bluuebunny/crossref_metadata_2025 huggingface dataset:
Hepatitis C virus infection in kidney transplant recipients
Authors: Lau Johnson Y. N. , Davis Gary L. , Brunson Mathew E. , Qian Ke-Ping , Lin Hsiang-Ju , Quan Stella , Dinello Robert , Polito Alan J. , Scornik Juan C.
Published: March 2007

To determine the prevalence and significance of hepatitis C virus infection in kidney transplant recipients, paired serum samples collected from 100 renal allograft recipients on admission for kidney transplantation and 1 yr after transplantation were tested for antibody to hepatitis C virus with second-generation enzyme immunoassay and recombinant immunoblot assay and for hepatitis C virus RNA with reverse transcription-polymerase chain reaction. Before kidney transplantation, hepatitis C virus antibody was detected with second-generation enzyme immunoassay in 18 patients (12 second-generation recombinant immunoblot assay-positive, 6 second-generation recombinant immunoblot assay-indeterminate). Nine of 12 second-generation recombinant immunoblot assay-positive and 2 of 6 second-generation recombinant immunoblot assay-indeterminate samples were hepatitis C virus RNA positive. In addition, 7 of 82 patients who had no detectable antibody on second-generation enzyme immunoassay or second-generation recombinant immunoblot assay were hepatitis C virus RNA positive. After kidney transplantation, hepatitis C virus antibody was detected in 19 patients (12 second-generation recombinant immunoblot assay-positive, 7 second-generation recombinant immunoblot assay-indeterminate, 14 seropositive for hepatitis C virus antibody). Eleven of 12 patients with second-generation recombinant immunoblot assay-positive results and 4 of 7 with second-generation recombinant immunoblot assay-indeterminate results were positive for hepatitis C virus RNA. Hepatitis C virus RNA was present in 28 patients 1 yr after kidney transplantation. Six patients appeared to have acquired active hepatitis C virus infection 1 yr after kidney transplantation (seroconverted to hepatitis C virus RNA positivity). We found no correlation between the presence of hepatitis C virus markers in the pretransplant and posttransplant sera and clinical/biochemical parameters or clinical outcome. Hepatitis C virus infection was clinically silent in most patients. Semiquantitation of serum hepatitis C virus RNA in 25 patients with hepatitis C virus infection showed that the hepatitis C virus RNA titer remained the same (n = 12) or increased at least 10-fold (n = 12), and only 1 patient had a decrease in serum hepatitis C virus RNA level. These data suggest that (a) the prevalence of hepatitis C virus infection is high in kidney transplant recipients, (b) active hepatitis C virus replication may be present in the absence of hepatitis C virus antibody, (c) many of these patients who are second-generation enzyme immunoassay-positive but second-generation recombinant immunoblot assay-indeterminate are hepatitis C virus RNA positive, (d) hepatitis C virus infection is usually clinically silent during the initial follow-up period and (e) antirejection immunosuppression may enhance hepatitis C virus replication. (HEPATOLOGY 1993;18:1027-1031).
Hepatic and extrahepatic HCV RNA strands in chronic hepatitis C: Different patterns of response to interferon treatment
Authors: Gil Beatriz , Qian Cheng , Riezu-Boj Jose I. , Civeira Maria P. , Prieto Jesús
Published: March 2007

We investigated the presence of positive (genomic) and negative (replicative intermediate) hepatitis C virus RNA strands in liver, peripheral mononuclear cells and serum from patients with chronic hepatitis C using a selective and semiquantitative polymerase chain reaction procedure. Negative and positive hepatitis C virus RNA strands were present in liver, serum and lymphoid cells in all untreated patients and in all those who did not respond to interferon therapy. In the latter group of patients, the titers of RNA strands in the liver and peripheral mononuclear cells at the end of the treatment were similar to those encountered in untreated patients, but the serum titers were about 100 times lower than pretreatment values. In patients who responded to interferon with normalization of serum aminotransferase levels (n = 10), the rate of detection and the titer of the two viral strands in liver, serum and mononuclear cells were markedly decreased at the end of the therapy. In the six responders who did not relapse after interferon withdrawal, both hepatitis C virus RNA strands were absent from the liver, serum and lymphoid cells. By contrast, the positive RNA strand was present in liver cells, mononuclear cells or both at the end of therapy in all patients who experienced posttherapy relapse. In conclusion, our results indicate that interferon can clear hepatitis C virus from hepatic and extrahepatic sites only in responder patients. Disappearance of genomic hepatitis C virus RNA from the liver and from mononuclear cells may predict complete response without posttherapy relapse. (HEPATOLOGY 1993;18:1050-1054).
Peripheral-blood mononuclear cell responses to recombinant hepatitis C virus antigens in patients with chronic hepatitis C
Authors: Schupper Haim , Hayashi Paul , Scheffel James , Acettuno Sherri , Paglieroni Teresa , Holland Paul V. , Zeldis Jerome B.
Published: March 2007

