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Background. A high incidence of thyroid autoantibodies and/or disorders was observed in subjects with hepatitis C virus-related chronic hepatitis during interferon-alpha therapy. Aim. To evaluate whether thyroid autoimmunity and dysfunction, induced by interferon-alpha therapy, could be viewed as predictors for treatment response and as valid prognostic markers of liver disease progression. Patients. A total of 136 subjects (96 males/40 females; median age 48 years; range 23–64) affected by biopsy-proven chronic hepatitis C (33.1 % with compensated liver cirrhosis). Methods. All subjects were treated with interferon-alpha therapy at 6 MU 3 times weekly for 12 months and then followed up for an average period of 60 months (range 12–108). Routine laboratory tests, virological assessment, liver ultrasound, thyroid function tests (serum free-triiodothyronine, free-thyroxine, serum thyrotropin), and autoimmunity were performed for all subjects. Results. Percentage of thyroid autoimmunity and thyroid dysfunction in long-term responders was not significantly different compared to that in non-responders (47.0% and 11.8% vs 35.3% and 5.9%, respectively; non significant). The multivariate model demonstrated that the absence of cirrhosis was the only factor significantly related to successful response to therapy (odds ratio: 14.9; 95% confidence interval: 1.9–115.0 for chronic hepatitis C vs presence of cirrhosis). Moreover, the occurrence of thyroid autoimmunity during interferon therapy was similar both in patients with or without worsening of liver disease (33.3% and 39.8%, respectively, p = not significant). No subject with ongoing liver disease developed thyroid dysfunction during treatment, as opposed to the 10 118 (8.4%) with a better course of liver disease; however, this difference was not statistically significant. The multivariate model showed that age was the only covariate significantly associated with unfavourable outcome of liver disease (odds ratio: 18.6; 95% confidence interval: 2.3–151.9, for those over 48 years vs younger patients). Conclusions. There is no evidence that the immune mechanism involved in the pathogenesis of thyroid autoimmune phenomena is the same as that regulating the therapeutic clearance of HCV or modulating the unfavourable course of HCV-related chronic hepatitis. However, our study confirmed that liver disease seems to progress more slowly in younger subjects.

Background. Quite often subjects affected by chronic hepatitis C virus infection have no clinical signs of liver disease and serum aminotransferase values never go beyond the upper limit of normal. Yet these subjects, defined “asymptomatic HCV carriers”, often have active viral replication and various degrees of histological damage. Aims. To verify, in a population of antibody to hepatitis C virus carriers, if normal serum aminotransferase values in hepatitis C virus-RNA positive differed considerably from those in hepatitis C virus-RNA negative subjects. Subjects/Methods. We followed 24 anti-hepatitis C virus-positive subjects (15 hepatitis C virus-RNA positive and 9 negative) by measuring alanine and aspartate aminotransferase levels at 3-month intervals for a median of 40 months (range 6–77). Results. Determinations resulted repeatedly and rigorously within the normal range in all participants. Alanine aminotransferase values were higher in hepatitis C virus-RNA positives than in negatives (mean ± SD: 0.609±0.172 vs 0.434±0.153 times the upper limit of normal; p

the AFP, the poorer the prognosis, but no good evidence in the literature exists on the minority of patients with HCC who do not secrete AFP. Our aim was to analyse this sub-group of patients from a cohort with advanced HCC treated with Sorafenib, and assess survival outcomes. Methods: Consecutive patients with a confirmed diagnosis of HCC as per AASLD criteria, treated with Sorafenib at Queen Elizabeth Hospital Birmingham UK, between April 2009 and March 2014 were included in the analysis. We took our local reference range of 400ng/ml (HR: 1.02, p = 0.89). The Kaplan–Meier curve presented shows the separation of survival curves for the 3 groups. Conclusions: There is very little published in the literature about the AFP non-secretor HCC sub-group. Our data shows that this sub-group has better prognosis compared to patients with AFP secretory HCC. We believe that this data is important for clinicians and patients for prognostication and decision making. P0340 CLINICAL PATTERNS OF HEPATOCELLULAR CARCINOMA (HCC) IN NON ALCOHOLIC FATTY LIVER DISEASE (NAFLD): A MULTICENTER CASE–CONTROL STUDY F. Piscaglia, G. Svegliati Baroni, A. Barchetti, A. Pecorelli, S. Marinelli, C. Tiribelli, S. Bellentani, HCC-NAFLD Italian Study (full list exceeds available space) Bolondi L, Zoli M, Trevisani F, Malagotti D, Bugianesi E, Vanni E, Mezzabotta L, Cabibbo G, Petta S, Fracanzani A, Fargion S, Marra F, Fani B, Sacco R, Morisco F, Caporaso F, Guarino M, et al.. Dpt of Medical and Surgical Sciences, University of Bologna, Uni of Internal Medicine, University of Bologna, Bologna, Gastroenterology, University of Ancona, Ancona, Liver Research Center, Science Park – Basovizza Campus, Trieste, Italy E-mail: fabio.piscaglia@unibo.it

