bluuebunny/crossref_metadata_2025 huggingface dataset:
Screening for hepatocellular carcinoma in chronic carriers of hepatitis B virus: Incidence and prevalence of hepatocellular carcinoma in a North American urban population
Authors: Sherman Morris , Peltekian Kevork M. , Lee Cindy
Published: March 2007

Objective: To prospectively determine the prevalence and annual incidence of hepatocellular carcinoma in hepatitis B carriers in a heterogeneous urban North American population and to assess the diagnostic accuracy of tests used for screening for this cancer. Design: Prospective cohort study of 1,069 chronic carriers of hepatitis B virus using screening with α-fetoprotein alone or in combination with ultrasonography every 6 months. Results: The mean age of the cohort was 39 ± 12 years (± SD), 65% were men, 71% were Asians. At the first screening visit, serum α-fetoprotein was ≥ 20 μg/L in 4%. In those subjects who were also screened by ultrasonography during the first visit, 9% were found to have focal lesions. Only 3 subjects were found to have hepatocellular carcinoma at the first screening, giving a prevalence of 281/100,000 chronic carriers of hepatitis B virus. The cohort was followed for 2,340 person-years (mean, 26 months follow-up, with a range from 6 to 60 months). During this period, 11 more subjects, 10 men and 1 woman, were diagnosed to have hepatocellular carcinoma (annual incidence, 470/100,000). In men only, the annual incidence was 657/100,000. During the study, 5 subjects died from hepatocellular carcinoma (annual mortality rate, 214/100,000). Sensitivity and specificity of serum α-fetoprotein >20 μg/L were 64.3% and 91.4%, respectively. For ultrasonography, sensitivity was 78.8% and specificity 93.8%. Conclusions: These data suggest that the incidence and prevalence of hepatocellular carcinoma in hepatitis B carriers in our area, an urban North American setting, are as high as in countries where hepatitis B is endemic. Current screening tests have significant false-positive and false-negative rates, raising questions about the cost-benefit of screening for hepatocellular carcinoma in our study population. (Hepatology 1995;22:432-438.)
Efficacy of screening donors for antibodies to the hepatitis C virus to prevent transfusion-associated hepatitis: Final report of a prospective trial
Authors: González Antonio , Esteban Juan I. , Madoz Pedro , Viladomiu Luis , Genesca Juan , Muñiz Eduardo , Enríquez Jaime , Torras Xavier , Hernández José M. , Quer Josep , Vidal Xavier , Alter Harvey J. , Shih James W. , Esteban Rafael , Guardia Jaime
Published: March 2007

Routine screening of blood donors for anti-hepatitis C virus (HCV) has been implemented in most developed countries. However, the independent efficacy of such screening has not been established in a controlled, prospective study. We tracked 478 patients transfused with anti-HCV-negative blood by first-generation enzyme-linked immunoassay (EIA) between July 1989 and May 1990 and compared the incidence of transfusion-associated hepatitis and HCV infections with that found among 280 patients transfused with blood unscreened for anti-HCV during the immediately preceding year. Of the 280 patients who had received transfusions before donors were screened for anti-HCV, 27 (9.6%) developed posttransfusion hepatitis and 1 additional patient sero-converted to anti-HCV without evidence of hepatitis, for a risk of posttransfusion HCV infection of 10.7% (28 of 262 recipients seronegative for anti-HCV before transfusion). Of the 478 patients transfused after July 1989 with blood screened for anti-HCV, only 9 (1.9%) developed posttransfusion hepatitis for a risk reduction of 80%. Seven of the 9 residual cases of hepatitis were caused by HCV (7 of 456 recipients seronegative before transfusion or 1.5%) for a risk reduction of transfusion-associated HCV infection of 86%. In retrospect, an anti-HCV positive donor was detected by second-generation immunoassay in 4 (57%) of the 7 HCV cases from the study cohort and in 19 of the 23 (83%) cases from whom all donor samples were available for testing in the historical cohort. No additional infectious donors were detected by third-generation immunoassay or serum HCV-RNA by polymerase chain reaction. Implementation of donor screening for anti-HCV with a first-generation immunoassay appeared to be independently associated with an 80% and 86% reduction in the risks of transfusion-associated hepatitis and HCV infection, respectively. Screening of donors with second-generation immunoassays might have further reduced the risk by 57%. An additional 0.4% may be at risk of developing non-A, non-B, non-C hepatitis. (Hepatology 1995;22:439-445.)
Increased liver iron stores in patients with hepatocellular carcinoma developed on a noncirrhotic liver
Authors: Turlin Bruno , Juguet Frédéric , Moirand Romain , Le Quilleuc Danielle , Loréal Olivier , Campion Jean-Pierre , Launois Bernard , Ramée Marie-Paule , Brissot Pierre , Deugnier Yves
Published: March 2007

Iron was systematically studied in the nontumorous liver of 24 patients with hepatocellular carcinoma (HCC) developed on a noncirrhotic liver compared with 4 control groups (cirrhosis with and without HCC, liver metastasis, and normal liver) matched according to age, sex, and presence of chronic alcoholism. Assessment of liver iron was made by (1) histology according to iron distribution and quantification (total iron score: 0 to 60), and (2) biochemistry (liver iron concentration-N < 36 μmol/g) with calculation of the hepatic iron index (liver iron concentration/age). Patients with hepatocellular carcinoma developed on a noncirrhotic liver presented with (1) histological iron in 83%; (2) parenchymal iron excess significantly more frequent (90%) than in controls; (3) total iron score (15 ± 12) and liver iron concentration (81 ± 96) significantly greater than in controls; and (4) hepatic iron index significantly increased (1.4 ± 1.5) when compared with control groups, except for the hepatocellular carcinoma complicating cirrhosis group (0.9 ± 1.1). This study (1) shows a mild but unquestionable parenchymal iron excess in the nontumorous liver of most patients presenting with hepatocellular carcinoma developed on a noncirrhotic liver and, at a lesser extent, on cirrhosis, (2) should incite others to study the putative role of iron in the development of liver cancer both in patients with cirrhosis and those without it, whatever the cause of the underlying liver disease, and (3) add argument to take into account and to treat any liver iron excess, even when mild. (Hepatology 1995;22:446-450.)
A prospective, randomized trial of sclerotherapy versus ligation in the management of bleeding esophageal varices
Authors: Lo Gin-Ho , Lai Kwok-Hung , Cheng Jing-Shiung , Hwu Jia-Huey , Chang Chia-Fu , Chen Sam-Ming , Chiang Hung-Ting
Published: March 2007

