Localization of hepatitis C virus RNA on human erythrocytes by RT in situ PCR technique.

BACKGROUND: Our previous results showed that hepatitis C virus (HCV) is detectable on erythrocytes by RT in situ PCR. The aims of the present study were to compare the sensitivity of this erythrocyte in situ PCR to routine serum solution phase HCV PCR as well as to obtain more data about the binding and cellular localization of HCV in the erythrocyte. METHODS: 105 previously HCV-infected patients and 20 control individuals were studied using RT in situ PCR on erythrocytes and solution phase RT-PCR from serum samples. Binding of HCV to erythrocytes was studied by in vitro inoculation. RT in situ PCR results were evaluated by fluorescence and confocal laser scanning microscopy. RESULTS: From 105 HCV cases, 78 gave positive, while 5-and all control cases-gave negative results by both PCR techniques. In 21 cases, only the in situ technique provided positive results, while in only I case did the solution phase method provide positive results. During in vitro inoculation, an early HCV-erythrocyte binding was detected followed by virus internalization. CONCLUSIONS: Erythrocyte-in situ PCR was found to show higher sensitivity for the detection of HCV compared to the generally applied serum PCR method. In vitro studies suggested a specific binding of HCV to erythrocyte and showed the virus to be capable of internalization.

Prevalence, genotype distribution and outcome of hepatitis C infections among the employees of the Hungarian Central Hospital for infectious diseases.

Blood samples from 477 hospital workers (HWs) at the Central Hospital for Infectious Diseases, Budapest, Hungary were tested for hepatitis C virus (HCV) antibodies by enzyme immunoassay (EIA), and 13 (2.7%) of these were found to be HCV antibody positive. Ten (2.7%) were from nursing/housekeeping staff and three (2.9%) from medical staff. HCV antibody positive HWs were detected in 10 of 17 work places, and the prevalence rates in these departments or units varied between 1.2% and 6.5%. The prevalence increased gradually with increasing age, being 0% in these under 21 years of age and 9.5% in those above 50 years of age. Eleven (85%) of 13 HCV antibody positive HWs had HCV RNA in their sera, four of them intermittently during the follow-up period. HCV genotype 1 was present in two HWs, 1b in six HWs, 3a in one HW and 4 in two HWs. Chronic hepatitis C has developed in six (46%) HCV antibody positive HWs. Although the source of infection through needlestick could only be traced directly in one case, circumstantial evidence indicated that the majority of infections were occupationally acquired, originating from percutaneous or mucocutaneous exposure.

Therapy for chronic hepatitis C.

BACKGROUND/AIMS: in Hungary, over the past 5 years more than 900 patients with chronic hepatitis C have been examined for treatment with interferon at 16 major hepatology centres, using unified diagnostic and therapeutical criteria. Authors give an account of their experiences on the clinical features of patients with chronic hepatitis C and report the results of the interferon therapy. METHODS: a total of 993 patients--virtually the entire Hungarian hepatitis C patient population who had been referred for interferon treatment--were included in the program. Actually, the sustained efficacy of the therapy was evaluated in 724 cases. Treatment protocols (dose of interferon and duration of therapy) have changed with time from a weekly dose of 3x3 MU IFN for 6 months in the first period, to 3x3-5 MU for 12 months in the second period, and finally in the third period a combination therapy with ribavirin has also been introduced. RESULTS: in the first period, the end-of-treatment response (ETR) was 35%, sustained response (SR) 13%, the second phase schedule resulted in 42% ETR and 22% SR, while in the third period, ETR was 49% and SR 36%, respectively. Fibrosis in histology and baseline pretreatment HCV-RNA level appeared as predictors of response. The duration of treatment and the total dose of interferon exerted a moderate effect on therapeutic efficacy. Neither age nor gender influenced the outcome. CONCLUSIONS: our results-obtained in a Central East European country-are in accordance with findings of suboptimal efficacy of traditional interferon monotherapy for chronic hepatitis C reported in the West, and suggest the benefit of the combination treatment of interferon with ribavirin.

