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Highlights • β-Blocker propranolol worsens liver injury in model of non-alcoholic steatohepatitis. • Mechanism of hepatic injury is via activation of apoptotic pathway in hepatocytes. • β-Blockers should be avoided or used with extreme caution in patients with NASH., Prazosin an α1-adrenoceptor (AR) antagonist has been shown to reduce liver injury in a mouse model of non-alcoholic steatohepatitis (NASH) and is suggested as a potential treatment of NASH especially given its concomitant anti-fibrotic properties. The effect however, of β-AR blockade in non-cirrhotic NASH is unknown and is as such investigated here. In the presence of the β-blocker propranolol (PRL), mice fed normal chow or a half methionine and choline deficient diet, supplemented with ethionine (HMCDE), to induce NASH, showed significantly enhanced liver injury, as evidenced by higher hepatic necrosis scores and elevated serum aminotransferases (ALT). Mechanistically, we showed that murine hepatocytes express α and β adrenoceptors; that PRL directly induces hepatocyte injury and death as evidenced by increased release of lactate dehydrogenase, FASL and TNF-α from hepatocytes in the presence of PRL; and that PRL activated the apoptotic pathway in primary hepatocyte cultures, as indicated by upregulation of Fas receptor and caspase-8 proteins. The β-AR antagonist PRL therefore appears to enhance liver injury through induction of hepatocyte death via the death pathway. Further studies are now required to extrapolate these findings to humans but meanwhile, β-AR antagonists should be avoided or used with caution in patients with non-cirrhotic NASH as they may worsen liver injury.

Background: We recently showed that human hepatic progenitor cells (HPCs) express muscarinic acetylcholine (Ach) receptor subtype 3 and that – following liver transplantation – HPC numbers are significantly reduced. To further elaborate on this, we examined whether HPC also express receptors for vasoactive intestinal peptide (VIP), which, besides Ach, also is an important parasympathetic neurotransmitter. VIP expressing nerves are known to be present in the liver. Methods: We performed immunohistochemistry for VIP receptor subtypes 1 and 2 (VIPR1 and 2), on sections of normal and diseased human liver (n=17), and double staining for VIPR2 and known HPC markers. We performed RT-PCR for VIPR1 and 2 on total RNA from purified rat HPC. To document the probability of direct interaction, we also performed double immunostaining for nerve markers and HPC markers on human liver sections. Results: VIPR2 immunostaining was clearly positive in HPC and reactive bile ductules on paraffin-embedded and frozen tissue sections. We could not demonstrate VIPR1 protein expression in the liver, with either of two VIPR1 antibodies tested. The presence of VIPR2 mRNA in HPC was confirmed by RT-PCR. Nerve endings were shown to abut on reactive bile ductules. Conclusion: We show here for the first time that HPC express VIPR2 and receive nerve endings. These features, and the fact that HPC numbers are influenced by the presence or absence of the autonomic innervation of the liver, suggest a direct interaction.

Background and aims: Hepatic stellate cells (HSC) are activated by liver injury to become proliferative fibrogenic myofibroblasts. This process may be regulated by the sympathetic nervous system (SNS) but the mechanisms involved are unclear. Methods: We studied cultured HSC and intact mice with liver injury to test the hypothesis that HSC respond to and produce SNS neurotransmitters to promote fibrogenesis. Results: HSC expressed adrenoceptors, catecholamine biosynthetic enzymes, released norepinephrine (NE), and were growth inhibited by α- and β-adrenoceptor antagonists. HSC from dopamine β-hydroxylase deficient (Dbh−/−) mice, which cannot make NE, grew poorly in culture and were rescued by NE. Inhibitor studies demonstrated that this effect was mediated via G protein coupled adrenoceptors, mitogen activated kinases, and phosphatidylinositol 3-kinase. Injury related fibrogenic responses were inhibited in Dbh−/− mice, as evidenced by reduced hepatic accumulation of α-smooth muscle actin+ve HSC and decreased induction of transforming growth factor β1 (TGF-β1) and collagen. Treatment with isoprenaline rescued HSC activation. HSC were also reduced in leptin deficient ob/ob mice which have reduced NE levels and are resistant to hepatic fibrosis. Treating ob/ob mice with NE induced HSC proliferation, upregulated hepatic TGF-β1 and collagen, and increased liver fibrosis. Conclusions: HSC are hepatic neuroglia that produce and respond to SNS neurotransmitters to promote hepatic fibrosis.

