bluuebunny/crossref_metadata_2025 huggingface dataset:
Molecular characterization and dynamics of hepatitis C virus replication in human fetal hepatocytes infected in vitro
Authors: Iacovacci S , Manzin A , Barca S , Sargiacomo M , Serafino A , Valli M B , Macioce G , Hassan H J , Ponzetto A , Clementi M , Peschle C , Carloni G
Published: July 2004

The molecular features of hepatitis C virus (HCV) replication in human fetal hepatocytes (HFHs) were addressed in this study. Using a competitive reverse-transcription polymerase chain reaction (RT-PCR) assay for the quantitation of HCV-RNA molecules, the highest level of viral replication was detected 30 days' postinfection. At this time point, viral particles of 41 to 45 nm in diameter accumulated in the cell cytoplasm. Their density in cell extracts and culture medium was distributed between heavy (1.180-1.360 g/cm3) and light fractions (1.105-1.050 g/cm3) of a sucrose gradient, while, in the serum inoculum, they had a positive fraction at 1.180 g/cm3. In infected HFHs, minus-strand HCV RNA was observed in fractions displaying a sedimentation coefficient of 28 S to 18 S, while plus-strand HCV RNA showed a peak restricted to the 21 S fraction; the HCV RNA of serum inoculum had a sedimentation coefficient of 38 to 40 S, which revealed the presence of HCV RNA of unique positive polarity. The 21 S RNA fraction of cell extracts was resistant to 20 minutes of RNase I digestion, while the same incubation time totally inactivated a comparable amount of HCV RNA purified from the serum inoculum, revealing the presence of completely and/or partially double-stranded HCV-RNA molecules in the infected cells. Detection in HFHs of replicative forms and replicative intermediates suggests that the dynamic profile of HCV replication in these cells is similar to that described in other flaviviruses.
Long-term outcome of hepatitis B e antigen-positive patients with compensated cirrhosis treated with interferon alfa
Authors: Fattovich G , Giustina G , Realdi G , Corrocher R , Schalm S W
Published: July 2004

The aim of this study was to evaluate whether interferon alfa (IFN-α) treatment-associated virological and biochemical remission improves survival in a cohort of 90 white patients with compensated cirrhosis caused by hepatitis B (Child A) followed for a mean period of 7 years. Inclusion criteria were biopsy-proven cirrhosis, hepatitis B e antigen (HBeAg) positivity, abnormal serum aminotransferase levels, exclusion of hepatitis delta virus, and absence of complications of cirrhosis. Of the 40 IFN-treated patients, 27 (67%) showed sustained HBeAg loss with alanine aminotransferase (ALT) normalization. Of the 50 untreated patients, 30 (60%) cleared HBeAg, but only 21 (42%) normalized ALT after HBeAg loss. Compared with the untreated patients, IFN-treated patients had similar cumulative rates of HBeAg clearance (P = .48), but higher rates of ALT normalization (P = .016) and of HBsAg loss (P = .028). During follow-up, liver-related death occurred in 8 treated patients, caused by liver failure in 5 and hepatocellular carcinoma (HCC) in 3; all 8 had continued to be HBeAg-positive with elevated ALT. None of the treated patients undergoing remission developed liver-related complications. At univariate analysis, life expectancy was longer in treated patients showing sustained remission than in those who did not (5-year survival: 100% vs. 81%; P = .048). Fourteen untreated patients died (from liver failure in 10 and HCC in 4); all but 3 had continued to be HBeAg-positive with elevated ALT. Cox's model identified age and ALT normalization as the only significant predictors of survival. In conclusion, in patients with HBeAg-positive compensated cirrhosis, virological and biochemical remission following IFN therapy is associated with improved survival.
Inhibition of hepatocellular carcinoma development in hepatitis B virus transfected mice by low dietary casein
Authors: Cheng Z , Hu J , King J , Jay G , Campbell T C
Published: July 2004

In a comprehensive human ecological study, primary liver cancer has been shown to be highly significantly associated with 1) the prevalence of persistent infection with hepatitis B virus (HBV) and 2) plasma cholesterol concentrations that are, in turn, associated with the consumption of animal based foods. In rat studies, aflatoxin-induced hepatocellular carcinoma is substantially prevented by decreasing the intake of animal based protein (casein), a hypercholesterolemic nutrient. Thus the development of primary liver cancer associated with persistent HBV infection or with aflatoxin exposure may be controlled by reduced intake of animal-based proteins. Transgenic mice transfected with an HBV gene fragment containing the viral transactivator of hepatis B virus, HBx, which induces the formation of hepatocellular carcinoma, were used to examine the ability of dietary casein to modify tumor formation. Reducing the concentration of dietary casein to 6% from the traditional level of 22% markedly inhibited (by 75%) hepatic tumor formation in these transgenic mice. Tumor development also was substantially altered by interchanging dietary casein concentration well after tumor development had begun (at 8 months), increasing by 173% from the expected yield when casein intake was increased and decreasing by 99% when casein was reduced. These findings suggest that the development of liver tumor formation among individuals persistently infected with HBV may be controlled by minimizing or eliminating the intake of animal protein-based foods.
Blunted responsiveness of the neuronal activation marker Fos in brainstem cardiovascular nuclei of cirrhotic rats
Authors: Breitman D R , Lee S. S.
Published: July 2004

