Chronic hepatitis C (HCV) and B (HBV) are major risk factors for the development of liver cirrhosis, favouring the development of hepatocellular carcinoma (HCC). The lipid metabolism plays a major role in the pathogenesis of HCV-infection, as HCV is known to require lipid droplets (LDs) for the production of infectious virus particles. Amphiphilic proteins of the perilipin/PAT-family decorate the surface of LDs and determine their formation, maintenance and degradation. In mammals, the perilipin-family consists of 5 members, with perilipins 1 and 2 being constitutively expressed at LDs. Aim of this project was to further the understanding of LD-histopathology in chronic hepatitis.

Introduction: Phenprocoumon (Marcumar®), a derivative of 4-hydroxycoumarin with similarity to warfarin, is the most frequently used oral anticoagulant drug in continental europe. Apart from bleeding disorders, other adverse reactions are rare due to its mechanism of action. In the last years, hepatotoxicity has been observed and several case reports have been published. Yet, so far, no systematic analyses of this potentially life-threatening complication have been performed.

Abstract Purpose: The aim of the present prospective European multicenter study was to demonstrate the non-inferiority of point shear wave elastography (pSWE) compared to transient elastography (TE) for the assessment of liver fibrosis in patients with chronic hepatitis C. Materials and Methods: 241 patients with chronic hepatitis C were prospectively enrolled at 7 European study sites and received pSWE, TE and blood tests. Liver biopsy was performed with histological staging by a central pathologist. In addition, for inclusion of cirrhotic patients, a maximum of 10 % of patients with overt liver cirrhosis confirmed by imaging methods were allowed by protocol (n = 24). Results: Owing to slower than expected recruitment due to a reduction of liver biopsies, the study was closed after 4 years before the target enrollment of 433 patients with 235 patients in the ‘intention to diagnose’ analysis and 182 patients in the ‘per protocol’ analysis. Therefore, the non-inferiority margin was enhanced to 0.075 but non-inferiority of pSWE could not be proven. However, Paired comparison of the diagnostic accuracy of pSWE and TE revealed no significant difference between the two methods in the ‘intention to diagnose’ and ‘per protocol’ analysis (0.81 vs. 0.85 for F ≥ 2, p = 0.15; 0.88 vs. 0.92 for F ≥ 3, p = 0.11; 0.89 vs. 0.94 for F = 4, p = 0.19). Measurement failure was significantly higher for TE than for pSWE (p = 0.030). Conclusion: Non-inferiority of pSWE compared to TE could not be shown. However, the diagnostic accuracy of pSWE and TE was comparable for the noninvasive staging of liver fibrosis in patients with chronic hepatitis C.

Acoustic Radiation Force Impulse (ARFI)-Imaging (Siemens Acuson S2000, Virtual TouchTM tissue quantification, Siemens) ist ein neues Ultraschall-basiertes Elastografie-Verfahren, welches in ein konventionelles Ultraschallgerät integriert ist und die genaue Platzierung des Messgebietes erlaubt. Es könnte eine alternative Methode zur bereits etablierten Transienten Elstografie (FibroScan®, Echosens) für die nicht-invasive Beurteilung der Leberfibrose darstellen. Das Ziel der vorliegenden Multicenter-Studie ist die Evaluation der ARFI für die Beurteilung der Leberfibrose bei Patienten mit chronischer Hepatitis B.

Hintergrund: Acoustic Radiation Force Impulse (ARFI)-Imaging (Siemens Acuson S2000, Virtual TouchTM tissue quantification, Siemens) ist ein neues Ultraschall-basiertes Elastografie-Verfahren, welches in ein konventionelles Ultraschallgerät integriert ist und die genaue Platzierung des Messgebietes erlaubt. Es könnte eine alternative Methode zur bereits etablierten Transienten Elstografie (FibroScan®, Echosens) für die nicht-invasive Beurteilung der Leberfibrose darstellen. Das Ziel der vorliegenden Multicenter-Studie ist die Evaluation der ARFI für die Beurteilung der Leberfibrose bei Patienten mit chronischer Hepatitis B.

Introduction: Acute liver failure (ALF) is associated with massive short term cell death, while chronic liver injury is accompanied by continuous cell death. Hepatic stellate cells (HSCs) contribute to tissue repair and liver fibrosis in chronic liver injury, although their role in ALF remains unclarified.

Purpose: Transient elastography (FibroScan, [TE]) and serum fibrosis markers such as the FibroTest (FT) are established methods for the noninvasive staging of liver fibrosis. A study using real-time elastography (HI-RTE), which is integrated in a conventional ultrasound system, was recently published with comparable results to transient elastography. The aim of the present study was to validate real-time elastography using the formulas calculated in previous studies and to compare the results to transient elastography and FibroTest for the noninvasive assessment of liver fibrosis.Materials and Methods: One hundred and thirty-four patients with chronic liver disease and either histological assessment of liver fibrosis (n = 112) or proven liver cirrhosis (n = 22) were included in the study. All patients received TE, HlRTE, and biochemical evaluation on the same day as presentation. The calculation of the elasticity score of real-time elastography was performed in accordance with the two previously published studies.Results: The Spearman correlation coefficient between transient elastography, real-time elastography and FibroTest with the histological Chevallier score was statistically significant with 0.78, 0.34, and 0.67, respectively (p < 0.01). The diagnostic accuracy expressed as areas under ROC curves was 0.84, 0.69 and 0.85 for the diagnosis of significant fibrosis (F >= 2), and 0.97, 0.65, and 0.83 for the diagnosis of cirrhosis, respectively.Conclusion: Real-time elastography in its present form cannot replace transient elastography for noninvasive assessment of liver fibrosis.

