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The epidemiological picture of hepatitis C virus (HCV) infection in the general population is largely unknown, even in developed countries. The aim of this study was to estimate the prevalence and genotype distribution of HCV amongst a large sample of the Italian general population. A total of 3,577 serum samples were collected and screened for anti-HCV antibodies. ELISA and RIBA tests were used to assess the presence of anti-HCV. NS5b region sequencing was performed for molecular characterization. Of 3,577 tested sera, 95 (2.7%) were anti-HCV positive and a genome was detected and sequenced in 50 sera. The age-adjusted prevalence was 4.4%. Seroprevalence increased with age, following a North-South gradient, and increased steeply between the 15 and 30 and 31-45 age groups. Subtype 1b showed the highest prevalence in all geographical areas and age groups, followed by subtypes 2c (detected mainly in the elderly population in Southern Italy), 4a/d, and 3a (detected exclusively in adults) and 1a. These findings confirm that Central and Southern Italy are hyperendemic areas. The high prevalence observed in adults over age 30 is mainly attributable to an increase in 1b-prevalence but also to subtypes 2c- and 3/4-infections. Age-specific prevalence data and molecular characterization of the virus suggest that two transmission patterns co-exist in Italy: one characterized by subtype 1b and 2c infections, mainly in adults older than 60 years, and the other by subtype 3 and 4 infections, mainly in the 31-60 year age group, and consistent with intravenous drug use and immigration., (Copyright (c) 2005 Wiley-Liss, Inc.)

Although the haemagglutination inhibition assay is considered the "gold standard" for antigenic characterisation of influenza viruses, some limitations of this technique are well known. A new microneutralisation assay, as a tool for antigenic characterisation of influenza B viruses, has been standardised and its performance evaluated in comparison with the haemagglutination inhibition test in the light of molecular characterisation of the haemagglutinin. Twelve B viruses belonging to the two lineages and the four sub-lineages discriminated by phylogenetic analysis of HA were tested. The microneutralisation assay clearly distinguishes viruses belonging to different lineages and, in addition, discriminates strains belonging to different sub-lineages that are poorly or not discriminated using the haemagglutination inhibition test. This new microneutralisation assay could provide a useful tool for antigenic characterisation of circulating influenza viruses and contribute, together with the haemagglutination inhibition test and sequence analysis of the haemagglutinin and neuraminidase, in the choice of the strain for use in vaccine composition.

Markers of viral replication are fundamental tools for understanding the mode of transmission, diagnosis and management of hepatitis C virus (HCV) infection. A new enzyme-linked immunosorbent assay for quantitative detection of free and complexed HCV core antigen (HCV Ag) has been developed. The aim of this study is to evaluate the clinical performance of the new test and compare it with the most widely used commercially available RT-PCR-based assay. To determine the cut-off value we tested 60 samples from anti-HCV negative samples and selected a qualitative cut-off value of 3 pg/ml. To evaluate the usefulness of the new assay in confirming serologically indeterminate results we collected 62 sera. To evaluate the HCV Ag and HCV-RNA relationship we tested 245 samples from patients with different clinical conditions. The results of 61 out of 62 (98.4%) anti-HCV indeterminate samples were found to agree, whereas only one serum was found to be RT-PCR positive and HCV Ag negative. We also found the results to agree in 77.6% (190/245) of the samples from infected patients, while we observed higher agreement in untreated patients, both with and without evidence of liver damage. The correlation coefficient (r) observed between HCV Ag and HCV-RNA was 0.88. The regression line meets the cut-off value at an HCV-RNA concentration of approximately 40,000 IU/ml. In conclusion, we found that the results from the total HCV Ag test agree with the RT-PCR results and therefore we believe that this test could become a useful tool for the diagnosis and management of HCV infection.

Hepatitis C virus (HCV) infection remains a major problem in haemodialysis units despite the risk decrease provided by anti-HCV screening of blood. The exact mode of transmission of HCV in the dialysis units remains unclear. To identify the route of the virus and the mechanisms of transmission an investigation into the outbreak of HCV infection in a haemodialysis unit on a molecular level was held: 12 newly infected patients and 14 already infected were investigated by sequencing the 5' untranslated region of the viral genome. The results showed that 3 strains infected new cases and these strains matched those sequenced in already infected patients. Transmission occurred between patients treated on the same shift and also between different rooms. Console and blood or blood product contamination was excluded. Our study gave molecular evidence of patient-to-patient transmission of HCV in a haemodialysis unit. Our data underline the importance of the strict enforcement of standard precautions to prevent HCV transmission and failure of the isolation of anti-HCV positive patients as preventive measure.