Cardiovascular function in cirrhosis is deranged, with indirect evidence of abnormal central cardiovascular regulation. We aimed to elucidate the role of brainstem cardiovascular nuclei in hemodynamic regulation by examining the protein product, Fos, of the immediate-early gene c-fos, in cirrhotic rats. Cirrhosis was induced by chronic bile duct ligation (BDL) of 25-days duration, while controls underwent a sham operation. To examine the effects of jaundice per se in the absence of cirrhosis, a third group of 5-day BDL rats was also studied. All rats were anesthetized with pentobarbital, and catheters were inserted to measure baseline blood pressure and heart rate. Separate groups were then subjected to volume manipulation by a hypotensive hemorrhage or isotonic saline infusion, or no challenge. Ninety minutes after the volume manipulation, the animals were killed and the medulla sectioned and stained for Fos by immunohistochemisty. The nucleus tractus solitarius (NTS) of the sham-operated unchallenged rats showed scant Fos immunoreactivity (27.8 ± 3.3 cells), but both hemorrhage and volume infusion significantly increased Fos staining (86.0 ± 3.7 and 95.2 ± 8.5, respectively). In contrast, the unchallenged cirrhotic rats showed markedly increased Fos in the NTS (154.6 ± 27.0), but neither hemorrhage nor volume infusion significantly changed the amount of Fos staining. Fos staining in the ventrolateral medulla (VLM) followed a similar pattern with low staining in the unchallenged sham rats and increased staining in the other groups, but no differences between the unchallenged and the volume-manipulated cirrhotic groups. The 5-day BDL jaundiced rats showed no baseline increase in Fos staining, nor any significant increase after hemorrhage. These results showing baseline activation of central neuronal regions responsible for blood pressure homeostasis, but completely blunted responsiveness in cirrhotic rats, confirm a central origin of disordered cardiovascular regulation. The presence of jaundice may also contribute to the central cardiovascular hyporesponsiveness.
Indomethacin normalizes intracranial pressure in acute liver failure: A twenty-three-year-old woman treated with indomethacin
Authors: Clemmesen J O , Hansen B A , Larsen F S
Published: July 2004

In patients with acute liver failure, cerebral herniation is a common cause of death. The present study reports the effect of indomethacin on four occasions of intracranial hypertension, in a 23-year old previously healthy woman with severe acetaminophen poisoning. During each episode of intracranial hypertension, the patient was treated with 25 mg of indomethacin, and each time the intracranial pressure normalized. We recommend further controlled studies to determine the exact effect of indomethacin on cerebral blood flow and metabolism before it is recommended for treatment of intracranial hypertension in patients with acute liver failure.
Modulation of liver canalicular transport processes by the tyrosine-kinase inhibitor genistein: Implications of genistein metabolism in the rat
Authors: Jager W , Winter O , Halper B , Salamon A , Sartori M , Gajdzik L , Hamilton G , Theyer G , Graf J , Thalhammer T
Published: July 2004

Rat liver cells express the multispecific organic anion transporter (cmoat, cmrp, mrp2) and P-glycoprotein (Pgp) in their canalicular membranes, proteins that are homologous to the multidrug-resistance related protein (MRP) and multidrug resistance (MDR) gene products in multidrug resistant tumor cells. We tested whether genistein, a modulator of drug resistance in tumor cells, affects biliary secretion of substrates of canalicular multispecific organic anion transporter (cmoat) (glucuronides of bilirubin and rhodamine, glutathione conjugate of bromsulphthalein) and of P-glycoprotein (Pgp) (rhodamine), respectively. Using the isolated perfused rat liver of control Wistar rats (TR+) and of a mutant strain (TR-) that expresses Pgp but not cmoat, we show that genistein effectively inhibits the secretion of anionic substrates of cmoat in Wistar rats but stimulates secretion of cationic rhodamine in TR-rats. Genistein is subject to glucuronidation and sulfatation and secretion of genistein and its metabolites stimulates bile flow in Wistar rats, but secretion is nearly absent in TR-rats. Because genistein and its metabolites are substrates for cmoat, inhibition of anion secretion by genistein is partially explained by competition for this transporter. Genistein is also a substrate of uridindiphosphate (UDP)-glucuronyltransferase isoenzyme(s). Inhibition of glucuronidation reduces the availability of bilirubin and rhodamine glucuronates for transport via cmoat, but unconjugated cationic rhodamine becomes available for transport via Pgp at an increased cellular concentration. Daidzein, a genistein analogue with no effect on protein tyrosine kinase (PTK) shows Similar effects on secretion of organic anions and cations supporting the conclusion that genistein affects transport in liver mainly through competition with other substrates at the sites of glucuronidation and transport via cmoat.
Hepatic expression of c-Myb in chronic human liver disease
Authors: Kitada T , Seki S , Nakatani K , Kawada N , Kuroki T , Monna T
Published: July 2004