Background/Aim: The aim of this study was to evaluate if plasma levels of alpha-glutathione-S-transferases (determined in basal conditions and monthly for 1 year during and 1 year after interferon therapy) could characterize patients who show only a primary response. Methods: We studied 48 patients with biopsy-proven, hepatitis C virus ribonucleic acid positive chronic hepatitis treated with interferon: 18 were "Sustained Responders", 12 "Relapsers" and 18 "Non-Responders". Results: Relapsers showed higher basal levels of alpha-glutathione-S-transferases, which remained higher than normal even when alanine aminotransferases normalized. No correlation was documented between alpha-glutathione-S-transferase levels and all other parameters examined (alanine aminotransferases, γ-glutamyl-transpeptidase, viremia, and histological activity index). Conclusions: These findings suggest that alpha-glutathione-S-transferase levels may be considered a predictive index of response to interferon therapy in chronic hepatitis C patients.

In recent years, a number of case reports and clinical studies have highlighted the risk of hepatitis B and C virus reactivation in patients with inflammatory bowel disease who are treated with immunosuppressive drugs. The cases of viral hepatitis reactivation that have been reported are characterized by a wide range of clinical manifestations, from viremia without clinically relevant manifestations to fulminant life-threatening hepatitis. The development and dissemination of biological immunosuppressive drugs have led to a significant increase in the number of reports of interest to physicians in a variety of clinical settings. On this topic, there have been a number of published guidelines and reviews that have collected the available evidence, providing recommendations on prophylactic and therapeutic strategies and methods for monitoring patients at risk. However, it should be noted that, to date, very few clinical studies have been published, and most of the recommendations have been borrowed from other clinical settings. The published studies are mostly retrospective and are based on very heterogeneous populations, using different therapeutic and prophylactic regimens and obtaining conflicting results. Thus, it seems clear that it is desirable to concentrate our efforts on prospective studies, not conducting further reviews of the literature in the continued absence of new evidence.

Hepatitis C virus (HCV) infection is one of the main causes of liver disease worldwide. Patients undergoing surgery are at risk of acquiring acute HCV infection and those undergoing surgical eradication of a neoplasia may be indicated for adjuvant treatment. Therefore, unlike chronic infection, these patients may simultaneously need antiviral therapy with interferon for acute hepatitis C and cytotoxic chemotherapy. To date, no data are available regarding the efficacy and tolerability of concomitant interferon treatment and antineoplastic chemotherapy in the setting of acute hepatitis C treatment. Here, we report the case of a 60-year-old man who developed acute hepatitis C after left hemicolectomy for an adenocarcinoma. He received concomitant antiviral treatment with interferon-α and adjuvant chemotherapy with capecitabine and oxaliplatin. Both treatments were well-tolerated and the patient completed the scheduled therapies. HCV infection was eradicated and the patient is free of neoplastic disease two years and 6 months after surgery.

Viral hepatitis reactivation has been widely reported in patients undergoing immunosuppressive therapy; however, few data are available about the risk of HBV and HCV reactivation in patients with inflammatory bowel disease, receiving immunosuppressive drugs. The aim of our study was to assess the prevalence of HBV and HCV infection in a consecutive series of patients with inflammatory bowel disease and to value the effects of immunosuppressive therapy during the course of the infection. Retrospective observational multicenter study included all consecutive patients with inflammatory bowel disease who have attended seven Italian tertiary referral hospitals in the last decade. A total of 5096 patients were consecutively included: 2485 Crohn's disease and 2611 Ulcerative Colitis. 30.5% and 29.7% of the patients were investigated for HBV and HCV infection. A total of 30 HBsAg positive, 17 isolated anti-HBc and 60 anti-HCV-positive patients were identified. In all, 20 patients with HBV or HCV infection received immunosuppressive therapy (six HBsAg+; four isolated anti-HBc+ and 10 anti-HCV+). One of six patients showed HBsAg+ and one of four isolated anti-HBc+ experienced reactivation of hepatitis. Two of six HBsAg patients received prophylactic therapy with lamivudine. Only one of 10 anti-HCV+ patients showed mild increase in viral load and ALT elevation. Screening procedures for HBV and HCV infection at diagnosis have been underused in patients with inflammatory bowel disease. We confirm the role of immunosuppressive therapy in HBV reactivation, but the impact on clinical course seems to be less relevant than previous reported.