To investigate whether it is appropriate to assume comparability of hepatitis virus C (HCV)-RNA results across laboratories in multi-centre studies, nine laboratories of the European Paediatric HCV Network participated in an international proficiency study of HCV-RNA assays. A panel of 12 samples of different dilutions and genotypes was sent to each laboratory and tested with qualitative and/or quantitative HCV-RNA assays according to local procedures. Commercial assays were used in seven laboratories and in-house assays in two. All six laboratories in which a commercial qualitative assay was used were proficient, as were four of six runs (in five laboratories) in which a commercial quantitative assay was used. The proficiency of the laboratories where in-house assays were used could not be assessed according to the VQC definition because of differences in the methods used. Overall, there were several false-negative results, but only one false-positive result with a quantitative assay and none with a qualitative assay. The false-negative results may have implications for the diagnosis of infection, and highlight the need for an antibody test to be performed at 18 months to confirm the absence of infection. The results of qualitative assays were generally consistent across laboratories but it was difficult to evaluate and compare the results of quantitative assays. Multivariate analysis of data collected in multi-centre studies should therefore allow for centre and/or assay used.

To assess the prevalence of human T cell leukemia virus type I (HTLV-I) and 2 (HTLV-II) infection and the associated risk factors among immigrants living in Northern Italy, we surveyed 3017 open-population subjects from three geographical areas and 371 prisoners. In the open population, the overall prevalence was 0.3% for HTLV-I and 0.1% for HTLV-II, while among prisoners, HTLV-I and HTLV-II infection were detected in 1.4 and 0.8% of subjects, respectively. HTLV-I prevalence was higher in subjects with multiple sexual partners or sexually transmitted diseases. This association was significant in the open-population group and close to significance in prisoners. Multivariate analysis showed that human immunodeficiency virus (HIV) seropositivity remained significantly associated with HTLV-I infection in both targeted populations (OR: 11.2 in the open population; OR: 9.9 among prisoners), whereas sexual exposure was associated with HTLV-I seropositivity only for prisoners (OR: 14.3). No independent variable was related to HTLV-II infection.

A clinical trial with four groups receiving either 0.6, 3.5, 10 or 20 micro g of purified non-adsorbed hepatitis B surface antigen (HBsAg) was performed to study the kinetics as well as the capacity of the immune memory to respond following exposure to HBsAg in responders to a complete course of hepatitis B vaccine, in whom anti-HBs titres had declined below the seroprotective level. The study population included 64 healthy individuals. All response parameters seropositivity, seroprotection rates, booster response rates and geometric mean titres (GMTs), consistently showed that the immune response was highly satisfactory and dose-dependent. A remarkable immune response was obtained even with a trace amount of HBsAg. This study further supports recent indication that booster hepatitis B vaccine doses may be unnecessary in healthy adult responders to a full course of hepatitis B vaccination.

Background/aims: Recent experiences suggest that interferon may significantly decrease the incidence of hepatocellular carcinoma. We conducted a randomized study with interferon versus no therapy in hepatitis C virus Child A cirrhosis with abnormal alanine aminotransferase and HCV-RNA positive serum with the aim to investigate the incidence of hepatocellular carcinoma, worsening of cirrhosis's stage and death or orthotopic liver transplantation., Methodology: A cohort of 122 patients prospectively followed was analyzed retrospectively to assess the effect of interferon therapy (mean follow-up: 96 +/- 18.3 months). We only chose patients with hepatitis C virus infection who had undergone blood transfusion before 1980. Hepatitis C virus serotype was determined by hepatitis C virus serotyping 1-6 assay (Murex Biothec Limited Temple Hill, Dartford, Kent, UK). HCV-RNA level was determined by bDNA, Chiron Corporation Emeryville, CA. Diagnosis of hepatocellular carcinoma was made on the basis of the appearance of local lesions at periodic ultrasound examination of the liver and confirmed with spiral computed tomography. Fine needle biopsy under sonographic guidance was effected. Fifty-nine patients (mean age: 55.3 +/- 7) received interferon (3MU three times a week for 12 months), 8 stopped therapy for side effects, 71 did not receive interferon (mean age: 56.8 +/- 8). Baseline characteristics were similar., Results: It emerges how interferon does not reduce the risk of hepatocellular carcinoma in compensated cirrhosis. In interferon treated patients an improvement in relation with worsening and death/orthotopic liver transplantation has been noted., Conclusions: The use of the interferon seems to be scarcely useful when structural alterations of the cirrhotic kind show up, as cirrhosis represents by itself a risk factor for hepatocellular carcinoma. Nevertheless, in relation to the worsening of cirrhosis's stage the interferon therapy can be useful in compensated cirrhosis.