C-Myb is a sequence-specific DNA binding protein that regulates the expression of genes involved in cell proliferation and differentiation. The present study was designed to elucidate the role of c-Myb in the pathogenesis of chronic liver disease by an immunohistochemical approach. In normal (control) livers, few or no hepatic cells were positive for c-Myb. In livers from patients with chronic viral hepatitis, positive staining for c-Myb was found not only in spindle-shaped mesenchymal cells in expanded portal areas, but also in perisinusoidal cells (PSCs) within liver lobules. In cirrhotic livers, a few PSCs within lobules were positive for c-Myb, while no staining was seen in fibrous septa. Immunoelectron microscopy revealed that c-Myb-positive PSCs displayed morphological features of hepatic stellate cells. Other sinusoidal lining cells including Kupffer cells and sinusoidal endothelial cells, as well as hepatocytes, were all negative for c-Myb. Dual c-Myb/α-smooth muscle actin (αSMA) staining revealed that more than 97% of c-Myb-positive cells were αSMA-positive. Moreover, dual c-Myb/proliferating cell nuclear antigen (PCNA) staining showed that approximately 70% of c-Myb-positive cells also expressed PCNA. The labeling index (LI) (number of c-Myb-positive cells/0.1 mm2) significantly correlated with serum transaminase concentrations and increased in parallel with the disease activity. However, the LI showed no correlation with the degree of fibrosis. These results suggest that c-Myb may be involved in stellate cell activation and proliferation in chronically diseased human livers, and that the level of c-Myb expression is associated with the activity of chronic hepatitis.
Interleukin-6 protects liver against warm ischemia/reperfusion injury and promotes hepatocyte proliferation in the rodent
Authors: Camargo C A , Madden J F , Gao W , Selvan R S , Clavien P
Published: July 2004

Interleukin-6 (IL-6) is an acute reactant cytokine with anti-inflammatory properties, which has been found to prevent injury in a model of acute hepatitis in mice through downregulation of tumor necrosis factor α (TNF-α); to correlate inversely with markers of hepatocellular injury in patients with liver ischemia; and to initiate liver regeneration in mice. In this study, we investigated the role of IL-6 in rodent models of hepatic warm ischemia/reperfusion (WI/Rp) injury. IL-6-deficient mice (-/-) were subjected to hepatic WI and compared with C57BL/6 mice, as well as IL-6 -/- mice pretreated with recombinant IL-6 (rIL-6). The effects of rIL-6 following various periods of ischemia were further studied in models of hepatic ischemia in rats. IL-6 -/- mice had increased reperfusion injury as assessed by transaminase levels and a tissue necrosis scoring system when compared with controls, an effect prevented by pretreatment with rIL-6. Similarly, rats pretreated with rIL-6 had reduced reperfusion injury and better survival than controls in each respective WI group. Tissue TNF-α expression measured by Northern blot analysis and serum C-reactive protein (CRP) levels, a marker of inflammation, were significantly reduced in animals pretreated with rIL-6. Administration of antibodies to TNF-α reproduced the beneficial effect of rIL-6. Hepatocyte proliferation, as assessed by a scoring method for mitotic index and proliferating nuclear cell antigen staining, was markedly increased in rIL-6-treated rats when compared with controls. In conclusion, this study suggests that IL-6 could play an important role in limiting hepatic warm ischemia/reperfusion (WI/Rp) injury, probably through its anti-inflammatory properties, modulation of TNF-α, and/or promotion of liver regeneration. rIL-6 might become an important cytokine in clinical situations associated with WI/Rp injury.
Dual expression of matrix metalloproteinase-2 and membrane-type 1-matrix metalloproteinase in fibrotic human livers
Authors: Takahara T , Furui K , Yata Y , Jin B , Zhang L P , Nambu S , Sato H , Seiki M , Watanabe A
Published: July 2004

We have previously reported increased expression of matrix metalloproteinase-2 (MMP-2) using a rat model of liver fibrosis. However we did not clarify how the precursor of MMP-2 (proMMP-2) was activated. Therefore, we used human liver specimens with chronic hepatitis (CH) and liver cirrhosis (LC) to examine expression of membrane-type-1-MMP (MT1-MMP), which has recently been determined to activate proMMP-2. Northern hybridization studies showed a 5.4- and 1.4- fold increase in MMP-2 expression in CH and LC, respectively, as compared with normal liver. MT1-MMP gene expression simultaneously increased 4.0- and 1.4-fold in CH and LC, respectively. In situ hybridization using 35S-cRNA probes of MMP-2 and MT1-MMP showed prominent silver granules in elongated cells found in the lobules, periportal areas, and fibrous septa of CH and LC samples. These elongated cells expressed α-smooth muscle actin by immunohistochemistry. Immunoelectron microscopic examination localized MMP-2 and MT1-MMP to the rough endoplasmic reticulum of stellate cells located in the lobules and periportal areas, or to fibroblasts in the fibrous septa, suggesting that MMP-2 and MT1-MMP were produced by these cells. In addition, cytoplasmic and membranous immunodeposits of both MMPs were found in endothelial cells, Kupffer cells, capillary endothelial cells, and lymphocytes, indicating that activation of proMMP-2 occurs locally. Increased expression of MMP-2 and MT1-MMP was detected in CH and LC, while dual over-expression was found in stellate cells and fibroblasts, possibly resulting in the increase of active MMP-2 in and around these cells. These findings suggest that activated MMP-2 may remodel liver parenchyma during the process of liver fibrosis.
Neonatal granulocytosis is a postpartum event which is seen in the liver as well as in the blood
Authors: Kawamura T , Toyabe S , Moroda T , Iiai T , Takahashi-Iwanaga H , Fukada M , Watanabe H , Sekikawa H , Seki S , Abo T
Published: July 2004