Peripheral blood mononuclear cell proliferative responses in vitro to recombinant yeast or Escherichia coli hepatitis C virus fusion proteins were evaluated in 20 patients with chronic hepatitis C who were reactive for antibody to hepatitis C virus (on enzyme immunoassay, version 2.0, and a four-antigen recombinant immunoblot assay). Twenty age-matched, healthy individuals negative for antibody to hepatitis C virus were used as a control group. Peripheral-blood mononuclear cells from all chronic hepatitis C patients with antibodies to hepatitis C virus antigens c22 and c100-3 proliferated in vitro in response to the corresponding recombinant hepatitis C virus fusion protein. Peripheral-blood mononuclear cells from 75% of patients infected with hepatitis C virus proliferated in response to cytidine monophosphate-keto-3-deoxyoctulosonic acid-core recombinant antigen but there was no proliferative response to cytidine monophosphate-keto-3-deoxyoctulosonic acid-EF (derived from the NS5 region). All hepatitis C virus-infected patients had 33c antibody, but peripheral-blood mononuclear cells from only 9 of 14 (64%) proliferated in vitro in response to 33c. Ninety-five percent of all hepatitis C virus-in-fected patients had peripheral-blood mononuclear cells that proliferated in response to at least one recombinant hepatitis C virus fusion protein. The numbers and percentages of CD3 T cells, CD19 B cells and natural killer cells from patients with chronic hepatitis C virus infection did not differ from those in the healthy control group. However, the number of non-major histocompatibility complex-restricted cytotoxic T cells (CD3-positive, CD56-positive, CD16-positive) was increased in patients with chronic hepatitis C virus infection (p < 0.05). (HEPATOLOGY 1993;18:1055-1060).
Positron-emission tomographic localization of abnormalities of brain metabolism in patients with minimal hepatic encephalopathy
Authors: Lockwood Alan H. , Murphy Brian W. , Donnelly Kerry Z. , Mahl Thomas C. , Perini Sean
Published: March 2007

Many patients with compensated cirrhosis without overt hepatic encephalopathy have deficits in visualspatial perception, a condition we call minimal hepatic encephalopathy. Five patients with alcohol-induced cirrhosis and nine control subjects underwent positron-emission tomographic imaging of the brain with 18F-fluorodeoxyglucose. Patients also underwent neuropsychological and clinical chemistry tests. The patients had mild arterial hyperammonemia (62 ± 13 μmol/L, range = 11 to 35 μmol/L) and other abnormalities typical of patients with cirrhosis. The patients’ mean percentile scores on the digit symbol and block design subtests, from the Wechsler Adult Intelligence Scale (revised), and Purdue pegboard test were 11 ± 7, 24 ± 7 and 7 ± 8 (right hand). Tests of vocabulary, memory, and new learning were normal. The technique of statistical parametric mapping was used to identify regions where cerebral 18F-fluorodeoxyglucose uptake and metabolism were abnormal. We noted significant reductions in the cingulate gyrus, a center mediating attention, target analysis and response formulation and significant increases in visual associative regions subserving motion and color perception and object orientation. We suggest that minimal hepatic encephalopathy is due to a deficit in the detection and formulation of responses to visual stimuli, a function of the cingulate, which is a part of the anterior attentional system of the brain. Increases in 18F-fluorodeoxyglucose metabolism may be compensatory. These studies show that brain regions differ in their sensitivity to the agents that cause hepatic encephalopathy and that positron-emission tomography is useful in studying the pathophysiology of this disorder. (HEPATOLOGY 1993;18:1061-1068).
Human monoclonal antibodies from a patient with primary biliary cirrhosis that recognize two distinct autoepitopes in the E2 component of the pyruvate dehydrogenase complex
Authors: Matsui Mieko , Nakamura Minoru , Ishibashi Hiromi , Koike Kichiko , Kudo Jiro , Niho Yoshiyuki
Published: March 2007

