bluuebunny/crossref_metadata_2025 huggingface dataset:
Liver Transplantation in Patients With Hepatitis B Virus Infection: Outcome in Asian Versus White Patients
Authors: Teo Eng-Kiong , Han Steven-Huy , Terrault Norah , Luketic Velimir , Jensen Donald , Keeffe Emmet B. , Lok Anna S.F.
Published: August 2002

Previous studies have found that Asian patients transplanted for hepatitis B virus (HBV) infection had worse outcomes than white patients. The aim of this study was to compare outcomes in Asian and white patients listed for liver transplantation for HBV infection. Data of all patients with HBV infection listed for liver transplantation between January 1996 and June 1998 from 20 centers in North America were collected using a survey. Total patients enrolled were 325 (171 whites, 126 Asians, 28 other races). There was no difference in demographics, liver biochemistry, and HBV replicative status between Asians and whites at the time of listing. More Asians had hepatocellular carcinoma and fewer Asians had hepatitis C or D virus coinfection. At the time of this survey, 70 Asians (55%) and 99 whites (58%) had been transplanted. Actuarial 2–year survival posttransplantation for Asians (88%) and whites (92%) was similar. Recurrent HBV infection occurred in 8 (11%) Asians and 12 (12%) whites. Five patients with recurrent HBV infection died, 4 of whom were Asian. Actuarial 2–year survival for Asians versus whites with recurrent HBV infection was 60% versus 90% (P = .04). In this large cohort of patients, overall survival and recurrent HBV infection posttransplantation were comparable between Asians and whites. However, Asians with recurrent HBV infection posttransplantation had significantly higher mortality.
Cigarette Smoking and Hepatic Lesions in Patients With Chronic Hepatitis C
Authors: Pessione Fabienne , Ramond Marie-José , Njapoum Corinne , Duchatelle Véronique , Degott Claude , Erlinger Serge , Rueff Bernard , Valla Dominique-Charles , Degos Françoise
Published: August 2002

A possible hepatotoxicity of cigarette smoke has been recently suggested by epidemiological and experimental studies. Our aim was to study the possible relationships between smoking and liver fibrosis and activity in patients with chronic hepatitis C. A cross–sectional study was performed in a group of 310 patients with chronic hepatitis C consecutively hospitalized for their first liver biopsy. The relationships between age, gender, alcohol consumption, route of contamination, tobacco consumption, and Knodell fibrosis and activity scores were examined in univariate, age–adjusted, and multivariate analyses. One hundred and seventy–six patients (57%) were current smokers. Smokers were younger (P < .001), more often of male gender (P = .001), more often alcohol consumers (P = .001), and more often had a history of intravenous drug use (P = .0001) than never smokers. Smoking was related to increased fibrosis and activity scores in age–adjusted (P = .009 and P = .005, respectively) and multivariate analyses (P = .03 and P = .04, respectively). Smoking increases the severity of hepatic lesions in patients with chronic hepatitis C.
Oxidative Stress in Fatty Livers of Obese Zucker Rats: Rapid Amelioration and Improved Tolerance to Warm Ischemia With Tocopherol
Authors: Soltys Kyle , Dikdan George , Koneru Baburao
Published: August 2002

Fatty livers in humans and rats are less tolerant of ischemia, endotoxin, and alcohol. We hypothesized that fatty livers of obese (Ob) Zucker rats are oxidatively stressed and oxidative stress could be relieved by antioxidant treatment, leading to improved tolerance to ischemia. Total glutathione (GSH), tocopherol (TOC), ascorbic acid (AA), catalase (CAT), superoxide dismutase (SOD), and selenium-glutathione peroxidase (Se–GPx) were measured in the livers of Ob and lean (Ln) Zucker rats before and after treatment with high–dose TOC and ascorbate. Also, survival in treated Ob rats following a lethal 90 minutes of partial in vivo warm ischemia was examined. Fatty livers of Ob rats contained significantly less GSH, TOC, and CAT, in comparison with livers of Ln rats. Immunoblotting showed significantly decreased CAT protein without changes in mRNA in fatty livers. There were no significant differences in AA, SOD, and Se–GPx between the 2 groups. Pretreatment with TOC and ascorbate over 48 hours completely corrected the decreases in GSH, TOC, and CAT. Most importantly, TOC with or without ascorbate pretreatment significantly improved survival in Ob rats following ischemia in a dose–dependent manner. In conclusion, TOC administration corrected the oxidative stress in fatty livers of Ob Zucker rats and improved survival following lethal ischemia. Additional studies are needed to determine the efficacy of TOC—a relatively inexpensive agent—in treating patients with fatty livers in a variety of clinical conditions, possibly including liver transplantation.
Temporal Changes in Insulin Sensitivity Following the Development of Acute Liver Failure Secondary to Acetaminophen
Authors: Clark Sarah J. , Shojaee-Moradie Fariba , Croos Premila , Seed Paul T. , Umpleby Margot A. , Wendon Julia A. , Miell John
Published: August 2002