In a recent series of studies, we demonstrated that stress in humans and animals, with resultant sympathetic nerve strain, induces severe granulocytosis, because granulocytes carry adrenergic receptors on the surface. Because activated granulocytes produce free radicals and superoxides, they sometimes induce tissue damage if the stress is too strong or continuous. Human neonates are also known to show high levels of granulocytes in the peripheral blood. In this study, we investigated whether such neonatal granulocytosis are a stress-associated response at birth. Both human and mouse materials, before and after birth, were used. The number of leukocytes in the blood, as well as some other factors in the serum, were measured. Although levels of granulocytes were found to be low in fetal humans and mice, they increased sharply after birth. In parallel with this postpartal granulocytosis, transaminases in sera increased transiently. In reference to results of a transient elevation in the levels of catecholamines at birth in mice, all these phenomena resemble stress-associated responses. Indeed, fatty liver and hematopoietic destruction in the liver were also observed in mice and humans. At this time, the production of inducible nitric oxide synthase (iNOS) by granulocytes in the liver was evident. These results suggest that neonatal granulocytosis is a postpartum event which results from various stresses (e.g., oxygen stress) at birth. This event may be responsible for such well-known neonatal phenomena as the termination of fetal hematopoiesis in the liver and as neonatal jaundice.
Nucleotide sequence variations in the internal ribosome entry site of hepatitis C virus-1b: No association with efficacy of interferon therapy or serum HCV-RNA levels
Authors: Yamamoto C , Enomoto N , Kurosaki M , Yu S , Tazawa J , Izumi N , Marumo F , Sato C
Published: July 2004

The extreme 5′-proximal sequences of the hepatitis C virus (HCV) genome including the 5′untranslated region (5′UTR) and the first 30 nucleotides of the core region are highly conserved, and serve as an internal ribosome entry site (IRES) that initiates the cap-independent translation of HCV polyprotein. Mutations in the IRES sequence have been shown to cause changes in the efficiency of protein translation in vitro. However, the significance of genetic variations in the IRES is not fully known in clinical settings. Pretreatment sera of 25 patients with HCV-1b infection who were treated with interferon were amplified by polymerase chain reaction (PCR), and the IRES sequence was directly sequenced. Correlation of interferon responses or other clinical features with IRES sequence variability was studied. Eleven of 25 patients were sustained responders (SR) of interferon treatment (negative serum HCV RNA and normal alanine transaminase levels for 6 months after the end of interferon treatment), and the other 14 patients were nonresponders ([NR], defined as any patient with positive serum HCV RNA within 6 months after the end of interferon therapy). In each patient, one to four nucleotide substitutions were found compared with the consensus sequence of HCV-1b genotype. There were no differences in the number of nucleotide substitutions between either SR and NR (mean, 1.8 in SR, 2.1 in NR; P = .30), and no specific variations associated with SR or NR were observed. Although NR had significantly higher serum levels of pretreatment HCV RNA than SR (median, 16 vs. <0.5 Meq/mL; P = .02), there was no correlation between the HCV-RNA level and the number of nucleotide substitutions in the IRES (mean, 1.9 nucleotide substitutions in 12 patients with HCV RNA <0.5 Meq/ mL vs. 2.1 nucleotide substitutions in 13 patients with HCV RNA >0.5 Meq/mL; P = .61). Sequence variability of the IRES has no influence on interferon efficacy or serum HCV-RNA concentrations in patients with chronic HCV-1b infection.
Frequent detection of hepatitis A viral RNA in serum during the early convalescent phase of acute hepatitis A
Authors: Fujiwara K , Yokosuka O , Ehata T , Imazeki F , Saisho H , Miki M , Omata M
Published: July 2004

The diagnosis of type A hepatitis is performed mainly by immunoglobulin M (IgM) anti-hepatitis A antibody assay, but it has not been established whether there is a correlation between changes in viremia and the clinical course of type A hepatitis. We examined hepatitis A virus (HAV) RNA in the sera from type A and non-A, non-B, non-C acute hepatitis and analyzed the relation of HAV viremia with alanine aminotransferase (ALT) and IgM-HA levels. Two hundred sera from 38 patients with type A acute hepatitis and 20 patients with non-A, non-B, non-C acute hepatitis were examined for the presence of HAV RNA. HAV RNA was detected by nested reverse transcription-polymerase chain reaction (RT-PCR) with primers located at the 5′ non-translated region of HAV. HAV RNA was detected in 35 of 38 (92%) type A hepatitis patients and in 60 of 156 (38%) serum samples. In contrast, it was detected in none of 44 serum specimens from 20 non-A, non-B, non-C acute hepatitis patients. In type A hepatitis, the mean ALT level in HAV RNA positive specimens was 1,481 ± 2,042 (range, 20-10,370) IU/L and that in HAV RNA negative specimens 186 ± 330 (range, 8-1,698). The positivity of HAV RNA was correlated with the level of transaminase at the time of sample collection. The mean duration from the onset of symptoms to disappearance of HAV RNA was 18 ± 14 days. The mean titer of IgM-HA in HAV RNA positive cases was 5.0 ± 1.4, in negative cases 5.7 ± 1.1, with no statistical difference. Our results indicate that HAV RNA in serum is detectable in the majority of type A hepatitis cases in their early convalescent phase by nested RT-PCR.
Prediction of response during interferon alfa 2b therapy in chronic hepatitis C patients using viral and biochemical characteristics: A comparison
Authors: Tong M J , Blatt L M , McHutchison J G , Co R L , Conrad A
Published: July 2004