Peripheral B lymphocytes from a patient with primary biliary cirrhosis were infected with Epstein-Barr virus, and Epstein-Barr virus-transformed B lymphocytes producing large amounts of IgG antibodies to pyruvate dehydrogenase complex were selected, expanded and fused with the human-mouse heteromyeloma cell line F3B6. The resulting Epstein-Barr virus-transformed B-cell hybrids were repeatedly cloned by limiting dilution, and three stable hybridoma clones producing human monoclonal antibodies to pyruvate dehydrogenase complex were generated. These monoclonal antibodies, designated M18GP8, M37GP11 and M82GP8, specifically bound to pyruvate dehydrogenase complex, and their dissociation constant with pyruvate dehydrogenase complex was calculated to be 2.4 × 10−11, 2.3 × 10−10 and 2.6 × 10−11 mol/L, respectively. These three monoclonal antibodies stained the mouse stomach/kidney cryostat sections in a typical immunofluorescence pattern of antimitochondrial antibody. Furthermore, the enzymatic activity of pyruvate dehydrogenase complex was almost completely inhibited by the three monoclonal antibodies. Western blotting analysis revealed that M18GP8 and M82GP8 reacted with only pyruvate dehydrogenase complex-E2 in contrast to M37GP11, which reacted with both pyruvate dehydrogenase complex-E2 and protein X. The binding of monoclonal antibody M37GP11 to solid-phase pyruvate dehydrogenase complex was partially inhibited by two different synthetic peptides corresponding to both the inner and outer lipoyl-binding domains of pyruvate dehydrogenase complex-E2. These monoclonal antibodies, which are the first human monoclonal antibodies to pyruvate dehydrogenase complex generated from a patient with primary biliary cirrhosis, will be a valuable tool for studying the B-cell autoepitopes in PDC and the mechanism of autoantibody production in primary biliary cirrhosis. (HEPATOLOGY 1993;18:1069-1077).
Factors predictive of liver histopathological appearance in chronic alcoholic pancreatitis with common bile duct stenosis and increased serum alkaline phosphatase
Authors: Lesur Gilles , Levy Philippe , Flejou Jean-François , Belghiti Jacques , Fekete François , Bernades Pierre
Published: March 2007

In the course of alcoholic chronic pancreatitis, increased serum alkaline phosphatase level is usually caused by common bile duct stenosis but may also be due to alcoholic liver disease. The aims of this prospective study were to investigate whether clinical, biochemical and radiological factors could predict liver histopathological appearance. The study comprised 48 patients with chronic alcoholic pancreatitis, common bile duct stenosis and increased serum alkaline phosphatase levels; clinical, biochemical, radiological and histological data were recorded in all cases. Liver biopsy examination (surgical [n = 45] or intercostal [n = 3]) showed (a) biliary obstructive liver abnormalities (n = 33), which were severe in 20 cases (biliary fibrosis in 15, secondary biliary cirrhosis in 3, secondary sclerosing cholangitis in 2) and moderate in 13 cases; (b) alcoholic liver disease in 9; and (c) normal liver in 6. Clinical, biochemical and radiological data were not statistically different between patients with biliary obstructive liver disease and those with alcoholic liver disease. Forty-five patients underwent surgery; two patients with alcoholic hepatitis died after surgery, at the beginning of this study. We conclude that in chronic alcoholic pancreatitis with common bile duct stenosis and increased serum alkaline phosphatase levels, clinical, biochemical and radiological data cannot be used to predict the type of liver lesions. Therefore liver biopsy is warranted to identify (a) alcoholic hepatitis, which increases operative risk; and (b) biliary obstructive liver disease, frequent and often severe, in which surgical biliary decompression should be considered. (HEPATOLOGY 1993;1078-1081).
Reduction of variceal pressure by propranolol: Comparison of the effects on portal pressure and azygos blood flow in patients with cirrhosis
Authors: Feu Faust , Bordas Josep María , Luca Angelo , García-Pagán Joan Carles , Escorsell Angels , Bosch Jaume , Rodes Joan
Published: March 2007

