Qualitative serum HCV-RNA detection was performed with reverse transcriptase-polymerase chain reaction in the 5��-noncoding region of the HCV genome (Roche COBAS Amplicor HCV Test, version 2.0: Roche Diagnostics, Basel, Switzerland). Quantification was performed using branched DNA with the Bayer��s VERSANT bDNA 3.0 assays (Bayer Diagnostics, Emeryville, CA, USA). selleck chemicals The detection threshold was 3200 copies (615 IU) per mL. HCV genotyping was performed with INNO-LIPA HCV II (Innogenetics, Gent, Belgium). Liver histology Following assessment of prothrombin time and platelet count, patients underwent a percutaneous, ultrasound-guided liver biopsy under local anaesthesia (lignocaine 1%). Specimens obtained by means of Menghini needles, diameter 1.

6 mm, had an average length of 20 �� 5 mm (range, 15�C25 mm), and representative according to accepted standards. Biopsy specimens were fixed with formalin, embedded in paraffin and stained with haematoxylin and eosin. All sections were reviewed by an expert pathologist blinded to patient clinical data and breath-test results. Necroinflammatory score was graded using the HAI score based on periportal or periseptal interface hepatitis (piecemeal necrosis) (0�C4), confluent necrosis (0�C6), focal (spotty) lytic necrosis, apoptosis, and focal inflammation (0�C4) and portal inflammation (0�C4) [13]. Fibrosis was staged using the Ishak (modified HAI) fibrosis score on a scale from 0 to 6 [13]. Table 3 shows selected patient data grouped by fibrosis score.

Table 3 HCV patient population grouped by modified HAI fibrosis stage for patient data and blood test results Noninvasive breath testing Following an overnight (>8 h) fast, patients and healthy volunteers were connected to the breath-testing unit��s BreathID? system (BreathID Ltd, Jerusalem, Israel) via nasal cannula (IDcircuitTM), and received 75 mg of N-(4-methoxy-13C-phenyl)acetamide (methacetin, Isotec) dissolved in 150 mL of water. Breath samples were collected using an automatic breath sampling unit under continuous capnographic control, before and for 60 min after the labelled substrate was administered to the patient. The 13CO2/12CO2 ratios in the breath samples were determined and mapped on the screen at a high frequency (once every 2�C3 min). During the test period, all patients and healthy volunteers continued fasting and were at rest to eliminate any variability in CO2 excretion due to the ingestion of food or physical activity.

Analysis of breath-test data Results obtained from the device were expressed as percentage of administered dose of 13C per cent dose Dacomitinib recovered (PDR) and the cumulative PDR (CPDR) percentage of 13C recovered over time at 10, 15, 20, 30 and 60 min after ingestion of methacetin, respectively, as well as the PDR peak and peak time. PDR refers to the rate at which the 13C substrate is metabolized and is expressed in %/h.