We conducted a prospective, randomized trial comparing sclerotherapy and ligation in 120 patients with acute bleeding of esophageal varices. All the patients were cirrhotic, 59 received sclerotherapy, and 61 received ligation. Treatment was repeated regularly until the varices were obliterated. The mean follow-up period was 295 ± 120 days and 310 ± 105 days for the sclerotherapy and ligation groups, respectively. The control of active bleeding was 12/15 (80%) in the sclerotherapy group and 18/19 (94%) in the ligation group (P = .23). The numbers of treatment sessions required to achieve variceal obliteration were 6.5 ± 1.2 in the sclerotherapy group and 3.8 ± 0.4 in the ligation group (P < .001). Recurrent bleeding from the gastrointestinal tract was 51% in the sclerotherapy group compared with 33% in the ligation group (P < .05). Recurrent bleeding from esophageal varices was 36% in the sclerotherapy group and 11% in the ligation group (P < .01). However, bleeding from ectopic varices and congestive gastropathy was less common in the sclerotherapy group (7%) than in the ligation group (18%) (P = .05). Significant complications were encountered in 19% of the sclerotherapy group and in 3.3% of the ligation group (P < .01). Comparison of Kaplan-Meier estimates of time to death of both groups showed a significantly lower mortality in the ligation group (P = .011). Both sclerotherapy and ligation can effectively arrest active bleeding from esophageal varices. However, ligation is more-effective than sclerotherapy in decreasing the risk of rebleeding from esophageal varices with fewer complications. Ligation can also achieve obliteration of esophageal varices more rapidly than sclerotherapy. Survival was significantly more improved after ligation than after sclerotherapy. However, there was a greater incidence of bleeding from sites other than the esophageal varices after ligation. (Hepatology 1995; 22:466-471.)
Effect of upright posture and physical exercise on endogenous neurohormonal systems in cirrhotic patients with sodium retention and normal supine plasma renin, aldosterone, and norepinephrine levels
Authors: Saló Joan , Ginès Angels , Anibarro Luis , Jiménez Wladimiro , Bataller Ramón , Clària Joan , Ginès Pere , Rirvera Francisca , Arroyo Vicente , Rodés Juan
Published: March 2007

It is well known that sodium retention occurs in a significant proportion of patients with cirrhosis despite normal supine plasma levels of renin, aldosterone (ALDO), and norepinephrine (NE). The current study was performed to assess whether this subset of patients also present normal activity of the renin-aldosterone and sympathetic nervous systems during upright posture in sitting position and moderate physical exercise. Nine healthy controls, 14 patients with compensated cirrhosis and 10 patients with cirrhosis, ascites, sodium retention, and normal supine plasma renin activity (PRA) and ALDO and NE concentration were sequentially studied after 60 minutes in supine rest, 30 minutes in sitting position, and 30 minutes of cycloergometric exercise (3-METs). Sitting position and exercise were associated with similar stimulation of the renin-aldosterone and sympathetic nervous systems in the three groups of subjects. Consequently, cirrhotic patients with ascites showed values of PRA and plasma concentration of ALDO and NE similar to healthy subjects and patients with compensated cirrhosis during supine rest (renin: 1.4 ± 0.3, 0.8 ± 0.2, and 0.8 ± 0.3 ng/mL.hr; aldosterone: 24.3 ± 4.7,20.2 ± 3.9 and 21.4 ± 3.4 ng/dL; norepinephrine: 252 ± 23, 250 ± 16, and 255 ± 23 pg/mL), sitting position (renin: 2.1 ± 0.5,1.1 ± 0.3, and 1.6 ± 0.4; aldosterone: 32.2 ± 7.3, 23.7 ± 5.3, and 26.2 ± 4.5; norepinephrine: 356 ± 38, 401 ± 63, and 420 ± 35), and exercise (renin: 2.9 ± 0.8, 1.6 ± 0.4, and 2.2 ± 0.5; aldosterone: 43 ± 6.4, 34.9 ± 8.5, and 38.2 ± 5.3; norepinephrine: 481 ± 35, 499 ± 54, and 534 ± 48). Plasma atrial natriuretic peptide concentration was significantly (P < .05) higher in cirrhotic patients with ascites than in healthy subjects during supine rest (5.69 ± 1.1 vs. 21.0 ± 3.3 fmol/L) and exercise (9.1 ± 2.2 vs. 28.3 ± 6.6). These results indicate that sodium retention in cirrhosis may occur in the setting of a normal activity of the endogenous neurohormonal antinatriuretic systems and high plasma levels of ANP. (Hepatology 1995; 22:479-487.)
Calcium carbonate in cholesterol gallstones: Polymorphism, distribution, and hypotheses about pathogenesis
Authors: Taylor Donald R. , Crowther Roger S. , Cozart John C. , Sharrock Pamela , Wu Jinguang , Soloway Roger D.
Published: March 2007