UnlabelledThe aim of the present work was to test the potential of Proton Transfer Reaction Time-of-Flight Mass Spectrometry (PTR-ToF-MS) in the diagnosis of liver cirrhosis and the assessment of disease severity by direct analysis of exhaled breath. Twenty-six volunteers have been enrolled in this study: 12 patients (M/F 8/4, mean age 70.5 years, min-max 42-80 years) with liver cirrhosis of different etiologies and at different severity of disease and 14 healthy subjects (M/F 5/9, mean age 52.3 years, min-max 35-77 years). Real time breath analysis was performed on fasting subjects using a buffered end-tidal on-line sampler directly coupled to a PTR-ToF-MS. Twelve volatile organic compounds (VOCs) resulted significantly differently in cirrhotic patients (CP) compared to healthy controls (CTRL): four ketones (2-butanone, 2- or 3- pentanone, C8-ketone, C9-ketone), two terpenes (monoterpene, monoterpene related), four sulphur or nitrogen compounds (sulfoxide-compound, S-compound, NS-compound, N-compound) and two alcohols (heptadienol, methanol). Seven VOCs (2-butanone, C8-ketone, a monoterpene, 2,4-heptadienol and three compounds containing N, S or NS) resulted significantly differently in compensate cirrhotic patients (Child-Pugh A; CP-A) and decompensated cirrhotic subjects (Child-Pugh B+C; CP-B+C). ROC (Receiver Operating Characteristic) analysis was performed considering three contrast groups: CP vs CTRL, CP-A vs CTRL and CP-A vs CP-B+C. In these comparisons monoterpene and N-compound showed the best diagnostic performance.ConclusionsBreath analysis by PTR-ToF-MS was able to distinguish cirrhotic patients from healthy subjects and to discriminate those with well compensated liver disease from those at more advanced severity stage. A breath-print of liver cirrhosis was assessed for the first time.

Background & aims: We previously demonstrated the efficacy of garlic extract (GE) in the prevention of rat liver fibrosis by inhibiting tissue transglutaminase (tTG) activity. In the present study we aimed to evaluate the potential of GE in the regression of liver fibrosis and the underlining mechanism. Methods: Male Wistar rats were i.p. injected, twice a week, for 7 weeks, with CCl4 to develop liver fibrosis. Successively, a group was immediately sacrificed, while the remaining two groups received the GE or the vehicle, respectively, over the following 2 wks. A group of normal rats was also included in the study. Liver function, histology, and collagen deposition in parallel with gene and protein expression of α-SMA, tTG, TGF-β1, SEMA-7A, and metalloproteinase inhibitor 1 (TIMP1) as well as measure of active by total TGF-β1 were assessed. Results: CCl4 administration increased alanine-aminotransferase (ALT) activity, hepatic collagen deposition and gene and protein expression of all monitored markers. GE, but not the sole vehicle, restored liver histology and function by decreasing fibrogenesis markers (α-SMA, tTG, TGF-β1, SEMA-7A and TIMP1). Active by total TGF-β1 was significantly reduced (p < 0.05) in GE treated rats compared to the CCl4 at 7 weeks, and vehicle rats. Conclusions: These findings concurrently suggested that GE elicited therapeutic effect against liver fibrosis. Regression of liver fibrosis occurred by reducing myofibroblasts (through modulation of HSCs activation mechanisms), remodelling extracellular matrix (through increase of its degradation) and regenerating liver tissue and functions: three processes regulated by fine mechanisms where active TGF-β1 and tTG play a central role.