Patients with chronic hepatitis C (n = 103) were treated for 24 weeks with interferon alfa 2b and followed up for 24 weeks after cessation of therapy (week 48). When hepatitis C virus (HCV) RNA at week 48 was used to assess interferon response, 15 (14.6%) were virological complete responders, and all have remained HCV RNA negative for a mean of 3 years. At week 48, 3 of 15 virological complete responders had elevated alanine transaminase (ALT) values. When serum ALT level was used at week 48 to determine response to interferon, 20 (19.4%) were biochemical complete responders. However, 8 of the 20 patients with normal ALT levels were HCV RNA positive at week 48, and 7 of these individuals have had a recurrence of elevated ALT levels within 3 years after cessation of treatment. These findings indicate that measurement of HCV RNA was more accurate than ALT in determining true responses to interferon therapy. Identification of nonresponders early during the course of interferon treatment showed that an elevated ALT level at week 12 was 92% predictive (odds ratio 3.7) but misidentified 33% (5 of 15) of the patients who were virological complete responders at week 48. In contrast, a positive HCV RNA at week 12 of treatment was 98% predictive (odds ratio 35.5) and misidentified only 6.7% (1 of 15) of the virological complete responders. Thus, positive HCV RNA at week 12 of therapy was more accurate in identifying eventual virological nonresponders than measurement of ALT at this time. Termination of interferon therapy in patients who were HCV RNA positive at week 12 would result in a 27% reduction in the direct medical costs and keep patients from undergoing unnecessary treatment. Therefore, testing for HCV RNA at week 12 to identify nonresponders and then discontinuing their treatment is practical, cost-efficient and beneficial both to patients and to third-party payers.
A case-control study on GB virus C/hepatitis G virus infection and hepatocellular carcinoma.
Authors: Tagger A , Donato F , Ribero M L , Chiesa R , Tomasoni V , Portera G , Gelatti U , Albertini A , Fasola M , Nardi G
Published: July 2004

A new hepatitis-associated RNA virus of the Flaviviridae family has been identified and named GB virus C/ hepatitis G virus (HGV). We carried out a case-control study to evaluate the association of HGV infection with hepatocellular carcinoma (HCC). We recruited 170 patients hospitalized for HCC (143 male and 27 female, mean age 64 years) and 306 patients hospitalized for nonliver diseases (controls) in Brescia, Italy. HGV RNA was detected by reverse transcription-polymerase chain reaction (RT-PCR) and antibodies against HGV E2 protein (anti-E2) by an immunoassay test. HGV RNA was found in 8 cases (4.7%) and 4 controls (1.3%). The relative risk (RR) for HGV RNA positivity adjusted for demographic variables and hepatitis B surface antigen (HBsAg), hepatitis C virus (HCV) RNA, and alcohol was 7.3 (95% confidence interval, 1.7-30.6; P = .009). No HGV RNA-positive subject was also positive for anti-E2. Anti-E2 prevalence did not differ significantly between cases (20%) and controls (15.3%), and no RR increase was found by this marker. Among subjects with HGV exposure (HGV RNA plus anti-E2 positive), a greater proportion of cases (40%) than controls (14%) had transfusion history. The possible role of HGV in HCC etiology seems modest because the population-attributable risk is lower (4%) than those for HBsAg (22%), HCV RNA (36%), and heavy alcohol intake (52%). This study supports the hypothesis of an association between HGV infection and HCC, although at present there are insufficient data on the causality of the association.
Hepatitis C and hepatocellular carcinoma
Authors: Di Bisceglie Adrian M
Published: July 2004

Hepatitis C virus (HCV) infection is now recognized to be a major risk factor for hepatocellular carcinoma (HCC), evidenced by finding both antibody to HCV (anti-HCV) and HCV RNA in serum of a substantial proportion of patients with HCC around the world and by the progression of liver disease to cirrhosis and HCC in individual patients infected with HCV. There seems to be an incubation period of two to three decades on average in most cases of HCV-related HCC. HCV infection usually results in development of HCC via cirrhosis, although the possibility of direct carcinogenic effects of HCV is still under study. Possible additional risk factors include infection with HCV genotype 1b, alcohol consumption, and co-infection with the hepatitis B virus. Estimates of the development of HCC among patients with cirrhosis of all types range between 1% and 4% per year. Assuming that 20% of patients with chronic hepatitis C go on to develop cirrhosis over a 10- year period, between 1.9% and 6.7% of all patients with chronic hepatitis C can be expected to develop HCC over the first two decades of infection. Although tests are available to screen for early HCC, the results of treating these small tumors have been disappointing. Thus, it is imperative that cost-effective means be developed for screening and prevention of HCV-related HCC.
Hepatitis C and alcohol
Authors: Schiff E R
Published: July 2004

Chronic alcoholism in patients with chronic hepatitis C appears to cause more severe and rapidly progressive liver disease leading more frequently to cirrhosis of the liver and hepatocellular carcinoma. The primary risk factor for acquiring hepatitis C among alcoholics is injection drug use. However, the epidemiology is not well defined, and other sources of spread must be important. Alcohol intake in excess of 10 g/d has been associated with increased serum hepatitis C viral RNA and aminotransferase levels, the mechanism of which is poorly understood. The histological picture of hepatitis C in patients with chronic alcoholism is typically indistinguishable from chronic hepatitis C in nonalcoholic patients. Interferon therapy is less effective among alcoholic than nonalcoholic patients, even after a period of abstinence. Patients with chronic hepatitis C should restrict their alcohol intake to less than 10 g/d, and if cirrhosis is present or interferon therapy is planned, abstinence from alcohol should be encouraged. Future research efforts should focus on the epidemiology and pathogenesis of combined chronic hepatitis C and alcoholism.
Diagnostic tests for hepatitis C
Authors: Gretch D R
Published: July 2004

