Fax +41 61 306 12 34 E-Mail karger@karger.ch www.karger.com Neonatology 2012;101:360–366 DOI: 10.1159/000337360 Abstracts 27th International Workshop on Surfactant Replacement, Lisbon, June 1–2, 2012 2 Synergistic Effect of Caffeine and Glucocorticoids on Surfactant Protein B Expression I. Bersani a, b , M. Fehrholz a , S. Kunzmann a , C.P. Speer a a University Children’s Hospital, University of Würzburg, Würzburg, Germany; b Department of Pediatrics, A. Gemelli Hospital, Catholic University of the Sacred Heart, Rome, Italy Background: Caffeine and cortisone administration represent a therapeutic intervention of high clinical relevance during the neonatal period. The aim of this study was to characterize the ef- fects and interactions between caffeine and different glucocorti- coids on the expression of surfactant protein B (SP-B), essential for the physiological function of pulmonary surfactant. Methods: The expression level of SP-B was measured by real time quantita- tive RT-PCR and immunofluorescence staining in the type II lung epithelial cell line H441 after induction with different doses of caffeine and glucocorticoids. Results: Dexamethasone (1 M) in- duced SP-B expression in a dose-dependent manner with a maxi- mal 37 8 12-fold increase, and caffeine (5 m M) with a 3.1 8 1-fold increase. However, induction by a combination of dexamethasone and caffeine reached levels that were 250 8 47-fold higher than in non-induced cells (p ! 0.0001). The synergistic induction of SP-B mRNA expression could also be observed by using caffeine with hydrocortisone (110 8 1), prednisolone (191 8 15), and be- tamethasone (504 8 118), which had the most accentuated effect. The maximal effect of caffeine and steroids on SP-B expression was reached after 12 h. These findings could be confirmed at the protein level by immunofluorescence staining. Conclusion: The present study demonstrated a synergistic upregulating effect on SP-B expression by glucocorticoids in combination with caffeine. Co-administration of caffeine together with steroids seems to be of benefit in surfactant homeostasis. 1 Molecular Basis of Sustained Inflammation after Neonatal Sepsis: A Matter of Life and Death? T. Orlikowsky, S. Dreschers, C. Gille Department of Neonatology, University Childrens’ Hospital, Aachen, Germany Background: Neonatal sepsis-related sequelae such as bron- chopulmonary dysplasia are characterized by parenchymal de- struction through prolonged inflammation. Crucial steps in ter- minating inflammation are elimination of effector cells by apop- tosis and their subsequent removal. Monocytes play a dual role: as first line defense with their dying after ingestion of pathogens (phagocytosis induced cell death, PICD) and as scavenger cells removing their dead counterparts. Objective: To determine if PICD and scavenger functions are altered in neonatal monocytes. Methods: Monocytes from cord blood (CBM) and from adults (PBM) were infected with E. coli and phagocytosis, apotopsis, TNF- , as well as CD95L-release were quantified, or co-cultured with apoptotic neutrophils, and phagocytosis and IL-8-release were assessed. Results: Despite equivalent phagocytotic activity upon E. coli, CBM compared to PBM showed reduced apoptosis, reduced TNF- - and reduced CD95L-production. Apoptosis was blocked by anti-TNF-  and anti-CD95L. Furthermore, CBM were impaired in phagocytizing apoptotic granulocytes com- pared to PBM. Phagocytosis of apoptotic neutrophils led to an up-regulation of IL-8 production in CBM and a down-regulation in PBM. Conclusion: CBM are less sensitive towards PICD be- cause of diminished secretion of TNF-  and CD95L, are impaired in eliminating apoptotic granulocytes and in down-regulating pro-inflammatory IL-8 secretion. These altered reactions may impair CBM in terminating inflammatory responses, and may play a role in the perpetuation of inflammatory organ damage in neonates. Published online: June 1, 2012 © 2012 S. Karger AG, Basel 1661–7800/12/1014–0360$38.00/0 Accessible online at: www.karger.com/neo