This study of sets of cholesterol gallstones collected consecutively from 222 patients in La Paz, Bolivia, and Mexico City, Mexico, has developed a reliable infrared (IR) spectroscopic method for the detection of calcium carbonate in cholesterol gallstones and provided the basis for simultaneous identification of each of its three polymorphs: calcite, vaterite, and aragonite. The peaks in the 854 to 876 cm−1 region demonstrated 98% sensitivity and specificity for carbonate detection. As little as 3% carbonate by weight could be detected using these peaks. The overall incidence of carbonate was 19% in these populations containing a high proportion of Amerinds. Infrared microspectroscopy of 10 to 50 μm particles, dissected from stones, allowed a ring-by-ring examination of 11 carbonate-containing stones. It was determined that different carbonate polymorphs, when present in the same gallstone, almost always occurred in separate rings. In approximately half of the gallstones, different polymorphs were present in successive layers in the same stone, indicating that conditions governing stone growth changed cyclically. Carbonates were usually precipitated in peripheral layers rather than in the center, supporting the theory that formation of calcium carbonates may be related to episodes of intermittent obstruction of the cystic duct, as opposed to being a major factor in stone nidation. (Hepatology 1995;22:488-496.)
Peroxisome mosaicism in the livers of peroxisomal deficiency patients
Authors: Espeel Marc , Mandel Hanna , Poggi Florence , Smeitink Jan A. M. , Wanders Ronald J. A. , Kerckaert Ingrid , Schutgens Rudolf B. H. , Saudubray Jean-Marie , Poll-The Bwee-Tien , Roels Frank
Published: March 2007

Peroxisomal deficiency disorders, which are genetically transmitted, are assumed to be expressed in all cells, and the use of cultured skin fibroblasts for diagnosis and research is based on this assumption. We describe three patients with clinical, biochemical, and microscopic evidence of a peroxisomal disorder. However, their liver displays mosaicism, i.e., parenchymal cells with peroxisomes are adjacent to cells without peroxisomes. Ten percent (volume), 8%, and less than 1% of the parenchyma possessed peroxisomes that can be identified in immunocytochemical tests for six matrix and membrane proteins performed by light and electron microscopy. In the bulk of the parenchyma, catalase is localized in the cytoplasm, and in such cells no peroxisomes are evident by electron microscopy and immunolabeling for the 43-kd peroxisomal membrane protein (PMP) in two patients; in the third case, peroxisomal membrane ghosts are present. Immunoblots of peroxisomal β-oxidation enzymes show a pattern similar to that from patients with a generalized peroxisomal deficiency. In contrast to the clinical and biochemical signs of peroxisomal dysfunction and hepatic histopathology, cultured fibroblasts from two patients demonstrate normal peroxisomal functions, including very-long-chain fatty acid oxidation and plasmalogen synthesis. (Hepatology 1995; 22:497-504.)
Autoantibodies to BCOADC-E2 in patients with primary biliary cirrhosis recognize a conformational epitope
Authors: Leung Patrick S. C. , Chuang David T. , Wynn Max R. , Cha Sanghoon , Danner Dean J. , Ansari Aftab , Coppel Ross L. , Gershwin Eric M.
Published: March 2007

Primary biliary cirrhosis (PBC) is an autoimmune disease of liver associated with a unique serologic response to mitochondrial autoantigens. Many of the autoantigens recognized by autoantibodies in PBC are members of the 2-oxo-acid dehydrogenase complex. The two major autoantigens are the E2 component of the pyruvate dehydrogenase complex (PDC-E2) and the E2 component of the branched chain 2-oxo-acid dehydrogenase complex (BCOADC-E2). The autoantibody response to PDC-E2 has been mapped to one immunodominant epitope, which consists of both linear and conformational components. The presence of a single immunodominant epitope in PDC-E2 is unusual when contrasted to the immune response to autoantigens in other human autoimmune diseases. We have mapped the epitope recognized by antimitochondrial autoantibodies (AMA) specific to BCOADC-E2 in patients with PBC by taking advantage of the full-length bovine BCOADC-E2 complementary DNA (cDNA) and a series of expression clones spanning the entire molecule. Reactivity to the various expression clones was studied by immunoblotting, enzyme-linked immunosorbent assay (ELISA), as well as selective absorption of patient sera by expressed protein fragments. Autoantibodies to BCOADC-E2 map within peptides spanning amino acid residues 1 to 227 of the mature protein; our data demonstrate that the epitope is dependent on conformation and includes the lipoic acid binding region. However, only the full-length clone (amino acid residue 1 to 421) is sufficient to remove all detectable BCOADC-E2 reactivity. Moreover, the absence of lipoic acid on the recombinant polypeptides used in this study indicates that antibody binding to BCOADC-E2 is not dependent on the presence of lipoic acid. (Hepatology 1995; 22:505-513.)
The liver in adolescents with α-antitrypsin deficiency
Authors: Sveger Tomas , Eriksson Sten
Published: March 2007

Of 200,000 Swedish infants screened for α1-antitrypsin deficiency (α1ATD), 184 (127 PiZ, 2 PiZ-, 54 PiSZ, and 1 PiS-) children have been followed prospectively, of whom 1 PiSZ and 5 PiZ children died in early childhood. We now report clinical and biochemical signs of liver disease in adolescence and the prognosis of neonatal liver disease up to the age of 18 years. The α1ATD subjects were offered a clinical checkup and liver tests at 16 and 18 years of age, 150 of 178 α1ATD subjects undergoing checkups at age 16 and 166 at age 18. Liver tests were performed in 121 adolescents at both the 16- and 18-year checkups. None of the PiZ and PiSZ subjects checked at the age of 16 and 18 years had any clinical signs of liver disease. Abnormalities of serum alanine aminotransferase (S-ALAT) or γ-glutamyl transferase (S-GT) were found at the 16-year checkup (all PiZ and PiSZ subjects tested included) in 17% of PiZ and 8% of PiSZ adolescents, and at the age of 18 years in 12% of PiZ and 15% of PiSZ subjects. In only two cases were both S-ALAT and S-GT concentrations abnormal at both the 16-year and 18-year follow-ups. Serum procollagen III peptide concentrations were normal in all those with abnormal liver test results. Of 127 PiZ subjects, 22 had manifested clinical signs of liver disease in infancy. Of these 22, two died early in life of cirrhosis. Another two children died of other causes; however, autopsy showed histological signs of cirrhosis in one of them, fibrosis in the other. All of the remaining 18 subjects were clinically healthy at the 16- and 18-year checkups. Marginal liver test abnormalities were found in two of them. To sum up, α1ATD children followed prospectively up to 18 years of age continue to have favorable prognosis. (Hepatology 1995; 22:514-517.)
Synthesis of interleukin-1β in primary biliary cirrhosis: Relationship to treatment with methotrexate or colchicine and disease progression
Authors: Miller Laurie C. , Sharma Archna , McKusick Augusta F. , Tassoni Joseph P. , Dinarello Charles A. , Kaplan Marshall M.
Published: March 2007

