A 34-year-old-man with short-bowel syndrome received an isolated small bowel graft. On postoperative day (POD) 11, ileal biopsy specimen demonstrated mild to moderate rejection that did not respond to corticosteroid bolus therapy. On POD 14, endoscopy and histologic examination revealed exfoliative rejection that was not controlled after 14 days of therapy with thymoglobulin. On POD 95, the patient underwent surgery again because of intestinal obstruction. The graft was removed 6 months after transplantation because of continuous severe abdominal pain with weight loss. After enterectomy, the patient developed multiple-organ failure and died on POD day 8. This case underlines the severity of exfoliative rejection and suggests that early enterectomy be performed when the diagnosis is made, before deterioration of clinical status and development of infectious and nutritional complications.

Le dépistage systématique du VHC n'est pas recommandé dans la population générale mais proposé à des populations à risque [1]. Le but de notre étude était de déterminer les facteurs associés à la pratique du dépistage du virus de l'hépatite C (VHC) chez les femmes jeunes. La population d'étude était issue des 300 femmes enceintes ayant participé à l'étude ALHICE [1] réalisée entre 1998 et 2002 dont le critère d'inclusion était la présence d'une sérologie VHC positive, connue ou pas avant leur participation. Le moment de la découverte du VHC, les antécédents médicaux et les facteurs de risque liés à leur contamination ont été recueillis par questionnaire réalisé en entretien. La population cible du dépistage a été identifiée selon les recommandations de l'ANAES (janvier 2001) qui regroupe les populations dites à risque élevé (prévalence VHC > 2 %) et celle à risque faible (prévalence VHC < 2 %). Les facteurs associés au dépistage ont été analysés en comparant les femmes ayant connaissance de leur séropositivité avant leur inclusion dans ALHICE à celles l'ignorant avant celle-ci. Le moment de la découverte du VHC n'étant pas renseigné pour 3 femmes, l'analyse a été menée chez 297 femmes. Parmi elles, 224 (75 %) avaient été diagnostiquées avant l'étude ALHICE ; 279 (94 %) présentaient un des critères définis dans les recommandations (population globale cible), 203 (68 %) appartenaient au groupe de risque élevé et 76 au groupe à risque faible. Parmi les 279 femmes de la population globale cible, 68 (24 %) n'avaient pas été diagnostiquées avant leur entrée dans l'étude. Chez les 18 (6 %) femmes ne réunissant pas un des critères de dépistage, 5 (28 %) ont découvert leur séropositivité au moment de leur participation à l'étude. Le dépistage du VHC avant la grossesse était significativement associé à un antécédent d'usage de drogues (62 % versus 28 %, p < 0,001), une séropositivité au VIH (25 % versus 13 %, p = 0,02), un partenaire sexuel infecté par le VHC (42 % versus 25 %, p = 0,01) ou à un partenaire sexuel usager de drogues (65 % versus 41 %, p = 0,01). En revanche, il n'a pas été observé d'association entre le dépistage avant la grossesse et un antécédent de transfusion sanguine avant 1992, une greffe avant 1992 ou une hémodialyse, ni avec l'année de début de grossesse. Le VHC a été découvert au moment de leur inclusion dans l'étude chez une proportion significativement plus élevée de femmes originaires d'une zone à forte prévalence de VHC (27 % versus 14 %, p = 0,01) ou sans emploi (59 % versus 45 %, p = 0,03). Après ajustement, le pays d'origine de la mère (OR : 2,1 IC95 % 1,1-4,1 ; p = 0,03) et l'absence d'emploi (OR : 1,8 IC95 % 1,0-3,1 ; p = 0,05), restaient indépendamment associés à l'absence dépistage du VHC. Pour améliorer le dépistage du VHC chez la population de femmes jeunes, les efforts devraient être concentrés vers les femmes nées dans une des zones à risque (Afrique, Amérique du Sud, Asie du Sud-Est, Moyen-Orient) ainsi que vers celles en situation de précarité. Étude financée par l'ANRS et la Fondation de France.