This study investigated the correlation between changes in hepatic hemodynamics and esophageal variceal pressure-measured with a noninvasive, pressure-sensitive endoscopic gauge-in 37 portal-hypertensive cirrhotic patients receiving propranolol (0.15 mg/kg, intravenously; n = 21) or placebo (n = 16) under strict double-blind conditions. Placebo administration had no effect on hepatic venous pressure gradient, azygos blood flow or variceal pressure. Propranolol caused a significant reduction in hepatic venous pressure gradient (from 19.6 ± 1 to 17.3 ± 1 mm Hg, p < 0.001), azygos blood flow (from 0.61 ± 0.06 to 0.39 ± 0.03 L/min, p < 0.001) and variceal pressure (from 13.1 ± 0.9 to 10.2 ± 0.9 mm Hg, p < 0.001). In eight patients (propranolol nonresponders) hepatic venous pressure gradient was not modified or decreased by less than 10% after propranolol (mean change, −4.1 ± 1.6%). However, we found no significant differences between propranolol responders and nonresponders with regard to the decrease in variceal pressure (3.3 ± 0.7 vs. 2.3 ± 1.4 mm Hg) and azygos blood flow (0.23 ± 0.07 vs. 0.21 ± 0.07 L/min). As expected, in most propranolol responders (11 of 13), reduction in hepatic venous pressure gradient was accompanied by a similar response in variceal pressure (>10% decrease). However, among propranolol nonresponders, in terms of reduction in hepatic venous pressure gradient, four out of eight patients had decreases greater than 10% in variceal pressure. The results of this study confirm that reduction in hepatic venous pressure gradient by propranolol is associated with a significant decrease in variceal pressure and azygos blood flow. These favorable effects are also observed in some patients in whom hepatic venous pressure gradient fails to decrease by more than 10% of the baseline value, suggesting that propranolol administration has beneficial effects in some of these patients. (HEPATOLOGY 1993;18:1082-1089).
Preoperative measurement of segmental liver volume of donors for living related liver transplantation
Authors: Kawasaki Seiji , Makuuchi Masatoshi , Matsunami Hidetoshi , Hashikura Yasuhiko , Ikegami Toshihiko , Chisuwa Hisanao , Ikeno Tatsuo , Noike Terumasa , Takayama Tadatoshi , Kawarazaki Hideo
Published: March 2007

Segmental liver volume determination by computed tomographic scan was carried out preoperatively in nine donors for living related liver transplantation. The calculated volume was compared with the graft size actually obtained by three types of donor hepatectomy. The volume of the left lateral segment (175 to 241 ml) and the left lobe (310 to 490 ml) varied markedly among the donors. The ratio of the left lobar to total liver volume also showed a wide range of values (23.2% to 35.9%). The value of the left lobar volume did not correlate positively with the donor’s body weight, suggesting that graft size cannot be predicted only on the basis of the donor’s body size. Segmental graft liver volume was estimated by use of computed tomographic scan, with acceptable accuracy on comparison with the graft volume actually obtained. In living related liver transplantation, the type of donor hepatectomy should be selected on the basis of the segmental liver volume of the donor in addition to the recipient’s body size so that liver failure can be prevented in recipients and the donor’s safety can be assured as far as possible. (HEPATOLOGY 1993;18:1115-1120).
Treatment of small hepatocellular carcinoma in cirrhotic patients: A cohort study comparing surgical resection and percutaneous ethanol injection
Authors: Castells Antoni , Bruix Jordi , Bru Concepció , Fuster Josep , Vilana Ramon , Navasa Miquel , Ayuso Carmen , Boix Loreto , Visa Josep , Rodés Joan
Published: March 2007

This study was intended to compare the survival rates of two contemporary cohorts of patients with solitary hepatocellular carcinomas ≤ 4 cm subjected to surgical resection (n = 33) or percutaneous ethanol injection (n = 30). Outcomes in a third cohort, 21 patients with hepatocellular carcinoma who underwent orthotopic liver transplantation, were also assessed. Surgical and ethanol-treated patients were similar with regard to age and tumor stage, differing only in liver function; 30 of the 33 surgical patients were of Child-Pugh class A, whereas only 7 of the 30 ethanol-treated patients were of class A (p < 0.05). Surgical resection was successful in 30 cases; ethanol injection achieved initial success in 23 patients. Tumor recurrence rate at 2 yr was 45% in the surgical group and 66% in the ethanol group. The difference was significant only for cases with tumors between 3 and 4 cm. Despite poorer liver function, the 1-, 2-, 3- and 4-yr survival rates of ethanol-treated patients (83%, 66%, 55% and 34%, respectively) were not different from those of surgically treated patients (81%, 73%, 44% and 44%, respectively). The 1- and 2-yr survival rates of patients given liver transplants were 81% and 66%, without tumor recurrence, after 16-mo follow-up. These data confirm that ethanol injection is a useful treatment for patients with solitary small hepatocellular carcinomas and suggest that surgical resection and liver transplantation may achieve better results only after strict candidate selection to reduce mortality and tumor recurrence during follow-up. (HEPATOLOGY 1993;18:1121-1126).
Primary sclerosing cholangitis and Hodgkin’s disease
Authors: Man Kevin M. , Drejet Anne , Keeffe Emmet B. , Garcia-Kennedy Richard , Imperial Joanne C. , Esquivel Carlos O.
Published: March 2007