[Adult-onset Kawasaki syndrome in the differential diagnosis of liver disease]. / Felnóttkori Kawasaki-szindróma a májbetegségek differenciáldiagnózisában.

The authors present a case of an adult with Kawasaki syndrome, who, due to jaundice, enlarged liver and abnormal liver function tests, was admitted the hospital with the suspicion of liver disease. The symptoms of Kawasaki syndrome appeared during the first nine days of the hospital stay. The authors emphasise, that liver function tests are frequently abnormal in adults presenting with this clinical entity, therefore Kawasaki syndrome should be taken into consideration in the differential diagnosis of liver diseases.

Hepatocyte proliferation and cell cycle phase fractions in chronic viral hepatitis C by image analysis method.

OBJECTIVE: Chronic hepatitis is characterized by necrosis of liver cells, accompanied by an inflammatory reaction and compensatory cell proliferation. The interaction of the core and non-structural proteins of hepatitis C virus (HCV) with several cellular factors suggests that cell proliferation may be influenced by HCV. The aim of this study was to investigate hepatocyte proliferation and DNA ploidy patterns in patients with chronic viral hepatitis C (CH-C) compared with chronic non-viral hepatitis (CH-N), using a TV image analysis method. METHODS: The DNA index (DI) and cell phase fractions (G1, S, G2) were measured by means of digital picture analysis method on nuclear suspensions of Feulgen stained hepatocytes. Cells were taken from the liver biopsy specimens of 71 patients with CH-C and 24 patients with CH-N. Twenty-six normal liver samples were used as controls. RESULTS: Significantly higher G1 (94 +/- 4) and lower S (3.56 +/- 3.16) phase fractions were measured in CH-C compared with CH-N (G1, 90 +/- 6; S, 6.4 +/- 5.99). The DI of moderate (1.12 +/- 0.05) and severe (1.12 +/- 0.05) CH-C showed near-aneuploid DNA content, while diploidy (DI < 1.10) was detected in cases of CH-N. CONCLUSION: The higher G1 and lower S cell cycle phase fractions in CH-C reflect decreased hepatocyte proliferation compared with CH-N. The near-aneuploid DNA content of the HCV-infected liver samples may be a sign of increased genetic instability, which may contribute to the carcinogenic potential of HCV.

C5b-9 and interleukin-6 in chronic hepatitis C. Surrogate markers predicting short-term response to interferon alpha-2b.

BACKGROUND: Available data and our observations suggest that elevated levels of interleukin (IL)-6 and -10 and some complement parameters may be associated with a poor response to IFN alpha. We evaluated how baseline levels of C5b-9, IL-6, and IL-10 influence the outcome of IFN alpha treatment. METHODS: Fifty-one patients with established chronic hepatitis C were enrolled and treated with IFN alpha-2b. Before and after a 12-week-IFN-treatment (3 MU or 5 MU tiw) serum levels of IL-6, IL-10, C5b-9 and RNA of hepatitis C virus (HCV) were assessed. Sera of 46 sex- and age-matched, healthy blood donors served as control. RESULTS: While two-thirds of patients was considered 'responder', 14 patients had no significant decrease either in HCV RNA or in ALT levels. In the responder's group lower baseline levels of IL-6 and C5b-9 were found than those in the 'non-responder' group. As a result of IFN therapy HCV RNA and C5b-9 levels significantly decreased. While the serum concentration of IL-6 increased during the follow-up period, regarding IL-10, no change was observed. In patients with 'low' baseline levels of C5b-9 (<2053 ng/ml) IFN alpha resulted in a significantly (P = 0.0005) higher decrease in HCV RNA level. Regarding 'low' IL-6 values (< 1.47 pg/ml) similar but somewhat less significant (P = 0.0039) difference was found if the change of HCV RNA was investigated. The odds ratio of patients with low IL-6 and/or C5b-9 to responding to IFN alpha treatment was almost 10 times (CI: 9.1 (1.8-50.9)) higher as compared with patients without 'low' levels of these parameters. CONCLUSION: Our data suggest that serum level(s) of IL-6 and/or C5b-9 taken prior to the initiation of IFN treatment may serve as surrogate marker(s) in evaluating patients with chronic hepatitis C whether to get IFN alpha in monotherapy or to consider having combination therapy in the form of IFN alpha-ribavirin.