Diagnostic tests for hepatitis C can be divided into the following two general categories: 1) serological assays that detect antibody to hepatitis C virus (anti-HCV); and 2) molecular assays that detect, quantify, and/or characterize HCV RNA genomes within an infected patient. Serological assays have been subdivided into screening tests for anti-HCV, such as the enzyme immunoassay (EIA), and supplemental tests such as the recombinant immunoblot assay (RIBA). Three generations of anti-HCV tests have been developed, and each generation has resulted in an improvement in the sensitivity of detecting anti- HCV. Supplemental anti-HCV tests are designed to resolve false-positive testing by EIA, and are appropriately used in low-prevalence settings in which false-positive anti-HCV tests remain a problem. Third- generation anti-HCV tests (EIA-3 and RIBA-3, respectively) contain antigens from the HCV core, nonstructural 3, nonstructural 4, and nonstructural 5 genes. Detection of HCV RNA in patient specimens by polymerase chain reaction (PCR) provides evidence of active HCV infection and is potentially useful for confirming the diagnosis and monitoring the antiviral response to therapy. Optimal HCV PCR assays at present have a sensitivity of less than 100 copies of HCV RNA per milliliter of plasma or serum. Standardization and proficiency testing of diagnostic laboratories performing HCV PCR remains an important problem for future study. Two main technologies exist for assessing HCV RNA levels or viral load. Quantitative PCR is the most sensitive test for determining hepatitis C viral load, whereas the branched-chain DNA test appears to be the most precise method. Major limitations of the current tests are inadequate dynamic range and high variability of PCR- based assays, and poor sensitivity of the branched-chain DNA test. Molecular tests have also been developed to classify HCV into distinct genotypes; the clinical importance of HCV genotype determination remains a subject for future investigation.
The role of liver biopsy in hepatitis C
Authors: Perrillo R P
Published: July 2004

This article reviews the ways in which liver biopsy provides a resource to the clinician when making management decisions for patients with chronic hepatitis C. Liver biopsy provides information about the extent and distribution of inflammation and allows grading and staging of the disease (the amount of fibrosis). Furthermore, the liver biopsy allows some assessment of the rate of disease progression whenever the date of onset of infection is known. The presence of diffuse fibrosis or cirrhosis correlates with a lower likelihood of response to antiviral therapy, and the finding of severe necroinflammatory and fibrotic changes is helpful in determining the relative importance of beginning treatment early rather than deferring therapy. The importance of liver biopsy in the selection of patients for treatment is underscored by the fact that hepatitis C is a condition in which therapy is currently unable to successfully treat the majority of patients. There are no strong correlations between serum viral RNA, alanine aminotransferase (ALT) levels, and liver histopathology; therefore, the utility of aminotransferases and hepatitis C virus (HCV) RNA as surrogate tests for measuring disease activity in individual cases is limited. It is concluded that liver biopsy should be performed before initiating interferon therapy. At the present time, it is not known if and when repeat histological assessment is necessary. Preferably, it should only be performed in instances when the findings from the biopsy are likely to lead to a change in management of the patient.
Sexual and perinatal transmission of hepatitis C
Authors: Dienstag J L
Published: July 2004

Such nonpercutaneous routes of hepatitis C virus (HCV) transmission as sexual and perinatal spread are relatively inefficient. Several observations have been cited to support a role for sexual transmission of hepatitis C. Approximately 10% of persons with reported cases of acute hepatitis C in the United States report a history of potential sexual exposure. Anecdotal cases of sexual transmission have been reported, and HCV nucleotide sequence homology has been observed in viral isolates from sexual partners. Similarly, the prevalence of HCV infection is increased in groups with a high risk of exposure to sexually transmitted viral infections. Other observations, however, weigh against sexual transmission of HCV infection. Sexual transmission is negligible in sex-partner studies; alternative risk factors account for many cases of apparent sexual transmission between sexual partners; the prevalence of HCV infection in high-risk groups is much lower than that of other sexually transmitted infections; and the risk of apparently sexually transmitted HCV infection does not always correlate with intensity and duration of sexual exposure. The United States Public Health Service has estimated that the risk of sexual transmission is approximately 5%, well below the risk of sexual transmission of hepatitis B or human immunodeficiency virus (HIV). Similarly, perinatal HCV infection, though documented to occur, is unusual, except in babies born to mothers with very high levels of HCV RNA, including mothers with concomitant HIV infection. Weighing many, often conflicting reports, the United States Public Health Service has estimated that the likelihood of perinatal infection is low, on the order of 5% to 6%, and that breast feeding does not increase the risk of HCV infection in infants of mothers with hepatitis C. Current data do not support household exposure as a risk for HCV infection.
Management of hepatitis C: A national survey of gastroenterologists and hepatologists
Authors: Everhart J E , Stolar M , Hoofnagle J H
Published: July 2004