Insulin resistance in chronic liver disease (CLD) is well documented. This study investigated whether similar changes occur in acute liver failure (ALF). Patients with ALF (n = 10) were recruited within 72 hours of their peak prothrombin time (range 42–120 seconds). All patients were ventilated for encephalopathy (grade III-IV). Peripheral and endogenous insulin sensitivity were assessed by a hyperinsulinemic euglycemic clamp (Human Actrapid 1.5 mU/min/kg) with an infusion of d–[6,6–2H2] glucose. The clamp was performed on day 0 and then on day 7 and day 14. During the insulin infusion, the mean total peripheral glucose uptake (area under the curve [AUC]) was 1,422 (SD, 1,253), 2,244 (SD, 1,392), and 4,500 (SD, 1,120) μmol/kg on days 0, 7, and 14, respectively. Significant changes occurred from day 0 to 14 (day 14-day 0: 3,078 [95% CI, 1,798 to 4,359]; P = .001) and day 7 to 14 (day 14-day 7: 2,256 [95% CI, 923 to 3,589]; P = .001). No significant difference in endogenous glucose production was demonstrated over time. Mean peripheral insulin sensitivity altered over time, increasing from 0.09 (SD, 0.09) μmol/kg/min/mU/L on day 0 to 0.24 (SD, 0.16) on day 7 and 0.5 (SD, 0.1) on day 14. Significant changes occurred between days 0, 7, and 14 (day 7-day 0: 0.15 [95% CI, 0.04 to 0.26], P = .006; day 14-day 0: 0.4 [95% CI, 0.28 to 0.5], P = .001; day 14-day 7: 0.2 [95% CI, 0.12 to 0.38], P = .001). This study demonstrates that in ALF, impaired peripheral uptake of glucose occurs, peripheral insulin sensitivity being restored at 2 weeks in subjects who survived.
A Cellular Gene Up–Regulated by Hepatitis B Virus-Encoded X Antigen Promotes Hepatocellular Growth and Survival
Authors: Lian Zhaorui , Liu Jie , Pan Jingbo , Satiroglu Tufan Lale N. , Zhu Minghua , Arbuthnot Patrick , Kew Michael , Clayton Marcy M. , Feitelson Mark A.
Published: August 2002

Polymerase chain reaction (PCR) select complementary DNA (cDNA) subtraction of hepatitis B x antigen (HBxAg)–positive compared with –negative HepG2 cells resulted in the up–regulated expression of a cellular gene that encodes a transcript of 745 bases and a polypeptide 99 amino acids long. GenBank analysis revealed extensive homology with the amino terminal domain of cellular multidrug resistant proteins (MRP), although overexpression of this gene did not confer an MRP phenotype. In situ hybridization and immunostaining showed colocalized expression with HBxAg in the liver of hepatitis B carriers. Overexpression of this protein stimulated the growth of HepG2 cells in serum–free medium, and partially protected cells from anti–Fas–mediated killing, but did not promote growth in soft agar or tumor formation in nude mice. Introduction of the dominant negative inhibitor of nuclear factor κB (IκBα) into HBxAg–positive HepG2 cells decreased the levels of messenger RNA (mRNA) and protein, suggesting that its up–regulation is nuclear factor κB (NF–κB) dependent. Hence, HBxAg activation of NF–κB may result in the up–regulation of a cellular protein that promotes growth factor–independent survival and protects against Fas–mediated killing. This factor may contribute to the persistence of infected hepatocytes during chronic infection, which is important for the later development of hepatocellular carcinoma (HCC).
Treatment of Hepatitis C Infection in Injection Drug Users
Authors: Backmund Markus , Meyer Kirsten , Von Zielonka Michael , Eichenlaub Dieter
Published: August 2002

Chronic hepatitis C is the most common infectious disease among injection drug users (IDUs). Because of the allegedly poor compliance of IDUs with treatment requirements and conditions, hepatologists recommend treatment only if former IDUs have spent 6 to 12 months drug free. The aim of this prospective study was to investigate whether opiate–dependent IDUs with chronic hepatitis C virus (HCV) infection can be treated successfully with interferon. Eligibility for the study meant IDUs had to be HCV–RNA positive by polymerase chain reaction. Subsequently 50 inpatients were enrolled during detoxification treatment. HCV treatment was started with interferon alfa–2a (through 1998) or a combined regimen consisting of interferon alfa–2a and ribavirin (begun in 1998). All patients were treated and supervised by specialized physicians in both hepatology and addiction medicine. The end point for this study was defined as a loss of detectable serum HCV RNA at week 24 after treatment. The rate of sustained virologic response was 36%. Sustained response rates were not significantly different for patients who relapsed and returned to treatment (53%), relapsed and did not return to treatment (24%), or did not relapse (40%; P > .05). During the 24 weeks after treatment, we were unable to detect any reinfection, even among patients who injected heroin during this period. This surprising result should be examined in further studies. In conclusion, HCV–infected drug addicts with chronic HCV infection can be treated successfully with interferon alfa–2a and ribavirin if they are closely supervised by physicians specialized in both hepatology and addiction medicine.
Navajo Neurohepatopathy: A Mitochondrial Dna Depletion Syndrome?
Authors: Vu Tuan H. , Tanji Kurenai , Holve Stephen A. , Bonilla Eduardo , Sokol Ronald J. , Snyder Russell D. , Fiore Stephany , Deutsch Gail H. , DiMauro Salvatore , De Vivo Darryl
Published: August 2002

Navajo neurohepatopathy (NNH) is an autosomal recessive disease of full–blooded Navajo children living in the Navajo Reservation of southwestern United States. Clinical features of NNH include peripheral and central nervous system involvement, acral mutilation, corneal scarring or ulceration, liver failure, and metabolic and immunologic derangement. The cause of NNH is unknown, but the clinical features of NNH are similar to those of patients with mitochondrial DNA (mtDNA) depletion. Therefore, we studied mtDNA concentration in the liver from 2 patients with NNH. Using histochemical, biochemical, and molecular techniques, we found evidence of mtDNA depletion, and we propose that the primary defect in NNH is in the nuclear regulation of mtDNA copy number.
Adult Hepatitis B Vaccination Using A Novel Triple Antigen Recombinant Vaccine
Authors: Young Michael D. , Schneider David L. , Zuckerman Arie J. , Du Wei , Dickson Brian , Maddrey Willis C.
Published: August 2002

