EBPβ) IS ESSENTIAL FOR GLUTAMINE-MEDIATED ACTIVATION OF HEAT SHOCK FACTOR-1 (HSF1) TRANSCRIPTION

Hormones, mediators and immunity II Methods: We performed a retrospective review of prospectively collected data on thirty-seven patients who underwent pancreaticoduodenectomy in our institution. Twelve patients in the perioperative group received IED enriched with arginine, omega-3 fatty acids, and RNA, for five days before surgery, prolonged after surgery by enteral infusion of IED. Twenty-five patients in the preoperative group received only preoperative oral feeding of IED and received postoperative enteral infusion of a standard formula. Results: The eicosapentaenoic acid/arachidonic acid ratio was significantly higher in perioperative group than in preoperative group on postoperative day (POD) 7 and 14. The level of plasma IL-6 was significantly lower in perioperative group than in preoperative group on POD 1. CRP level was significantly lower in perioperative group than in preoperative group on POD 1 and 3. On POD 7 and 14, Con A- or PHA-stimulated lymphocyte proliferation was significantly higher in perioperative group than in preoperative group. Although the rate of infectious complications was lower in perioperative group (8.3%) than in preoperative group (28%), there were no statistically significant differences between the two groups. Conclusion: These results indicate that postoperative prolongation with IED may have some additional benefits in patients undergoing pancreaticoduodenectomy. Disclosure of Interest: None Declared.

PP023-MON LIVER DYSFUNCTION IN TRAUMA CRITICALLY ILL PATIENTS RECEIVING EPA AND GLA ENRICHED ENTERAL FEEDING M. Theilla1 , I. Kagan1 , R. Anbar1 , O. Tirosh2 , J. Singer3 , M. Grunev1 , P. Singer1 . 1 Intensive Care and Institute for Nutrition Research, Rabin Medical Center and Tel Aviv University, Petah Tikva, 2 Nutrition, Faculty of Agriculture, Rehovot, 3 Endocrinology Institute, Rabin Medical Center and Tel Aviv University, Petah Tikva, Israel Rationale: Critically ill patients exhibit frequently liver function tests abnormalities. Fish oil enriched nutrition has been shown to decrease these disturbances. We compared enteral feeding enriched in EPA and GLA (Oxepa, Aboott, USA) or not (Pulmoncare, Abbott, USA), administered to trauma ventilated patients, in terms of liver disturbancies and membrane lipid composition. Methods: A PRDB study was performed in multiple trauma ventilated patients receiving enriched in EPA (4.5 g/L) and GLA (4 g/L) or isocaloric iso protein diet targeted to resting energy expenditure. A glucose control was kept, liver function tests taken at day 1, 3, 7, 10 and lipid membrane composition was expressed by Individual fatty acid methyl esters by comparing retention times with authentic standards. Values were expressed as wt.% of total identified fatty acids. Results were compared using T test and ANOVA analysis of variance. Results: Sixty patients (30 in each group) were included and age, APACHE II and ISS were indentical. Glucose control was kept below 120 mg/dL in mean. ASAT, ALAT were elevated from baseline and remained elevated with-

147 out difference between the 2 groups, Alk Phosph, GGT were normal at admission but increased in the following days (NS). Total bilirubin was increased from baseline (1.7±3.3 in the EPA GLA group vs 0.8±0.4 in the control, P < 0.04) during the all week of observation. While in the control group the lipid membrane omega-3:omega-6 ratio remained rather constant from day 1 to day 7 (0.2±0.04), this ratio in the EPA+GLA group increased significantly (day 1: 0.19±0.04, day 4: 0.22±0.05, day 8: 0.25±0.06, p < 0.05). Conclusion: The addition of EPA GLA in enteral nutrition of trauma ventilated patients is safe, could be beneficial in terms of cytolytic enzymes but is not preventing from cholestasis. EPA is efficiently absorbed and integrated in the cell membrane. Disclosure of Interest: None Declared.