Background: Hepatitis C virus (HCV) infection is a worldwide problem of public health. Epidemiological studies have shown a significant higher prevalence of infection in the elderly. Amantadine is an antiviral agent active against the influenza A virus that has been used in cases of chronic hepatitis C., Objectives: To evaluate the antiviral activity and the safety of amantadine (200 mg daily for 6 months) in elderly patients with chronic hepatitis C., Methods: The study group consisted of 23 consecutive patients over 65 years suffering from chronic hepatitis C. Aminotransferase (ALT) levels were tested at baseline, at 15 days and then monthly until the end of therapy. HCV genotype was determined at baseline. A quantitative HCV-RNA measurement was performed at baseline, at 15 days and at the 1st, 3rd and 6th month of treatment., Results: 13 males and 10 females were enrolled (mean age 70.1 +/- 3.4 years; range: 65-75). The mean ALT levels did not change significantly during therapy except in 1 patient subsequently returned normal. The HCV-RNA remained detectable in all patients, but a significant difference in response was observed in patients infected by genotype 1b., Conclusions: Our results confirm the antiviral activity of amantadine against HCV, mainly for genotype 1b with initial high viral load. No consistent effects on aminotransferases were observed., (Copyright 2001 S. Karger AG, Basel)

Background: Host and viral factors have been suggested as possible causative factors for the presence of liver iron accumulation in chronic hepatitis C. However, there is no agreement regarding the influence of liver iron accumulation on the biochemical and histological severity of chronic hepatitis C. Moreover, data concerning the relationships between both viral load and genotype and liver iron accumulation are scanty., Aims: To evaluate the biochemical, histological and virological assessment of a group of chronic hepatitis C patients without risk factors for iron overload, on the basis of the presence, degree and distribution of liver iron accumulation., Methods: Fifty-three chronic hepatitis C patients (34 men, 19 women; age 44 +/- 11 years) with no risk factors for liver iron accumulation and showing no HFE mutations were chosen from a broader cohort of chronic hepatitis C patients. The presence, degree and distribution of liver iron accumulation were assessed using Deugnier's score. Relationships between the presence of liver iron accumulation and grading and staging were carried out separately. Hepatitis C virus RNA serum levels and viral genotype were compared in patients with or without liver iron accumulation. Alpha glutathione S-transferase serum levels were assessed in all patients., Results: Overall, liver iron accumulation was mild and was present in 19 patients (36%). It was associated with male gender (P = 0.0358), and was reflected by high serum iron levels (P = 0.001) and high ferritin levels (P < 0.0001). Hepatitis C virus RNA levels and genotype were not associated with the presence of liver iron accumulation. In multivariate analysis, ferritin was the only variable significantly associated with liver iron accumulation (P < 0.0001). Grading was higher in patients with liver iron accumulation regardless of the site of iron deposition. Fibrosis was present in all patients with iron overload; these patients were more frequently cirrhotic. Moreover, patients with mesenchymal or mixed deposition had higher staging than patients with hepatocytic or no iron deposition. This feature was reflected by higher alpha-glutathione S-transferase levels., Conclusions: Liver iron accumulation is mild in chronic hepatitis C patients without HFE mutations and is mainly reflected by serum ferritin levels. Viral characteristics do not seem to play a role in iron deposition. Liver iron accumulation is associated with higher grading, advanced fibrosis and cirrhosis. Moreover, higher staging is associated with mesenchymal or mixed iron deposition. In these patients, higher alpha-glutathione S-transferase levels seem to reflect more complex damage.

The window period in hepatitis C virus (HCV) infection is still a major problem in ensuring blood safety. HCV RNA detection by nucleic acid amplification technology-based tests has contributed to reduce the infectivity of blood products, but it is expensive, time-consuming and affected by a high prevalence of false-positive results. The aim of this study was to assess the performance of a newly developed enzyme immunoassay for the detection of HCV core antigen and its suitability for use in the screening of blood units in order to identify infecting samples that do not contain specific antibodies. For evaluation of laboratory performance, different samples were selected: to evaluate specificity, we tested 2,586 sera from blood donors, 500 general population samples, and 58 "difficult sera". All samples were tested by two screening assays, and results were negative. To estimate clinical sensitivity, 103 HCV RNA-positive, anti-HCV-negative samples, 6 natural seroconversion panels, and 9 commercial seroconversion panels were tested. Intra- and interassay precision were determined on two HCV-RNA-positive, anti-HCV-negative sera. Seventeen (0.66%) blood donor samples, 2 (0.4%) general population samples, and 2 (3.44%) difficult sera were initially reactive; 3 sera were positive on repetition. These 21 samples tested by reverse transcription-PCR were negative. The clinical sensitivity calculated with seroconversion panels and seroconverted patient samples was very similar to PCR sensitivity: 95% of PCR-positive, antibody-negative samples contained detectable HCV antigen. Data on intra- and interassay precision showed dispersion indices with values of less than 10%. In conclusion, the HCV antigen assay showed high sensitivity and specificity and could become a useful means of improving the safety of blood and blood products.