Present hepatitis B vaccines use multidose prolonged regimens, which even healthcare workers at risk do not always complete. Moreover, when vaccination is completed there remain some who fail to achieve adequate protection. The protection of adults at risk could be improved if there were a more potent vaccine and/or a shorter vaccination regimen available. Vaccine–naive adults were randomized to vaccination with either Engerix–B (SmithKline Biologicals, Rixensart, Belgium) or a novel triple antigen (S, pre–S1, and pre–S2) recombinant vaccine (Hepacare; Medeva Pharma Plc, Speke, UK). The primary efficacy parameter was the degree of seroprotection 6 or 7 months (26 ± 2 weeks) after beginning vaccination. A total of 304 adults entered the study. Of these, 16 failed to complete the study (9 on Hepacare and 7 on Engerix–B). With the Engerix–B standard (0, 1, 6) regimen, 88% of subjects were protected by month 7, whereas with the triple antigen vaccine a 2–dose regimen (0, 1) provided equivalent protection (91%) within 6 months and a 3–dose (0, 1, 6) regimen was significantly superior (98% seroprotected by 7 months after starting vaccination P < .001). With adults at risk for a suboptimal response (i.e., older adults, the obese, men, and smokers) the triple antigen vaccine produced a greater degree of protection. The vaccines had similar safety profiles. Both vaccines were well tolerated. In healthy normal adults, a triple antigen hepatitis B vaccine containing S and pre–S antigens produced an enhanced immunologic response and was as effective as a 2– and 3–dose regimen.
Up–Regulation of Basolateral Multidrug Resistance Protein 3 (Mrp3) in Cholestatic Rat Liver
Authors: Donner Markus Georg , Keppler Dietrich
Published: August 2002

Cholestasis induces down–regulation of multidrug resistance protein 2 (Mrp2, symbol Abcc2), which is localized to the canalicular membrane. Given the overlapping substrate specificities of Mrp2 and multidrug resistance protein 3 (Mrp3, symbol Abcc3), we examined the hypothesis of a different subcellular and lobular localization of these members of the Mrp family in rat liver after bile duct ligation. We raised a polyclonal antibody against rat Mrp3 and detected this protein in the basolateral plasma membrane of hepatocytes surrounding the central veins and of cholangiocytes. The Mrp3 protein level was less than 2% of the expression observed after 72 hours of obstructive cholestasis. After 48 hours of bile duct ligation, the Mrp3 protein was increased and was further enhanced after 72 hours. In 72–hour–cholestatic rat liver Mrp3 was expressed, in addition, in periportal hepatocytes. However, there was a preponderance of Mrp3 in the pericentral area of the liver lobule. In Mrp2–deficient mutant rat liver, the Mrp3 protein expression was most enhanced and its zonation was lost. The Mrp3 immunostaining of cholangiocytes was preserved in cholestatic and in Mrp2–deficient mutant liver. Canalicular Mrp2 decreased and amounted to 34% of normal after bile duct ligation for 72 hours. We conclude that the hepatocellular up–regulation of Mrp3 in cholestasis together with cholangiocellular Mrp3 may compensate for the biliary obstruction and impaired canalicular Mrp2 function by clearing cholephilic anionic substances into the blood.
Studies of Murine Schistosomiasis Reveal Interleukin–13 Blockade As A Treatment for Established and Progressive Liver Fibrosis
Authors: Chiaramonte Monica G. , Cheever Allen W. , Malley James D. , Donaldson Debra D. , Wynn Thomas A.
Published: August 2002

In several allergic, autoimmune, and infectious diseases, fibrosis is a major cause of morbidity and mortality. Here, using a model of infection–induced liver fibrosis, we show that interleukin (IL)–13 is required at all stages of Schistosomiasis mansoni infection to induce fibrosis. IL–4 production was preserved in IL–13-deficient mice, yet failed to significantly contribute to the fibrotic response in either acute or chronic infection. Significant fibrosis develops in all infected mice, although the magnitude of the response varies widely in inbred mice. C3H/HeN, BALB/c, and C57BL/6 mice develop high, intermediate, and low levels of fibrosis, respectively. Despite these differences, IL–13 antagonism resulted in a marked amelioration of fibrosis in all strains. The fibrotic mechanism in the high– and low–responder strains was unrelated to their tissue eosinophil or mast cell responses, but did correlate with their patterns of IL–13, IL–10, and interferon gamma (IFN–γ) mRNA expression. Indeed, severe fibrosis correlated with a high IL–13 and low IFN–γ/IL–10 mRNA response. Because fibrotic diseases are typically progressive disorders, an important issue was to determine whether IL–13 inactivation might be used to treat an established and ongoing fibrotic disease. Here, IL–13 antagonism was highly efficacious, even after fibrosis and the Th2 cytokine response were firmly established. These studies demonstrate the central role played by IL–13 in fibrogenesis and suggest that therapeutic approaches aimed at disrupting the IL–13 pathway will be highly effective at preventing fibrotic disease caused by chronic Th2–mediated inflammatory reactions.
Indomethacin Prevents the Development of Experimental Ammonia–Induced Brain Edema in Rats After Portacaval Anastomosis
Authors: Chung Chuhan , Gottstein Jeanne , Blei Andres T.
Published: August 2002