Ziel: Verringerung der Gewebetraumatisierung durch Reduktion der benötigten Stanzzylinder zur Diagnosesicherung durch Einsatz einer 16G Stanzbiopsienadel mit Koaxialkanüle im Vergleich zu standardmäßig (gem. S3-Leitlinie 2008) eingesetzten 14G Stanzbiopsiesystemen.

Ziel: Analyse der Treffsicherheit und Materialausbeute der Stanzbiopsie mit minimal-invasivem 16G-Coax-System anstelle der in der S3-Leitlinie geforderten großkalibrigen 14G-Systeme.

OBJECTIVE Recently, transient elastography (FibroScan) has been introduced for noninvasive staging of liver fibrosis. Here, we investigated a novel approach for noninvasive assessment of liver fibrosis using sonography-based real-time elastography, which can be performed with conventional ultrasound probes during a routine sonography examination. MATERIALS AND METHODS Real-time elastography was performed in 79 patients with chronic viral hepatitis and known fibrosis stage and in 20 healthy volunteers. A specially developed program was used for quantification of tissue elasticity. Stepwise logistic regression analysis was performed to define an elasticity score using variables with high reproducibility in a preceding analysis of data from 16 different patients. In addition, aspartate transaminase-to-platelet ratio index (APRI) and routine laboratory values were included in the analysis. RESULTS The Spearman's correlation coefficient between the elasticity scores obtained using real-time elastography and the histologic fibrosis stage was 0.48, which is highly significant (p < 0.001). The diagnostic accuracy expressed as areas under receiver operating characteristic (ROC) curves were 0.75 for the diagnosis of significant fibrosis (fibrosis stage according to METAVIR scoring system [F] > or = F2), 0.73 for severe fibrosis (F > or = F3), and 0.69 for cirrhosis. For a combined elasticity-laboratory score, the areas under the ROC curves were 0.93, 0.95, and 0.91, respectively. DISCUSSION Real-time elastography is a new and promising sonography-based noninvasive method for the assessment of liver fibrosis in patients with chronic viral hepatitis.

MicroRNAs (miRNAs) are small, endogenously expressed, noncoding RNAs, which regulate gene expression through RNA interference by binding to a defined set of target transcripts. Recent data have shown that miRNAs are involved in cancer initiation and progression by affecting tumorigenic pathways with impact on cell proliferation and apoptosis. Since miRNA expression pattern can be used for the classification, prognosis and therapeutic approaches of human malignancies, we studied the miRNA expression profiles by quantitative PCR in hepatocelluar carcinoma (HCC).

Epithelial cell adhesion molecule (Ep-CAM) is expressed in a several epithelial tissues and carcinomas, but not on mature hepatocytes. Here, we analysed the expression of Ep-CAM in 230 patients suffering from various liver diseases like chronic hepatitis B and C (HBV and HCV infection), chronic autoimmune hepatitis (AIH), chronic alcoholic liver disease (ALD), primary biliary cirrhosis (PBC), primary sclerosing cholangitis (PSC), hereditary hemochromatosis and dysplastic nodules (DNs) as well as hepatocellular carcinomas (HCCs) and cholangiocellular carcinomas (CCCs) by immunohistochemistry. De novo hepatocellular Ep-CAM expression was found in 75.9% of ALD (22/29), 63.6% of HCV (21/33) and 55.6% of each AIH and HBV cases (5/9 and 15/27, respectively). Lower Ep-CAM expression levels were observed for primary sclerosing liver diseases (PBC and PSC) with 25% (3/12) and 7.7% (1/13) of cases. Moreover, only 14.3% of HCCs (9/63) manifested expression, while all CCCs showed strong Ep-CAM expression (5/5). For DNs and hereditary hemochromatosis, Ep-CAM expression was found in 10 and 50% (3/30 and 2/4), respectively. In HBV and HCV, Ep-CAM expression correlated significantly with inflammatory activity as assessed by histological parameters and to the extent of fibrosis. In addition, for HCV also transaminase levels correlated significantly with Ep-CAM expression. Our results indicate that de novo Ep-CAM expression in hepatocytes is frequent in inflammatory liver diseases and is potentially linked to regenerative activity. CCCs and Ep-CAM positive HCCs may represent an attractive target group for Ep-CAM-directed immunotherapies, yet unwanted toxicity may limit the use of such strategies due to Ep-CAM expression in biliary epithelium and several chronic liver diseases such as HBV-and HCV-hepatitis.

