Christophe Renou

A randomized controlled trial of high-dose ursodesoxycholic acid for nonalcoholic steatohepatitis

BACKGROUND & AIMS Nonalcoholic steatohepatitis (NASH) is a prevalent liver disease associated with increased morbidity and mortality. Ursodeoxycholic acid (UDCA) may have antioxidant, anti-inflammatory, and antifibrotic properties and may reduce liver injury in NASH. To date, no studies have assessed the efficacy and safety of high-dose UDCA (HD-UDCA) in patients with NASH. METHODS We conducted a 12-month, randomized, double-blind, placebo-controlled multicenter trial to evaluate the efficacy and safety of HD-UDCA (28-35 mg/kg per day) in 126 patients with biopsy-proven NASH and elevated alanine aminotransferase (ALT) levels. The primary study end point was reduction in ALT levels from baseline in patients treated with HD-UDCA compared with placebo. Secondary study end points were the proportion of patients with ALT normalization, relative reduction in the scores of serum markers of fibrosis and hepatic inflammation, and safety and tolerability. RESULTS HD-UDCA significantly reduced mean ALT levels -28.3% from baseline after 12 months compared with -1.6% with placebo (p<0.001). At the end of the trial, ALT levels normalized (≤35 IU/L) in 24.5% of patients treated with HD-UDCA and in 4.8% of patients who received placebo (p=0.003). Both results were not accounted for by changes in weight during the trial. HD-UDCA significantly reduced the FibroTest® serum fibrosis marker (p<0.001) compared with placebo. HD-UDCA also significantly improved markers of glycemic control and insulin resistance. There were no safety issues in this population. CONCLUSIONS Treatment with HD-UDCA was safe, improved aminotransferase levels, serum fibrosis markers, and selected metabolic parameters. Studies with histologic end points are warranted.

Independent Prospective Multicenter Validation of Biochemical Markers (Fibrotest-Actitest) for the Prediction of Liver Fibrosis and Activity in Patients with Chronic Hepatitis C: The Fibropaca Study

OBJECTIVES:Fibrotest (FT) and Actitest (AT) are biochemical markers of fibrosis and activity for use as a non-invasive alternative to liver biopsy in patients with chronic hepatitis C virus (HCV). The aim of this study was to perform an external validation of FT and AT and to study the discordances between FT/AT and liver biopsy in patients with chronic hepatitis C.METHODS:A total of 519 consecutive patients with chronic HCV were prospectively included in five centers, with liver biopsy and biochemical markers taken at the same day. Fifteen patients were excluded because their biopsies could not be interpreted. Diagnostic accuracies were assessed by receiver operating characteristic (ROC) curve analysis.RESULTS:Median biopsy size was 15 mm (range: 2–58), with 9 portal tracts (1–37) and 1 fragment (1–12). 46% (230/504) were classified F2–F4 in fibrosis and 39% A2–A3 in activity. FT area under ROC curve for diagnosis of activity (A2–A3), significant fibrosis (F2–F4), and severe fibrosis (F3–F4) were 0.73 [0.69–0.77], 0.79 [0.75–0.82], and 0.80 [0.76–0.83], respectively. Among the 92 patients (18%) with 2 fibrosis stages of discordance between FT and biopsy, the discordance was attributable to FT in 5% of cases, to biopsy in 4%, and undetermined in 9%.CONCLUSIONS:This prospective independent and multicenter study confirms the diagnostic value of FT and AT found in the princeps study and suggests that FT and AT can be an alternative to biopsy in most patients with chronic HCV.

Accuracy of hyaluronic acid level for predicting liver fibrosis stages in patients with hepatitis C virus.