Patients with fulminant hepatic failure (FHF) die with brain edema, exhibiting an increased cerebral blood flow (CBF) at the time of cerebral swelling. Mild hypothermia prevents brain edema in experimental models and in humans with FHF, an effect associated with normalization of CBF. To study the effects of alterations of CBF on the development of brain edema, we administered intravenous (IV) indomethacin to rats receiving an ammonia infusion after portacaval anastomosis. This model predictably develops brain edema and a marked increase in CBF at 3 hours of infusion. Brain water was measured with the gravimetry technique; CBF was monitored with both laser Doppler flowmetry and radioactive microspheres, whereas intracranial pressure (ICP) was monitored with a cisterna magna catheter. Coadministration of indomethacin prevented the increase in CBF seen with ammonia alone (110 ± 19% vs. -2 ± 9%) as well as the increase in brain water (80.86 ± 0.12% vs. 80.18 ± 0.06%) and the increase in ICP. Plasma ammonia and brain glutamine levels were markedly elevated in the ammonia–infused group and unaffected by indomethacin. However, ammonia uptake by the brain was significantly reduced by indomethacin. Levels of 6–keto–PGF1α, a stable metabolite of prostacyclin, were reduced in the cerebrospinal fluid (CSF) of indomethacin–treated animals. As with mild hypothermia, avoiding cerebral vasodilatation with indomethacin will prevent the development of brain edema in this hyperammonemic model. Cerebral vasoconstriction reduces cerebral ammonia uptake and, if selective to the brain, may be of benefit in FHF.
Proliferative Activity of Intrahepatic Colorectal Metastases After Preoperative Hemihepatic Portal Vein Embolization
Authors: Kokudo Norihiro , Tada Keiichiro , Seki Makoto , Ohta Hirotoshi , Azekura Kaoru , Ueno Masashi , Ohta Keiichiro , Yamaguchi Toshiharu , Matsubara Toshiki , Takahashi Takashi , Nakajima Toshifusa , Muto Tetsuichiro , Ikari Takaaki , Yanagisawa Akio , Kato Yo
Published: August 2002

Although hemihepatic portal vein embolization (PVE) has been used preoperatively to extend indications for hepatectomy in patients with colorectal metastases, the effects of this procedure on tumor growth and outcome remain controversial. To address this issue, we assessed the proliferative activity of intrahepatic metastases after PVE and the long–term outcome of this procedure. Eighteen patients with colorectal metastases underwent preoperative PVE between 1996 and 2000 (PVE group). Twenty–nine patients who underwent major hepatic resection without PVE served as control (non–PVE group). The hepatic parenchymal fraction of the left lobe had significantly increased from 38.1 ± 3.2% to 45.9 ± 2.9% 3 weeks after PVE (+20.5%, P < .0001). Tumor volume and percent tumor volume had also significantly increased from 223 ± 89 mL to 270 ± 97 mL (+20.8%, P = .016) and from 13.7 + 4.3% to 16.2 + 4.9% (+18.5%, P = .014), respectively. There was no apparent correlation between the increase in parenchymal volume and that in tumor volume. The Ki–67 labeling index of metastatic lesions was 46.6 ± 7.2% in the PVE group and 35.4 ± 12.6% in the non–PVE group (P = .013). Long–term survival was similar in the PVE and non–PVE groups, however, disease–free survival was significantly poorer in the PVE group than in the non–PVE group (P = .004). We conclude that PVE increases tumor growth and probably is associated with enhanced recurrence of disease. Although PVE is effective in extending indications for surgery, patient selection for PVE should be cautious.
Leptin Augments Inflammatory and Profibrogenic Responses in the Murine Liver Induced by Hepatotoxic Chemicals
Authors: Ikejima Kenichi , Honda Hajime , Yoshikawa Mutsuko , Hirose Miyoko , Kitamura Tsuneo , Takei Yoshiyuki , Sato Nobuhiro
Published: August 2002

Lines of evidence suggested a possible link between leptin and hepatic fibrosis; however, whether leptin modulates the fibrogenesis in the liver remains unclear. The purpose of this study, therefore, was to evaluate the effect of leptin on inflammatory and profibrogenic responses in the liver caused by hepatotoxic chemicals. Male C57Bl/6 mice were given carbon tetrachloride (CCl4) (0.1 μL/g body wieght [BW], intraperitoneally [IP]) and/or recombinant murine leptin (1 μg/g BW, IP) simultaneously, and sacrificed up to 72 hours later. Further, some mice were given thioacetamide (TAA; 200 μg/g BW, IP) and leptin 3 times per week for 4 weeks to evaluate the effect of leptin on chronic fibrogenic responses. A simultaneous injection of leptin enhanced acute CCl4–induced necroinflammatory and subsequent fibrotic changes in the hepatic lobules. The steady–state messenger RNA (mRNA) levels of α1(I) procollagen and heat shock protein 47 (HSP47) in the liver were potentiated when leptin was injected together with CCl4. Expression of α smooth muscle actin (α–SMA) in the liver after CCl4 treatment was also augmented markedly in combination with leptin. Further, leptin increased transforming growth factor β1 (TGF–β1) mRNA in the liver 24 hours after acute CCl4 about 4–fold higher than CCl4 alone. Moreover, leptin enhanced hepatic fibrosis and induction of α1(I) procollagen mRNA caused by chronic TAA administration. Collectively, these findings indicated that leptin augments both inflammatory and profibrogenic responses in the liver caused by hepatotoxic chemicals. It is postulated that the increase in systemic leptin levels enhances up–regulation of TGF–β1, leading to activation of stellate cells, thereby augmenting the fibrogenic response in the liver.
Cc–Chemokine Receptor 6 and Its Ligand Macrophage Inflammatory Protein 3α Might Be Involved in the Amplification of Local Necroinflammatory Response in the Liver
Authors: Shimizu Yukihiro , Murata Hiroyuki , Kashii Yoshiro , Hirano Katsuharu , Kunitani Hitoshi , Higuchi Kiyohiro , Watanabe Akiharu
Published: August 2002