Primary biliary cirrhosis (PBC) is a chronic, progressive, cholestatic liver disease. Interleukin-1β (IL-1β) may play a role in the pathogenesis of PBC by contributing to altered immune function and fibrosis. Colchicine or methotrexate has some beneficial effects in the treatment of PBC, and also affects interleukin-1 (IL-1). Therefore, we prospectively studied the synthesis of IL-1β by peripheral blood mononuclear cells (PBMC) from 42 patients with PBC entered into a randomized, doubleblind, double-dummy controlled trial of colchicine and methotrexate. PBMC obtained at entry, 6, 12, 18, and 24 months were stimulated to produce IL-1β with phytohemagglutinin (PHA), lipopolysaccharide (LPS), Staphylococcus epidermidis, recombinant IL-2, or mitochondrial antigen. Patients in the two treatment groups did not differ at entry in biochemical measures or liver histological stage. Over 24 months in both groups, serum bilirubin and histologic stage remained stable and alkaline phosphatase decreased significantly. For all patients, synthesis of IL-1β increased constitutively and in response to immune-mediated stimulants (PHA, IL-2, and mitochondrial antigen) but not the bacterial stimulants LPS or S epidermidis. Compared with levels of IL-1β at entry, PHA induced increases for patients treated with methotrexate (12, 18, and 24 months) or colchicine (18 and 24 months). At 24 months, IL-2-induced IL-1β synthesis was increased in patients treated with methotrexate, whereas S epidermidis-induced IL-1β was enhanced in colchicine-treated patients. Before treatment, IL-1β production did not relate to severity of disease except in response to S epidermidis. Notably, in patients with stable or improving liver histological stage, IL-1β synthesis increased dramatically at 12 months in response to IL-2 and S epidermidis, and also to LPS by 24 months. Alterations in IL-1β synthesis in patients with PBC may reflect underlying immunoregulatory changes that contribute to disease progression or stabilization. (Hepatology 1995;22:518-524.)
Serum concentration of intercellular adhesion molecule-1 in patients with hepatocellular carcinoma is a marker of the disease progression and prognosis
Authors: Shimizu Yukihiro , Minemura Masami , Tsukishiro Takashi , Kashii Yoshirou , Miyamoto Megumi , Nishimori Hiroshi , Higuchi Kiyohiro , Watanabe Akiharu
Published: March 2007

Serum levels of soluble forms of intercellular adhesion molecule-1 (sICAM-1) and lymphocyte function-associated antigen-3 (sLFA-3) in 122 patients with chronic liver disease including hepatocellular carcinoma (HCC) were measured by enzyme-linked immunosorbent assays. Serum levels of sICAM-1 in patients with HCC were significantly higher than those of chronic hepatitis (CH) and cirrhosis. On the other hand, serum levels of sLFA-3 in patients with HCC were almost the same as those of cirrhosis. Western blot analyses showed that molecular sizes of sICAM-1 and sLFA-3 detected in the sera were 90 kd and 50 kd, respectively, indicating that both molecules include whole extracellular domains. In patients with HCC, circulating sICAM-1 levels were significantly (P < .001) correlated with tumor volume (r = .50), total bilirubin (r = .38), serum aspartate aminotransferase levels (r = .51), and γ-globulin (r = .63). Furthermore, serum sICAM-1 levels were significantly elevated in patients with multiple HCC (tumor number >3) or HCC with tumor embolus in the first branch or trunk of portal vein. Survival periods were analyzed in relation to serum sICAM-1 levels in patients with HCC who had been treated by transcatheter arterial chemoembolization. The HCC patients with <1,000 ng/mL of serum ICAM-1 showed significantly (P = .0005) longer survival than those with higher levels of the molecule. The same results were obtained when only patients with moderately differentiated HCC were analyzed (P = .02). Analyses by Cox's proportional hazard model showed that sICAM-1 is a significant (P = .032) prognostic factor for patients with HCC. These data show that circulating sICAM-1 in the sera of patients with HCC is a marker for tumor progression and prognosis of the patients. (Hepatology 1995; 22:525-531.)
Relationship between cardiomyopathy and liver disease in chronic alcoholism
Authors: Estruch Ramon , Fernández-Solá Joaquim , Sacanella Emilio , Paré Carles , Rubin Emanuel , Urbano-Márquez Alvaro
Published: March 2007