BackgroundIn patients with chronic hepatitis C virus, liver biopsy is the gold standard for assessing liver disease stage; nevertheless, it is prone to complications, some of them serious. Non-invasive methods have been proposed as surrogate markers for liver fibrosis. It was shown that serum hyaluronic acid (HA) level increases with the development for liver fibrosis. The aim of this study was to evaluate the diagnostic value of HA as well as to determine the HA level cut-off for predicting the presence or absence of fibrosis, severe fibrosis, and cirrhosis.Results405 patients with chronic hepatitis C were prospectively included with biomarker measurement and liver biopsy done the same day: 151 in the training set (only biopsy lengths of 25 mm or more) and 254 in the validation set. For the discrimination of significant fibrosis, severe fibrosis, and cirrhosis in the training set, the areas under curve (AUCs) were 0.75 ± 0.03, 0.82 ± 0.02, and 0.89 ± 0.03, respectively. Absence of significant fibrosis, severe fibrosis, and cirrhosis can be predicted by HA levels of 16, 25, and 50 μg/l, respectively (with negative predictive values of 82%, 89%, and 100%, in the same order). Presence of significant fibrosis, severe fibrosis, and cirrhosis can be predicted by HA levels of 121, 160, and 237 μg/l, respectively (with positive predictive values of 94%, 100%, and 57%, in the same order).ConclusionIn the validation set, HA was accurate in predicting significant fibrosis, severe fibrosis, and cirrhosis with AUCs of 0.73, 0.77, and 0.97, respectively. Moreover, accurate HA level cut-offs were defined for predicting significant fibrosis, severe fibrosis, and cirrhosis. Thus, the study supports that HA level may be clinically useful as a non-invasive marker for liver fibrosis and/or cirrhosis.

A national survey of acute hepatitis E in France

Background  Few data are available on the incidence, risk factors and contamination pathways involved in acute indigenous hepatitis E in developed countries.

Quantitative study of digestive enzyme secretion and gastrointestinal lipolysis in chronic pancreatitis

BACKGROUND & AIMS The contribution of human gastric lipase (HGL) to the overall lipolysis process in chronic pancreatitis (CP), as well as the relative pancreatic enzyme levels, rarely are addressed. This study was designed to quantify pancreatic and extrapancreatic enzyme output, activity, and stability in CP patients vs. healthy volunteers. METHODS Healthy volunteers (n = 6), mild CP patients (n = 5), and severe (n = 7) CP patients were intubated with gastric and duodenal tubes before the administration of a test meal. HGL, human pancreatic lipase (HPL), chymotrypsin, and amylase concentrations were assessed in gastric and duodenal samples by measuring the respective enzymatic activities. Intragastric and overall lipolysis levels at the angle of Treitz were estimated based on quantitative analysis of lipolysis products. Similar analyses were performed on duodenal contents incubated ex vivo for studying enzyme stability and evolution of lipolysis. RESULTS Although HPL, chymotrypsin, and amylase outputs all were extremely low, HGL outputs in patients with severe CP (46.8 +/- 31.0 mg) were 3-4-fold higher than in healthy controls (13.3 +/- 13.8 mg). Intragastric lipolysis did not increase, however, in patients with severe CP, probably because of the rapid decrease in the pH level of the gastric contents caused by a higher gastric acid secretion. HGL remains active and highly stable in the acidic duodenal contents of CP patients, and, overall, can achieve a significant lipolysis of the dietary triglycerides (30% of the control values) in the absence of HPL. CONCLUSIONS Although all pancreatic enzyme secretions are simultaneously reduced in severe CP, gastric lipase can compensate partly for the loss of pancreatic lipase but not normalize overall lipolytic activity.

Hepatitis C Virus Genotyping Based on 5′ Noncoding Sequence Analysis (Trugene)

ABSTRACT Hepatitis C virus (HCV) genotyping of samples from 184 patients with chronic HCV infection by the Trugene 5′NC genotyping kit, based on sequence analysis of the 5′ noncoding region (5′ NCR), and the InnoLiPA assay was evaluated. In addition to these methods, the 184 samples were also analyzed by sequencing of part of the NS5B of the HCV genome after in-house PCR amplification, as a means of validating results obtained with the 5′ NCR. The distribution of the genotypes typed by NS5B sequence analysis was as follows: 1a, 41 samples; 1b, 58 samples; 1d, 1 sample; 2a, 5 samples; 2b, 2 samples; 2c, 7 samples; 3a, 46 samples; 4a, 7 samples; 4c, 1 samples; 4e, 9 samples; 5a, 6 samples; 6a, 1 sample. The Trugene and InnoLiPA assays gave concordant results within HCV types in 100% of cases. The ability to discriminate at the subtype level was 76 and 74% for the Trugene and the InnoLiPA assays, respectively.