It is not fully understood how antigen–specific or–nonspecific T cells migrate into the liver in various liver diseases. Macrophage inflammatory protein (MIP)–3α is a chemokine that is expressed mostly in the liver, and the receptor CCR6 is reported to be expressed on memory T cells. In the present study, we focused on the expression of CCR6 on T cells from peripheral blood (PB) and inflamed liver, and analyzed the involvement of MIP–3α and CCR6 in the immunopathogenesis of liver diseases. Intrahepatic T cells showed significantly (P < .0001) higher percentages of CCR6+CD4+ T cells compared with PB, which is significantly (P = .037) correlated with the necroinflammatory response in the liver. Most of intrahepatic CCR6+CD4+ cells were also positive for CCR5, which is known to express on T–helper 1 cells. MIP–3α was stained in the cells located near piecemeal necrosis, where dendritic cells (DCs) are often observed, and coculture of activated DCs with apoptotic cells induced MIP–3α production from the DCs. These data suggest that MIP–3α is produced by periportal DCs and/or macrophages after necroinflammatory response, leading to the recruitment of activated T cells into the liver. This process could be important to augment the local immune response in the livers of various liver diseases. The finding might be important not only for the understanding of immunopathogenesis of liver diseases but also for the therapeutic strategy to control the local immune response in the liver.
Occurrence of Identical Hypervariable Region 1 Sequences of Hepatitis C Virus in Transfusion Recipients and Their Respective Blood Donors: Divergence Over Time
Authors: Lin Hsiang Ju , Seeff Leonard B. , Barbosa Luiz , Hollinger Blaine F.
Published: August 2002

A total of 240 stored serum specimens from 30 transfusion recipients and 120 blood donors from the Transfusion–Transmitted Viruses Study (TTVS) were evaluated with the objective of establishing transmission of hepatitis C virus (HCV) by specific blood donors. Phylogenetic analysis of hypervariable region 1 (HVR1) and HCV genotyping were performed on the genomic region encoding amino acids 329 to 410. Amino acid distances between HVR1 sequences were calculated by the Kimura formula. Bootstrap analysis of HVR1 sequences provided support for linking recipients to specific donors. Linear regression analysis showed no differences between donor and recipient HVR1 sequences 7.9 weeks posttransfusion, but donor and recipient sequences diverged thereafter (r = 0.690). The initial lag phase in the evolution of HVR1 in the infected recipient was attributed to the time required to mount host immunologic defenses against the virus. Within–recipient divergence in HVR1 was determined from analyses of serial specimens collected within 2 weeks after the alanine transaminase peak, at the end of the original study (1974–1979), and in the follow–up study (1987–present). HVR1 remained invariant over a period of 6.7 to 9.5 days (95% CI) during acute infection. Within–patient divergence in HVR1 increased over a period of 11 to 15 years (r = 0.771), reaching the degree of divergence observed between unlinked subjects. In cases in which transfusion involved more than one HCV subtype, only one of the HCV subtypes established infection in the recipient. Subtype–specific differences in HVR1 were shown.
Acquisition of susceptibility to hepatitis C virus replication in HepG2 cells by fusion with primary human hepatocytes: Establishment of a quantitative assay for hepatitis C virus infectivity in a cell culture system
Authors: Ito Takayoshi , Yasui Kotaro , Mukaigawa Jun , Katsume Asao , Kohara Michinori , Mitamura Keiji
Published: August 2002

Hepatitis C virus (HCV) replicates in human and chimpanzee hepatocytes. To characterize the nature of HCV and evaluate antiviral agents, the development of an HCV replication system in a cell culture is essential. We developed a cell line derived from human hepatocytes by fusing them with a hepatoblastoma cell line, HepG2, and obtained several clones. When we tested the clones for their ability to support HCV replication by nested RT-PCR, we found 1 clone (IMY-N9) that was more susceptible to HCV replication than HepG2. The negative-strand HCV RNA was detected in IMY-N9 by strand-specific RT-PCR, and viral RNA was identified in culture supernatant during the culture. Then we monitored HCV RNA titers in IMY-N9 and HepG2, respectively, by real-time detection PCR throughout the culture. A significant increase in the HCV RNA titer was observed only in IMY-N9. Serial passages of HCV culture supernatant were shown in the culture system. Furthermore, we tested several infectious materials for viral infectivity by monitoring HCV RNA titers and/or 50% tissue culture infectious dose (TCID50) of HCV on IMY-N9. In each material, HCV showed various growth patterns and a different TCID50 even though the PCR titer in each material was identical. The results showed that HCV in each material served various growth patterns and different TCID50 even though PCR titer in each material was identical. This cell line is useful for estimating viral activity and for studying cellular factors that may be necessary to HCV replication in human hepatocytes. (Hepatology 2001;34:566-572.)
Increased production of vascular endothelial growth factor in peritoneal macrophages of cirrhotic patients with spontaneous bacterial peritonitis
Authors: Cejudo-Martín Pilar , Ros Josefa , Navasa Miguel , Fernández Javier , Fernádez-Varo Guillermo , Ruiz-del-Arbol Luis , Rivera Francisca , Arroyo Vicente , Rodés Juan , Jiménez Wladimiro
Published: August 2002

Spontaneous bacterial peritonitis (SBP) is a common complication of cirrhotic patients with ascites that usually results in renal failure and death despite the efficacy of the current antibiotic therapy. The pathogenesis of these phenomena is poorly known but it has been related to the production of vasoactive cell mediators locally acting on the splanchnic vasculature. Because previous studies showed that peritoneal macrophages of cirrhotic patients may produce high quantities of vascular endothelial growth factor (VEGF), a powerful vessel permeabilizing agent, when stimulated by cytokines and bacterial lipopolysaccharide, the present study was aimed to seek whether peritoneal macrophages of SBP patients are induced to produce increased amounts of VEGF. Our results indicate that the production rate and the messenger RNA (mRNA) and protein expression of this substance are increased in macrophages of patients with SBP in comparison with those of noninfected cirrhotic patients. This characteristic feature is absent in circulating monocytes of these patients. Moreover, enhanced endothelial cell proliferation induced by conditioned medium of macrophages isolated from the ascites of patients with SBP is abolished by anti-VEGF antibody, and peritoneal tissue of cirrhotic patients expresses both VEGF receptors, Flt-1 and KDR. These results, therefore, are consistent with the concept that locally released macrophage-derived VEGF may result in increased vascular permeability and plasma leakage in the peritoneal vessels of cirrhotic patients with SBP. (Hepatology 2001;34:487-493.)
Activation of caspases occurs downstream from radical oxygen species production, Bcl-xL down-regulation, and early cytochrome C release in apoptosis induced by transforming growth factor β in rat fetal hepatocytes
Authors: Herrera Blanca , Fernández Margarita , Álvarez Alberto M. , Roncero César , Benito Manuel , Gil Joan , Fabregat Isabel
Published: August 2002