Three patients with primary sclerosing cholangitis and Hodgkin’s disease, a previously unrecognized association, are reported. All three patients were men, and one patient had Crohn’s disease of the colon. Primary sclerosing cholangitis was diagnosed 2, 11 and 17 yr before diagnosis of Hodgkin’s disease in the three patients, and all three had advanced biliary cirrhosis prompting referral for liver transplantation. The symptoms of Hodgkin’s disease were often masked by similar manifestations of primary sclerosing cholangitis, particularly symptoms of recurrent biliary sepsis. Hodgkin’s disease is another disorder that may occur in patients with primary sclerosing cholangitis, particularly in the setting of advanced disease, and may be masked by the underlying hepatobiliary disease. (HEPATOLOGY 1993;18:1127-1131).
Circulating interleukin-1 and tumor necrosis factor antagonists in liver disease
Authors: Tilg Herbert , Vogel Wolfgang , Wiedermann Christian J. , Shapiro Leland , Herold Manfred , Judmaier Gert , Dinarello Charles A.
Published: March 2007

The proinflammatory cytokines interleukin-1 and tumor necrosis factor-α are thought to play important roles in the pathophysiology of liver disease. Specific antagonists of these cytokines have been found in recent years. Interleukin-1 receptor antagonist is a specific interleukin-1 antagonist. The soluble receptor derived from the cell-surface p55 tumor necrosis factor receptor p55 is a naturally occurring substance that inhibits the biological effects of tumor necrosis factor. We used specific radioimmunoassays to detect circulating interleukin-1 receptor antagonist and tumor necrosis factor soluble receptor p55 levels in 14 patients with acute viral hepatitis and in 160 patients with various chronic liver diseases. Levels of interleukin-1 receptor antagonist and, especially, tumor necrosis factor soluble receptor were markedly increased in most patients with chronic liver disease regardless of pathogenesis and in viral hepatitis. Patients with chronic liver disease and cirrhosis showed significantly higher levels of both cytokine antagonists than did noncirrhotic patients. Correlations between interleukin-1 receptor antagonist and tumor necrosis factor soluble receptor were more significant than those of either antagonist with C-reactive protein or blood sedimentation rate. Interleukin-1 receptor antagonist and tumor necrosis factor soluble receptor levels were also positively correlated with bilirubin and AST levels. We conclude that circulating levels of interleukin-1 receptor antagonist and tumor necrosis factor soluble receptor may reflect ongoing disease activity and probably modulate some effects of endogenous interleukin-1 and tumor necrosis factor. (HEPATOLOGY 1993;18:1132-1138).
Oxygen free radical production mediated by cocaine and its ethanol-derived metabolite, cocaethylene, in rat hepatocytes
Authors: Boelsterli Urs A. , Wolf Armin , Göldlin Christian
Published: March 2007