Parenteral transmission is the major route of hepatitis C virus transmission in adults; however, vertical transmission is most common in children. There are several factors that have been shown to be associated with vertical transmission of hepatitis C virus, including hepatitis C virus RNA, human immunodeficiency virus coinfection, and peripheral blood mononuclear cell infection. As there is no effective vaccine to prevent hepatitis C virus infection, and there are no human data describing the safety of the new direct acting antiviral agents in pregnancy, the only preventive strategy for vertical transmission is to treat the hepatitis C virus infection before becoming pregnant. Direct acting antiviral agents are interferon-free, and many are also ribavirin-free. Based on animal studies, sofosbuvir plus ledipasvir may be the best safety profile during pregnancy for now; however, it is too early to recommend treating hepatitis C virus-infected pregnant women with these direct acting antiviral agents currently.

This cross-sectional study aimed to investigate, during a short period between 2000 and 2001, in a large population of patients with chronic hepatitis C, the epidemiological characteristics of hepatitis C virus (HCV) genotypes in France. Data from 26 referral centres, corresponding to 1769 patients with chronic hepatitis C were collected consecutively during a 6-month period. HCV genotyping in the 5'-non-coding region (NCR) was performed in each center using the line probe assay (LiPA, in 63% of cases), sequencing (25%) or primer-specific polymerase chain reaction (PCR) (12%). HCV genotypes 1a, 1b, 2, 3, 4, 5, non-subtyped 1 and mixed infection were found in 18, 27, 9, 21, 9, 3, 11 and 1% of our population, respectively. HCV genotype distribution was associated with gender, age, source and duration of infection, alanine aminotransferase (ALT) levels, cirrhosis, alcohol consumption, hepatitis B virus (HBV) and human immunodeficiency virus (HIV) coinfection. In multivariate analysis, only the source of infection was the independent factor significantly associated with genotype (P = 0.0001). In conclusion, this study shows a changing pattern of HCV genotypes in France, with i.v. drug abuse as the major risk factor, an increase of genotype 4, and to a lesser extent 1a and 5, and a decrease of genotypes 1b and 2. The modification of the HCV genotype pattern in France in the next 10 years may require new therapeutic strategies, and further survey studies.

Recent attention has focused on the liver profibrogenic role of leptin in animal models. The purpose of this study was to evaluate the role of leptin and TNF-alpha in the severity of liver fibrosis in patients with chronic hepatitis C (CHC). We used a radioimmunoassay to determine serum leptin concentrations in 77 consecutive patients with CHC and 22 healthy controls. Leptin was correlated with liver histological (METAVIR) and metabolic indices. Sixty five patients had none to moderate liver fibrosis (F0-F2) and twelve severe fibrosis (F3-F4). Steatosis was observed in all but 27 patients. Leptin was significantly increased in patients compared with controls and was significantly more elevated in females both in patients and controls. The age, age at infection, prothrombin index, body mass index (BMI), triglycerides, glycaemia, ferritin, leptin and TNF-alpha, were associated with severe fibrosis. Steatosis was significantly more pronounced in patients with severe than those without or moderate fibrosis (P = 0.04). Only leptin was significantly and independently associated with severe fibrosis (OR = 1.2, CI 95%: 1.1-1.4, P = 0.03). Leptin was significantly associated with BMI (r = 0.64, P < 0.001) and glycaemia (r = 0.43, P < 0.001). Significant correlations were found between steatosis and BMI (r = 0.30, P < 0.01) and glycaemia (r = 0.30, P < 0.01). In patients with CHC and higher BMI and glycaemia levels, the severity of liver fibrosis is associated with serum leptin. TNF-alpha is a putative candidate involved in the mechanism.

