Diagnostic performance of FibroTest, SteatoTest and ActiTest in patients with NAFLD using the SAF score as histological reference

BACKGROUND: Blood tests of liver injury are less well validated in non‐alcoholic fatty liver disease (NAFLD) than in patients with chronic viral hepatitis. AIMS: To improve the validation of three blood tests used in NAFLD patients, FibroTest for fibrosis staging, SteatoTest for steatosis grading and ActiTest for inflammation activity grading. METHODS: We pre‐included new NAFLD patients with biopsy and blood tests from a single‐centre cohort (FibroFrance) and from the multicentre FLIP consortium. Contemporaneous biopsies were blindly assessed using the new steatosis, activity and fibrosis (SAF) score, which provides a reliable and reproducible diagnosis and grading/staging of the three elementary features of NAFLD (steatosis, inflammatory activity) and fibrosis with reduced interobserver variability. We used nonbinary‐ROC (NonBinAUROC) as the main endpoint to prevent spectrum effect and multiple testing. RESULTS: A total of 600 patients with reliable tests and biopsies were included. The mean NonBinAUROCs (95% CI) of tests were all significant (P < 0.0001): 0.878 (0.864–0.892) for FibroTest and fibrosis stages, 0.846 (0.830–0.862) for ActiTest and activity grades, and 0.822 (0.804–0.840) for SteatoTest and steatosis grades. FibroTest had a higher NonBinAUROC than BARD (0.836; 0.820–0.852; P = 0.0001), FIB4 (0.845; 0.829–0.861; P = 0.007) but not significantly different than the NAFLD score (0.866; 0.850–0.882; P = 0.26). FibroTest had a significant difference in median values between adjacent stage F2 and stage F1 contrarily to BARD, FIB4 and NAFLD scores (Bonferroni test P < 0.05). CONCLUSIONS: In patients with NAFLD, SteatoTest, ActiTest and FibroTest are non‐invasive tests that offer an alternative to biopsy, and they correlate with the simple grading/staging of the SAF scoring system across the three elementary features of NAFLD: steatosis, inflammatory activity and fibrosis

Endothelial cell apoptosis in the context of quantification of angiogenesis in solid human adenocarcinomas:A novel double immunolabelling technique to identify endothelial cell apoptosis

Standardised methods of microvessel density quantification have been published. However, a reliable and reproducible method to visualise endothelial cell apoptosis is lacking, which is a shortcoming in assessing vascular remodelling during angiogenesis. The aim of this study was to validate a newly developed technique to demonstrate endothelial cell apoptosis by double immunolabelling with anti-CD34 and anti-activated caspase-3 in human adenocarcinomas. Double immunolabelling was used to identify apoptotic endothelial cells in six tumours of eight different human adenocarcinomas. Microvessel density and rate of apoptotic endothelial cells were quantified. The technique revealed endothelial cell apoptosis simultaneously with the identification of microvessel density on one slide. These characteristics were reproducible in adenocarcinomas of various sites. In conclusion, apoptotic endothelial cells and microvessel density can now be evaluated simultaneously within one and the same area, allowing a more reliable histological quantification of angiogenesis. (c) 2005 Elsevier Ltd. All rights reserved

Precision-cut bile duct slices as a model to study regeneration of bile ducts of human donor livers after ischemic preservation injury

Introduction Clinical studies have shown a high incidence of biliary epithelial injury after cold ischemic preservation of donor livers. Insufficient epithelial regeneration from the peribiliary glands (PBGs) has been proposed as a critical mechanism in the pathogenesis of biliary strictures after transplantation. Severe biliary epithelial injury requires proliferation and mobilization of biliary progenitor cells nested in PBGs. Studies on the pathogenesis and prevention of biliary strictures are hampered by the lack of suitable laboratory and animal models. Precision-cut tissue slices are a widely used ex vivo technique, in which thin slices of human tissue are cultured and kept viable with intact intercellular and cell-matrix interactions for up to several days. Aims The aim of this study was to generate precision-cut bile duct slices and study their suitability as a model to study the regenerative capacity of PBGs in donor bile ducts. Methods Large bile ducts of cold preserved livers declined for transplantation (n=10) were isolated and sliced, using a Krumdieck slicer. Bile duct slices were cultured in Williams’ medium E up to 144 hr. Histology and immunohistochemistry (HE, Ki-67, and K19) were performed to assess cell morphology. Results Viable K19-positive PBG cells were observed at all time points. Cell proliferation rate, as assessed by Ki-67 staining, increased from 0% at 24 hr to 25% at 96 hr of culture. Proliferation rate was significantly higher in bile ducts of donors <60 years, compared to donors ≥60 years (21% vs. 11%, p = 0.026) Conclusions Precision-cut bile duct slices are a feasible model to study biliary epithelial regeneration of ischemic injured donor liver bile ducts. This model provides a new ex vivo tool to develop pharmacological strategies to prevent post-transplant biliary strictures

Long-term Outcome of Asymptomatic Patients With Graft Fibrosis in Protocol Biopsies After Pediatric Liver Transplantation

Background: The histological prevalence of allograft fibrosis in asymptomatic children after liver transplantation (LT) is well documented. However, long-term graft and patient survival remain unclear. This retrospective multicenter study aims to determine the prevalence of allograft fibrosis and analyze the long-term outcome for patients transplanted in childhood. Methods: We reviewed clinical data of children who had undergone 10-y protocol liver biopsies. We excluded patients with autoimmune hepatitis, primary sclerosing cholangitis, hepatitis B or C, and retransplantation. In total, 494 patients transplanted in childhood across 12 international transplant centers were included. We evaluated the development of fibrosis by comparing the results with biopsies obtained 5 and 15 y post-LT. Histological findings were correlated with graft and patient survival up to 20 y post-LT. Results: In the 10-y biopsies, periportal or pericentral fibrosis was observed in 253 patients (51%), 87 (18%) had bridging fibrosis, 30 (6%) had cirrhosis, and 124 (25%) had no fibrosis. The prevalence and stage of graft fibrosis significantly progressed from 5 to 10 y. At 10 y, the severity of fibrosis correlated significantly with inflammation. Patients with graft cirrhosis in the 10-y biopsy were more likely to die or require retransplantation subsequently (P = 0.027). Conclusions: At 10 y post-LT, most patients transplanted in childhood developed fibrosis, based on the protocol liver biopsies. Although mild-to-moderate graft fibrosis did not largely affect patient or graft survival up to 20 y post-LT, this progressive fibrosis finding has substantial implications for developing cirrhosis and portal hypertension in adult care