Background: Recent evidence indicate that gut microbiota plays a key role in the development of NAFLD through the gut-liver axis. An altered gut permeability induced by alterations of tight junction (TJ) proteins allows the passage of bacteria and substances leading to liver inflammation, hepatocyte damage and fibrosis. This study aims to evaluate the influence of decaffeinated coffee on gut permeability in a rat model of fat liver damage induced by a high fat diet (HFD). Methods: Twelve Male Wistar rats were assigned to 3 groups. The first group received HFD for 5 months and drank water; the second group received HFD for 5 months and drank water added with 1.2mL decaffeinated coffee/day starting from the 4th month; the third group received standard diet (SD) and drank water. Protein and mRNA expression levels of Toll-Like Receptor-4 (TLR-4), Occludin and Zonula occludens-1 (ZO-1) were assessed in rat intestines. Results: A significant reduction of Occludin and ZO-1 was observed in HFD fed rats (0.97±0.05 vs 0.15±0.08 p˂0.01, and 0.97±0.05 vs 0.57±0.14 p˂0.001, respectively); this reduction was reverted in HFD+COFFEE rats (0.15±0.08 vs 0.83±0.27 p˂0.01, and 0.57±0.14 vs 0.85±0.12 p˂0.01, respectively). The TLR-4 expression up-regulated by HFD was partially reduced by coffee administration. Conclusions: HFD impairs the intestinal tight junction barrier integrity. Coffee increases the expression of tight junctions proteins reverting the altered gut permeability, and reduces TLR4 expression. Key Words: NAFLD, coffee, Occludin, Zonulin-1, rat model, TLR4.

Breath testing has been largely used as a diagnostic tool, but the difficulties in data interpretation and sample collection have limited its application. We developed a fast (20 s), on-line, non-invasive method for the collection and analysis of exhaled breath in awake rats based on proton transfer reaction time of flight mass spectrometry (PTR-ToF-MS) and applied it to investigate possible relationships between pathologies induced by dietary regime and breath composition. As a case study, we investigated rats with dietary induced non-alcoholic steatohepatitis (NASH) and modifications induced by coffee addition to the diet. We considered two different diets (standard and high fat) complemented with two different drinking possibilities (water or decaffeinated coffee) for a total of four groups with four rats each. Several spectrometric peaks were reliable markers for both dietary fat content and coffee supplementation. The high resolution and accuracy of PTR-ToF-MS allowed the identification of related compounds such as methanol, dimethyl sulphide, dimethyl sulphone and ammonia. In conclusion, the rapid and minimally invasive breath analysis of awake rats permitted the identification of markers related to diet and specific pathologic conditions and provided a useful tool for broader metabolic investigations.

Non-alcoholic fatty liver disease (NAFLD) is a rapidly emerging chronic liver disorder and will increasingly be a critical global health problem. Diet is an important pathogenic factor of NAFLD and it is well documented that the increased prevalence of NAFLD during the last decades was associated with deep modification of dietary habits, especially increased intakes of fats and simple carbohydrates. As the disease stems from excess calorie intake and lack of physical activity, the correction of unhealthy lifestyles is the basis of any prevention and treatment strategy while drugs should remain a second-line of treatment. NAFLD patients should receive counselling for a low carbohydrate and low saturated fat diet, avoidance of fructose-enriched soft drinks and increased consumption of fruits and vegetables. The use of functional foods in NAFLD has been poorly studied and up to day only few reports are encouraging and incite the promotion of functional food approach on NAFLD prevention on the basis of food and nutritional genomics.

Background and aim: Tissue transglutaminase contributes to liver damage in the development of hepatic fibrosis. In a model of neurodegeneration, the therapeutic benefit of cystamine has been partly attributed to its inhibition of transglutaminase activity. Garlic extract contains many compounds structurally related to cystamine. We investigated the anti-fibrotic effect of garlic extract and cystamine as specific tissue transglutaminase inhibitors. Methods: Rat liver fibrosis was induced by intraperitoneal injection of carbon tetrachloride (CCl4) for 7 weeks. Cystamine or garlic extract was administrated by daily intraperitoneal injection, starting from the day after the first administration of CCl4. Hepatic function, histology, tissue transglutaminase immunostaining and image analysis to quantify Red Sirius stained collagen deposition were examined. Reverse transcription-polymerase chain reaction to detect alpha-SMA, IL-1 and tissue transglutaminase expression and Western blot for tissue transglutaminase protein amount were performed. Transglutaminase activity was assayed on liver homogenates by a radio-enzymatic method. Results: Transglutaminase activity was increased in CCl4 group and reduced by cystamine and garlic extract (p < 0.05). Treatment with cystamine and garlic extract reduced the liver fibrosis and collagen deposition, particularly in the garlic extract group (p < 0.01). Moreover, the liver damage improved and serum alanine aminotransferase was decreased (p < 0.05). Tissue transglutaminase immunolocalised with collagen fibres and is mainly found in the ECM of damaged liver. Alpha-SMA, IL-1, tissue transglutaminase mRNA and tissue transglutaminase protein were down-regulated in the cystamine and garlic extract groups compared to controls. Conclusion: These findings concurrently suggest that transglutaminase may play a pivotal role in the pathogenesis of liver fibrosis and may identify garlic cystamine-like molecules as a potential therapeutic strategy in the treatment of liver injury.