Influence of HLA-DR phenotype on the risk of hepatitis C virus-associated mixed cryoglobulinemia.

OBJECTIVE Circumstances predisposing hepatitis C virus (HCV)-infected patients to develop mixed cryoglobulinemia (MC), which may manifest as a small-vessel systemic vasculitis (MC vasculitis), remain unclear. Previous studies have failed to demonstrate a clear role of either viral factors (genotype, viral load) or host factors (lymphocytes or immunoglobulin subsets). This study was undertaken to examine a possible role of HLA class II alleles in HCV-associated MC. METHODS One hundred fifty-eight HCV-infected patients, of whom 76 had MC (56 with type II MC and 20 with type III MC) and 82 did not have MC, were studied prospectively. MC vasculitis was noted in 35 HCV-infected patients with type II IgMkappa-containing cryoglobulins. HLA-DRB1 and HLA-DQB1 polymorphism was analyzed by hybridization using allele-specific oligonucleotides, after gene amplification. The odds ratio (OR) was calculated with Woolfs method. Then, using multivariate analysis, demographic, biologic, immunologic, virologic, and liver histologic factors associated with the presence of MC and MC vasculitis were investigated. RESULTS HLA-DR11 was significantly more frequent in patients with type II MC than in those without MC (41.1% versus 17.1%; OR 3.4, corrected P [Pcorr] = 0.017), regardless of the presence of vasculitis accompanying the MC (37.1% of those with MC vasculitis, 34.1% of those with MC but no vasculitis). HLA-DR7 was less frequent in HCV-infected patients with MC than in those without MC (13.2% versus 30.5%; OR 0.34, P = 0.012, Pcorr not significant), with a particularly lower frequency in those with type II MC and those with MC vasculitis (12.5% and 8.6%, respectively). There was no significant difference in HLA-DQB1 distribution between the different patient groups. By univariate and multivariate analysis, HLA-DR11 was the only positive predictive factor, besides female sex and advanced age, for the presence of MC and HCV-associated MC vasculitis (OR 2.58). CONCLUSION Our results indicate that the presence of the DR11 phenotype is associated with a significantly increased risk for the development of type II MC in patients with chronic HCV infection. In contrast, HLA-DR7 appears to protect against the production of type II MC. These results suggest that the hosts immune response genes may play a role in the pathogenesis of HCV-associated MC.

Possible Zoonotic Transmission of Hepatitis E from Pet Pig to Its Owner

Hepatitis E is transmitted mainly by water or food, but in industrialized countries, all routes of transmission have not been identified. We describe possible zoonotic transmission of hepatitis E virus that involved direct contact between a pet pig and its owner.

Molecular Evidence of Male-to-Female Sexual Transmission of Hepatitis C Virus after Vaginal and Anal Intercourse

ABSTRACT Hepatitis C virus (HCV) was transmitted from a chronic carrier to his female partner during unprotected anal and vaginal intercourse. Based on HVR1 and phylogenetic tree analysis, the couple had closely related isolates. These findings confirm sexual transmission of HCV without other risk factors.

Foodborne transmission of hepatitis E virus from raw pork liver sausage, France.