Fungal colonization of the esophagus in alcoholic liver disease.

OBJECTIVES: Little is known about the fungal colonization of the esophagus. Since alcoholic liver disease (ALD) patients are prone to fungal esophagitis, we have investigated the esophageal fungal colonization of this patient group. METHODS: One hundred consecutive ALD patients were enrolled in this prospective study. 22 patients with dyspeptic symptoms acted as controls. After taking an oropharyngeal swab, patients underwent an upper gastrointestinal endoscopy and surface material was obtained from the esophagus for direct smears and culture. RESULTS: In the ALD group pseudohyphae were found in 21.5% and yeast forms in 6.4% of the direct smears. The culture was positive in 40.8% of the ALD patients, the isolated strains were: 30 C. albicans, 2 C. kefyr, 2 C. krusei, 1 C. zeylanoides and in 3 cases the species could not be identified. 41.9% of ALD patients and 13.6% of control patients (p = 0.013) had fungi in their esophagus. Significantly more ALD patients had fungal esophagitis than in the control group (19.3% vs. 0%, p = 0.021), the rate of fungal colonization was also higher, but the difference was not significant (22.5% vs. 13.6%). A significantly higher rate of fungal esophagitis and esophageal colonization was found in patients with fungi in their oropharyngeal swabs (p = 0.00001). CONCLUSIONS: Fungal colonization of the esophagus is frequent in ALD patients. Its presence might have clinical significance in the case of liver transplantation.

[Symptoms of fungal esophagitis in alcoholic liver disease]. / Túnetek alkoholos májbetegek gombás oesophagitisében.

Symptoms related to fungal esophagitis were studied in patients with alcoholic liver disease who underwent upper gastrointestinal endoscopy. Data of 517 patients were studied retrospectively (group I) and 100 alcoholic liver disease patients, that were successively admitted to hospital, were enrolled in the prospective part (group II). Out of the 41 cases with fungal esophagitis found in group I, data of 38 could be evaluated. In group II 13 of the 93 evaluable patients had fungal esophagitis; according to Kodsi's grading 10 patients had grade 1., one patient grade 2. and two patients grade 2-3. oesophagitis. There was no case with grade 4. esophagitis. The rate of symptoms among the 51 patients with fungal esophagitis was: anorexia 23 (45.0%), abdominal pain 22 (43.1%), vomiting 17 (33.3%), nausea 15 (29.4%), occult gastrointestinal bleeding 12 (23.5%), weight loss 9 (17.6%), melena 7 (13.7%), bloating 6 (11.7%), acidic regurgitation 3 (5.8%), haematemesis 2 (3.9%), thoracic pain 2 (3.9%), singultus 1 (1.9%), odynophagia 0 and dysphagia 0. In 7 patients (13.7%) none of the studied symptoms could be identified. Despite the relatively high frequency of symptom free fungal esophagitis reported in the literature, the total lack of odynophagia and dysphagia in our patient group was remarkable. In the lack of deglutition disorders the other symptoms do not raise the suspicion of esophagitis. The diagnosis in such cases can be established only by endoscopy.

Changes in the acute phase complement component and IL-6 levels in patients with chronic hepatitis C receiving interferon alpha-2b.

In order to study the effect of interferon alpha on the levels of acute phase complement proteins in vivo, serum concentrations of C9 and C1-inhibitor (C1-INH) were measured in patients with chronic hepatitis C before and 3 months after the beginning of interferon alpha2b therapy. Serum levels of the activation product of terminal complement pathway, C5b-9, HCV RNA and IL-6 were also determined. IFN alpha treatment significantly (P<0.0001) increased the serum concentrations of both complement proteins. C5b-9 levels were found to significantly decrease during the same period of time. When the patients were divided into responders or non-responders (more or less than 50% decrease in plasma HCV RNA concentrations) C9 and C1-INH levels were elevated only in the responder patients. There was no correlation between the changes of IL-6 levels or the amounts of IFN alpha administrated on one hand, and the changes in the complement protein levels on the other. These findings suggest that the marked increase in the serum concentrations of the acute phase complement proteins is a secondary phenomenon due to the IFN alpha-caused diminution of the viral load and the resulting immune complex-induced complement activation.