A survey on the management of hepatitis C virus (HCV) infection was conducted by the American Digestive Health Foundation among United States physicians who are most familiar with the disease. The two-page survey was completed by 57% of a random sample of 2,500 members of the American Gastroenterological Association and the American Association for the Study of Liver Diseases. Analysis was limited to the 1,249 responses from physicians who spent at least 1 day per week in patient care. These physicians frequently managed patients with HCV, and nearly three quarters treated patients with interferon. To prevent transmission, the large majority of physicians recommended measures to avoid blood exposures, were uncertain or disagreed about the importance of sexual contact, and did not caution patients about casual contact. More than 70% of physicians told their patients to stop or minimize alcohol consumption. In the management of a patient with antibody to HCV but normal serum aminotransferase activities, 87% of physicians would have ordered a supplemental test, and if HCV were confirmed, 46% would have obtained a liver biopsy, but only 15% would have treated the patient with interferon. For a patient with chronic HCV infection and elevated serum aminotransferase activities, more than 90% of physicians would have obtained a liver biopsy and approximately 60% would have treated with interferon. Physicians who are most familiar with the management of patients with HCV generally agreed with the recommendations of the Consensus Development Conference Panel regarding prevention of transmission, minimizing alcohol consumption, and managing patients with typical presentations. Controversies remain regarding some issues of general management, the value of molecular testing, and the need to treat certain patients with interferon.
Therapy of hepatitis C: Interferon alfa-n1 trials
Authors: Farrell G C
Published: July 2004

Interferon alfa-n1 is produced from a lymphoid cell line and consists of multiple α interferon subtypes. Early studies indicated that interferon alfa-n1 was effective against hepatitis C, and a meta- analysis of published trials indicated that it was equally likely as recombinant α interferons to produce an end-of-treatment biochemical and histological response. However, there appeared to be a lower rate of posttreatment relapse after a 6-month course of interferon alfa-n1, so that the sustained response rate was 25% compared with 16% for recombinant α interferons. Subsequently, the efficacy of interferon alfa-n1 has been compared with recombinant α interferon directly in two large multicenter studies. The 096 International Hepatitis Comparative Study compared alfa-n1 with interferon alfa-2b in doses of 3 million units (MU) three times weekly for 24 weeks in a total of 1,071 patients. Biochemical end-of-treatment response rates were similar (35% for alfa-n1; 38% for alfa-2b), as were virological and histological responses. Relapse occurred more frequently after interferon alfa-2b, so that sustained biochemical (10.3% vs. 6.7%) and virological responses at 12 months were higher for alfa-n1. In a second study from Italy, interferon alfa-n1 was compared with interferon alfa-2a, using higher doses (6 MU three times weekly) until a biochemical response was obtained, and then using 3 MU three times weekly for a total of 12 months. The combined biochemical and virological sustained response rates were higher than reported with 6 months of treatment and were similar for alfa-n1 (17%) as for alfa-2a (16%). The superiority of 12 versus 6 months of treatment was also confirmed in the large multicenter 091 European Comparative Treatment Schedules study of 440 patients given four different regimens of therapy. Sustained biochemical response rates were 6% in patients who received 3 MU three times weekly for 6 months and 19% in those who received this dose for 12 months. When given for 12 months, there was no advantage to a slightly higher dose of interferon alfa-n1 (5 MU three times weekly). Thus, therapy with interferon alfa-n1 produces sustained response rates equivalent to the best obtained with recombinant α interferon. The optimal treatment regimen appears to be 3 MU given three times weekly for 12 months.
Factors predictive of a beneficial response to therapy of hepatitis C
Authors: Davis G L , Lau J Y
Published: July 2004

Alpha interferon is the only drug that has been shown to be effective in the treatment of chronic hepatitis C, but only half of patients respond, either transiently or permanently. Pretreatment features that are associated with a greater likelihood of response to short courses of interferon include low hepatitis C virus (HCV) RNA levels, viral genotypes 2 or 3, and the absence of fibrosis or cirrhosis on liver biopsy. Each of these features is more predictive of sustained response (SR) than the end-of-treatment response (ETR). However, the accuracy of these features in predicting response in individual patients is poor. Furthermore, there are several limitations to using these factors in the clinical management of patients. Most importantly, they were identified in 6-month treatment trials. Longer treatment or combination of interferon with ribavirin reduces relapses and will therefore lessen the association of these factors with long-term response. In addition, changes in the definition of treatment end points and the technology used to measure HCV RNA might change the association between these predictive factors and response. The best predictor of a treatment response is the early normalization of the serum alanine aminotransferase (ALT) level during interferon treatment. HCV RNA loss during treatment may also be helpful in predicting response, but it is probably no better than serum ALT levels and is expensive. In summary, several clinical and virological features are associated with higher response rates to interferon treatment. Although pretreatment factors do not accurately predict treatment outcome in individuals, they may be helpful in counseling patients and making treatment decisions.
Therapy of hepatitis C: Patients with cirrhosis
Authors: Schalm S W , Fattovich G , Brouwer J T
Published: July 2004