In the rat the hepatic branch of the nervus vagus stimulates proliferation of hepatocytes after partial hepatectomy and growth of bile duct epithelial cells after bile duct ligation. We studied the effect of hepatic vagotomy on the activation of the hepatic progenitor cell compartment in human and rat liver. The number of hepatic progenitor cells and atypical reactive ductular cells in transplanted (denervated) human livers with hepatitis was significantly lower than in innervated matched control livers and the number of oval cells in vagotomized rat livers with galactosamine hepatitis was significantly lower than in livers of sham-operated rats with galactosamine hepatitis. The expression of muscarinic acetylcholine receptors (M1-M5 receptor) was studied by immunohistochemistry and reverse transcriptase-polymerase chain reaction. In human liver, immunoreactivity for M3 receptor was observed in hepatic progenitor cells, atypical reactive ductules, intermediate hepatocyte-like cells, and bile duct epithelial cells. mRNA for the M1-M3 and the M5 receptor, but not the M4 receptor, was detected in human liver homogenates. In conclusion, the hepatic vagus branch stimulates activation of the hepatic progenitor cell compartment in diseased liver, most likely through binding of acetylcholine to the M3 receptor expressed on these cells. These findings may be of clinical importance for patients with a transplant liver.

The aim of this study is to investigate the feasibility of a 'holistic' approach, using near infrared (NIR) spectroscopy and electronic devices (electronic nose and electronic tongue), as instrumental tools for the classification of different coffee varieties. Analyses were performed on green coffee, on ground roasted coffee and on coffee beverage. Principal component analysis was applied on spectral and sensory data to uncover correlations between samples and variables. After variable selection, linear discriminant analysis was used to classify the samples on the basis of the three coffee classes: Robusta, natural Arabica and washed Arabica.Linear discriminant analysis demonstrates the practicability of this approach: the external test set validation performed with NIR data showed 100% of correctly classified samples. Moreover, a satisfying percentage of correct classification in cross-validation was obtained for the electronic devices: the average values of correctly classified samples were 81.83% and 78.76% for electronic nose and electronic tongue, respectively.NIR spectroscopy was shown to be a very reliable and useful tool to classify coffee samples in a fast, clean and inexpensive way compared to classical analysis, while the electronic devices could assume the role of investigating techniques to depict the aroma and taste of coffee samples.

The serum levels of soluble β2-μ-associated and β2-μ-free HLA class I heavy chains were determined in 28 interferon-α nonresponder chronic hepatitis C patients retreated with interferon-α plus ribavirin and in 70 healthy subjects. The baseline levels of β2-μ-associated and β2-μ-free HLA class I heavy chains were significantly higher in patients than in healthy controls(P = 0.001). The levels of β2-μ-associated HLA class I heavy chains significantly increased in responder patients with respect to nonresponders at the third month of treatment(P = 0.03). At the sixth month of treatment and after 6 months of follow up the levels of β2-μ-associated HLA class I heavy chains decreased in responder patients and increased in nonresponders. The levels of β2-μ-free HLA class I heavy chains showed only minor changes during and after treatment. We suggest that the determination of hepatitis C virus RNA levels combined with soluble β2-μ-associated HLA class I heavy chains, as a marker of immune activation, could identify interferon-α non responder chronic hepatitis C patients most likely to respond to a retreatment with interferon-α plus ribavirin.

UNLABELLED OBJECTIVE; To evaluate the results of a large cohort of non-responder or relapsing responder patients with chronic hepatitis C retreated with various schedules of interferon (IFN). METHODS Our study included 276 patients (158 non-responders and 118 relapsing responders) who underwent IFN retreatments. Among the non-responder group, 158 patients underwent further courses of IFN. In particular, 108 patients underwent one course of IFN retreatment, 40 patients underwent two courses, eight patients underwent three courses, and two patients underwent four courses. Regarding the relapsing responder group, the 118 patients were retreated with the same dosage for varying periods. In particular, 50 patients were treated for 6 months, 43 patients for 12 months, and 25 for 24 months. Patients in the subgroups of IFN retreatment were homogeneous as far as age and gender distribution, as well as virological and histological characteristics, are concerned. Qualitative and quantitative HCV-RNA was evaluated at baseline, at the end of treatment and at the last check-up of follow-up. HCV genotype was determined on baseline serum samples. Alanine transaminase (ALT) levels were tested monthly. RESULTS Long-term biochemical (normal ALT levels) and virological (HCV-RNA negative) response was obtained in 2.6% of non-responder retreated patients, and in 33.9% of relapsing responder retreated patients. Evaluation of response on the basis of the duration of treatment showed that 48%, 19% and 16% of relapsing responder patients retreated for 24, 12 and 6 months, respectively, obtained long-term biochemical and virological response. CONCLUSION Non-responder patient retreatment is inefficient especially in cirrhotic and/or genotype 1 b patients. IFN retreatment is warranted in relapsing responder patients. In particular, 24-month therapy induces significant long-term biochemical and virological response.