Therapeutic failures due to in vivo loss of drug sensitivity are still a major problem in AIDS care. Currently, the role of and methods for detecting resistant mutant strains in patients before therapeutic choices are still under debate. To investigate the relevance of screening for key mutations alone the commercial INNO-LiPA HIV-1 RT method was applied retrospectively to analyzing several HIV codons correlated with resistance to RTI (reverse-transcriptase inhibitors) in sera from 62 patients before starting HAART protocols, selected on the basis of clinical parameters. INNO-LiPA detected several resistant mutant strains, which were strictly consistent with previous selective pressure in the patients. A significant correlation between genotype pattern and response to HAART was found. The presence of key mutations associated with resistance to one or two RTI included in the protocol correlated with a decrease in treatment benefits, whereas patients with wild-type or non-resistant viral strains exhibited better response to HAART. Even if this information had been available when treatment was started, 45 of the patients would not have received different treatment. When compared with the total number of patients, the subgroup receiving a treatment that was considered retrospectively as consistent with the key mutation pattern exhibited a significantly better outcome. Although the interpretation of resistance-related key mutations needs improvement, this surrogate LiPA method seems to maintain a predictive role in the management of HIV infection, and is less expensive., (Copyright 2001 Wiley-Liss, Inc.)

The determination of HCV genotypes, subtypes and isolates has been helpful in understanding the evolution and the epidemiology of the virus, and is an important factor in the pre-treatment evaluation. A new simpler and automated sequencing based system has been developed recently, the Visible Genetics TruGene Hepatitis C Assay. The aim of the study was to compare this new genotyping assay with reverse hybridization based Innogenetics INNO-LiPA HCV II assay that is used most commonly. Eighty-eight HCV-RNA positive patients were enrolled and divided in four groups: 26 hemodialysed patients, 30 untreated patients with chronic HCV hepatitis, 12 IFN non-responder patients with chronic HCV hepatitis, 20 asymptomatic HCV positive subjects. The 5'-UTR region was amplified by RT-PCR and the nucleotide sequences determined by the TruGene assay. In parallel, the amplicons were also tested by INNO-LiPA. Concordant results were obtained in 80 out of 88 cases (90.9%). The new assay allowed to genotype 2 samples not typed by LiPA as 1b and 2a/c. The new system also allowed the subtyping of 3 untypable samples, classified as genotype 1 by INNO-LiPA, as genotype 1b (1 sample) and, as genotype 4 (2 samples). The difference between these genotype 4 isolates and the closest genotype 1 isolate was 6 nucleotides. One LiPA genotype 1a sample was typed as 1b and 2 genotype 1b samples were all typed as 1a by the sequence analysis. In conclusion, the new assay is a sensitive and rapid method that is suitable for accurate large-scale genotyping.

Objective: The presence of steatosis is a common histological finding in patients with chronic hepatitis C (CHC). The causes of the severity of this condition are not yet clear, although both metabolic and viral factors supposedly are involved. In this study our aim was to examine the possible influence that leptin levels, hepatitis C virus (HCV) RNA levels, and hepatitis G virus (HGV) infection have on the severity of steatosis and on the presence and degree of fibrosis in patients with CHC., Methods: One hundred eighty-two CHC patients with histological findings of steatosis were chosen from among a cohort of patients referred to our center for staging of liver disease. Among them 48 CHC patients were accurately selected so as to rule out possible confounding factors for the presence of steatosis (diabetes mellitus, hyperlipemia, obesity, alcohol). Leptin levels, HCV RNA levels, and HCV genotype, and the presence of HGV RNA were assessed in these patients and related to histological findings., Results: We found that leptin levels in CHC patients were similar to those in healthy subjects. No relationship was found between leptin levels and severity of steatosis. HCV RNA levels, HCV genotype, and the presence of HGV infection were no different among CHC patients with various degrees of steatosis. Leptin was not related to different degrees of fibrosis, whereas higher viral load was the only parameter associated to higher fibrosis scores., Conclusions: These findings suggest that the degree of steatosis in patients with CHC does not seem to depend on serum leptin levels or on viral factors, at least as far as HCV viremia and genotype and HGV infection are concerned. The severity of fibrosis does not seem to be influenced by leptin levels, whereas HCV viral load does seem to play some role.

Background/aims: About 50% of patients with chronic hepatitis C do not respond to interferon therapy and this failure is expensive. The aim of this study was to identify possible predictive factors of biochemical non-response during interferon therapy among biochemical, virological (HCV genotype), histological (Knodell's score) and pharmacokinetic (monoethylglycinexylidide formation test) pre-treatment parameters., Methodology: Our study included 60 patients with chronic hepatitis C undergoing a course of Interferon therapy. Patients whose serum ALT levels were normal at the 3rd month of therapy and remained so until the end of treatment were regarded as responders., Results: In univariate analysis, only the gamma-glutamyltransferase (gamma-GT) and the gamma-GT/alanine aminotranferase ratio were significantly higher in non-responder patients. Multivariate logistic analysis showed that high gamma-GT levels, high histological activity index, low monoethylglycinexylidide formation rate and viral genotype 1 were the best combination for the identification of non-responder patients (16.7% error rate). By adding alanine aminotranferase modification at the 1st month of therapy the probability error was reduced to 5%., Conclusions: These results show that the combination of biochemical, histological, virological and pharmacokinetic pre-treatment variables, associated with alanine aminotranferase modification at the 1st month of therapy, can predict non-response to interferon and allow therapeutic modifications.