Most of the morphologic changes that are observed in apoptotic cells are caused by a set of cysteine proteases (caspases) that are activated during this process. In previous works from our group we found that treatment of rat fetal hepatocytes with transforming growth factor β1 (TGF-β1) is followed by apoptotic cell death. TGF-β1 mediates radical oxygen species (ROS) production that precedes bcl-xL down-regulation, loss of mitochondrial transmembrane potential, release of cytochrome c, and activation of caspase-3 (Herrera et al., FASEB J 2001;15:741-751). In this work, we have analyzed how TGF-β1 activates the caspase cascade and whether or not caspase activation precedes the oxidative stress induced by this factor. Our results show that TGF-β1 activates at least caspase-3, -8, and -9 in rat fetal hepatocytes, which are not required for ROS production, glutathione depletion, bcl-xL down-regulation, and initial cytochrome c release. However, caspase activation mediates cleavage of Bid and Bcl-xL that could originate an amplification loop on the mitochondrial events. An interesting result is that transmembrane potential disruption occurs later than the initial cytochrome c release and is mostly blocked by the pan-caspase inhibitor Z-VAD.fmk, indicating that the decrease in mitochondrial transmembrane potential (ΔΨm) may be a consequence of caspase activity rather than the mechanism by which TGF-β induces cytochrome c efflux. Finally, although Z-VAD.fmk completely blocks nucleosomal DNA fragmentation, it only delays cell death, which suggests that activation of the apoptotic program by TGF-β in fetal hepatocytes inevitably leads to death, with or without caspases. (Hepatology 2001;34:548-556.)
Chronic ethanol consumption stimulates hepatitis B virus gene expression and replication in transgenic mice
Authors: Larkin Jonathan , Clayton Marcia M. , Liu Jie , Feitelson Mark A.
Published: August 2002

Epidemiologic observations show a higher frequency of hepatitis B virus (HBV) serologic markers in chronic alcoholics compared with the general population. This may be the result of an increased susceptibility of alcoholics to infection and/or to an ethanol-mediated stimulation of HBV gene expression and replication. To test the latter hypothesis, HBV transgenic SCID mice, which support consistent levels of virus replication, were fed with a standard Lieber-DiCarli or isocaloric diet for 5 weeks. In ethanol-fed mice, the levels of hepatitis B surface antigen (HBsAg) and viral DNA in serum increased by up to 7-fold compared with mice fed the control diet. Ethanol-treated mice also had elevated HBV-RNA levels, and increased expression of surface, core, and X antigens in the liver, especially in the pericentral regions. None of these changes were observed in transgenic mice fed isocaloric diets. Thus, chronic alcohol consumption alters the patterns of HBV gene expression and replication in the serum and liver of HBV transgenic SCID mice, and may provide a partial explanation for the increased frequency of HBV markers among alcoholics.
The risk of liver and bile duct cancer in patients with chronic viral hepatitis, alcoholism, or cirrhosis
Authors: Kuper Hannah , Ye Weimin , Broomé Ulrika , Romelsjö Anders , Mucci Lorelei A. , Ekbom Anders , Adami Hans-Olov , Trichopoulos Dimitrios , Nyrén Olof
Published: August 2002

No prospective study has analyzed simultaneously chronic viral hepatitis and alcoholism as risk factors for liver carcinogenesis, while taking into consideration the role of cirrhosis. Nor has the risk for hepatocellular carcinoma among patients with chronic viral hepatitis been prospectively evaluated in a low-risk Western population. Last, the relationship between hepatocellular carcinoma risk factors and bile duct cancer remains to be clarified. We analyzed prospectively the risk for primary liver and extrahepatic biliary tract cancer among 186,395 patients hospitalized with either chronic viral hepatitis, alcoholism, cirrhosis, or any combination of these conditions through linkages between national Swedish registers. Compared with the general population, the relative risk of hepatocellular carcinoma was 34.4 for chronic viral hepatitis alone, 2.4 for alcoholism alone, and 40.7 for cirrhosis alone. Among patients with combinations of these risk conditions, the relative risk of hepatocellular carcinoma was 27.3 for chronic viral hepatitis and alcoholism, 118.5 for chronic viral hepatitis and cirrhosis, 22.4 for alcoholism and cirrhosis, and 171.4 for all 3 conditions. We found limited evidence for an excess risk of intrahepatic, but not for extrahepatic, biliary duct cancer. Cirrhosis amplifies the risk of hepatocellular carcinoma among patients with chronic viral hepatitis, but it is not a prerequisite for liver carcinogenesis. In contrast, cirrhosis may be a necessary intermediate for the development of hepatocellular carcinoma among alcoholics.
Augmented hepatic interferon gamma expression and T-cell influx characterize acute hepatitis progressing to recovery and residual lifelong virus persistence in experimental adult woodchuck hepatitis virus infection
Authors: Hodgson Paul D. , Michalak Tomasz I.
Published: August 2002

