center, Torrance, CA. Background: Prenatal inflammation results in a paradoxical decrease in respiratory distress syndrome but an increase in bronchopulmonary dysplasia. We have implicated epithelial-mesenchymal signaling in mediating this response. Cyclooxygenase 2 (COX-2) and its metabolites are known mediators of lung inflammation, yet their role in parathyroid hormone–related protein (PTHrP)-mediated epithelial- mesenchymal paracrine signaling is not known. We hypothesize that COX-2 plays an integral role in inflammation-induced lung injury via PTHrP-driven signaling. Objectives: (1) Determine the dose response and time course of COX-2 expression and prostaglandin E 2 synthesis in lipopolysaccharide (LPS)-induced lung injury; (2) determine the specificity of COX-2 in mediating inflammation-induced effects on PTHrP signaling by examining (a) the effect of the nonselective COX inhibitor indomethacin on the expression of PTHrP signaling pathway-related markers and (b) the effect of the PTHrP signaling pathway agonist PGJ 2 on COX-2 expression. Design/Methods: Fetal rat lung explants (FRLE), alveolar type II cells (ATII), and lipofibroblasts (LF) from embryonic day 19.5 Sprague-Dawley rat fetuses were treated with LPS (0–50 mg/mL) with or without I (1 or 10 mM) and PGJ2 (5–50 mM) for up to 72 hours. FRLE, LF, and ATII cells were maintained in standard culture media. Using RT-PCR and Western hybridization, the expression of COX-2 and markers of the PTHrP pathway were analyzed, including PTHrP, PTHrP receptor, peroxisome proliferator–activated receptor c, adipocyte differentiation–related protein, surfactant protein B, and cholinephosphate cytidyltransferase a. Results: LPS treatment of FRLE, ATII, and LF increased COX-2 expression in a time- and dose- dependent manner. Increased expression of COX-2 and markers of the PTHrP signaling pathway were blocked by pretreatment with indomethacin. Treatment with PGJ2 blocked the effect of LPS on PTHrP signaling pathway markers but did not block the increase in COX-2 expression. Conclusion: COX-2 may play an integral role in LPS-induced prenatal lung inflammation and its modulation may attenuate the consequent lung injury. COX-2 does not seem to be centrally involved in LPS-induced effects on PTHrP-driven epithelial-mesenchymal paracrine signaling. We speculate that in combination, COX-2 inhibitors and PTHrP pathway agonists might have an additive effect on blocking LPS-induced lung injury. NIH grants HL55268 and HL075405. 131 WNT5A: A POTENTIAL LINK BETWEEN INFLAMMATION AND MORPHOGENESIS IN BRONCHOPULMONARY DYSPLASIA. M.F. Villosis, C. Li, M. Durand, R. Ramanathan, K.Y.C. Kwong, P. Minoo. Division of Newborn Medicine, Keck School of Medicine, University of Southern California, Los Angeles, CA. Introduction: Wnt5a is a signaling molecule that is expressed in both the mesenchyme and the epithelium of the embryonic lung and plays a key role in its morphogenesis. Recently, Wnt5a was found in adult mononuclear cells. Whether inflammatory cells from the lungs of preterm neonates at risk for BPD also express Wnt5a is unknown. Objective: To examine Wnt5a expression in lung inflammatory cells from preterm neonates. Methods: Inflammatory cells (macrophages and neutrophils) were isolated from lungs of human preterm neonates. Cells were incubated in the presence or absence of LPS and Wnt5a mRNA was measured by real-time RT-PCR. Results: Wnt5a was constitutively expressed by lung inflammatory cells from preterm infants. LPS stimulated Wnt5a mRNA in some but not all samples. The magnitude of response correlated with gestational age, birth weight, and positive endotracheal tube and blood cultures. Conclusion: Expression of Wnt5a by lung inflammatory cells may impact on inflammatory cells functions and/or a morphogenetic impact on lung structural development. Supported by NHLBI and the Hastings Foundation. 