We evaluated the etiology and risk factors for transient and persistently elevated aspartate and/or alanine aminotransferase levels in virus-free blood donors.HBsAg/HBV-DNA and anti-HCV/HCV-RNA negative blood donors with elevated aspartate aminotransferase and/or alanine aminotransferase, observed in 5 blood transfusion centres in Italy from 2004 to 2005. Aspartate aminotransferase/alanine aminotransferase levels were measured at entry and every 2 months during a period of 6 months.291 individuals were evaluated (144 with persistent and 147 with transient abnormal aminotransferases). High body mass index was the most frequent (75.5%) etiological factor and was more common in the persistent elevated levels group, compared to the transient elevated levels group (82.0% vs 65.3%; p0.01). Excessive alcohol intake (2 units/day) was reported in 23.6%, with no differences between the two groups. Instead, recent use of medication or paint exposure were most frequently associated with transient elevated levels than persistent elevated levels (61.6% vs 23.3% for drugs and 13.7% vs 4.3% for paint, p0.001). Considering the participants with transient elevated levels as controls, the multivariate analysis showed that high body mass index was the only independent predictor of persistent elevated aminotransferase levels (OR=5.3; 95%CI=1.88-13.42 for those with body mass index29.9).In virus-free blood donors, excessive body mass index is the most frequent etiological factor of abnormal aminotransferases and it is the sole risk factor associated with persistently elevated aminotransferases.

Background and Aim: Several in vitro studies have demonstrated the ability of pure trans-resveratrol (t-Res) to act as an anti-oxidant, but the scientific literature is lacking in in vivo studies dealing with dietary t-Res bioavailability in oxidative stress models. Our aim was to investigate the bioavailability of t-Res from dietary sources and its effect on an animal model of carbon tetrachloride (CCl4)-induced liver lipid peroxidation. Methods: Ten rats were intragastrically administered for 14 days with a grape-stalk extract determining a daily t-Res dosage of 3 mg/kg. The control group (10 rats) was daily injected with the vehicle solvent without the t-Res extract. After 1 week, the induction of liver lipid peroxidation by CCl4 injection was carried out. Serum and liver samples, at different time intervals, were collected to evaluate t-Res content, by high-performance liquid chromatography (HPLC) and liquid chromatography–mass spectometry–mass spectometry (LC-MS-MS). Liver malondialdehyde (MDA) as marker of oxidative stress was measured. Results: t-Res accumulates in the liver reaching 49.8 ± 10.2 ng/g after 7 days and 191.8 ± 15.3 ng/g after 14 days. No t-Res was detected in serum. The increase of MDA liver concentration due to CCl4 injection after 24 h and 1 week was reduced by 38% and a 63%, respectively, by the treatment with the t-Res extract. Conclusions: A moderate consumption of t-Res from a dietary source resulted in a time-dose-dependent liver accumulation. It was able to counteract in vivo CCl4-induced liver lipid peroxidation thus demonstrating the hepatoprotective property of t-Res.

Background and objective Loco-ablation therapy (LAT) has become standard treatment for patients with HCC who are candidates for liver transplantation (LT). The aim of this study was: to evaluate if LAT was able to induce complete necrosis of tumour mass; to determine the tumour recurrence rate after LT and factors associated with recurrence. Patients The percentage and the distribution of necrosis in 116 HCC nodules of 61 patients with (26 patients) and without (35 patients) previous types of LAT were examined in explanted livers. Results Total necrosis was found only in 7% of treated nodules, and 42% of these showed absence of necrosis. The amount of necrosis was significantly related to the gross appearance of HCC: a single nodule with smaller adjacent satellite nodules showed a higher percentage of necrosis. No relation was found between the amount of necrosis and the type of LAT. Recurrence was observed in 11.5% and 8.5% of patients with and without previous LAT, respectively (P = ns). Conclusions LAT very rarely induces complete necrosis; the amount of necrosis seems to depend on the growth pattern of the tumour and not on the type of previous LAT; the tumour size, measured at explant, is the only variable significantly related to recurrence.