Cocaine ethyl ester (“cocaethylene”) is a cocaine metabolite formed by hepatic carboxylesterases in the presence of ethanol. In the human liver, this pharmacologically active cocaine analog may reach concentrations similar to those of cocaine. To further investigate the intrinsic hepatotoxic potential of cocaethylene and to compare its effects with the known hepatocyte-damaging effects of cocaine, primary short-term cultures of hepatocytes derived from phenobarbital-pretreated rats were exposed to cocaine or cocaethylene (10 to 1000 μmol/L). Both drugs caused time- and concentration-dependent release of lactate dehydrogenase into the extracellular medium in a congruent pattern. Spontaneous hydrolysis of cocaethylene was negligible at pH 7.4, and the rate of 2-diethylaminoethyl-2,2-diphenylvalerate (SKF-525A) and phenylmethylsulfonylfluoride-sensitive enzymatic degradation in hepatocyte cultures was similar to that of cocaine (half-time, ≈ 3 hr). Furthermore, both cocaine and cocaethylene induced massive NADPH-dependent reactive oxygen species production in hepatic microsomal suspensions and in homogenates of cultured hepatocytes, as directly demonstrated by superoxide dismutase- and catalase-sensitive luminol chemiluminescence. The integrated photon emission and the maximal slopes of the luminol chemiluminescence displayed similar concentration-response curves for cocaine and cocaethylene. These data suggest that reactive oxygen species, generated during the interaction of cocaine and cocaethylene with cytochrome P-450, may be the common mediators of hepatotoxicity induced by these compounds. Moreover, they indicate that the potential for cocaethylene to induce acute lethal cell injury in hepatocytes through oxidative mechanisms is similar to that of its parent compound, cocaine. (HEPATOLOGY 1993;18:1154-1161).
Vasoactive effects of bile salts in cirrhotic rats: In vivo and In vitro studies
Authors: Pak Jung-Min , Lee Samuel S.
Published: March 2007

To clarify a possible pathogenic role for bile salts in the hyperdynamic circulation of cirrhosis, we studied the vasoactive effects of three different bile salts-tauroursodeoxycholic acid, taurochenodeoxycholic acid and taurodeoxycholic acid-in cirrhotic rats. Cirrhosis was induced with bile duct ligation; controls underwent sham surgery. In vivo, the bile salts were intravenously infused at one of three doses (1.2 × 10−7, 1.2 × 10−6 and 6.0 × 10−5 mol · 100 gm−1 · min−1) for 5 min. Taurochenodeoxycholic acid and taurodeoxycholic acid infusions increased mesenteric arterial blood flow and conductance and induced systemic arterial hypotension, whereas tauroursodeoxycholic acid had no significant effect. At similar plasma levels of bile salts, the responses in cirrhotic rats were attenuated compared with those of controls. In vitro, isolated rings of superior mesenteric and carotid arteries and portal vein were precontracted with phenylephrine; then dilatory responses to cumulative doses of bile salts (10−6 to 10−3 mol/L) were measured. In all three vessels, taurodeoxycholic acid produced stronger dilatory effects than did taurochenodeoxycholic acid, whereas tauroursodeoxycholic acid showed no significant effect. Vessels from cirrhotic and control rats did not differ in degree of response. These results indicate that bile salts are directly vasoactive and can induce splanchnic vasodilation at the pathophysiological plasma levels seen in cirrhosis. Bile salts may be involved in the pathogenesis of splanchnic hyperemia and hyperdynamic circulation in cirrhosis. (HEPATOLOGY 1993;18:1175-1181).
Expression of multidrug resistance genes in rat liver during regeneration and after carbon tetrachloride intoxication
Authors: Nakatsukasa Harushige , Silverman Jeffrey A. , Gant Timothy W. , Evarts Ritva P. , Thorgeirsson Snorri S.
Published: March 2007

We analyzed expression of multidrug resistance (mdr) genes in rat liver during regeneration after partial hepatectomy or carbon tetrachloride-induced necrosis. In situ hybridization revealed that in the normal liver the cellular distribution of mdr transcripts and protein is restricted to hepatocytes and that a gradient, highest in zone 1 and lowest in zone 3, exists in the level of the mdr transcripts in the liver acinus. Increased levels of mdr1a and mdr1b transcripts were observed 3 hr after administration of carbon tetrachloride and remained increased for the next 5 days. In contrast, increased expression of mdr1a and mdr1b was first observed 24 hr after partial hepatectomy. Use of gene-specific probes to compare the time courses of mdr1b and mdr2 expression after carbon tetrachloride administration showed distinctly different patterns of expression; mdr1b reached a maximum level of expression at 12 hr, whereas increased mdr2 expression was first observed 48 hr after administration. Nuclear run-on analysis at 12 and 24 hr after carbon tetrachloride administration demonstrated 10-fold and eightfold increases in mdr transcription, respectively. However, 72 hr after carbon tetrachloride treatment the rate of mdr transcription was back to the control level. The cellular patterns of mdr expression after partial hepatectomy and carbon tetrachloride administration were similar; the increase was first observed in zone 1 and gradually extended into zone 3. These data strongly suggest that the physiological roles of mdr1b and mdr2 are different and that liver regeneration is an appropriate model for elucidating these differences. (HEPATOLOGY 1993;18:1202-1207).
Modulators of the protein kinase C system influence biliary excretion of cationic drugs
Authors: Steen Herman , Smit Hans , Nijholt Almar , Merema Marjolijn , Meijer Dirk K. F.
Published: March 2007