132 STANDARDIZED NASAL CANNULA ORDER SETS DECREASE OXYGEN DAYS IN INFANTS , 1,500 G. M. Nelson, J. Zagorski, D. Regan, F. McGovern, Emanuel Children’s Hospital, Portland, OR. A standardized approach to weaning supplemental nasal cannula oxygen more quickly and consistently was undertaken in an attempt to minimize oxygen exposure in infants convalescing in the NICU at Emanuel Children’s Hospital. All nasal cannula support is administered with blended oxygen at this institution, so two parameters can be weaned: flow and oxygen tension. There is often confusion among the staff as to which parameter to wean consistently, leading to prolonged days on supplemental oxygen. Standardized order sets with weaning guidelines illustrated by flow diagram were developed. A 3-month pilot study was undertaken. All infants admitted to the NICU requiring nasal cannula oxygen were eligible. Infants with cyanotic heart disease were excluded. Data were prospectively collected on infants managed with nasal cannula order sets. Infants in the experimental group (NCO) were then compared with infants placed on nasal cannula without order sets (controls) from the same time period the year before. Infants were stratified by birth weight (# 1,500 g $ 1,501 g). There were 31 patients in the control group and 23 patients in the NCO group. There were 12 control patients # 1,500 g and 11 NCO patients # 1,500 g. Data were analyzed using Student t tests (p , .05 significant). Groups were well matched for demographics (under 1,500 g group: 6/12 male controls vs 5/11 male NCO; median bwt 5 1,080 g control vs 1,000 g NCO; mean GA 28.4 wk 6 2.7 wk control vs 28.5 wk 6 2.0 wk). Infants under 1,500 g were also well matched for number who received antenatal steroids (9/12 controls vs 9/11 NCO). Results: Nasal cannula oxygen days were much lower for the NCO group (4.5 d 6 2.6 d) versus the control group (8.4 d 6 4.8 d) for all infants in the study (big and small), but the difference was not statistically significant (p , .06). There was no significant difference for oxygen days (2.1 d 6 1.4d NCO vs 3.5 d 6 2.8 d control) for the large infants (p , .11). Upon comparison of small infants (# 1,500 g), a strongly significant difference was found between groups (7.2 d 6 3.4 d NCO vs 15.9 d 6 5.2 d controls; p , .02). These preliminary data indicate that days on nasal cannula oxygen may be significantly shortened if standardized nasal cannula order sets are used to direct consistent weaning. 133 VIDEO-ASSISTED ENDOTRACHEAL INTUBATION IN THE NEWBORN: ROLE IN TEACHING, SKILL ACQUISITION, AND TUBE PLACEMENT VERIFICATION. A.L. Vanderhal, G. Berci, C.F. Simmons, Cedars-Sinai Medical Center, Los Angeles, CA. Background: Endotracheal intubation (EI) in newborns is difficult, and skill acquisition may become more problematic as less time is spent in DR and NICU during training. The advantages of endoscopy lie in a wider angle of view, enlarged imaging and improved illumination. The Newborn Resuscitation Program (NRP) recommends EI attempts be limited to 20 seconds. NRP training includes EI on newborn manikin heads. Purpose: This study investigates if video-assisted techniques help in acquiring these skills and if time to verification of successful EI is shortened using this technology. Method: After standard NRP presentation, trainees attempted EI serially with three laryngoscopy techniques: standard SL, video-supervised VL, and video-guided modified VLM. VL implies intubation with the video laryngoscope, with assistant verbally guiding from screen images. In VLM, the ET is advanced via the blade groove and inserted under screen image viewing by the operator. Miller #1 blades and Storz videolaryngoscopy equipment were used for VL and VLM; images were recorded on a laptop computer. Number of EI attempts, success of EI, and time to verification of EI were recorded. Time to correct EI was recorded at chest movement on bag ventilation in SL by stopwatch, and until the double-line ET marker passed through the vocal cords in VL and VLM by image timer. Trainees completed questionnaires and comments were solicited. Results: SL first EI attempt was successful in 18/23 participants; 5/23 needed three attempts. In VL and VLM, first EI attempt succeeded in 20/23 and 21/ S98 Journal of Investigative Medicine