The aim of this study was to detect hepatitis G virus RNA (HGV RNA) and antibodies against the virus envelope protein E2 (anti-E2) in 107 patients either on maintenance haemodialysis (n = 78) or peritoneal dialysis (n = 29) to evaluate the prevalence of HGV infection and to establish its role in liver disease. The total prevalence of HGV infection was of 15.4% among haemodialysis patients, whereas it was 10.3% among peritoneal dialysis patients. HGV RNA was detected in 2 haemodialysis patients (2.6%) and in 3 peritoneal dialysis patients (10.3%). Anti-E2 was found in 10 haemodialysis patients (7.8%), whilst all peritoneal dialysis patients resulted negative. In only 1 patient the alanine aminotransferase level was elevated. This patient underwent liver biopsy that did not reveal evidence of chronic hepatitis. The lower HGV prevalence in haemodialysis patients, when compared with data reported by other European authors, should be related to the lower rate of polytransfused patients in our series (29.5%). Multiple blood transfusions should be considered as the main factor to explain the different prevalence of HGV infection among various European dialysis centres. Detection of both antibody and viraemia is important to establish the real rate of the infection.

We have attempted to correlate the outcome of interferon (IFN) therapy with circulating soluble intercellular adhesion molecule-1 (sICAM-1) and the level of viremia in a sample of patients with chronic hepatitis C virus (HCV) infection. Forty-two patients were studied. Eighteen patients were maintained in long-term remission following IFN therapy, whereas 24 did not respond or relapsed. Serum concentrations of sICAM-1 were measured by enzyme-linked immunoassay. Viremia was measured by branched DNA signal amplification assay. Basal sICAM-1 was significantly higher in long-term responders than in nonresponder/relapsing patients. It was found that very high levels (1000 ng/ml) were closely associated with long-term clinical response. A quantitative evaluation of viremia in basal conditions, which was significantly lower in long-term responders, gave completely opposite results. During treatment, sICAM-1 concentrations significantly decreased in the group of long-term responders but not in the nonresponders. sICAM-1 reduction was apparent as early as 1 month after treatment started. Serum sICAM-1 may be a useful parameter in evaluating the outcome of patients with chronic hepatitis C infection treated with IFN.

Serum alpha-fetoprotein (AFP) levels were determined monthly for 1 year in 30 patients with biopsy-proven chronic hepatitis C virus (HCV) infection before, during, and after recombinant interferon alfa-2b (IFN) therapy. On the basis of their response to therapy, as evaluated by serum alanine aminotransferase levels, patients were classified as responders (10 patients), partial responders (10), and nonresponders (10). Ten healthy subjects comprised the control group. In patients with chronic HCV infection, baseline serum AFP levels (7.2 ± 1.0 ng/mL, 5.2 ± 0.4 ng/mL, and 6.2 ± 0.7 ng/mL in responders, partial responders, and nonresponders, respectively) were slightly but significantly higher than those recorded in healthy subjects (2.2 ± 0.3 ng/mL). Furthermore, IFN therapy caused a decrease in serum AFP levels that was more evident and prolonged in responders but that also occurred in partial responders and nonresponders. However, in the latter groups, this reduction in serum AFP levels was evident only during IFN therapy. These data suggest that IFN therapy may affect AFP secretion, possibly throughout the improvement of hepatic necrosis and, consequently, of hepatic regeneration.

Sixty patients of both sexes with biopsy-proven chronic hepatitis C were randomized to receive recombinant alpha-2b interferon 3 million IU or 10 million IU three times a week for 4 months. A 6-month follow-up period was scheduled for the end of therapy. Eighteen patients (30.0%) had normalized alanine aminotransferase (ALT) levels at the end of therapy and during the follow-up period. Of these, 10 received 3 million IU (33.3%) and 8 (26.7%) received 10 million IU. Twenty-three (38.3%) of the 60 patients relapsed during the follow-up period. No significant difference in relapse rate was observed between the two groups. The overall percentage of nonresponder patients was 31.7%. Treatment was discontinued because of noncompliance and/or side effects in 7 patients (11.7%), 1 in the 3-million IU group and 6 in the 10-million IU group. All patients were positive for hepatitis C virus—RNA at the beginning of the study, and it became undetectable in almost all responder patients. No correlation between viremia and biochemical signs of liver disease was observed in some cases. No virologic differences were found between the two treatment groups. Our study shows that 10 million IU does not increase the response or the relapse rate compared with 3 million IU in the treatment of patients with chronic hepatitis C.

