E. Del Ninno

PREVALENCE OF ANTIBODIES TO HEPATITIS C VIRUS IN ITALIAN PATIENTS WITH HEPATOCELLULAR CARCINOMA

A sensitive radioimmunoassay was used to detect antibodies to hepatitis C virus (HCV) in patients with hepatocellular carcinoma and chronic hepatitis. HCV antibodies (anti-HCV) were detected in 86 of 132 patients with hepatocellular carcinoma with no relation to the presence or absence of hepatitis B surface antigen (HBsAg). The prevalence of anti-HCV was also high in patients with diseases thought to predispose to hepatocellular carcinoma, such as non-A, non-B chronic hepatitis and cirrhosis (74%). In HBsAg-negative patients with hepatocellular carcinoma the prevalence of anti-HCV was lower than that in patients with non-A, non-B chronic hepatitis (16% vs 55%); the prevalence of serum antibodies to hepatitis B core antigen (anti-HBc), a marker of hepatitis B virus infection, was 70% and 28%, respectively. In HBsAg-negative patients with hepatocellular carcinoma, anti-HCV and anti-HBc occurred together nearly three times as often as in patients with chronic hepatitis (54% vs 19%). These data indicate that, in Italy, HCV is an important factor associated with hepatocellular carcinoma and non-A, non-B chronic hepatitis.

Asymptomatic liver disease in haemophiliacs.

The incidence of jaundice and of abnormal liver function tests has been assessed in 91 multitransfused patients with severe haemophilia A and B. Tests of hepatocyte function were within the normal range in the majority of patients. On the contrary, tests of biliary cell function, liver cell damage, and bromsulphthalein retention gave high rates of abnormal values, which tended to increase with age. Hepatitis B surface antigen was present in 8% and the corresponding antibody in 66% of the cases; 18% had a history of jaundice. All patients were asymptomatic and only a minority showed clinical signs of liver involvement. These data suggest that in haemophilacs repeated and prolonged contact with the agent(s) responsible for post-transfusion hepatitis may cause chronic liver damage not associated with overt illness.

Hepatitis C reactivation in patients with chronic infection with genotypes 1b and 2c: a retrospective cohort study of 206 untreated patients

Background: We previously described hepatitis reactivation in two carriers of the hepatitis C virus (HCV) genotype 2c. Aim: To assess the relationship between HCV genotypes and risk of hepatitis reactivation, we studied the course of aminotransferases in patients infected with the two relevant genotypes in Italy. Patients: A cohort of 100 patients with genotype 2c chronic hepatitis and 106 with genotype 1b were subjected to surveillance. Methods: Hepatitis reactivation was defined as an alanine aminotransferase (ALT) value ⩾400 IU/l or a maximum/minimum ALT ratio value of ⩾8. Results: Over a period of 71 (24–144) months, one or more flares of ALT (201–2200 IU/l, 6–90 months’ duration) occurred in 31 patients with genotype 2c and in eight patients with genotype 1b (rates of flares: 55.6 per 1000 person years for genotype 2c v 15.0 for genotype 1b; p = 0.001). On repeat biopsy, hepatic fibrosis increased by more than 2 points in 10/16 patients examined either during or after an ALT flare compared with 7/36 flare free patients (63% v 19%; p = 0.003). Hepatitis flares were significantly associated with genotype 2c (odds ratio 6.48 (95% confidence interval 2.57–16.35)) but not with sex, age, modality or duration of infection, baseline ALT values or histological severity of hepatitis, hepatitis other than HCV, or reinfection. Conclusions: Genotype 2c carriers are at high risk of hepatitis reactivation, suggesting that virus genetic heterogeneity is important in the natural history of HCV, questioning the linearity of hepatic fibrosis progression during hepatitis C.