Hormones, mediators and immunity II PP024-MON Outstanding abstract CCAAT ENHANCER BINDING PROTEIN-b (C/EBPb) IS ESSENTIAL FOR GLUTAMINE-MEDIATED ACTIVATION OF HEAT SHOCK FACTOR-1 (HSF1) TRANSCRIPTION H. Xue1 , D. Slavov2 , P. Wischmeyer1 . 1 Anesthesiology, 2 Cardiology, University of Colorado, Aurora, United States Rationale: Heat shock factor-1 (HSF1) is the master regulator for cytoprotective heat shock protein (Hsp) expression. The key control point of HSF1-regulated HSP expression is thought to be regulation of HSF1’s transactivation activity. The role of enhanced HSF1 expression and how HSF1 expression is regulated remains unknown. Exogenous glutamine (GLN) upregulates Hsp expression, however it’s potential to regulate HSF1 expression is unstudied. Methods: HSF1 protein expression, mRNA expression kinetics and promoter activity in response to GLN were examined in YAMC colonic epithelial cells exposed to GLN at 0, 0.5 or 2mM. Deletion analysis of HSF1 promoter was used to identify the key cis-element(s) involved in GLN-induced HSF1 transcription activation. SiRNA specific for CCAAT enhancer binding protein-b (C/EBPb) and transfection of expression plasmids bearing cDNAs for the active and inhibitory isoforms of C/EBPb were used to identify the potential role of C/EBPb in GLN-mediated HSF1 response. Results: GLN enhanced total HSF1 protein and mRNA expression and HSF1 gene promoter activity. Within the 281/ 200 region of the HSF1 promoter, deletion of the putative C/EBP binding site abolished GLN’s HSF1 response. C/EBPb was shown to bind this 82bp sequence. GLN deprivation increased the relative abundance of C/EBPb inhibitory isoform as compared to its active isoforms, which repressed HSF1 expression; whereas a relative enrichment of the active isoform resulted from adequate GLN supply activated HSF1 transcription and expression. Conclusion: We show for the first time GLN induces HSF1 expression by activating its transcription in a

148 C/EBPb-dependent manner. Here, we describe a novel amino acid-mediated signaling pathway, which can activate HSF1 gene expression and elicit a robust Hsp response to stress. Disclosure of Interest: None Declared.

PP025-MON EXENDIN-4 INHIBITS INOS EXPRESSION AT THE PROTEIN LEVEL IN LPS-STIMULATED RAW264.7 MACROPHAGE BY THE ACTIVATION OF CAMP/PKA PATHWAY S.-Y. Chang1 , J.-M. Cho1 , Y.-H. Jo1 , M.-J. Kim1 . 1 Department of Physiology, College of Medicine, The Catholic University of Korea, Seoul, Korea, Republic Of Rationale: Glucagon-like peptide-1 (GLP-1) and its potent agonists have been widely studied in pancreatic islet b-cells. However, GLP-1 receptors are present in many extrapancreatic tissues including macrophages, and thus GLP-1 may have diverse actions on these tissues and cells. Therefore, we examined the mechanism by which exendin-4 (EX-4), a potent GLP-1 receptor agonist, inhibits lipopolysaccharide (LPS)-induced iNOS expression in Raw264.7 macrophage cells. Methods: The expression of iNOS, p-IúBa and p65 were detected by Western blot analysis and content of nitrite was measured using Griess reagent. Additionally, iNOS mRNA expression and promoter activity were observed by Northern blot analysis and luciferase assay, respectively. To observe the stability of iNOS mRNA and protein, actinomycin D chase and cycloheximide chase studies were performed, respectively. Also, we examined using adenylate cyclase inhibitor, PKA inhibitor and PKA gene silencing. Results: EX-4 significantly inhibited LPS-induced iNOS protein expression and nitrite production. However, EX-4 did not inhibit LPS-induced iNOS mRNA expression and iNOS promoter activity. Consistent with the result of iNOS promoter, LPS-induced IúBa phosphorylation and nuclear translocation of p65 were not inhibited by EX-4. Also, actinomycin D chase study showed that EX-4 did not affect iNOS mRNA stability. Meanwhile, cycloheximide chase study demonstrated that EX-4 significantly accelerated iNOS protein degradation. The EX-4 inhibition of LPS-induced iNOS protein was significantly reversed by adenylate cyclase inhibitors (MDL-12330A and SQ 22536), a PKA inhibitor (H-89) and PKAa gene silencing. Conclusion: These findings suggest that EX-4 inhibited LPS-induced iNOS protein expression at protein level, but not at transcriptional mechanism of iNOS gene and this inhibitory effect of EX-4 was mainly dependent on cAMP/PKA system. Disclosure of Interest: None Declared.