[Severe relapse of hepatitis C following liver transplantation. Successful treatment with a combination of interferon-alpha and ribavirin]. / Hepatitis C súlyos relapsusa májátültetés után. Sikeres kezelés interferon-alpha és ribavirin kombinációval.

A 38-year-old, male patient, with end-stage HCV cirrhosis, underwent liver transplantation (OLT). After a sufficient recovery a rapid elevation of ALT and profound jaundice developed 3 months after OLT, together with a 15-fold rise of pre-transplant HCV RNS level. Liver biopsy was carried out, histology excluded rejection and signs of acute hepatitis were observed. Interferon alpha 2b 3 MU TIW and ribavirin 800 mg/day resulted in normalization of ALT, se. bilirubin and decrease of viral load by 90 per cent at the 3rd month of treatment. Improvement of hepatitis and no rejection was shown by control histology. A 6-month combination therapy followed by continuous ribavirin monotherapy maintains a permanent good condition with normal ALT, no icterus, a continuously low HCV RNA level and a mild chronic hepatitis with fibrosis in the liver histology 18 months after OLT. Danger of HCV reactivation after OLT, difficulties of diagnosis, interactions of immunostimulant and immunosuppressive drugs, advantages of combination therapy are discussed.

[Interferon therapy of chronic viral hepatitis in Hungary: 5-year experience. A multicenter study]. / Krónikus vírushepatitisek interferon kezelése Magyarországon: 5 éves tapasztalatok. Multicentrikus tanulmány. Hepatológiai Centrumok Munkacsoportjai.

In Hungary over the past 5 years more than thousand patients with chronic viral hepatitis have been examined and included in a treatment program with interferon (IFN) at 16 major hepatology centers, using unified diagnostic and therapeutical criteria. Authors give an account of their experiences on the clinical features of patients with chronic viral hepatitis and report the results of the treatment with IFN. According to the rules and availability of IFN for patients with chronic viral hepatitis in the country, virtually the entire Hungarian population with this diseases who required IFN therapy have been included. A total of 94 patients suffered from hepatitis B virus (HBV) infection, in addition 11 HBV + hepatitis Delta virus (HDV), 24 HBV + Hepatitis C virus (HCV) related liver disease, and 993 had chronic hepatitis C. IFN therapy for chronic HBV hepatitis consisted of IFN 5 MU thrice weekly for 6 months, and resulted in 33% seroconversion and sustained remission with 14% HBsAg clearance. For chronic hepatitis C treatment protocols (dose of IFN and duration of therapy) have changed with the time (from a weekly dose of 3 x 3 MU IFN for 6 months, to 3 x 3 MU for 12 months), and even a combination with ribavirin has been introduced. Although the therapeutic results showed a gradual improvement form a 13% sustained response over 22% in the first and second periods, respectively, differences were most significant with the advent of the combination therapy, that resulted in 36% remission rate. Only fibrosis in histology and baseline pretreatment HCV-RNA level appeared as predictors of response in chronic hepatitis C. Neither age nor gender did influence the outcome, but longer duration of treatment and higher total dose of IFN resulted moderately higher sustained remission rates. The experiences are in accordance with findings of suboptimal efficacy of IFN monotherapy reported worldwide and emphasize the need of seeking for newer and combination therapeutic modalities for these chronic viral diseases.

[Liver changes in HIV/AIDS]. / Májelváltozások HIV/AIDS-betegségben.