Sixty men and women with biopsy-proven chronic hepatitis C were randomized to receive interferon (IFN) subcutaneously 3 million units three times a week (n=30) or 3 million units three times a week with tauroursodeoxycholic acid (TUDCA) 250 mg BID (n=30) for 6 months. A follow-up period of 3 months at the end of therapy was scheduled. Thirty-four patients (56.7%) had normalized serum alanine aminotransferase levels at the end of therapy; 16 received IFN alone and 18 received IFN plus TUDCA. Sixteen of these 34 patients (47.1%) relapsed in the follow-up period. No significant difference in relapse rate was observed between the two treatment groups. The overall percentage of nonresponders was 36.6%. Treatment was discontinued for noncompliance and/or side effects in four patients (6.7%), two in the IFN group and two in the IFN plus TUDCA group. Hepatitis C virus-ribonucleic acid was evident at the start of treatment in all patients and became undetectable in almost all responders. In some cases there was no correlation between viremia and biochemical signs of liver disease. No virologic differences were found between treatment groups. The tendency toward a reduced, but not significant, relapse rate in patients treated with IFN plus TUDCA must be confirmed in studies using a larger sample size.

Recovery from liver damage might be enhanced by encouraging repopulation of the liver by endogenous hepatic progenitor cells. Oval cells are resident hepatic stem cells that promote liver regeneration and repair. Little is known about the mediators that regulate the accumulation of these cells in the liver. Parasympathetic nervous system inhibition reduces the number of oval cells in injured livers. The effect of sympathetic nervous system (SNS) inhibition on oval cell number is not known. Adrenergic inhibition mobilizes hematopoietic precursors into the circulation and has also been shown to promote liver regeneration. Thus, we hypothesized that SNS inhibition would promote hepatic accumulation of oval cells and reduce liver damage in mice fed antioxidant-depleted diets to induce liver injury. Our results confirm this hypothesis. Compared with control mice that were fed only the antioxidant-depleted diets, mice fed the same diets with prazosin (PRZ, an alpha-1 adrenoceptor antagonist) or 6-hydroxydopamine (6-OHDA, an agent that induces chemical sympathectomy) had significantly increased numbers of oval cells. Increased oval cell accumulation was accompanied by less hepatic necrosis and steatosis, lower serum aminotransferases, and greater liver and whole body weights. Neither PRZ nor 6-OHDA affected the expression of cytokines, growth factors, or growth factor receptors that are known to regulate progenitor cells. In conclusion, stress-related sympathetic activity modulates progenitor cell accumulation in damaged livers and SNS blockade with alpha-adrenoceptor antagonists enhances hepatic progenitor cell accumulation.

Leptin’s actions on certain cells require a leptin-inducible neurotransmitter, norepinephrine (NE). NE modulates hepatic fibrosis. Therefore, decreased NE may explain why leptin deficiency inhibits hepatic fibrosis. We manipulated adrenergic activity in leptin-deficient ob/ob mice, leptin-sufficient, dopamine β-hydroxylase deficient (Dbh−/−) mice, and HSC cultures to determine if leptin requires NE to activate HSC and induce hepatic fibrosis. ob/ob mice have chronic liver injury, but reduced numbers of HSC. Supplemental leptin increases HSC, suggesting that leptin-dependent, injury-related factors permit expansion of HSC populations. NE also increases HSC numbers and activation, normalizing fibrogenesis. When fed hepatotoxic diets, NE-deficient Dbh−/− mice fail to accumulate activated HSC and have impaired fibrogenesis unless treated with adrenergic agonists. NE acts directly on HSC to modulate leptin’s actions because leptin increases HSC proliferation and prazosin, an α-adrenoceptor antagonist, inhibits this. Thus, leptin permits injury-related increases in adrenergic activity and requires NE to activate HSC and induce hepatic fibrogenesis.