Hintergrund: Realtime-Elastographie ist ein Verfahren mit welchem die physikalischen Eigenschaften von Gewebe untersucht werden können. Hierbei werden Verschiebungen von Echofrequenzmustern vor und nach einer Kompression des zu untersuchenden Gewebes abgeleitet und das aus den Verschiebungswerten rekonstruierte Dehnungsfeld dem B-Bild farbkodiert überlagert.

Einleitung: Die Sonographie-basierte Realtime-Elastographie (SonoElastographie Modul, Hitachi EUB-8500) ist ein Verfahren mit welchem die physikalischen Eigenschaften von Gewebe untersucht werden können. Hierbei werden Verschiebungen von Echofrequenzmustern vor und nach einer Kompression des zu untersuchenden Gewebes abgeleitet und das aus den Verschiebungswerten rekonstruierte Dehnungsfeld dem B-Bild farbkodiert überlagert. Bisher wurde die Realtime-Elastographie zur Beurteilung von Tumoren in der Mamma, Schilddrüse und Prostata evaluiert. Die Wertigkeit der Realtime-Elastographie zur Einschätzung des Leberfibrosestadiums wurde bisher nicht untersucht.

BACKGROUNDApoptosis, programmed cell death, is involved in a broad range of pathological processes. Dysregulation of apoptosis plays a key role in the pathogenesis of hepatitis, toxic liver disease and also liver tumor development. For the study of apoptosis in liver diseases, different in vivo models and different in vitro approaches have been developed. They include cell culture models based on hepatocellular carcinoma cell lines or isolated primary hepatocytes.MATERIALS AND METHODSWe have established precision cut tissue slices (PCTS) of the liver as a morphological tool for the study of apoptosis. From porcine livers, PCTS were prepared and incubated in a static system with different types and amounts of media. Viability, morphology, spontaneous apoptosis and proliferation were investigated. Apoptosis was induced with actinomycin D and tumor necrosis factor (TNF) alpha.RESULTSMorphology and viability was well preserved for at least 24 h. After 48 h, deterioration with single and group cell autolysis was seen. There was a low rate of spontaneous apoptosis and proliferation. Using a combination of TNF alpha and actinomycin D, a significant amount of apoptosis occurred.CONCLUSIONPCTS can be used to directly analyse apoptosis at the tissue level in a qualitative and quantitative manner.

BACKGROUND:Transcription mediated amplification (TMA) is known to be one of the most sensitive detection assays for hepatitis C virus (HCV) RNA in serum but has not yet been evaluated in liver tissue. It is unknown whether the higher sensitivity of TMA in comparison with polymerase chain reaction (PCR)-based assays is related to a higher efficiency of the extraction and/or amplification step. OBJECTIVES:The sensitivity of a TMA-based assay (Versant HCV RNA Qualitative assay, Bayer Diagnostics) and a standard RT-PCR-based assay (Cobas Amplicor HCV 2.0, Roche Diagnostics) was compared in formalin-fixed paraffin-embedded liver biopsy specimens of patients with chronic hepatitis C. STUDY DESIGN:After deparaffinization of 7.5 microm liver sections HCV RNA was extracted by standard phenol/chloroform. HCV RNA dilution panels were transferred in parallel to cDNA synthesis and amplification steps of PCR and TMA. Furthermore, TMA amplification from stepwise diluted HCV sera was performed following RNA extraction by either microcentrifuge colums (QIAmp Viral RNA spin Kit, Qiagen, Hilden, Germany) or magnetic microparticles (VERSANT HCV RNA Qualitative assay). RESULTS:The total number of HCV RNA positive liver specimens detected by TMA was higher compared with those detected by RT-PCR (P=0.032). The total number of TMA positive serum samples was higher when HCV RNA was extracted using magnetic microparticles in comparison with multicentrifuge column extraction (P=0.019). CONCLUSION:Our results suggest that both the extraction and amplification step of the TMA-based assay contribute to the higher sensitivity compared with standard RT-PCR.

Metastases are the most common malignant tumors of the liver. In the files of the Institute of Pathology of the University of Cologne 12,161 liver tissue cases are registered. Of them, 1,357 cases (11.2%) showed tumors or tumor like masses. Liver metastases of solid tumors were the largest group of the neoplasias with 611 cases (5.0%) followed by hepatocellular carcinoma (380 cases; 3.1%). Other entities were rare and include cholangiolar carcinoma (0.5%), vascular tumors (0.4%), lymphomas (0.4%), focal nodular hyperplasias (0.36%) and liver cell adenomas (0.23%). Adenocarcinomaa are the largest group of metastases with 400 cases (65.5%). 48.2% of this group were metastases of colorectal cancer, 13.5% of pancreatic cancer, 13% of breast cancer, 6.2% of gastric cancer, 4.5% of lung cancer and 3.7% of esophageal cancer. Neuroendocrine carcinomas are the second largest group with 16% of liver filiae. Other entities were rarely found. Metastases in cirrhotic livers are seldom. The gross findings, the histology, the differential diagnosis including immunohistochemistry and the value of the liver biopsy is discussed.