PP026-MON HEPCIDIN STORM AFTER ABDOMINAL SURGERY K.H. Park1 , T. Sawada1 , K. Kubota1 . 1 Second Department of Surgery, Dokkyo Medical University, Tochigi, Japan Rationale: Hepcidin plays a crucial role in iron homeostasis and is also a marker of acute inflammation. In this study, we investigated the changes in the serum hepcidin level and correlations between hepcidin and other

Poster presentations markers of acute inflammation during the perioperative period in patients after abdominal surgery. Methods: Serum hepcidin, hemoglobin (Hb), hematocrit (Ht), white blood cell (WBC) count, and C-reactive protein (CRP) were measured preoperatively (Pre), and on postoperative days (PODs) 1, 3, 7, and 14. Results: In patients undergoing gastrectomy, the median levels of hepcidin on Pre, and on POD 1, 3, 7, and 14 were 6.5, 53.1, 31.7, 15.6, and 4.0 ng/dL, respectively (P < 0.0001). The corresponding levels in colectomy patients were 8.5, 78.3, 60.1, 49.7, and 8.4 ng/dL, respectively (P = 0.0002), those in hepatectomy patients were 6.6, 16.3, 3.5, 13.4, and 3.4 ng/dL, respectively (P = 0.0022), and those in patients undergoing surgery for diffuse peritonitis were 24.8, 50.1, 43.1, 31.2, and 31.7 ng/dL, respectively (P = 0.4933). There were no significant decreases in Hb and Ht in the patients undergoing gastrectomy, colectomy, or surgery for diffuse peritonitis. The level of hepcidin was significantly correlated with the WBC count, and CRP level during the perioperative period for all four types of operation. Conclusion: Like other inflammatory markers, an increase in the level of hepcidin (i.e. a hepcidin storm) occurs in the acute phase after gastrectomy, colectomy, hepatectomy, and surgery for diffuse peritonitis. Because hepcidin is known to divert iron to storage-type ferritin rather than to erythropoiesis, iron administration intended for erythropoiesis during this period may be ineffective. Disclosure of Interest: None Declared.

PP027-MON ENHANCEMENT OF THE LPS-INDUCED ACTIVATION OF NATURAL KILLER CELLS BY ORAL ADMINISTRATION OF CYSTINE AND THEANINE IN MICE K.A. Tanaka1 , S. Kurihara1 , T. Shibakusa1 , Y. Chiba2 . 1 Institute for Innovation, Ajinomoto Co., Inc., Kawasaki, 2 Nutrition Care Dept., Ajinomoto Co., Inc., Tokyo, Japan Rationale: Inhibition of surgery-induced immunosuppression might play a critical role in preventing infectious complications after surgery. We reported that oral administration of the amino acids cystine and theanine (CT) enhanced immune functions (Clin Nutr 2012). To apply CT as an immune modulator during perioperative periods in the future, we first investigated the role of CT on the immune system of lipopolysaccharide (LPS)treated mice. Methods: Twelve-week-old BALB/c mice were intraperitoneally injected with LPS (100 mg/mouse) 2 hours after a single oral administration of CT (280 mg/kg) or vehicle control (V), and interferon (IFN)-g levels in the plasma were measured. The expression levels of INF-g in isolated splenic natural killer (sNK) cells 6 hours after the LPS injection in CT- or V-administered mice were analysed by quantitative RT-PCR. The LPS-induced IL-18 (INF-g inducer) secretion in THP-1 monocytes was detected by ELISA. Results: We found that oral administration of CT significantly increased IFN-g levels in the plasma compared with control mice (CT: 382.4±20.5 pg/ml, V: 158.6±79.5 pg/ml, P < 0.05) and that the LPS-induced expression of IFN-g in the isolated sNK cells was signif-