Background: Hepatitis C virus infection accounts for varying severity of chronic liver disease. Clinical manifestations of infection have been related to different virus genotypes, with conflicting results., Design: We performed a cross-sectional study on a Northern-Italian group of patients with chronic hepatitis, cirrhosis and hepatocellular carcinoma related to hepatitis C virus infection in order to verify the association of different viral strains and the outcomes of viral disease., Methods: Two hundred and seventy-one patients referred to our unit for liver disease were studied and clinical, biochemical, histological, and functional parameters were investigated., Results: Different viral genotypes were not associated with peculiar findings in any of the degrees of liver disease. However, a progressive age increase was associated with disease severity, although clinical and functional staging of cirrhotic patients with hepatocellular carcinoma was better compared to tumour-free cirrhotic patients. There was an increased prevalence of genotype 1b related to the age of the patients. In multivariate regression analysis the patients' age and apparent duration of infection were independently associated with the presence of cirrhosis and only the age of patients was associated to hepatocellular carcinoma., Conclusions: In the population we studied age of the patients seemed to be a determinant conditioning disease severity, likely reflecting older infections and long-standing liver disease. The prevalence of certain genotypes in varying degrees of liver disease could be an epiphenomenon which might also be explained by the changing prevalence of infecting strains over the past decades.

The performance of a new automated analyzer for the processing and interpretation of the RIBA Strip Immunoblot Assay (SIA), used in the diagnosis of hepatitis C virus (HCV) infection, was evaluated. Laboratory performance of the RIBA SIA was compared with that of two manually processed supplementary anti-HCV tests (RIBA HCV 3.0 SIA and INNO-LIA HCV Antibody III). Specificity of the automated processing of SIA was 100% for 90 selected anti-HCV-negative samples. On the other hand, 119 of 120 (99.2%) previously confirmed anti-HCV-positive samples were also positive when assayed on the automated processor. Results for all specimens except one (51 of 52) were concordant for manual and automated RIBA, while 15 of 68 sera tested with automated RIBA and the INNO-LIA assay showed different patterns of reactivity. Three HCV sensitivity panels and one seroconversion panel were also compared. The results show a high sensitivity for SIA NS3- and NS5-encoded antigens. Moreover, data obtained for the anti-HCV seroconversion panel and for samples with borderline or discordant anti-HCV enzyme-linked immunosorbent assay results suggest that bands with a relative intensity of >0.5 on the automated analyzer (theoretically negative) should be evaluated with care. Coefficients of variability ranged from 9 to 14.8% in an interassay reproducibility study. Overall, the performance of the automated analysis of SIA is comparable to that of the manual RIBA assay. The new automated processor for SIA bands proved to be sensitive and specific. Its use makes the optical scoring of bands unnecessary by indicating relative intensity values, which could be particularly useful in the follow-up care of anti-HCV-positive patients receiving antiviral therapy.

An analytical and laboratory evaluation of a newly-developed fully-automated third generation ELISA for the detection of anti-HCV (Cobas Core Anti-HCV EIA, Roche) was undertaken. Coefficients of variation (CVs) calculated on positive control and serum samples ranged from 5.9 to 9.8% in the intra-assay precision test and from 3.9 to 11.3% in the inter-assay evaluation. With regard to the study of clinical laboratory performance, five groups of sera pre-screened with two third generation ELISA (Ortho HCV 3.0 ELISA; Innotest HCV Ab III) were assayed: anti-HCV negative samples (n = 932); anti-HCV positive samples (n = 449); difficult sera of different origin (n = 113); sera with discrepant results in the two ELISAs (n = 50); sera with an indeterminate result in one or more confirmatory test (n = 34). The overall concordance between the Roche anti-HCV EIA and the two reference assays was 97.5 and 97.8% for the Ortho and for the Innogenetics assays, respectively. Although it is not possible to provide absolute figures for clinical sensitivity and specificity, the results of the study on discrepant samples show that the Cobas Core Anti-HCV gives a number of negative results with positive or indeterminate confirmatory anti-HCV tests, which is intermediate between the Ortho and the Innogenetics assay. In contrast, only 5% Cobas Core Anti-HCV reactive sera are not positive or clear-cut single band reactive by supplemental assays. The results show that the new fully-automated third generation anti-HCV test is a valid alternative to other commercially available assays for screening of antibodies to the hepatitis C virus.