Based on anecdotal impressions, there is a common clinical perception that alcoholics with liver disease do not develop cardiomyopathy and that those with alcohol-induced cardiac disease are spared cirrhosis. To determine the relationship between alcoholic cardiomyopathy and cirrhosis, we carried out a prospective crosssectional study that included: (1) 30 alcoholic men with cardiomyopathy; (2) 30 alcoholic men without cardiomyopathy (left ventricular ejection fraction > 55%); (3) 20 actively drinking alcoholics with cirrhosis; (4) 15 abstaining alcoholics with cirrhosis; and (5) 15 nonalcoholics with cirrhosis of other etiologies. Cirrhosis was observed in 13 of 30 patients with alcoholic cardiomyopathy (43%), compared with 2 of 30 alcoholics without cardiomyopathy (6%) (P < .001). Ten of the 20 active alcoholics with cirrhosis (50%) showed evidence of dilated cardiomyopathy. Actively drinking alcoholics with cirrhosis had a significantly lower mean ejection fraction and shortening fraction, as well as a greater mean enddiastolic diameter and left ventricular mass than abstaining alcoholics with cirrhosis. Cardiac studies of patients with nonalcoholic cirrhosis were normal. We conclude that a positive correlation exists between alcoholic cardiomyopathy and cirrhosis. Alcoholics admitted solely for cardiomyopathy have a higher prevalence of cirrhosis than unselected alcoholics without heart disease. Actively drinking alcoholics admitted only for cirrhosis show impaired cardiac performance, whereas abstaining alcoholics with liver disease tend to manifest normal cardiac function. (Hepatology 1995; 22:532-538.)
Protective effects of N-acetylcysteine on hypothermic ischemia-reperfusion injury of rat liver
Authors: Nakano Hiroshi , Boudjema Karim , Alexandre Eliane , Imbs Pierre , Chenard Marie Pierre , Wolf Philippe , Cinqualbre Jacques , Jaeck Daniel
Published: March 2007

We investigated whether intraportal injection of 150 mg/kg N-acetylcysteine (NAC) into rats reduced hepatic ischemia-reperfusion injury after 48 hours of cold storage and 2 hours of reperfusion. The organ was isolated and perfused to evaluate liver function. The control group received an intraportal injection of 5% dextrose. NAC increased L-cysteine concentrations 15 minutes after injection (1.29 ± 0.11 μmol/g vs. 2.68 ± 0.4 μmol/g,P < .05). However, neither treatment modified glutathione liver concentrations relative to preinjection values. After 48 hours of cold storage and 2 hours of reperfusion, livers from NAC-treated rats produced larger amounts of bile than those in the control group (5.04 ± 1.92 vs. 0.72 ± 0.37 μL/g liver; P < .05), and showed a significant reduction in liver injury, as indicated by reduced release of lactate dehydrogenase (679.4 ± 174.4 vs. 1891.3 ± 268.3 IU/L/g; P < .01), aspartate transaminase (AST) (13.94 ± 3.5 vs. 38.75 IU/L/g; P < .01), alanine transaminase (ALT) (14.92 ± 4.09 vs. 45.91 ± 10.58 IU/L/g; P < .05), and acid phosphatase, a marker of Kupffer cell injury (344.4 ± 89.6 vs. 927.3 ± 150.8 IU/L/g; P < .01) in the perfusate. Reduced glutathione concentrations in the perfusate were similar in the two groups (805 ± 69 vs. 798 ± 252 nmol/L/g), whereas oxidized glutathione (GSSG) concentrations were higher in the control group (967 ± 137 vs. 525 ± 126 nmol/L/g; P < .05). Reduced glutathione (GSH) concentrations in liver tissue collected at the end of perfusion were significantly higher in the NAC group (7.3 ± 0.9 vs. 4.1 ± 1.0 μmol/g; P < .05). The protective effect of NAC on cold ischemia-reperfusion liver injury persisted when animals were pretreated with buthionine sulfoximine (BSO), a specific inhibitor of glutathione synthesis. Our results suggest that NAC enhances the concentrations of cysteine within hepatocytes, providing a substrate for glutathione synthesis during reperfusion. They also indicate that NAC has a direct protective effect on Kupffer cells, which are the first source of reactive oxygen intermediates during reperfusion. (HEPATOLOGY 1995; 22:539-545.)
Improved hepatocyte in vitro maintenance in a culture model with woven multicompartment capillary systems: Electron microscopy studies
Authors: Gerlach Jörg C. , Schnoy Norbert , Encke Jens , Smith Mark D. , Müller Christian , Neuhaus Peter
Published: March 2007

Primary pig hepatocytes form a tissuelike structure in an in vitro culture model that has provision for threedimensional cell orientation, cell aggregation, decentralized cell perfusion with low metabolite gradients, integral oxygenation, and nonparenchymal cell coculture. Scanning electron microscopy (SEM) has shown that hepatocytes spontaneously form aggregates in a three-dimensional structure between and on the surface of artificial capillaries. Transmission electron microscopy (TEM) has shown that after 7 weeks of in vitro perfusion, the cell ultrastructure remains similar to that of the parenchyma in vivo. Golgi complexes, active membrane processes, reorganization of cell junctions, and bile canaliculi-like intercellular spaces were demonstrated.(Hepatology 1995; 22:546-552.)
Loss of angiotensin-II receptors in portal hypertensive rabbits
Authors: Sitzmann James V. , Wu Yuping , Acuilera Greti , Cahill Paul A. , Burns Cartland R.
Published: March 2007