Loss of cellular immunity accompanying the progressive HIV/AIDS disease, results in altered clinical picture and outcome of traditional infections as well as severe and often lethal illness caused by facultative pathogens. Unusual infections may call the attention to the underlining HIV disease. Liver disease appears in the great majority of HIV/AIDS patients. The authors review the viral, bacterial, fungal and protozoal infections involving the liver in AIDS. Liver biopsy has a diagnostic value beside the serology and bacteriology and may give therapeutic consequences.

Preliminary report of a controlled trial of MTH-68/B virus vaccine treatment in acute B and C hepatitis: a phase II study.

Eighty four patients with viral hepatitis attributed to infection with hepatitis B virus (HBV) (n = 43) or hepatitis C virus (HCV) (n = 41) were included in this study employing the MTH-68/B vaccine, an attenuated variant of Bursal Disease Virus. Twenty of the 43 patients in the HBV group, and 22 of the 41 HCV patients were treated with MTH-68/B. The remaining patients received conventional therapy. Significantly more patients progressed into active chronic hepatitis on conventional therapy (13% of HBV and 26% of HCV cases respectively) than in the vaccine treated groups (0% and 9%). Relapses occurred less frequently in the vaccine treated groups (5% of HBV and 32% of HCV) than in the control groups (9% and 79%), while remissions within one month of treatment were observed more often in the vaccine treated groups (both 50% respectively) than in the control groups (26% of HBV and 21% of HCV patients respectively). The duration of hepatitis was also considerably shortened by MTH-68/B treatment in both HBV (from 7.5 +/- 3.7 to 5.9 +/- 3.0 weeks) and HCV patient groups (from 8.9 +/- 7.4 to 5.3 +/- 4.4 weeks). The data presented suggest that attenuated, non-pathogenic viruses may be of significant benefit for patients with viral hepatitis B and C infections.

[Management of chronic hepatitis B]. / A krónikus B hepatitis kezelése.

The features of chronic hepatitis B virus (HBV) related liver diseases and the aim of their therapy have briefly discussed, then treatment modalities are listed. In Hungary, between 1994 and 1996, a total of 68 patients with chronic hepatitis B have been treated with interferon (IFN). IFN resulted in complete clinical-biochemical remission in 50% of the patients, and in 32% the HBV replication was also eliminated. There are various nucleoside analogues, among them mostly famciclovir and lamivudine have been intensively studied as potentially effective treatment for HBV infection, and controlled clinical trials are in progress with these drugs. Nucleoside analogues in combination with IFN possibly improve treatment results in this disease. Various immunomodulatory agents--such as levamisole, thymosine, interleukin-2, and other cytokines--as well as the prednisolon-withdrawal induced rebound phenomenon have also been tested in HBV infection, but with no generally established benefit. A recombinant HBsAg vaccine is under investigation for therapeutic use. For end-stage HBV liver cirrhosis, liver transplantation is the only treatment, but the problem of reinfection is not still solved for more reasons.

[Screening for hepatitis C of hospital personnel at the Szent László Hospital of Budapest]. / Egészségügyi dolgozók C hepatitis szürövizsgálata a Szent László Kórházban.

UNLABELLED: The health-care workers are known to be at risk of occupational transmission of blood-borne viruses. The goal of the investigation was to determine the prevalence of hepatitis C virus (HCV) antibody and the occupational risk of HCV transmission among personnel at the Central. Hospital for Infectious Diseases, Budapest, Hungary. Serum samples of 409 health-care workers were tested for antibody to HCV with second and third generation ELISA-s and anti-HCV positive samples were confirmed with Western Blot Line EIA. A total of 10 (2.4%) of the health-care workers were confirmed to be anti-HCV positive. The prevalence of anti-HCV increased with advancing age: zero under 20 yr age group (N = 0/15), 0.9% in 21-30 yr age group (N = 1/112), 1.8% in 31-40 yr age group (N = 2/111), 3.1% in 41-50 yr age group (N = 3/96) and 4.0% in above 50 yr age group (N = 3/75). We found anti-HCV positive hospital worker in 9 out of 17 departments. The prevalence of hepatitis C antibody was 7.1-1.9% among the personnel of internal departments, pathology, intensive care unit and pediatric departments. No anti-HCV positive health-care worker was found in the surgery and laboratories. None of the physicians tested was seropositive for HCV. Eight of the nurses, one of the sanitary personnel and one pathological technician were seropositive for HCV. Two nurses developed a chronic C hepatitis after a needlestick accident. CONCLUSIONS: 1. The hospital personnel is at risk for HCV infection. 2. The occupational risk of HCV infection increases with age but the risk is considerable lower than that of hepatitis B infection. 3. The occupational risk is highest among the workers of the chronic internal department, pathology and intensive care unit. 4. The nurses are at higher risk of HCV infection than the physicians. 5. The needlestick injury is associated with an increased risk for acquiring HCV infection.