The relationship of hepatitis A virus (HAV) isolates associated with an outbreak in Genoa, Italy, in 1993 was examined using direct sequencing of amplicons derived by antigen capture PCR (AC/PCR) from faecal samples of the infected persons. Forty samples recovered from 38 primary and two secondary cases were examined. The latter were household contacts of the primary cases. In addition, faecal material of 2 unrelated persons infected simultaneously with hepatitis A in Genoa were tested. The PCR products derived from the P1/P2 junction of the HAV genome were analysed. A 100% nucleotide identity was detected between the viral isolates originating from the primary as well as the secondary cases. The viral isolates recovered from the faecal samples of the two unrelated cases differed from the virus causing the outbreak as well as from each other. These results indicate that a single HAV strain caused the outbreak. The virus might have been transmitted by ingestion of contaminated food or water since all hepatitis A infected employees of the factory had eaten in the same canteen. Definitions of HAV genotypes are based on numerous genetic comparisons of different strains. The sequence comparison of the investigated isolates with published HAV sequences of the P1/P2 genome region revealed that the virus associated with the outbreak belongs to HAV subgenotype IA, whereas the strains recovered from the viral isolates of the unrelated cases belong to subgenotype IB.

Objective: To assess the role of IgM antibodies to hepatitis C virus core protein (anti-HCV IgM) as a marker of chronic HCV infection and as a predictor of successful interferon (IFN) treatment., Design: Anti-HCV IgM levels were evaluated at baseline, during IFN therapy and during a follow-up period., Methods: Anti-HCV IgM levels were evaluated in 62 patients (47 men and 15 women, aged 25-65 years) with biopsy-proven chronic active hepatitis C. Fifty-one of the patients received alpha-IFN 3 MU three times a week for 6 months and 11 received the same therapy for 12 months. Twenty patients showed a long-term response; fourteen responded but subsequently suffered a relapse; twenty-eight did not respond to the treatment. Follow-up in all patients lasted for at least 6 (mean +/- SD 9.8 +/- 5.4, range 6-29) months after the end of the therapy., Results: Anti-HCV IgM were detected in 35 patients (56.4%) at baseline; no significant differences were observed between the three groups studied. Almost all members of the groups showing a relapse or no response remained positive at the end of therapy and follow-up. In contrast, we observed a progressive disappearance of anti-HCV IgM in patients responsive to interferon therapy over the long term., Conclusion: The loss of anti-HCV IgM positivity in patients positive at baseline can predict the long-term response to IFN therapy.

Background: Pre and post treatment perfusion scintigraphy with Tc-99m-sestamibi is an accurate method to evaluate the effectiveness of thrombolytic therapy in patients with acute myocardial infarction. In the present study this method was used to verify whether thrombolytic therapy performed late (6 to 24 hours) after symptoms onset was still able to produce significant salvage of jeopardized myocardial tissue., Methods: Fifteen patients (mean age 60.9 +/- 9 years) admitted to CCU 8-23 hours (mean 13.2 +/- 4 hours) after symptoms onset of their first acute myocardial infarction were studied. Both myocardial perfusion and left ventricular function were assessed before and after thrombolysis. Myocardial perfusion was studied using Tc-99m-sestamibi Single Photon Emission Tomography (SPET) before (pre-lysis SPET), 4.4 +/- 0.9 days (post-lysis SPET A) and 32 +/- 6 days (post-lysis SPET B) after thrombolysis. Uptake defects were graded from 0 to 4 using a 20-segment scheme. Regional ventricular function was evaluated using two-dimensional echocardiography before (pre-lysis ECHO), simultaneously with post-lysis SPET A (post-lysis ECHO A) and 32 +/- 4 days after treatment (post-lysis ECHO B); asynergy was graded from 1 to 4. Global left ventricular function was estimated using gated blood pool imaging and measuring the ejection fraction (EF) 5.3 +/- 1 (GBP 1) and 32 +/- 4 days after treatment (GBP 2). Within 6-9 days of admission all patients underwent coronary angiography., Results: In pre-lysis SPET all patients had uptaken defects (score range 17-52, mean 34.6 +/- 12). According to the comparison between pre-lysis SPET and post-lysis SPET A, patients were divided into two groups: 7 patients with perfusion recovery (Group I) and 8 patients with absent or minimal perfusion recovery (Group II). In Group I the mean uptaken score decreased significantly (from pre-lysis SPET 34.2 +/- 12 to post-lysis SPET A 21.7 +/- 7, p < 0.05); in Group II the uptaken score remained unchanged (from pre-lysis SPET 34.8 +/- 12 to post-lysis SPET A 33.6 +/- 12, ns). In post-lysis SPET B both groups showed a defect reduction compared to post-lysis SPET A; however, the defect score of SPET B in Group I was significantly lower than that in Group II (18.7 +/- 16 vs 30.1 +/- 10, p < 0.05). The asynergic score in pre-lysis ECHO was the same in the two groups (19.5 +/- 4); in post-lysis ECHO A no significant decrease was observed in the two groups. In post-lysis ECHO B a significant decrease was seen in Group I (14.8 +/- 5, p < 0.05 vs pre-lysis ECHO and vs post-lysis ECHO A), but not in Group II (19.8 +/- 5, ns); the asynergic score in post-lysis ECHO B was significantly lower in Group I than in Group II (p < 0.05). In GBP 1 the EF was 38% +/- 6 in Group I and 40% +/- 6 in Group II (ns). In GBP 2 a significant increase was registered in Group I (47% +/- 10, p < 0.05), whereas in Group II the EF remained unchanged (40% +/- 6, ns). Coronary angiography showed a patent infarct related vessel in all Group I patients and in 2 Group II patients (p < 0.05)., Conclusions: These results show that in patients with acute myocardial infarction treated with thrombolysis late after symptoms onset it is still possible to obtain an effective reperfusion and the consequent salvage of jeopardized tissue, as demonstrated by the recovery of both regional and global left ventricular function.