Background/Aims : A simple and reproducible hepatic immunohistochemical staining (IHS) for hepatitis C virus (HCV) is not available. We aimed to validate hepatic IHS with monoclonal antibody (Mab) IG222, directed against the HCV-envelope 2 (E2) protein. Methods : A three-step indirect immunoperoxidase method was used for frozen sections and a two-step indirect EnVision technique was used for paraffin-embedded sections. Results : Naturally or in vitro HCV infected primary human hepatocytes were immunoreactive to HCV-E2. In the patient study ( n =253), IHS had a sensitivity of 96% and a specificity of 91%. Six patients who showed positivity in the liver with Mab IG222, but remained anti-HCV and HCV-RNA negative, had hepatitis C-like changes in their liver biopsy. In one patient HCV-RNA could be detected in the liver biopsy. We confirmed early graft reinfection in patients transplanted for HCV-related disease (34 patients with serial biopsies). Treatment for acute cellular rejection with steroids was associated with an increase in staining intensity. In nine patients with clearance of HCV-RNA during antiviral therapy, seven achieved negativation of immunoreactivity and two a marked reduction. Conclusions : The IHS with Mab IG222 is an accurate tool for diagnosis and clinical management of chronic hepatitis C.

Interferon alpha (IFN) has been the standard treatment for hepatitis C virus (HCV) infection. Using the kinetic curves of viral clearance, this study compared three treatment regimes based on IFN alone or in combination with Amantadine or Ribavirin to determine the mechanisms of action and the most suitable way to use these drugs. The early clearance kinetics of HCV were studied in 22 patients with chronic hepatitis C under different antiviral treatments: IFN 3 MU daily (7 pts); IFN 3 MU daily plus Amantadine 200 mg (7 pts); and IFN 3 MU daily plus Ribavirin 1–1.2 gr (8 pts), for 6 months. HCV-RNA was assessed qualitatively and quantitatively on serial samples. The HCV-RNA decay curves suggested a different behaviour of viral clearance induced by the three treatments. While no significant differences were present in the first 6 hours, between 6 to 12 hours Ribavirin induced a rapid decline in the viral load. Amantadine seemed to accelerate it in the third phase (12 to 30 hours) and to provoke a more pronounced viral decline when compared to IFN alone (P

Background. The recent discovery of a new parenterally transmitted DNA virus called TT virus (TTV ) led us to investigate its prevalence in haemodialysis patients, a high-risk group for blood-borne infection, and to evaluate its role in liver disease. Moreover, we compared the TTV prevalence with the prevalence of other hepatitis virus coinfections. Methods. Serum samples of 78 patients on maintenance haemodialysis were tested for TTV-DNA, hepatitis G virus (HGV)-RNA, anti-E2, anti-hepatitis C virus ( HCV ) and HCV-RNA. TTV-DNA was detected by semi-nested PCR using the primers from open reading frame 1 (ORF). HGV-RNA was detected by PCR using specific primers for the NS3 and the 5'-UTR genome regions while anti-E2 were checked by an enzyme immunological test. Anti-HCV was tested by the second generation Chiron RIBA HCV test system. HCV-RNA was evaluated by nested PCR with primers directed to the highly conserved 5' non-coding region of the HCV genome. Results. TTV prevalence in our patients was 19% (15/78) while the prevalence of HCV and HGV infection proved to be 20 and 15.4%, respectively. Among TTV positive patients HGV co-infection was present in five cases (33%), HCV in six cases (39.9%), while HBV co-infection was not present in any of the patients. Only three patients proved positive for all three viruses. ALT levels were normal in most cases (13/15; 86%). In particular, patients with TTV infection alone showed normal ALT levels and HCV coinfection was found in the two patients with moderate ALT increases. Conclusions. TTV prevalence in haemodialysed patients is significant though the real clinical impact is still unclear. However, we must keep in mind that the epidemiological relevance of TTV infection is probably underestimated due to the impossibility in detecting the corresponding antibody.

Background/Aimds: Jin Bu Huan and other Chinese herbal products are widely taken remedies. They have been developed as a natural alternative to traditional drugs in the treatment of various ailments. Their ability to induce several side effects such as acute hepatitis has already been described. We report a case of chronic hepatic damage following administration of Jin Bu Huan Anodyne tablets. Methods: The patients, a 49-year-old man, developed biochemical signs of liver damage 2 months after beginning Jin Bu Huan intake (3 tablets/daily) including biopsy-proven chronic hepatitis with moderate fibrosis. Virological, autoimmune, metabolic or other hepatotoxic causes were excluded. Liver function impairment was resolved by discontinuing Jin Bu Huan intake. Conclusions: This case reinforces the already known hepatotoxicity of this product and should make us think more about the uncontrolled use of alternative products.