Decreased splanchnic vascular response to exogenous angiotensin-II (A-II) infusion in portal hypertension has recently been documented. A-II receptor density and binding affinity in the mesenteric artery, portal vein, and adrenal gland of normal and portal hypertensive rabbits were studied. Portal hypertension was induced by partial portal vein ligation 3 weeks before study. There were no significant differences in serum concentrations of sodium, potassium, A-II, serum osmolality, or hematocrit between normal and portal hypertensive rabbits. The portal hypertensive portal vein exhibited a 60% fall in A-II receptor number from 65.1 ± 0.3 fmol/mg in normal to 27.0 ± 8 fmol/mg (P < .05) in portal hypertension. A significant decrease in receptor number occurred in the portal hypertensive mesenteric artery, 224 ± 39 fmol/mg compared with 345 ± 45 fmol/mg in normal rabbits, and in the adrenal cortex 6.8 ± 1.3 pmol/mg compared with 12.1 ± 2.5 pmol/mg in normal controls (P < .05). No significant difference in A-II receptor affinity was observed in tissues studied between normal and portal hypertensive rabbits. Autoradiographic study on A-II receptors was consistent with data from membrane binding assays. Receptor subtype analysis showed exclusive type I receptor binding in the mesenteric artery and portal vein. We conclude there is a global reduction in the A-II receptor number in portal hypertension that may mediate much of the decreased response to A-II seen in this disorder. This loss of the A-II receptor may partially explain hemodynamic derangements peculiar to portal hypertension. (Hepatology 1995; 22:559-564.)
Ascorbic acid inhibits chemically induced uroporphyria in ascorbate-requiring rats
Authors: Sinclair Peter R. , Gorman Nadia , Sinclair Jacqueline F. , Walton Heidi S. , Bement William J. , Lambrecht Richard W.
Published: March 2007

Ascorbate was previously shown to suppress accumulation of uroporphyrin (URO) in cultured chick embryo hepatocytes and to competitively inhibit microsomal oxidation of uroporphyrinogen catalyzed by cytochrome P4501A2. Here we used the Osteogenic Disorder Shionogi (ODS) mutant rat, which cannot synthesize ascorbic acid, to examine the in vivo effect of ascorbic acid on hepatic URO accumulation caused by treatment with 3-methylcholanthrene (MC) and 5-aminolevulinate (ALA). Female mutant rats maintained on three levels of dietary ascorbate (15,200, and 800 ppm) were treated for a total of 24 days. On the 11th and 16th days, rats were administered 3-methylcholanthrene, and 5-aminolevulinate was present continuously in the drinking water from day 14. Hepatic URO accumulated at the two lowest ascorbate levels, but not at 800 ppm ascorbate. The latter dose produced normal hepatic ascorbate levels. Plasma ascorbate levels were proportional to the hepatic values. Male rats also accumulated URO at the low dietary dose of ascorbic acid. The methylcholan-threne-induced increase in microsomal levels of CYP1A1 and CYP1A2, total cytochrome P450, and activities of uroporphyrinogen oxidation and ethoxyresorufin deethylase were not affected by the dietary level of ascorbate. Neither male nor female Fischer 344 rats accumulated URO when treated with the MC/ALA regime. Hepatic ascorbate concentrations in these rats were five-fold to seven-fold higher than they were in mutant rats that developed uroporphyria on 150 ppm dietary ascorbate. In ODS rats fed ascorbate at 90 but not 900 ppm in the diet, hexachlorobenzene caused hepatic URO accumulation, indicating that the effect of ascorbic acid is not unique to the regimen using methylcholanthrene. These results are consistent with a role for ascorbate in preventing chemically induced URO accumulation and suggest that ascorbate might have a role in preventing URO accumulation in humans. (Hepatology 1995; 22:565-572.)
Regulation of the α2(I) collagen gene transcription in fat-storing cells derived from a cirrhotic liver
Authors: Inagaki Yutaka , Truter Sharada , Greenwel Patricia , Rojkind Marcos , Unoura Masashi , Kobayashi Kenichi , Ramirez Francesco
Published: March 2007

Fat-storing cells (FSC) are the main producers of type I collagen in both normal and fibrotic livers. In order to elucidate the molecular mechanisms controlling collagen expression in FSC, we examined the transcription of the α2(I) collagen gene (COL1A2) in two distinct FSC clones, CFSC-2G and CFSC-5H, derived from a single CCl4-induced cirrhotic liver. The phenothpe of CFSC-2G resembles freshly isolated FSC, whereas that of CFSC-5H mimics activated myofibroblasts. Cell transfection experiments showed that the upstream sequence between nucleotides −378 and −183 is essential for COL1A2 transcription in both FSC clones. This is the same promoter region that is transcriptionally active and contains the binding site of a multimeric protein complex that mediates TGF-β stimulation of COL1A2 expression in dermal fibroblasts. We therefore examined the relative levels of endogenous and transfected COL1A2 transcription and their response to TGF-β treatment in the two FSC clones. The results showed that CFSC-5H expresses a significantly higher level of the COL1A2 mRNA than CFSC-2G. They also showed that TGF-β treatment increases both endogenous and transfected COL1A2 transcription in CFSC-2G but not in CFSC-5H. Interestingly, nuclear proteins from both FSC clones bind to the TGF-β-responsive element more strongly than those from dermal, fibroblasts. Altogether, the data suggest that collagen production in CFSC-5H cells has been already activated by the autocrine stimulation of TGF-β. In contrast, CFSC-2G cells are only partially activated but can be easily recruited to produce collagen when stimulated by exogenous TGF-β. Thus, we conclude that FSC activation during hepatofibrogenesis is probably a multistep process that involves autocrine and paracrine stimulation by TGF-β. (Hepatology 1995; 22:573-579.)
Intracellular pH regulation in hep G2 cells: Effects of epidermal growth factor, transforming growth factor-α, and insulinlike growth factor-II on Na+/H+ exchange activity
Authors: Strazzabosco Mario , Poci Carlo , Spirlì Carlo , Zsembery Akos , Granato Anna , Massimino Maria Luisa , Crepaldi Gaetano
Published: March 2007

