O0009 FUNCTIONAL SIGNATURE UNDERLYING LIVER CANCER PROGRESSION IN CELL LINES ESTABLISHED FROM A STEM CELL DERIVED HEPATOCHOLANGIOCARCINOMA

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Abstracts from 8th Congress of the European Federation of Internal Medicine / European Journal of Internal Medicine 20S (2009), S1–S283

O0009 FUNCTIONAL SIGNATURE UNDERLYING LIVER CANCER PROGRESSION IN CELL LINES ESTABLISHED FROM A STEM CELL DERIVED HEPATOCHOLANGIOCARCINOMA

Anna Chiara Piscaglia 1 , Nathalie Saulnier 1 , Valentina Tesori 2 , Marta Barba 1 , Mariachiara Campanale 1 , Flaminia Purchiaroni 1 , Emanuela D’angelo 1 , Giovanni Cammarota 1 , Giovanni Pani 2 , Antonio Gasbarrini 1 . 1 Department of Internal Medicine; Catholic University of Rome – College of Medicine, “a. Gemelli” Hospital, Italy; 2 Inst. of General Pathology – Catholic University of Rome – College of Medicine, “a. Gemelli” Hospital, Italy Introduction: We established several cancer cell lines (named LCSCs) from a new model of hepatocholangiocarcinoma, based upon induction of oval cell proliferation prior to carcinogen exposure. Objectives: Aim of this study was to compare the gene expression profile of two of these lines: LCSC-5 and LCSC-2. Materials & methods: LCSCs were observed daily and their immunophenotype was assessed on cytospins and chamber-slides. The clonogenic potential of LCSCs was evaluated by soft-agar colony formation assay, and their tumorigenicity was investigated upon transplantation into nude mice. The growth kinetics was assessed by MTT assay. Gene expression profiling was performed on Affymetrix platform. The cellular response to HGF was measured by western blot analysis for c-Met, p-ERK, pS6, p-Akt, and by functional assays. Finally, the effects of a novel and promising anticancer drug, Sorafenib, were investigated on both cell lines. Results: Although the immunophenotype and morphology of the two lines were similar, LCSC-2 had higher proliferative capacity and clonogenicity in vitro; moreover, upon transplantation, LCSC-2 induced macroscopic tumors in less than 30 days, while LCSC-5 required approximately 80 days. Transcriptional analysis revealed that genes overrepresented in LCSC-2 were mainly associated with tumor progression/growth (S100A4, ALDH3A1, ARMCX2), drug resistance (ABCC1) and invasion/cytoskeleton-remodelling (TPM2, PDLIM1, CAPG). Conversely, several liver-associated transcripts (IL6ST, ZFP36, IGF2, CYB5, LISCH7) and genes commonly downregulated in invasive cancers (GRO1, CEACAM1, NFKBIA, MGP) were reduced in LCSC-2. Interestingly, the expression level of the HGF receptor c-Met and its downstream signaling cascade appeared downregulated in LCSC-2 cells, compared to LCSC-5. Sorafenib had cytotoxic effects and, at cytostatic concentrations, potentiated HGF responses in both cell lines, this effect being again less pronounced in LCSC-2, which was also less sensitive to the drug. Discussion & conclusion: LCSC-2 displayed a more aggressive, less differentiated, and more drug-resistant phenotype when compared to LCSC-5. The HGF/c-Met pathway may play a causative role in this difference, likely by promoting hepatic differentiation. The gene expression analysis allowed the identification of a panel of genes underlying liver cancer invasiveness and aggressiveness and which might offer novel insights to study the effects of molecularly targeted agents against hepatic tumors. Keywords: gene expression profile; liver cancer; stem cells

were diagnosed as celiacs. The sensitivity, specificity, and positive and negative predictive values of the immersion technique in detecting marked villous atrophy patterns, irrespectively of the endoscopist who performed the exam, was always 100%. Discussion & conclusion: The under water evaluation of the duodenum can be efficiently and regularly performed during routinely EGD by endoscopists with a 2-years of experience and after a specific, brief training period. This procedure is simple, feasible and can accurately detect marked villous atrophy patterns. Keywords: water immersion; villous evaluation

O0011 AN ASSESSMENT OF RISK FACTORS FOR PROGNOSIS AND LENGTH OF HOSPITAL STAY IN COMMUNITY ACQUIRED PNEUMONIA – A DISTRICT GENERAL HOSPITAL, U.K. EXPERIENCE