Seventy mother-newborn pairs were studied for hepatitis C viremia to evaluate the risk of vertical transmission of hepatitis C virus from human immunodeficiency virus-negative mothers. Forty-five mothers were hepatitis C virus-RNA positive: 4 to 45 children were positive at birth and during follow-up. The level of viremia plays an important role in vertical transmission.

Objective To evaluate the relationship between hepatitis C virus (HCV)-RNA levels and genotypes in order to establish their potentially predictive role in interferon (IFN) response. Design To detect HCV genotype at baseline and HCV viraemia levels before and during IFN treatment in three groups of patients with different IFN response. Methods Our study included 85 patients with biopsy-proven chronic hepatitis C who underwent IFN therapy at standard schedule (3 MU thrice weekly for 6 months). On the basis of IFN response they were subdivided into three groups as follows: non responders (NR: 27 cases) when alanine aminotransferase (ALT) values (normal value: 0-40 IU) at the end of treatment were abnormal (101.7 +/- 10.4); responders relapsing (RR: 29 cases) when normal ALT values at the end of therapy (28.14 +/- 1.7) increased during follow-up; sustained (long-term) responders (LTR: 29 cases) when ALT values remained normal for at least 12 months of follow-up (ALT values at the end of therapy: 21.8 +/- 1.4). ALT activity was monitored monthly during therapy and each month during 12 months of follow-up. HCV genotype was evaluated before starting treatment whereas HCV-RNA viraemia was checked at baseline and at the 1st and 6th months of therapy. Results The baseline viral load was higher in the NR group than in the RR and LTR groups independently of genotype; HCV-RNA levels progressively decreased during therapy independently of response but the levels remained significantly higher in the NR group. Genotype 1b was prevalent in the NR group. However, levels of viraemia in genotype 1b LTR patients are significantly lower than in genotype 1b NR patients. Conclusion These results suggest that among viral-related parameters viraemia alone seems to play an important role in predicting response to IFN independently of genotype.

Interferon-alpha (IFN-alpha) may affect lipid metabolism by stimulating hepatic fatty acid synthesis. The aim of this study was to evaluate serum lipid levels during IFN-alpha therapy in patients with biopsy-proven chronic active hepatitis C. A total of 22 patients (18 males and 4 females; age 25-55 years) received 3 MU of recombinant IFN-alpha 2b 3 times a week for 6 months. Serum lipids were determined at baseline and then every month until the end of therapy. All patients had normal serum lipid levels at baseline. No significant level of modification occurred in patients during the therapy. An increase in serum lipid levels during low-dose IFN-alpha therapy seems to be uncommon in hepatitis C virus-infected patients with baseline normal levels.

Aim: To evaluate: I) TTV infection prevalence in a group o f patients (pts) with high risk o f parenteral exposure such as intravenous drug users 0DUs) and haemodialysed pts (HDs); 2) the clinical relevance o f TTV and the correlation with other hepatitis virus coinfection~ Mater ia l s and Methods: We studied 118 pts:40 with previous or ongoing history o f IDU and 78 HDs. All IDUs were anti-HCV+ve and 20 out them were HIV ceinfected. TTV-DNA was tested by semi-nested PCR using primers open reading flame 1 (ORF). HGV-RNA analysis was performed by PCR using specific primers for the NS3 and 5'UTR.The presence o f anti-E2 Ab was tested by ELISA assay. Qualitative HCV-RNA was performed by nested (in h o u s e ) R T PCR. R~ n l t s : Our data showed a lower prevalence o f TTV (21/118; 17.8%) as compared to HGV infection (38/118; 32.2%). In particular TTV-DNA was detected in 6 cases (15%) among IDUs and in 15 cases (19%) among HDs, while an higher prevalence (65%) o f HGV was present in IDUs group. TI 'V infection alone was present in few pts (4 l i d s ) which showed normal ALT levels while in all TTV+ve cases with ALT increase HCV-RNA was detected. Rarely we observed TTV and HGV coinfection in both groups. Conclusions: The prevalence o f TI 'V infection was lower when compared to HGV. These data suggest the lower importance o f parenteral route for TTV. As far as the clinical impact o f TTV is concerned, the data showed that in the TTV+ve pts ALT increase may be related to HCV activity, [ eo o3s I Viral hepatitis." clinical aspects