Lack of association between type of hepatitis C virus, serum load and severity of liver disease

SUMMARY. Chronic infection with the hepatitis C virus (HCV) may lead to a variety of hepatic lesions from benign inflammation to liver cancer, but the relationships between infection and development of liver disease are poorly understood. To assess whether virus type and load are of pathogenetic importance, 197 Italian carriers with various hepatic lesions were investigated consecutively. Of these, 187 (95%) patients had serum HCV RNA, by reverse transcription‐polymerase chain reaction (RT‐PCR) with a median level of 1003 × 103 genomic equivalents ml‐1 according to the branched‐DNA assay (b‐DNA). One hundred and seven patients (54%) had serotype 1, 22 (11%) had serotype 2, 9 (5%) had serotype 3, 17 (9%) had mixed serotypes and 42 (21%) had no specified serotype. One hundred and thirty four patients were also tested for genotype. The genotype distribution was as follows: 17 (13%) had genotype 1a; 67 (50%) 1b; 29 (22%) 2a; 12 (9%) 3a; 3 (2%) had genotype 1 not classified (NC); 3 (2%) had genotype 2 NC; 2 (1.4%) had genotype 4 and 1 (1%) had mixed genotype la + 3a. No virus type was associated with any particular histological diagnosis and all were equally distributed between progressive and non‐progressive liver disease groups. Serum HCV‐RNA levels were similar in the liver diseases groups. By analogy to hepatitis B, there was no direct correlation between type and level of viraemia and the severity of the underlying liver damage.

Progressive hepatic fibrosis in healthy carriers of hepatitis C virus with a transaminase breakthrough.

In short‐term studies, patients with chronic hepatitis C virus (HCV) infection, consistently normal serum aminotransferase (ALT) levels, and minimal or mild necro‐inflammatory changes in the liver, did not progress to histologically severe hepatitis. There are no data on longer term outcome of liver disease in these patients. We describe two patients with HCV infection (genotype 2c) with a rise in serum ALT values greater than 10 times the upper normal value that occurred after an 8‐ and 15‐year period of persistently normal or minimally elevated ALT levels. In both patients, the rise in ALT values lasted more than 16 weeks and was not associated with any symptom or risk factor for acute hepatitis. A liver biopsy performed 4 and 18 months after the ALT flare showed clear‐cut progression from chronic hepatitis with mild activity to chronic hepatitis with severe activity and central to portal septal fibrosis (Ishak score: grading 14 and 6; staging: 4 and 5, respectively). Hence, extended surveillance of HCV carriers with consistently normal or minimally elevated ALT values is warranted as these patients are at risk of ALT flares and may develop progressive liver disease.

Hepatitis C antibody in patients with chronic liver disease and hepatocellular carcinoma

Hepatitis C virus (HCV) is the major etiologic agent of parenterally transmitted non-A,non-B hepatitis. To determine whether there is a relationship between this virus agent and hepatocellular carcinoma (HCC), the sera of patients with HCC and chronic hepatitis were assessed using a sensitive immunoassay for HCV antibody. Anti-HCV was detected in 65% of 132 patients with HCC, without any relationship with the presence of the hepatitis B surface antigen (HBsAg). The prevalence (74%) of anti-HCV was high, as expected in patients with putative non-A,non-B cirrhosis also. The prevalence of anti-HCV was less in patients with HBsAg-positive cirrhosis (28%) and in patients with disease not related to viral hepatitis and healthy controls (8%). These data suggest, but do not prove, that HCV is an important factor associated with HCC.

Idiopathic haemochromatosis and HLA antigens in Italy: is A3 Bw35 HLA haplotype a marker for idiopathic haemochromatosis gene in north east regions?

Thirty two unrelated Italian subjects with idiopathic haemochromatosis were studied. HLA-A3 was present in 26 of them (81% v 22% in controls; p less than 0.001) and HLA B7 in eight (28% v 9%; p less than 0.01). There was no important association between idiopathic haemochromatosis and HLA B14. Subdividing the patients on the basis of their regional origin a noticeably higher prevalence of HLA Bw35 in patients with idiopathic haemochromatosis from north eastern Italy was found than in those from Lombardy, or in the controls; there were no differences in the incidence of HLA A3 and B7 between patients with idiopathic haemochromatosis from different areas. A high prevalence of A3, Bw35, and A3, B7 haplotypes was found in our patients with idiopathic haemochromatosis. A3, Bw35 could be the haplotype most commonly linked to the idiopathic haemochromatosis gene in north eastern Italy.