ABSTRACT Before initiating new large-scale therapeutic trials for hepatitis C virus (HCV)-infected patients, the French Health Authorities for HCV research decided to organize an evaluation of the expertise of laboratories that could be engaged to undertake molecular biology assays in such trials; 21 experienced laboratories participated in this national evaluation of laboratory expertise, which was performed in two successive rounds. The first round evaluated the laboratories for their abilities to detect HCV RNA in serum, determine genotypes, and quantify HCV RNA loads. The results observed by qualitative assays for HCV RNA detection were 100% sensitivity and 100% specificity for all laboratories. The genotyping results were 100% concordant for 9 laboratories and greater than 90% for 10 laboratories. By contrast, large coefficients of variation were observed for quantitative determination of HCV RNA loads, leading to a second round with standardized quantitative assays only. The dispersion of the results was larger by the AMPLICOR HCV Monitor assay than by the branched-DNA assay (mean coefficients of variation, 57.4 and 16.9%, respectively). In the majority of cases, discrepancies between the results of the two tests were found for samples with high viral loads. These results indicate the usefulness of validating, by controlling for expertise, both the reliabilities of laboratories involved in multicenter work and the standardized assays chosen for use in the evaluation of the biological impacts of new therapies.

Background and aims: Fatigue is a frequent and disabling symptom reported by patients with chronic hepatitis C (CHC). Its mechanism is poorly understood. Recent attention has focused on the role of leptin and energy expenditure in CHC. Our aims were to analyse fatigue in CHC and to determine its relationship with disease activity, resting energy expenditure (REE), circulating leptin, and tumour necrosis factor α (TNF-α). Methods: Seventy eight CHC patients, 22 healthy controls, and 13 primary biliary cirrhosis (PBC) patients underwent measurements of REE, body composition, leptin, and TNF-α. All subjects completed the fatigue impact scale (FIS) questionnaire. A liver biopsy and viral load measurements were performed in all patients. Results: Thirty eight of 78 CHC patients considered fatigue the worst or initial symptom of their disease. The fatigue score of patients was significantly higher than that of controls (53.2 (40.1) v 17.7 (16.9); p<0.0001) and was more pronounced in females (p=0.003). Leptin was increased significantly in CHC patients compared with controls (15.4 (20.7) v 6.4 (4.1) ng/ml; p<0.05). In CHC patients, the fatigue score correlated significantly with leptin corrected for fat mass (r=0.30, p=0.01). This correlation increased when the physical domain of fatigue was included (r=0.39, p=0.0009). Furthermore, a similar positive correlation was found in PBC patients (r=0.56, p=0.04). No correlation was found between fatigue and age, REE, liver function tests, viral load, or the METAVIR score in CHC patients. Conclusions: Fatigue is present in CHC patients and is more pronounced in females. The FIS questionnaire is clinically relevant and may be useful for future therapeutic trials aimed at reducing fatigue. Fatigue may be partly mediated by leptin.

Background: Certain chronic hepatitis C carriers have persistently normal transaminase activity. The aims of this study were to determine the virologic and histologic effects of 1 year of interferon-alpha treatment in such patients. Methods: Thirty-one patients were followed up in our Liver Unit. Eleven accepted interferon-alpha therapy; the 20 others were not treated and served as controls. Interferon-alpha, 3 MU, was given thrice weekly for 1 year. Serum was examined for hepatitis C virus (HCV)-RNA before, at the end of, and 6 months after treatment. Liver biopsy was performed 6 months after the cessation of treatment in 10 of 1 1 treated patients (one refused biopsy) and after a mean of 30.6 +/- 22.7 months in the 20 untreated patients. Results: At the end of follow-up two of the treated patients had undetectable serum HCV-RNA and five had increased alanine aminotransferase (ALAT) values. In contrast, only one of the untreated patients had abnormal ALAT activity. All 20 untreated patients were constantly viremic. No significant histologic improvement was observed in the treated patients evaluated by means of posttreatment liver biopsy. The mean annual progression rate of fibrosis was very slow and similar in the treated and untreated patients (0.09 (range, 0-0.62) versus 0.07 (range, 0-0.60) fibrosis units). Conclusions: One year of interferon-alpha treatment can suppress HCV-RNA in patients with chronic hepatitis C and persistently normal ALAT values followed up over long periods. The rate of fibrosis progression in such patients is very slow, and therapeutic strategies should take this fact into account. Antiviral treatment is debated for patients without fibrosis in initial biopsy specimens.