s / Digestive and Liver Disease 47S (2015) e226–e235 e235 genetic inhibition of IDO-1 on the development of steatohepatitis induced by amethionine and choline-deficient (MCD) diet in mice. Methods: C57Bl/6 mice fed a MCD diet for 4 weeks, were treated with the specific IDO inhibitor, 1-methyl-d-triptophan (1MT, 5mg/ml) or its vehicle in drinking water. WT and IDO-1 KO C57Bl/6 mice were fed MCD or its vehicle for 4 weeks. Intrahepatic gene expression was assayed by quantitative real time PCR. Histological examination was assessed by formalin-fixed, paraffinembedded liver sections stainedwithhematoxylinandeosin (H&E). Results: No changes were observed in ALT levels in both pharmacological and genetic interference with IDO-1. Genetic deletion of IDO-1 resulted in an amelioration of the inflammatory phenotype. These effects were associated both with reduced intrahepatic gene expression of proinflammatory factors, including CD11b, CCL2, IL-1 beta and iNOS and with reduced necroinflammation score. Pharmacological inhibition of IDO by 1-MT administration did not affect the phenotype of dietary experimental steatohepatitis in terms of proinflammatory markers or inflammation score. Conclusions: Interference with IDO pathway by 1-MT administration did not affect the phenotype of dietary experimental steatohepatitis. On the other hand, genetic deletion of IDO-1 ameliorates liver injury induced by experimental steatohepatitis and modulates the expression of proinflammatory factors in this context. http://dx.doi.org/10.1016/j.dld.2015.07.044

s / Digestive and Liver Disease 46S (2014) e136–e141 e137 all 248 patients had reached HBV undetectability

s / Digestive and Liver Disease 47S (2015) e1–e18 e7 (months; 95% CI): CPT 0 62 (52.9–71.1), A 44 (41.6–46.4), B 22 (19.7–24.3), C 9 (6.6–11.3), p

Background: Fresh fruit and vegetables exert multiple biological effects on the gastrointestinal mucosa. Aim: To assess whether apple extracts counteract oxidative or indomethacin induced damage to gastric epithelial cells in vitro and to rat gastric mucosa in vivo. Methods: Apple extracts were obtained from freeze dried apple flesh of the “Annurca” variety. Cell damage was induced by incubating MKN 28 cells with xanthine-xanthine oxidase or indomethacin and quantitated by MTT. In vivo gastric damage was induced by indomethacin 35 mg/kg. Intracellular antioxidant activity was determined using the (2,2′-azinobis (3-ethylbenzothiazolin-6-sulfonate) method. Malondialdehyde intracellular concentration, an index of lipid peroxidation, was determined by high pressure liquid chromatography with fluorometric detection. Results: (1) Apple extracts decreased xanthine-xanthine oxidase or indomethacin induced injury to gastric epithelial cells by 50%; (2) catechin or chlorogenic acid (the main phenolic components of apple extracts) were equally effective as apple extracts in preventing oxidative injury to gastric cells; and (3) apple extracts (i) caused a fourfold increase in intracellular antioxidant activity, (ii) prevented its decrease induced by xanthine-xanthine oxidase, (iii) counteracted xanthine-xanthine oxidase induced lipid peroxidation, and (iv) decreased indomethacin injury to the rat gastric mucosa by 40%. Conclusions: Apple extracts prevent exogenous damage to human gastric epithelial cells in vitro and to the rat gastric mucosa in vivo. This effect seems to be associated with the antioxidant activity of apple phenolic compounds. A diet rich in apple antioxidants might exert a beneficial effect in the prevention of gastric diseases related to generation of reactive oxygen species.