Woodchucks infected with woodchuck hepatitis virus (WHV) have profiles of liver disease and age-dependent rates of progression to chronic hepatitis (CH) comparable with those seen in human hepatitis B. The mechanism of recovery from acute hepadnaviral infection or its evolution to chronicity remains unknown, although the liver immune responses are expected to play an important role. To determine the dynamics of intrahepatic cytokine expression and T-cell involvement, and to assess their value in predicting the outcome of acute hepatitis (AH) in the adult onset of WHV infection, we evaluated liver transcription of interferon gamma (IFN-γ); tumor necrosis factor α (TNF-α); interleukins (IL)-2, -4, and -6; and the T-cell influx in relation to disease histologic severity and virus load in serial liver biopsies collected during the life span of experimentally infected woodchucks. Our results show that recovery from viral AH in adulthood is preceded by a significantly greater hepatic expression of IFN-γ and CD3, an increased TNF-α transcription, lower hepatic WHV load, and a greater degree of liver inflammation than those in acute infection with CH outcome. Furthermore, we have learned that the elevated IFN-γ, TNF-α, and CD3 expression in the liver endures for years not only in CH, but also, although to a lesser extent, in apparently completely resolved infection. This is consistent with our previous findings that residual WHV replication and remnant liver inflammation continue for life after recovery from AH. This study indicates that antiviral cytokines, in particular IFN-γ, may play a central role in the long-term control of occult hepadnavirus persistence in the liver.
Immunohistologic study on the expressions of α-fetoprotein and protein induced by vitamin K absence or antagonist II in surgically resected small hepatocellular carcinoma
Authors: Fujioka Miwako , Nakashima Yutaka , Nakashima Osamu , Kojiro Masamichi
Published: August 2002

Sixty-eight cases of single hepatocellular carcinoma (HCC) with less than 3 cm of diameter were immunohistochemically examined for the expressions of α-fetoprotein (AFP) and protein induced by vitamin K absence or antagonist II (PIVKA-II). In cancerous tissues, the expression rate was significantly higher for PIVKA-II (34 cases [50%]) than AFP (21 cases [31%]) (P < .05), suggesting a higher specificity of PIVKA-II to small HCC. Sixteen of the 68 cases (24%) were positive to both AFP and PIVKA-II, and in 8 of the 16 cases, AFP and PIVKA-II expressing areas within a nodule were clearly divided by a fibrous septum. According to histologic grades, PIVKA-II expression was confirmed in 2 of the 15 well-differentiated HCCs, and in the well-differentiated component of 6 of the 12 “nodule-in-nodule”-type well-differentiated HCCs. AFP expression was not found in well-differentiated HCCs, but found in 16 of the 40 moderately differentiated HCCs (40%) and in the moderately differentiated component of 3 of the 12 “nodule-in-nodule”-type well-differentiated HCCs. The positive rate in the tissues was correlated to the serum levels for both AFP and PIVKA-II. In addition, frequency of tissue-PIVKA-II expression was higher than tissue-AFP expression in the cases whose serum protein level was within the normal range. This indicates that AFP and PIVKA-II have different patterns of tissue expression and of secretion to the blood. In comparison with tissue-AFP-negative cases, tissue-AFP-positive HCCs had a larger tumor size, higher frequencies of portal vein invasion and intrahepatic metastasis, a high Ki-67 labeling index, and a lower rate of recurrence-free survival. Thus, tissue-AFP-positive HCCs are suggested to be biologically more malignant than those HCCs that are AFP-negative and PIVKA-II-positive.
Prevalence and risk factors for hepatitis C virus infection at an Urban veterans administration medical center
Authors: Briggs Megan E. , Baker Christiane , Hall Robert , Gaziano Michael J. , Gagnon David , Bzowej Natalie , Wright Teresa L.
Published: August 2002

This study was designed to determine the seroprevalence and risk factors for hepatitis C virus (HCV) infection in veterans. Anti-HCV testing was performed in 1,032 patients and a questionnaire regarding sociodemographic characteristics and potential risk factors was administered. Adjusted prevalence of unique HCV-positive patients using outpatient services was 17.7% (95% confidence interval [CI] 17.2%, 18.2%). The following risk factors were associated with HCV infection: a history of injection drug use (IDU), receipt of blood transfusion prior to 1992, history of tattoo (odds ratio [OR], 2.93; 95% CI, 1.70-5.08), combat job as a medical worker (OR, 2.68; 95% CI, 1.25-5.60), history of incarceration over 48 hours (OR, 2.56; 95% CI, 1.52-4.32), greater than 15 lifetime sexual partners (OR, 1.61; 95% CI, 0.94-2.76) and sexual relations with a prostitute (OR, 0.46; 95% CI, 0.25-0.82). We concluded that HCV is common in veterans. Risk factors independently associated with infection are IDU, prior transfusion, prior tattoo, combat medical work, incarceration, and multiple opposite sex partners. Infection with HCV among veterans is strongly associated with traditional risk factors for infection and less strongly associated with combat-related risk.
Different hepatitis C virus nonstructural protein 3 (Ns3)-DNA-expressing vaccines induce in HLA-A2.1 transgenic mice stable cytotoxic T lymphocytes that target one major epitope
Authors: Brinster Carine , Muguet Stéphanie , Lone Yu-Chun , Boucreux Delphine , Renard Nathalie , Fournillier Anne , Lemonnier François , Inchauspé Geneviève
Published: August 2002