Munish Batra, Poonam Batra. Victoria Hospital Aim: To assess factors associated with prolonged hospital stay and mortality in patients of community acquired pneumonia. Methods: Retrospective case notes review of 100 patients with pneumonia and data collection on demography, investigations and treatment. We used Pearson,s coefficient to identify independent risk factors associated with duration of hospital stay. A comparison of several factors, including demographic findings, clinical signs, underlying diseases, severity of diseases, blood biochemistry and causative pathogens was performed in both survival and fatal groups. We used t–test to compare differences in these risk factors in patients who survived and in those dead. Results: The mean age of patients were 72.8. The average length of hospital stay was 10.2 days with standard deviation of 8.6 days. 88 patients were discharged whereas12 patients died of severe pneumonia. We found that 24 patients had mild, 36 had moderate and 40 had severe pneumonia on CURB65 score. There was a positive correlation of age, urea levels with duration of hospital stay. There was a negative correlation of blood pressure levels and albumin levels with length of hospital stay. The mean age, urea and albumin were 82.5 years, 16.4mmol/l and 32.4mmol/l respectively in patients who died as compared to 72 years, 8.2 mmol/l and 36.5mmol/l in those survived. The differences were significant for age and urea (p value=0.01, 0.02). The prevalence of Gram negative bacteraemia was higher in patients who died than in patients who survived.Interestingly, the average length of hospital stay for patients who died was 3 days as compared to 11.8 days for patients who discharged from hospital after treatment. Conclusions: The duration of hospital stay tends to be prolonged in older patients with high urea and low albumin levels and hypotension. The prognosis of the patients with community acquired pneumonia tends to get worse with advancing age and renal impairment.

O0012 RISKY BUSINESS: COMPARISON OF SEVERITY OF ILLNESS IN EUROPEAN HOSPITALS

O0010 A SINGLE CENTRE, 2-YEARS EXPERIENCE ON WATER IMMERSION TECHNIQUE FOR DUODENAL VILLOUS EVALUATION

Giovanni Cammarota, Alessia Cazzato, Anna Chiara Piscaglia, Rossella La Mura, Gianluca Ianiro, Emanuela D’angelo, Antonio Gasbarrini, Giovanni Gasbarrini. Department of Internal Medicine, Endoscopy Unit; Catholic University of Rome – College of Medicine, “a. Gemelli” Hospital, Italy Introduction: The immersion technique was proved to offer a possibility to ameliorate the sensitivity of endoscopy in detecting duodenal villous atrophy patterns. Objectives: We investigated learning parameters of the immersion technique for the assessment of marked villous atrophy during routinely EGD. Materials & methods: Nine endoscopists, with at least 2-years of experience and who were previously trained to perform the immersion technique, searched for duodenal villi during routinely EGD. In a 2-years study, duodenal villi were evaluated under water duodenoscopy in 432 patients in whom duodenal biopsies were, for various reasons, obtained. Endoscopic findings were compared with histology. Learning parameters of the water immersion technique and the accuracy in detecting subjects with marked villous atrophy were considered. Results: A total of 28 (6.5%) patients with marked villous atrophy were identified during endoscopy by the trained endoscopists. All these patients

Will Gauntlett 2 , Christian P Subbe 2 , John Kellett 1 . 1 Nenagh Hosptial, Nenagh, Ireland; 2 Wrexham Maelor Hospital, Wrexham, UK Introduction: Kellet et al developed a Simple Clinical Score (SCS) [1] based on 16 leading physiological and social parameters affecting hospital mortality at 30 days in a large single centre study. Objectives: Pilot study examining the practical application of the SCS in 21 centres in Europe and New Zealand. Materials and method: Collection of data on admission from 12 or more consecutive acute medical in-patients. Outcomes collected were SCS, hospital discharge, mortality and admission to intensive care (ICU) or coronary care (CCU). Results: Data from 282 patients was submitted for analysis. Mean age was 64 years (SD 21). 48% of patients were male. Median SCS was 6 (IQR 3 to 9). 55% of patients were discharged within 6 days, 9% were discharged within 24 hours, 25% within 48 hours. 11 patients died, 18 patients were admitted to ICU and 18 were admitted to CCU. Severity of illness as estimated by SCS was related to risk of admission to ICU and CCU. Low scores were associated with earlier discharge and lower risk of mortality (Table 1). There were differences in the pattern of discharges in different hospitals with comparables SCS data (Figs. 1, 2). Clinicians reported no significant problems with the collection of data for the score in a number of different health care settings.