Sera from 82 haemodialysis patients were tested for anti-HCV, HCV-RNA, and HBsAg. Alanine aminotransferase (ALT) activity was monitored weekly for 2 months. Anti-HCV was positive in 31 patients (37.8%), showing different single-peptide patterns. HCV-RNA was detected in 26 anti-HCV-positive patients (84%) and also in two of 21 anti-HCV-negative patients. Twenty-seven (87%) of the 31 anti-HCV-positive patients had persistently normal ALT values; 22 of these patients were HCV-RNA positive. The four patients with elevated ALT values had HCV viraemia. HBsAg was positive in nine anti-HCV-negative patients. The close correlation between HCV viraemia and HCV status, independently of ALT values, requires that anti-HCV dialysis patients must be considered potentially infective and dialysed with reserved machines and/or in separate shifts.

The use of a single-day protocol for 99mTc MIBI myocardial scintigraphy is particularly well-suited for the nuclear medicine departments that are the referral centres for the cardiology departments of several community hospitals. The aim of the study was to compare, in the same patient population, the diagnostic accuracy of planar and SPECT imaging with 99mTc MIBI using a single-day protocol. Thirty-nine patients (31 males and 8 females, age range 32-69 years) were studied because of effort chest pain. Of them, 7 had a pre-test probability of coronary artery disease less than 2.5%; coronary angiography demonstrated significant coronary obstructions in the remaining 32 subjects and 21 of them also had a history of previous myocardial infarction. All patients underwent 99mTc MIBI myocardial scintigraphy with acquisition of planar and SPECT images after injection of a low dose (370 MBq) at rest and after a second higher dose (1110 MBq), injected approximately four hours later during exercise stress testing. The normalcy rate in the 7 patients with low pre-test probability was 100% using both imaging techniques. For the diagnosis of previous infarction, the sensitivities of planar and SPECT images were 71% and 96% respectively. For the diagnosis of effort ischemia the sensitivity values were 34% and 88%, respectively, using planar and SPECT imaging. With regard to the classification of the diseased coronary arteries, the sensitivities of planar and SPECT studies were 42% and 79% respectively, and the related specificities were 91% and 77%. We may conclude that using a single-day protocol planar imaging seems to allow lower levels of diagnostic accuracy both for the diagnosis of effort ischemia and the classification of the diseased vessels as compared to SPECT, which appears therefore in a similar setting the imaging technique of choice.

This paper reports the results of a study organized by the Istituto Superiore di Sanità (Rome, Italy) and carried out in collaboration with several national laboratories. Its aim was to ascertain whether diagnostic kits for detecting anti-HIV antibodies in human serum can also be used to detect the same antibodies in human immunoglobulins. Thirty-three lots of immunoglobulins supplied by six pharmaceutical companies present in Italy were examined using different procedures. On the basis of the results of this study it can be concluded that i. anti-HIV antibodies can be detected in immunoglobulins by means of commercial reagents; ii. a preliminary dilution of immunoglobulin samples should not be made; iii. Western blot method appears to be the 'reference' test; however, competitive EIA tests are equally valuable if a negative control made up of anti-HIV-negative Igs is employed; iv. Igs examination for anti-HIV antibody represents an indirect control on a correct donors' screening procedure.

The reaction of 2-amino-2-deoxy-D-glucose with lysine in water under simulated physiological conditions gave several browning products, with characteristic optical (lambda max 340 nm) and fluorescent properties (emission at 430 nm for excitation at 362 nm). The major product was isolated and characterized by mass spectrometry and n.m.r. spectroscopy as 2,5-bis(tetrahydroxybutyl)pyrazine derived by the condensation of two molecules of 2-amino-2-deoxy-D-glucose.

In a study comparing the safety and immunogenicity of a recombinant DNA yeast-derived hepatitis B vaccine with that of a plasma-derived vaccine in young female adults, 50 subjects were vaccinated with the former and 29 with the latter vaccine according to a 0, 1, and 6 month vaccination schedule. Results indicated that the yeast-derived vaccine was safe and highly immunogenic. Two months after the second vaccine dose, 86% of subjects seroconverted, a rate which increased to 100%, 30 days after the booster dose. Moreover, anti-HBs geometric mean titres increased progressively after the first two doses and rose markedly to 1098 IU/l after the booster dose. Although similar rates of seroconversion were obtained with both vaccines, the anti-HBs GMT of the plasma-derived vaccine was higher (P less than 0.05) than that elicited by the yeast-derived vaccine.