The immunogenicity of the Hepatitis C virus (HCV) nonstructural protein 3 (NS3) was investigated using different DNA-based strategies and a preclinical mouse model transgenic for the HLA-A2.1 molecule. Plasmids expressing NS3 either as a wild-type protein, as a fusion with murine lysosome-associated-membrane protein-1 specific sequences, or under the control of the Semliki Forest virus replicase were evaluated in vitro and in vivo. All plasmids were shown to express the expected size protein. These 3 NS3-expressing vaccines induced overall comparable levels of CTLs when measured at different times postvaccination although mice injected with the NS3-LAMP expressing plasmid showed a particularly homogeneous and overall vigorous response (specific lysis ranged from 60% to 90 % for an E:T ratio of 33.3:1 with a mean CTL precursor frequency of 1:2.105 cells). Out of the four HLA-A2.1-restricted NS3 epitopes previously described in HCV infected patients (aa 1073-1081, aa 1406-1415; aa 1169-1177 and aa 1287-1296), the NS3-DNA generated CTLs were predominantly targeted at the aa 1073-1081 epitope. Peptide-based immunization showed that the mouse repertoire was intact for all epitopes tested except one (aa 1287-1296). In conclusion, the 3 NS3-DNA vaccines although based on different mode of action, shared a comparable efficacy at inducing CTL. Surprisingly, the breadth of such response was restricted to a single, major epitope.
Hepatitis B virus X protein transactivates inducible nitric oxide synthase gene promoter through the proximal nuclear factor κB-binding site: Evidence that cytoplasmic location of X protein is essential for gene transactivation
Authors: Majano Pedro , Lara-Pezzi Enrique , López-Cabrera Manuel , Apolinario Arantxa , Moreno-Otero Ricardo , García-Monzón Carmelo
Published: August 2002

Nitric oxide appears to play a central role in the pathogenesis of many inflammatory disorders. We have previously shown that there is an enhanced intrahepatic expression of the inducible nitric oxide synthase (iNOS) gene during chronic hepatitis B virus infection, and that viral X protein (HBx) transcriptionally activates this cellular gene, but the molecular basis for this activation remains to be defined. We aimed to explore the involvement of different cis-acting elements of the human iNOS promoter in the HBx-mediated up-regulation as well as the effect of the intracellular distribution of the HBx on its transacting function. Cotransfection of human hepatocyte-derived cells with wild-type or mutated iNOS promoter and with an HBx expression vector showed that functional inactivation of the proximal nuclear factor κB (NF-κB)-binding site of the iNOS promoter markedly reduced the HBx-mediated transcriptional activity. Mobility shift assays showed increased DNA-protein complexes comprising mainly the p50 and p65 members of NF-κB family in the nuclear extracts from HBx-transfected cells. Transient transfection experiments with HBx-expressing plasmids containing distinct cellular localization sequences showed that cytoplasmic location of this viral protein activated the iNOS promoter but when HBx was targeted to the nucleus, the HBx-induced luciferase activity was almost completely abrogated. In conclusion, cytoplasmic location of HBx protein is essential for the transcriptional activation of the iNOS gene through the nuclear translocation of p50-p65 heterodimers.
The geographical distribution of primary biliary cirrhosis in a well-defined cohort
Authors: Prince Martin I. , Chetwynd Amanda , Diggle Peter , Jarner Mikala , Metcalf Jane V. , James Oliver F. W.
Published: August 2002

The incidence of primary biliary cirrhosis (PBC) varies widely between regions. However, little is known about variation within regions and the degree to which this may reflect environmental risk factors. The aim of this study was to describe the spatial distribution of cases of PBC in a defined region of Northeast England over a defined period, and to assess the magnitude of any departure from random spatial distribution. Seven hundred seventy patients with established PBC were identified in a previous comprehensive case finding study. A total of 3,044 control locations were randomly selected from postcode (zip code) data weighted for number of drop off points per postcode. Geographical analysis was performed by testing both for spatial variation in risk and local clustering by using previously described point process methods. Both tests used the same null hypothesis that risk of disease does not vary spatially and cases occur independently of each other. Statistically significant spatial variations in risk were found in the whole study region (P < .001) and in the major urban area within the region (P < .004). Risk was higher in the urban area of Tyneside than in the surrounding rural area. Within the rural area, spatial variation in risk was equivocal (P = .012), but there was significant (P = .001) clustering of cases (estimated average cluster effect approximately 10 excess cases within a 7-km radius). PBC occurred to a density of 10.7 cases/km2 in the highest risk areas. In conclusion, PBC is unevenly distributed in Northeast England. This may reflect one or more environmental risk factors in its etiology.
Interleukin-6 from intrahepatic cells of bone marrow origin is required for normal murine liver regeneration
Authors: Aldeguer Xavier , Debonera Fotini , Shaked Abraham , Krasinkas Alyssa M. , Gelman Andrew E. , Que Xingyi , Zamir Gideon A. , Hiroyasu Shungo , Kovalovich Kellen K. , Taub Rebecca , Olthoff Kim M.
Published: August 2002

Interleukin-6 (IL-6) is required for normal liver regeneration, but the specific cellular source of this growth factor is unknown. We investigated whether this signal originates from the resident macrophage, the Kupffer cell. Using a murine model of bone marrow transplantation, we replaced recipient bone marrow-derived cells, including Kupffer cells, with cells of donor genetic phenotype. Recipients deficient in IL-6 (IL-6−/−) were lethally irradiated, then rescued with 107 donor bone marrow cells capable of expressing IL-6 (IL-6+/+). Conversely, IL-6+/+ recipients received IL-6−/− marrow. Successful engraftment was measured by the presence of the Y chromosome SRY locus in the livers of female recipients receiving male marrow, in situ IL-6 expression by Kupffer cells, and up-regulation of IL-6 in splenocytes after activation with lipopolysaccharide (LPS). Kupffer cell isolation in IL-6−/− females receiving IL-6+/+ male marrow clearly showed the presence of the SRY locus and IL-6 disrupted allele, whereas males receiving female marrow demonstrated no SRY or IL-6 signals, confirming the extent of replacement. Replacement of these cells in IL-6−/− mice with IL-6+/+ bone marrow successfully restored the regenerative response after partial hepatectomy (PHx) as indicated by signal transduction and activator of transcription 3 (STAT3) activation and hepatocyte DNA replication. Alternatively, complete replacement of Kupffer cells in IL-6+/+ mice by transplantation with IL-6−/− cells significantly inhibited liver regeneration and was partially restored by administration of IL-6. This investigation demonstrates a paracrine mechanism by which cells of bone marrow origin, most likely Kupffer cells, regulate the regenerative capacity of the hepatocyte through IL-6 expression.