The management of both compensated and decompensated cirrhosis caused by hepatitis C must be viewed in the context of the natural history of the disease. The prognosis of compensated cirrhosis caused by hepatitis C is relatively good. In a recent retrospective study, after 5 years of follow-up evaluation, 18% of patients had developed hepatic decompensation and 7% hepatocellular carcinoma. Overall 5-year survival rate was 91%. Treatment with α interferon appears to decrease the incidence of hepatocellular carcinoma in patients who achieve a sustained remission. However, on an intention-to-treat basis and after adjustment for differences in clinical and serological features at entry, interferon therapy does not correlate with a reduced incidence of liver cancer or improved survival. Combined analysis of multiple large studies on patients with cirrhosis caused by hepatitis C indicates that current regimens of α interferon (3 to 6 million units three times weekly for 6 to 12 months) result in a sustained biochemical response in 9% of patients. The rates of sustained virological responses are less well documented. Virological measurements during therapy show that only 22% of patients become hepatitis C virus (HCV) RNA negative by 4 weeks and, thereafter, there is a high rate of breakthrough. In small studies, the combination of interferon and ribavirin leads to sustained biochemical and virological response rates of 21%, more than twice the rates achieved with interferon alone. The prognosis of decompensated cirrhosis caused by hepatitis C is poor, with a 5-year survival rate of only 50%. The efficacy of interferon in patients with decompensated cirrhosis is not well documented and its use cannot be recommended. In contrast, 5-year survival rates after liver transplantation for cirrhosis caused by hepatitis C is excellent, in the range of 70% to 80%. Recurrence of HCV infection occurs in more than 95% of patients, but in short-term follow- up studies, recurrence of cirrhosis and graft failure occurs in only 10% of patients.
Therapy of hepatitis C: Patients with normal aminotransferase levels
Authors: Marcellin P , Levy S , Erlinger S
Published: July 2004

An important group of patients with chronic hepatitis C have normal serum alanine aminotransferase (ALT) levels despite having hepatitis C virus (HCV) RNA detectable in serum. These patients are typically identified after donating blood and being found to be positive for antibody to HCV (anti-HCV). A strict definition of this patient population is needed, which should include presence of anti-HCV, HCV RNA detectable by polymerase chain reaction (PCR), and persistently normal ALT levels. These patients are usually asymptomatic, but almost all have histological evidence of chronic hepatitis on liver biopsy. The liver histological lesions are generally mild, and cirrhosis is rare. The long-term outcome of this group of patients with chronic HCV infection is not known, but the prognosis is probably good. In small, uncontrolled trials of α interferon in this group of patients, end- of-treatment virological responses occurred in approximately 50% and sustained responses 6 to 12 months afterwards in 20%. These rates of response are not very different from those reported in patients with abnormal ALT levels. Importantly, in most studies, serum ALT levels became abnormal during therapy in approximately half of patients, and levels remained abnormal in a proportion after therapy. These findings suggest that α interferon therapy is not usually beneficial and may be harmful in patients with chronic hepatitis C who have normal ALT levels, perhaps as a result of the immunomodulatory actions of α interferon that may alter the balance of host immune reactions and viral replication, which seem to be responsible for the liver injury in chronic hepatitis C.
Therapy of hepatitis C: Cost-effectiveness analysis
Authors: Koff R S
Published: July 2004

The natural history of untreated chronic hepatitis C is controversial, and direct knowledge of the long-term clinical and economic outcomes of current α interferon treatment regimens remains limited. Decision analytic models using available information on outcome probabilities and associated health care costs in the United States have been developed but are available only in abstract form. They suggest that chronic hepatitis C is a life-shortening disease and that α interferon treatment, for 6 or 12 months, despite its up-front costs and failure to induce a prolonged therapeutic response in most patients, increases life expectancy (which nevertheless is still reduced). It does so with a marginal cost-effectiveness well within the acceptable range of medical interventions in the United States. Even empiric therapy, without regard to viral level, genotype, and baseline histology, is within an acceptable cost-effectiveness range. Improving the response rate is likely to make treatment even more cost-effective and possibly cost-saving. Discounting at 3% would also lower the marginal cost-effectiveness; treatment of younger patients would likely lead to cost-savings. Future needs include the development of better databases and cost data for estimating outcomes.
Dysregulation of proteoglycan production by intrahepatic biliary epithelial cells bearing defective (Delta-f508) cystic fibrosis transmembrane conductance regulator
Authors: Bhaskar Ramakrishnan K. , Turner Bradley S. , Grubman Shelley A. , Jefferson Douglas M. , LaMont J.Thomas
Published: July 2004

Hepatic dysfunction in cystic fibrosis (CF) has been attributed to accumulation of viscous mucoid secretions in intrahepatic bile ducts. The purpose of our study was to compare glycoconjugate secretion by intrahepatic biliary epithelial (IBE) cells derived from normal livers and livers of CF patients with the delta F508 mutation of the cystic fibrosis transmembrane conductance regulator (CFTR). Confluent cells were incubated with 3H-glucosamine (GlcN) for 16 hours, and radiolabeled macromolecules were analyzed for the amount and type of glycoconjugates. Incorporation of 3H-GlcN into macromolecular glycoconjugates was two- to threefold higher in CF cells versus normals, as was uptake of 3H-Glcn into the cytoplasm of CF cells. Gel exclusion chromatography on Sepharose Cl 4B revealed that the secreted glycoconjugates from CF cells eluted entirely in the excluded fraction (molecular weight > 2 × 106), while, in the normal cells, 60% of the glycoconjugates eluted as lower-molecular-weight species. The high-molecular-weight glycoconjugates in both CF and normal cells were identified as chondroitin sulfates, as evidenced by susceptibility to beta elimination, chondroitinase digestion, and amino acid composition. Western blotting of IBE cell secretions with a polyclonal antibody to chondroitin sulfate revealed proteoglycan bands at 100 and 210 kd. Our results indicate that secretion of chondroitin sulfate is markedly increased in CF biliary epithelium in vitro compared with non-CF cells. Increased uptake of precursor 3H-GlcN may contribute to enhanced glycosylation of chondroitin sulfate in CF cells.