In this prospective study, we investigated whether the development of interferon (IFN)-alpha-related autoimmune thyroiditis (IFN-AT) was correlated with the sequential changes of cytokine pattern induced by IFNalpha in the peripheral lymphocytes.We enrolled 18 hepatitis C virus (HCV)-positive patients who developed IFN-AT, eight patients with euthyroidism [IFN-AT(Eu)] and 10 with thyroid dysfunction [IFN-AT(Dy)]. Twenty HCV-positive patients without IFN-AT acted as control group (Co-HCV+). Intracellular expression of IFNgamma and IL-4 was evaluated by multicolor flow-cytometry analysis in peripheral lymphocytes in vitro stimulated by phorbol-12-myristate-13-acetate (PMA) (25 ng/ml) and ionomycin (1 mug/ml) in presence of monensin (5 microm).At the appearance of thyroid disease, both IFN-AT(Eu) and IFN-AT(Dy) patients showed a significant increase of IFNgamma expression in CD3+CD56+ and CD3-CD56+ cells but not in CD4+ and CD8+ cells. At this time point, IFN-AT(Eu) but not IFN-AT(Dy) patients also showed an increase of IL-4 expression in CD3+CD56+ cells and CD4+ cells. Six months later, IFN-AT(Eu) patients maintained high expression of IL-4 in CD4+ and CD3+CD56+ cells without any further increase in IFNgamma expression. By contrast, IFN-AT(Dy) patients showed an increase of IFNgamma expression in CD4+ and CD8+ cells, with a concomitant decrease of IL-4 expression in CD4+ cells.Type 2 immune response is activated early and specifically in patients with IFN-AT who remain euthyroid throughout the follow-up. Predominant in patients developing thyroid dysfunction, by contrast, is the type 1 immune response that seems to occur earlier in innate than acquired immune system.

The effect of α-interferon (α-IFN) and ribavirin (RBV) treatment on oxidative status in chronic hepatitis C (CHC) is unknown. Aim: To study the time course of oxidative status in patients with CHC during α-IFN and RBV administration, and to evaluate the role of oxidative status in order to predict the therapeutic response. Patients and methods: Fifty one patients with CHC were studied. All received a combination of α-IFN and RBV for 6 or 12 months in relation to the type of response. The hydroperoxides concentration in serum test samples by D-ROM test was measured in all of the patients before therapy. In 27 patients, hydroperoxides were also measured during the treatment and during the 12 subsequent months. Results: Cross-sectional analysis demonstrates that patients with a successive long-term response had a lower basal serum hydroperoxide concentration than non-responders (280 ± 40.8 vs 337 ± 83 CARR Units, p < 0.05). This resulted to be an independent factor predictive of long-term response in the multi-varied analysis. Longitudinal observation on 27 patients showed that the mean hydroperoxide concentration decreased significantly during treatment (T0 329 ± 79.2 vs T12 272 ± 34.5 CARR Units) and that the decrease in the mean values was mainly due to variations in the relapsers group. Conclusions: Normal basal hydroperoxide concentration helps to predict long-term response to combination therapy. The D-ROM test may be used for screening patients before treatment. © 2004 Taylor & Francis Ltd.

GOALS: The authors conducted a study to verify whether supplementation with an antioxidant-rich tomato-based functional food reduces anemia during pegylated interferon and ribavirin therapy for chronic hepatitis C. BACKGROUND: Oxidative stress plays a major role in the physiopathology of hemolytic anemia during ribavirin therapy. The efficacy of antioxidant supplementation with vitamins C and E as pure compounds, is still controversial. METHODS: A functional food with a high content of natural antioxidants and with high carotenoid bioavailability was developed. The authors enrolled 92 patients with chronic hepatitis C, treated with standard combination therapy. Forty-six of them received a daily dose (100 g) of functional food (group 1), and 46 did not (group 2). The effect of antioxidant activity was assessed comparing compliance with the full dose of ribavirin and hemoglobin levels during the first 3 months of treatment. RESULTS: Only 8.7% of patients in group 1 had to reduce their daily ribavirin dose, whereas ribavirin reduction was necessary for 30.4% of patients in group 2 (P = 0.09). Hemoglobin levels showed significant differences at 15, 30, and 90 days during the observation time. CONCLUSION: Results demonstrated that the authors' functional food reduces the severity of ribavirin-related anemia and improves the tolerance to the full dose of ribavirin in patients with chronic hepatitis C.