A prospective study of blood alpha-fetoprotein messenger RNA as a predictor of hepatocellular carcinoma in patients with cirrhosis

Summary. Blood alpha‐fetoprotein messenger RNA (AFP mRNA) is thought to be a marker of hepatocellular carcinoma (HCC). Its value as a predictor of HCC in patients at risk is not known. A series of 201 patients with compensated cirrhosis (114 men, mean age 58 years) underwent surveillance with semi‐annual ultrasound and serum alpha‐fetoprotein measurements. Total RNA was extracted from peripheral blood mononuclear cells collected at different intervals and AFP mRNA was retrotranscribed and amplified by nested polymerase chain reaction. Ten patients with HCC and 30 blood donors were used as controls. Three patients with HCC, 39 with cirrhosis under surveillance and four blood donors circulated AFP mRNA (30, 20 and 13%, NS). During 50 months of surveillance, 27 patients with cirrhosis developed HCC: the tumour was detected more often in patients with higher than normal baseline serum AFP (≥7 IU/L) than in those with normal AFP levels (21%vs 9%, P = 0.02). The incidence of HCC was the same in patients with and without AFP mRNA at baseline (15%vs 14%). In 53 patients, AFP mRNA was re‐tested after 6–25 months of surveillance. HCC developed in two of 11 (18%) who were initially AFP mRNA positive and later became negative, in none of those who were initially negative and later became positive and in two of 39 (5%) who remained persistently negative. In conclusion, blood AFP mRNA is not a sensitive predictor of HCC in patients with compensated cirrhosis.

Serum levels of hepatitis C virus RNA predict non-response to interferon therapy: comparison of two commercial assays.

We compared two commercial assays for quantification of serum hepatitis C virus (HCV) RNA to investigate whether pretreatment levels of serum HCV RNA could predict the outcome of interferon (IFN) therapy. The Amplicor HCV Monitor test is based on a single, combined reverse transcription and amplification reaction carried out by the Tth DNA polymerase using specific primers for the 5′ untranslated (UTR) region. The Quantiplex HCV RNA 2.0 assay is based on specific hybridization of viral RNA by synthetic oligonucleotides complementary to the 5′‐UTR and core regions of the genome, allowing equal quantification of the six major genotypes. Receiver‐operating characteristic (ROC) analysis was employed to identify the best cut‐off value (predicting patients who were non‐responsive to treatment) with corresponding sensitivity and specificity values. Logistic regression analysis was performed using these cut‐off values. We studied 133 consecutive patients with chronic hepatitis C enrolled in a prospective trial of IFN‐α therapy (18 of whom were sustained complete responders). The median viraemia of the 18 sustained responders was 5322 copies ml–1 by the Monitor test and less than 0.2million equivalents ml–1 (MEq ml–1) by the Quantiplex assay; for the 115 non‐responders/relapsers, the median viraemia was 83125 copies ml–1 and 1.128 MEq ml–1 for the Monitor test and Quantiplex assay, respectively. Spearman’s rank test gave a correlation of 0.63 between assays. The best predicting cut‐off values were 22134 copies ml–1 for the Monitor test and 0.330 MEq ml–1 for the Quantiplex assay; their respective sensitivities and specificities were 72% and 75% for Monitor and 67% and 83% for Quantiplex. By logistic regression analysis, the age and gender‐adjusted odds ratios of high vs low HCV RNA levels, defining the risk of non‐response, were 10.6 (CI 3.1–35.7) for Monitor and 14.3 (CI 4.3–47.3) for Quantiplex. The two assays had comparable sensitivity for serum HCV RNA but they identified different predictive cut‐offs for non‐response to therapy.

Effect of carbon tetrachloride poisoning on the plasma levels of aspartate-aminotransferase isoenzymes in the rat

Mediante frazionamento cromatografico su colonna di resina a scambio anionico, abbiamo potuto documentare nel siero di ratti intossicati con dosi diverse di CCl4, incrementi assai significativi sia della componente mitocondriale che citoplasmatica della aspartato-aminotransferasi.