Intracellular pH (pHi) plays an important role in the metabolic activation of quiescent cells after a proliferative stimulus, and Na+/H+ exchange activity is required for growth in some extrahepatic tumors. To investigate intracellular acid/base homeostasis in hepatoma cells and the effects of putative liver growth factors on Na+/H+ exchange activity, we have studied intracellular pH (pHi) regulation in Hep G2 cells, a well-differentiated hepatoma cell line, both in resting conditions and after administration of epidermal growth factor (EGF), transforming growth factor-α (TGFα), and insulinlike growth factor-II (IGF-II). The effects of fetal calf serum, TGFα, and amiloride on 3H-Thymidine incorporation were also studied. Amiloride (1 mmol/L) and external Na+ removal decreased baseline pHi in both HEPES and KRB. In HEPES, cells recovered from an acid load (20 mmol/L NH4Cl) by an amiloride inhabitable Na+/H+ exchange. In KRB, an additional, DIDS-inhibitable, Na+- and HCO3-dependent, but Cl−-independent acid extruder (Na:HCO3 cotransport) was activated. No evidence was found for a C1/HCO3 exchange acting as acid loader. Administration of EGF and TGFα, but not of IGF-H, induced a dose-dependent, amiloride-inhibitable increase in baseline pHi, together with an increase in Na+/H+ exchange activity, shifting to the right the JH/pHi curve. Finally, 3H-thymidine incorporation in Hep G2 cells, in the presence of FCS or TGFα, was strongly inhibited by amiloride. In conclusion, in Hep G2 cells, pHi is mainly regulated by Na+/H+ exchange, which activity can be stimulated by EGF and TGFα, but not by IGF-II. Administration of TGFα stimulates DNA synthesis, an effect that is blocked by amiloride, an inhibitor of Na+/H+ exchanger. These data suggest that Na+/H+ exchange activation may play a critical role in the growth of some hepatic tumors. (Hepatology 1995; 22:588-597.)
Enhanced nitric oxide synthase activity in portal hypertensive rabbits
Authors: Cahill Paul A. , Foster Cory , Redmond Eileen M. , Gingalewski Cindy , Wu Yuping , Sitzmann James V.
Published: March 2007

Portal hypertension (PHT) is characterized by splanchnic hyperemia caused by a reduction in mesenteric vascular resistance. Mediators of this hyperemia include nitric oxide (NO). This is based on several reports indicating a marked splanchnic hyporesponsiveness in PHT to vasoconstrictor stimuli, both in vitro and in vivo, and a subsequent reversal using specific inhibitors of NO synthase (NOS). The objective of this study was to determine directly if the generation of NO is altered in PHT vasculature. Thus, we compared NOS activity in the hyperemic vasculature of normal rabbits and rabbits with PHT (after undergoing partial portal vein ligation). Nicotinamide adenine dinucleotide phosphate diaphorase staining indicated the presence of NOS within the vascular endothelium. Ca2+-dependent NOS activity was significantly increased (P < .05) in PHT particulate fractions from the superior mesenteric artery and thoracic aorta, but not from the portal vein. There was no change in NOS activity within the cytosolic fractions. Arterial wall cyclic guanosine monophosphate (cGMP) levels and plasma nitrite levels were both significantly increased in PHT. These results show enhanced NOS activity in PHT hyperemic vessels concurrent with increased tissue cGMP levels. We conclude that enhanced NO synthesis contributes to the hyperdynamic circulation of PHT. (Hepatology 1995; 22:598-606.)
Activation of Na+/H+ exchanger by hepatocyte growth factor in hepatocytes
Authors: Kaneko Akira , Hayashi Norio , Tanaka Yuji , Horimoto Masayoshi , Ito Toshifumi , Sasaki Yutaka , Fusamoto Hideyuki , Kamada Takenobu
Published: March 2007

The effect of the hepatocyte growth factor (HGF) on the Na+/H+ exchanger was studied using primary cultured hepatocytes. HGF induced intracellular pH (pHi) elevation of 0.10 pH units in hepatocytes cultured for 4 to 7 hours; the response was lower after other culture periods. Even with the same culture period, intercellular heterogeneity was found in the responsiveness to HGF. This heterogeneity may be partially accounted for by the weak but significant correlation observed between the basal pHi level and the degree of pHi elevation caused by HGF in hepatocytes. The pHi elevation caused by HGF was blocked on pretreatment of the hepatocytes with amiloride, suggesting that HGF activates the Na+/H+ exchanger. This hypothesis was confirmed by the fact that HGF increased the initial rapid rate of cell alkalization of acid-loaded hepatocytes. The tyrosine kinase inhibitor, genistein, also blocked the elevation, consistent with the fact that HGF receptor/c-met has a tyrosine kinase domain. To clarify the signal transduction pathway from tyrosine kinase to the Na+/H+ exchanger, we examined the effects of inhibitors of other kinases (H-7, H-8, and W-7) on the HGF-induced pHi elevation and found that only W-7 blocked it. This pHi elevation was also prevented on preincubation of the hepatocytes with thapsigargin, which blocks the calcium response caused by HGF. These results suggest that HGF activates the Na+/H+ exchanger in hepatocytes through a tyrosine kinase-calcium/calmodulin-dependent pathway. (Hepatology 1995;22:629-636.)
Transport proteins and molecular biology: Is cloning the beginning or the end?
Authors: Stump Decherd , Weisiger Richard A. , Berk Paul D.
Published: March 2007

Long chain fatty acids (LCFAs) are an important energy substrate used by cardiac myocytes and other cells, but the mechanism whereby these molecules cross the plasma membrane is poorly understood. We used an expression cloning strategy and a cDNA library from 3T3-L1 adipocytes to identify a cDNA that, when expressed in cultured cells, augments uptake of LCFAs. This cDNA encodes a novel 646 amino acid fatty acid transport protein (FATP) with six predicted membrane spanning regions and that is integrally associated with membranes. Immunocytochemistry and subcellular fractionation of 3T3-L1 adipocytes show that FATP is localized to the plasma membrane. We propose that FATP is a plasma membrane transporter for LCFAs.
Uridine diphosphate glucoronosyl transferases, candidate antigens of liver-kidney microsomal antibodies in hepatitis delta virus-infected patients
Authors: Bonino Ferruccio , Brunetto Maurizia Rossana
Published: March 2007