BACKGROUND Although experimental studies have demonstrated an important role of insulin-like growth factor I (IGF-I) in hepatocarcinogenesis, the clinical data about IGF-I in patients with hepatocellular carcinoma (HCC) are scarce and controversial. To the authors' knowledge, this is the first prospective study investigating the longitudinal correlation between modifications in serum IGF-I levels and the development of HCC in a cohort of patients with hepatitis C virus (HCV)-related cirrhosis. METHODS One hundred fourteen consecutive patients with HCV-related Child Grade A cirrhosis were followed prospectively at the Second University of Naples for 56.4 ± 12.0 months with ultrasound examinations of the liver and serum α-fetoprotein determination every 6 months. At each clinical evaluation, the severity of disease was graded according to the established Child–Pugh scoring system. Serum IGF-I levels were measured prospectively at the study entry and at least every 12 months throughout follow-up. RESULTS Twenty patients (19.2%) developed HCC during follow-up. Eleven of these patients had persistent Child Grade A cirrhosis for the whole study, whereas the other 9 patients developed HCC after their cirrhosis progressed from Child Grade A to Grade B. In patients who remained free of HCC for the whole study, serum IGF-I concentrations did not modify significantly during follow-up. Conversely, in patients who developed HCC, IGF-I levels decreased significantly during follow-up (from 72.6 ± 29.9 μg/L to 33.8 ± 14.5 μg/L; P = 0.001). In these patients, the significant decrease occurred both in patients with persistent Child Grade A cirrhosis and in patients with cirrhosis that progressed from Child Grade A to Grade B. The reduction in IGF-I level preceded the diagnosis of HCC by 9.3 ± 3.1 months. CONCLUSIONS This prospective study demonstrates that, in patients with HCV-related cirrhosis, 1) the development of HCC is accompanied by a significant reduction of serum IGF-I levels independent of the grade of impairment of liver function; and 2) modification of the IGF-I level precedes the morphologic appearance of HCC, permitting a precocious diagnosis of the tumor. Cancer 2002;95:2539–45. © 2002 American Cancer Society. DOI 10.1002/cncr.11002

Oxidative stress has been clearly implicated in human disease by a growing body of scientific evidences. There is no ideal method for the measurement of this parameter. A possible strategy would be to measure simultaneously several biomarkers representing damage to different cellular components or, alternatively, a method able to evaluate the hydroperoxides, intermediate products of oxidation originating from different classes of molecules, such as lipids, peptides, amino acids, etc. can be used. We are introducing a simple, rapid and inexpensive assay to measure the oxidative status of human plasma. It is based on the properties of N,N-dimethyl-p-phenylene-diamine (DMPD), a compound able to produce a fairly long-lived radical cation. The absorbance at 505 nm of a DMPD solution in the presence of plasma, which is proportional to the amount of hydroperoxyl compounds, is related to the oxidative status of the sample and could be expressed as hydrogen peroxide equivalents (HPE). This assay was not influenced by freezing-thawing and storage time of the plasma samples. The assay can be automated, performed in a kinetic mode, and used for routine analyses. The DMPD assay alone or in combination with analytical methods for assessing antioxidant capacity is suggested as a reliable tool to obtain information in pathologies related to oxidative stress.

OBJECTIVE: The aim of this study was to investigate whether the addition of ribavirin (RIBA) to interferon-alpha (IFN-alpha) therapy increases the risk of developing thyroid autoimmunity and/or dysfunction. DESIGN AND METHODS: The study group (group A) included 72 patients undergoing treatment with IFN-alpha (3-6 million units three times weekly) plus RIBA (1.0-1.2 g/day) for chronic hepatitis C (CHC), as first line therapy (30 cases) or as a second therapeutic attempt after a previous ineffective IFN-alpha treatment (42 cases). The control group (group B) encompassed 75 age- and sex-matched patients affected by CHC, undergoing treatment with IFN-alpha alone as first line therapy (35 cases) or as a second therapeutic attempt (40 cases). Thyroid autoimmunity and function were retrospectively evaluated on frozen aliquots, drawn before, after 6 months, and at the end of the antiviral treatment. In patients receiving two antiviral treatments (42 cases in group A and 40 cases in group B) thyroid parameters were also assayed on serum samples drawn before and at the end of the first IFN-alpha therapy. RESULTS: Thyroid autoimmunity rate (17/72 for group A and 17/75 for group B) as well as anti-thyroglobulin antibodies (TgAb) and anti-thyroid peroxidase antibodies (TPOAb) serum levels were comparable between the two groups. Similarly, in patients undergoing two consecutive antiviral treatments (42 cases in group A and 40 cases in group B) the percentage of positivity for thyroid autoantibodies did not change significantly from the first to the second therapeutic schedules in both groups, with no significant increase of median TgAb and TPOAb levels. By the same token, all but one patient negative for thyroid autoantibodies at the end of the first treatment remained so also during the subsequent treatment. In group A patients, the rate of hypothyroidism (11/72) was significantly higher than that observed in group B (3/75). Similarly, in patients undergoing two consecutive antiviral treatments the percentage of hypothyroidism increased significantly from the first to the second therapeutic schedule in group A (from 4.8% to 19.0%; P