To the Editor: The number of sporadic autochthonous cases of acute hepatitis E is increasing in many industrialized countries (1). These cases involve hepatitis E virus (HEV) genotypes 3 and 4, which are zoonotic. Although risk for foodborne transmission from pork is now recognized, we report here direct HEV transmission through ingestion of raw pig liver sausages (figatellu [plural: figatelli]) in southeastern France. The index case-patient was a 45-year-old woman from Hyeres (southeastern France) who had no underlying medical condition. She visited her general practitioner on December 17, 2013, reporting 3 days of weakness. Acute hepatitis was diagnosed 2 days later on the basis of elevated liver enzymes (alanine aminotransferase 1,265 IU/L [reference <35 IU/L]) and bilirubin (65 μmol/L [reference <17 μmol/L]). Serum markers for acute hepatitis A, B, and C; cytomegalovirus; and Epstein-Barr virus were negative. Jaundice appeared on December 19, and the patient was referred to the Medical Unit of Hyeres for additional investigations. A serum sample collected on December 20 tested positive for HEV RNA; viral load was 3.3 log10 IU/mL (Ceeram, La Chapelle sur Erdre, France), and IgM and IgG against HEV were found (Wantai, Beijing, China), which led to the diagnosis of acute hepatitis E. The HEV genotype was 3f, as determined from the phylogenetic analysis of a portion of the open reading frame (ORF) 2 (2). The index case-patient recovered by the end of January; HEV viremia was undetectable on January 17, 2014. The index case-patient and her family regularly ate figatelli (raw pork liver sausages) made in Corsica. The patient had most recently eaten figatelli at a lunch with 8 family members on October 28, 2013, seven weeks before illness onset. After receiving informed consent, we conducted laboratory investigations of samples from the other family members; tests included HEV serology and HEV RNA detection in serum and fecal samples. Samples were obtained from family members during January 8–21, 2014 (41–54 days after the lunch). Positive HEV IgM and detectable HEV RNA were found in the serum of the index case-patient’s daughter, who was asymptomatic. Because the sample was tested 10 weeks after the family lunch, the daughter’s HEV viral load was too low to enable sequence characterization and clustering of HEV strains. Three other family members were IgG positive for HEV, indicating previous HEV infection. Leftover sausages had been kept frozen and were available for HEV testing. HEV RNA was detectable from the leftover sausages, and HEV sequences were amplified in 2 different genomic regions (ORF1: RNA-dependent RNA polymerase and ORF2), as described previously (2). Comparison with the index case-patient’s sequences showed 100% nt identity for both regions (Figure). Samples of food and samples from the index case-patient were analyzed in 2 independent laboratories to avoid any cross-contamination. The level of contamination of the figatellu was ≈4.8 104 copies of HEV RNA/g of sausage (3). Figure Phylogenetic analysis of partial open reading frame (ORF) 2 and ORF1 sequences of hepatitis E virus (HEV). Phylogenetic trees were constructed in MEGA6 software (http://www.megasoftware.net) by using the neighbor-joining method from a Kimura 2-parameter ... Figatellu, a dried sausage, contains 30% pork liver and no heating step occurs during its manufacture. Usually deep cooking is recommended on the package, but consumers might not follow the cooking recommendation; also, figatelli can be sold in small local shops with no label. In the instance reported here, the figatellu was sold without any warning label and was eaten raw. That HEV was transmitted through ingestion of contaminated food is supported by the following evidence. First, 3 case reports have provided direct evidence of HEV transmission through ingestion of contaminated animal food products with identical or near identical sequences between the patients and the contaminated food they ate. Two cases occurred in the early 2000s in Japan through consumption of grilled wild boar (4) or sashimi of Sika deer (5); the third, reported recently in Spain, was transmitted through ingestion of pig meat (6). Second, HEV widely infects domestic pigs and wild boar (7). Third, swine and human HEV strains have genetic similarities and, in some cases, are indistinguishable (1). Fourth, in Marseille, France, a case–control study identified ingestion of figatellu as a risk factor for HEV infection. Genetic similarities were found between sequences isolated from patients with autochthonous hepatitis E and nonrelated figatelli purchased in the same region (8). Finally, infectious virus, replicating in a 3-dimensional culture system, was identified in a HEV RNA-positive figatellu (9). In the present study, the homology between sequences recovered from the index case-patient and those recovered from leftovers of figatellu provides additional proof of HEV foodborne transmission in a Western country. In France, information about the risk for HEV transmission through the ingestion of such delicatessen was published by French authorities in 2010 (10), but the present case demonstrates that public education and warning, or larger and more explicit labels on the package, must be improved to reduce the risk for HEV exposure.