Patients with chronic hepatitis D often have liver-kidney microsomal antibodies type 3 (LKM-3). These antibodies react with several microsomal antigens that have a molecular weight of 55 HDa and an isoelectric point of about 8. We studied the molecular nature of the antigen and, by immunoscreening a human liver cDNA expression library with KM-3 sera, found that uridine diphosphate glucuronosyl transferase (UGT) appeared as candidate antigens. We confirmed the identity of UGT as an antigen by reacting the sera with recombinant rabbit liver UGT proteins. Some sera reacted with rabbit UGT-2 proteins, but UGT-1 proteins were more sensitive and specific in detecting LKM-3 autoantibodies in patient sera. Anti-UGT-1 antibodies were detected in all LKM-3 positive sera from patients with hepatitis D and 1 out of 11 patients with autoimmune hepatitis type 2. Sera from patients who had hepatitis B only did not react with UGT proteins.
In vitro activation of woodchuck lymphocytes measured by radiopurine incorporation and interleukin-2 production: Implications for modeling immunity and therapy in hepatitis B virus infection
Authors: Cote Paul J. , Gerin John L.
Published: March 2007

Cellular immune responses to hepatitis B virus (HBV) play an important role in the resolution of acute infection. They also influence the course of chronic infection and disease but are inadequate to completely clear the infection. Woodchuck hepatitis virus (WHV) infection of the woodchuck can provide a model to study these processes. Lymphocyte responses of woodchucks were assessed by in vitro proliferation and/or interleukin (IL)-2 assays using mitogen (Concanavalin A [ConA]), cytokine (IL-2), superantigen (Staphylococcus aureus enterotoxin B [SEB]), major histocompatibility complex (MHC) allo-antigen (mixed lymphocyte reaction [MLR]), and viral antigens (woodchuck hepatitis virus core antigen: [WHcAg] and woodchuck hepatitis virus surface antigen [WHsAg]). ConA-stimulated woodchuck lymphocytes underwent cell division based on cell counting experiments and produced IL-2 as detected using an IL-2-dependent murine cell line but failed to incorporate sufficient tritiated thymidine; however, they did incorporate sufficient tritiated adenosine and deoxyadenosine to permit development of a meaningful proliferation assay. The IL-2 assay was sensitive and specific for detection of woodchuck IL-2 induced by mitogen, superantigen, and MLR, as shown by quantitative titration analysis and anti-body neutralization of ConA-supernatant activity. Cyclosporin A and FK506 specifically inhibited ConA-and SEB-induced IL-2 production by woodchuck lymphocytes. Positive two-way MLRs were detected by IL-2 production and proliferation assay between woodchucks from different geographic regions, thus indicating divergence among MHC molecules; however, occasional negative MLR reactions among indigenous pairs of woodchucks indicated that some woodchucks were mutually immunocompatible to some degree. The radioadenosine proliferation assay was sensitive for detecting peripheral blood lymphocyte responses to WHcAg and WHsAg in adult woodchucks with recently resolved acute infections. The above systems should facilitate the design of adoptive therapy and liver transplantation experiments in the woodchuck, and also enable modeling of immune responses that promote and maintain chronic hepadnavirus infection. (Hepatology 1995;22:687-699.)
Randomized trial comparing three different regimens of alpha-2a-interferon in chronic hepatitis C
Authors: Chemello Liliana , Bonetti Paola , Cavalletto Luisa , Talato Fabiola , Donadon Valter , Casarin Pietro , Belussi Fabio , Frezza Mario , Noventa Franco , Pontisso Patrizia , Benvegnù Luisa , Casarin Carla , Alberti Alfredo
Published: March 2007

Alpha-interferon (IFN-α) is an effective treatment for chronic hepatitis C, but only 20% to 30% of patients are apparently cured with the currently recommended schedule of 3 MU given three times a week for 6 months. To evaluate the efficacy of more aggressive treatment regimens, we have conducted a randomized trial in 174 patients with chronic hepatitis C using three different schedules: (1) 12-month treatment starting with 6 MU/three times a week and decreasing the dose on the basis of serum alanine transaminase (ALT) activities (group A: 59 cases); (2) fixed dose of 3 MU three times a week for 12 months (Group B: 61 cases), (3) fixed dose of 6 MU three times a week for 6 months (Group C: 54 cases). Patients were evaluated during therapy for biochemical and virological response and followed for at least 12 months after therapy to assess long-term efficacy and liver histological outcome. The genotype of infecting HCV was also analyzed in all patients, and predictors of response were determined by multivariate analysis. Serum ALT became normal during therapy in 76% of patients (95% confidence interval [CI]: 63 to 86), 65% (CI: 52 to 77), and 74% (CI: 60 to 85) in groups A, B, and C, respectively (P = NS). The corresponding figures for sustained response 12 months after therapy were 49% (CI: 36 to 62), 31% (CI: 20 to 44), and 28% (CI: 16 to 42) (A vs. B, P = .06; A vs. C, P = 0.03). Eighty-six percent of patients with sustained response cleared HCV-RNA from serum, and 72% improved histologically. Patients infected with HCV genotypes 2a and 3 had higher sustained response rates than those with Ib, independent of treatment schedule. In patients infected with genotype 1b, the rate of sustained response was related to dose and duration of therapy being 28% with schedule A, 16% with schedule B, and 9% with schedule C. Multivariate analysis indicated that younger age (P = .016), shorter disease duration (P = .003), and infection with HCV genotypes 2a (P = .0017) and 3 (P = .0083) were independent predictors of sustained response. These results indicate that sustained response to IFN-α in chronic hepatitis C is affected by dose and duration of therapy, particularly in patients infected with HCV genotype 1b. (Hepatology 1995; 22:700-706.)