Thirty neonates to HBsAg carrier mothers, two of whom were HBeAg positive, received passive-active immunoprophylaxis. Within 48 hours after birth and at the first month of life these newborns were given HBIG (0.5 ml/Kg) and at the third month of life they were given first vaccination (10 micrograms). "HB-Vax" (M.S.D.) has been employed and its schedule has been performed (2 degrees and 3 degrees injections respectively at 4th and 9th month of life). Blood specimens have been collected in infants at birth and at each HBIG and vaccine administration, two months after the 2nd vaccination, one and six months after the third one. All specimens have been tested for HBV markers: HBsAg, anti-HBs, anti-HBc, anti-HBe and anti-HBc IgM. The anti-HBs titration has been performed according to WHO International Reference anti-HBs Standard. ALT and AST levels were determined in all infants specimens. The newborns did not show clinical signs or lab tests expressing HBV infection during the follow-up period. The anti-HBs geometric mean titres have been the following: 125 mIU/ml at 1 month, 176.4 mIU/ml at 3 months, 64.4 mIU/ml at 4 months, 119 mIU/ml at 6 months, 321 mIU/ml at 9 months, 2580.8 mIU/ml at 10 months and 715.3 mIU/ml at 15 months of life. Finally, side effects were observed for no infants.

Anti-HTLV III prevalence has been investigated in serum samples of 638 intravenous drug users collected over May 1981 - March 1985 and stored at -20 degrees C. Separately, in a prospective way, we have studied 68 IV drug abusers (53 Genoese and 15 of Sanremo area) of whom we have collected, at least, one serum specimen for each year, starting with 1981. We have also tested for anti-HTLV III presence serum samples of: 91 subjects of Hospital staff (Infectious Diseases Department and Laboratory workers); 32 workers in Therapeutic Communities for drug users and 24 family contacts of anti-HTLV III positive drug users. And then serum samples of two groups of general population collected for other seroepidemiological investigations in 1982 (256 subjects) and 1984 (538 subjects) were tested. No IV drug user was positive in 1981 whilst from 1982 up to 1984 there was a strong rising of the prevalence of anti-HTLV III positive subjects: 2, 22, and 39 per cent, respectively. The prevalence remained about 40% in the first months of 1985. The investigations carried out also show that HTLV III spread in Sanremo area slightly before than in Genoa and neighbourhood. No subject positive for anti-HTLV III has been detected among the Hospital staff and in workers of Therapeutic communities who have more probability to get in contact with infected subjects or their blood, as well as in the general population. A positive case has been discovered in a family contact (the wife of a positive for anti-HTLV III IV drug user). Some epidemiological and public health questions linked to the situation observed, at present, in Liguria, are discussed.

Clofibrate (a very effective lipid-lowering drug) may cause serious muscular damage in uraemic patients, even if used at appropriately low dose. Some authors have reported that D,L-Carnitine and Thiadenol can reduce lipid levels in non uraemic subjects, without adverse effects. In our study D,L-Carnitine and Thiadenol were administered for 3 months to respectively 21 and 15 patients on chronic haemodialysis with hypertriglyceridaemia. With D,L-Carnitine 52% of patients showed lower plasma triglycerides (TG) (-40.6%, p less than 0.002); after Thiadenol 66.6% of patients showed reduced plasma TG concentration (-42.9%, p less than 0.001). No dietary changes were recorded during the study. No major side effects nor biochemical changes were observed. D,L-Carnitine and Thiadenol appear to be less effective, but also less dangerous than Clofibrate in chronic uraemic patients.

The Authors carried out a methodological study about the way of expressing in mIU/ml the antibody to HBV surface antigen (anti-HBs) titer. The results showed that Hollinger's formula seems to provide a good accuracy when enzyme immunoassay is employed (AUSAB EIA, Abbott). Moreover, moderate variations in the commercial anti-HBs kits studied (AUSAB, AUSAB EIA, Abbott) have been verified. Considering that and the progress of the calibration curve of the WHO Reference International Standard, it is also advised to include always in the test a calibration curve of the standard serum when a higher precision is required.

Prevalence of positive subjects to anti-HTLV III and HBV markers (HBsAg; anti-HBc; anti-HBs) has been studied both among jailed people and wardens of Sanremo Jail. Out of 92 subjects in custody, 11 were anti-HTLV III positive and 44 had acquired HBV infection markers (antigen and/or antibodies). One of the wardens resulted anti-HTLV III positive whilst 14 appeared to have been infected by HBV. All anti-HTLV III positive subjects, but the warden, were intravenous drug users. The study of prevalence was the first step of a perspective monitoring program in Ligurian Jails.