The first and second generations of the Cobas Amplicor HCV assay were compared among patients at risk of hepatitis C virus (HCV) infection. The second-generation test was found to be of greater sensitivity and of good specificity among clinical specimens containing HCV RNA of different genotypes. Finally, this new test is shown to predict the outcome of interferon therapy better.

There is an increasing demand for the genotyping of hepatitis C virus (HCV), since it has been shown that different HCV genotypes are associated with distinct profiles of pathogenicity and responses to antiviral treatment. Hence, there is a need for a simple and precise genotyping assay for routine diagnosis of HCV types and subtypes. Here we show that direct sequencing, considered as the reference method, can provide an accurate and rapid method for large-scale screening of HCV genotypes. PCR-amplified cDNAs of the HCV 5' non-coding region (5' NCR) were obtained from the widespread "Amplicor" HCV detection system. Semi-purified PCR products were directly cycle-sequenced in a single tube using multicolour dye terminator chemistry. Sample loading, electrophoresis and sequence analysis were automatically achieved by a capillary electrophoresis-based genetic analyser. Out of a total of 500 samples, HCV subtype 1b accounted for the majority of the infections (41%), followed by HCV 3 (31%) and HCV 1a (22%). This procedure failed to identify a genotype in only 3 samples. In addition, several cases of mixed HCV infection were also documented. The combination of direct cycle sequencing of PCR products with capillary electrophoresis provides a simple and rapid method convenient for routine HCV genotyping analysis.

Complementary DNA encoding a human Gal(beta1-3)GalNAc alpha2,3-sialyltransferase type II (hST3Gal II) was cloned from a CEM T-cell cDNA library using a 23-base oligonucleotide probe. The sequence of this probe was established on the basis of a slightly divergent sialylmotif L that was obtained by polymerase chain reaction with degenerate oligonucleotide primers based on the conserved sialylmotif L of mammalian Gal(beta1-3)GalNAc alpha2,3-sialyltransferases. It was thus confirmed that a short oligonucleotide probe may be sensitive and highly specific. The nucleotide and amino acid sequences of hST3Gal II show, respectively, 56.3% and 49.3% similarity to hST3Gal I [Kitagawa, H. & Paulson, J. C. (1994) J. Biol. Chem. 269, 17872-17878] and 88.1% and 93.7% similarity to murine ST3Gal II [Lee, Y. C., Kojima, N., Wada, E., Kurosawa, N., Nakaoka, T., Hamamoto, T. & Tsuji, S. (1994) J. Biol. Chem. 269, 10028-10033]. hST3Gal II mRNA was highly expressed in heart, liver, skeletal muscle and various lymphoid tissues but not in brain and kidney. A soluble form of hST3Gal II expressed in COS-7 cells was tested in vitro for substrate specificity and kinetic properties. Asialofetuin and asialo-bovine submaxillary mucin appeared better substrates for hST3Gal II than for its murine counterpart as previously reported [Kojima, N., Lee, Y.-C., Hamamoto, T., Kurosawa, N. & Tsuji, S. (1994) Biochemistry 33, 5772-5776]. In previous studies, we have shown hyposialylation of O-glycans attached to two major lymphocyte CD43 and CD45 cell surface molecules in human-immunodeficiency-virus-1(HIV-1)-infected T-cell lines. Since comparable levels of hST3Gal I and hST3Gal II mRNA and enzymatic activity were observed in parental and HIV-1-infected CEM T-cell lysates, the sialylation defect associated with HIV infection of this cell line is probably due to a mechanism different from a simple altered catalytic activity of these sialyltransferases.

Several bacterial and viral agents have been implicated in the pathogenesis of the Guillain-Barré syndrome, an acquired immune-mediated disorder. A case of Guillain-Barré syndrome associated with acute Q fever is described. Coxiella burnetii should therefore be added to the list of microorganisms capable of inducing the Guillain-Barré syndrome.