[Experience gained from the organization of the first group of patients who benefited from liver transplantation]. / Tapasztalatok a májátültetéssel gyógyítható betegek elsö csoportjának szervezésével.

The authors report in this ciency in the majority of the cases was alcohol abuse or article about their experiences, how could they select a small group of patients awaiting liver transplantation since September 1994. The cause of the liver insuffiHCV infection. During 37 weeks 3 liver transplantations were performed and 5 patients died on the waiting list. (15,6% of the patients on the waiting list.)

Comparison of single-dose and 7-day fluconazole treatment of fungal esophagitis in alcoholic liver disease.

The aim of the study was to assess the effectiveness of a single-dose fluconazole treatment of fungal esophagitis in patients with alcoholic liver disease. Twenty-two alcoholic liver disease patients with fungal esophagitis were randomly assigned to receive either a single-dose of 150 mg fluconazole or a 7-day treatment of daily 50 mg fluconazole. Control esophagoscopy was performed in both groups on days 9-11. Direct smears and cultures on Sabouraud's medium were performed at both endoscopies. Patients' sera were tested for Candida antigens and for antibodies against Candida albicans on days 1, 8 and 15. Twenty patients (18 C. albicans, 1 C. tropicalis, 1 C. pseudotropicalis) completed the study, there were two drop-outs from the single-dose group. Antibodies against C. albicans were found in four cases, Candida antigens in five. There were no significant differences in the treatment outcome between the two groups, clinical cure was recorded in eight out of nine patients in the single-dose group and nine out of eleven patients in the 7-day group, mycological eradication in four out of nine, and in three out of eleven, respectively. Single-dose fluconazole treatment seems to be an effective therapy of fungal esophagitis in alcoholic liver disease patients.

[Diagnostic value of abdominal ultrasonography and endoscopic retrograde cholangiopancreatography in obstructive jaundice]. / A hasi ultrahangvizsgálat és az endoscopos retrograd cholangiopancreatographia diagnosztikus értéke elzáródásos sárgaságban.

The diagnostic value of ultrasonography (US) and endoscopic retrograde cholangio-pancreatography (ERCP) was compared, in 66 patients with bile duct obstruction, who underwent subsequent biliary surgery. The level of the blockage was diagnosed by US with a 80%, by ERCP with a 95.4% sensitivity, while the cause of the obstruction was determined with a 50% and a 89% sensitivity, respectively. Predictive value of these examinations is over 90%. Based on the clinical results and on the high diagnostic value of the above mentioned examinations, it is essential to fill up the bile ducts with direct contrast material.

Polyphasic and protracted patterns of hepatitis A infection: a retrospective study.

The present study was performed in order to evaluate the frequency and clinical features of polyphasic and protracted forms of HA infection. Out of 297 adult HA patients admitted to our hospital, 12.5% and 8.4% were polyphasic and protracted, respectively, in clinical course. 21.6% of the polyphasic patients had more than one relapses. The rates of symptomatic relapse during the follow-up of the polyphasic and protracted HA infections were 51% and 56%, respectively, One % of all HA patients presented a cholestatic pattern. Three % of all the patients had to be readmitted for hospital treatment. The outcome of disease was benign in all HA patients, though, complete recovery sometimes needed six months.