To investigate whether hepatobiliary transport of organic cations is under regulatory control, we studied transport of tri-n-butylmethylammonium in the isolated perfused rat liver and in isolated rat hepatocytes. Transport was investigated in the presence of modulators of the protein kinase C and the cyclic AMP second-messenger system. In the isolated perfused rat liver, it was observed that compounds modulating protein kinase C activity clearly affected the biliary excretion process of the cation tri-n-butylmethylammonium. Phorbol 12-myristate 13-acetate, a compound that directly stimulates protein kinase C, elevated the biliary excretion rate of tri-n-butylmethylammonium in a concentration-dependent manner, reaching a twofold increase at 60 nmol/L of the phorbol ester. The inactive derivative 4α-phorbol 12, 13-didecanoate (60 nmol/L) did not show any effect. Vasopressin (48 nmol/L), a receptor-mediated activator of protein kinase C, stimulated the excretion rate of the cation by about 50%. Staurosporin (1 μmol/L), an inhibitor of protein kinase C, clearly decreased the biliary excretion rate of the cation and also blocked its stimulation by phorbol 12-myristate 13-acetate. Neither phorbol 12-myristate 13-acetate nor vasopressin (at concentrations ranging from 10−9 to 10−6 mol/L) affected the initial uptake velocity of tri-n-butylmethylammonium in isolated hepatocytes and isolated perfused livers, whereas staurosporin (1 μmol/L) showed only a modest inhibition of the uptake of the cation. It is inferred that the effect of protein kinase C modulators on hepatobiliary transport of organic cations occurs at the level of carrier-mediated transport in the canalicular membrane. Because bile flow was only slightly affected by these agents, effects on biliary excretion rate of the cation are unlikely to be caused by changes in bile flow. With regard to the cyclic AMP second-messenger system, neither glucagon (concentration range of 10−9 to 10−6 mol/L), a receptormediated activator of adenylate cyclase, nor forskolin (100 μmol/L), a direct activator of adenylate cyclase and dibutyryl cyclic AMP (100 μmol/L), affected the biliary excretion rate and the hepatic uptake rate of the cation in these preparations. In conclusion, cell-to-bile transport of the organic cation tri-n-butylmethylammonium at the canalicular level is directly or indirectly regulated by protein kinase C. Neither the protein kinase C nor the cyclic AMP second-messenger systems seem to be involved in the hepatic uptake process of the cation. (HEPATOLOGY 1993;18:1208-1215).
Membrane-associated C-reactive protein on rat liver macrophages is synthesized within the macrophages, expressed as Neo-C-reactive protein and bound through a C-reactive protein-specific membrane receptor
Authors: Egenhofer Christine , Alsdorff Kathrin , Fehsel Karin , Kolb-Bachofen Victoria
Published: March 2007

We could recently show that rat liver macrophages (Kupffer cells) express a membrane-bound form of C-reactive protein on their surface. Because it is removed by washing the cells in buffers containing Ca++-chelators, membrane-bound C-reactive protein is a peripheral protein rather than an integral part of the Kupffer cell membrane. This Kupffer cell membrane-bound C-reactive protein is identical to the galactose-specific particle receptor previously characterized. We now present evidence that Kupffer cells do not acquire soluble serum C-reactive protein but synthesize their own membrane-bound C-reactive protein. By RNA-RNA in situ hybridization, it was found that hepatocytes are not the only sort of liver cells synthesizing C-reactive protein, but C-reactive protein-specific mRNA is present also in Kupffer cells. During acute-phase response C-reactive protein mRNA is found in increased amounts within liver macrophages too. Furthermore, by labeling experiments with antisera against native, pentameric soluble serum C-reactive protein and monoclonal antibodies against a neoepitope present on C-reactive protein subunits only, we could establish that the membrane-bound C-reactive protein expressed on the liver macrophage is not the pentameric molecule of soluble serum C-reactive protein, but rather consists of C-reactive protein subunits. Finally, we present evidence that liver macrophages contain a binding protein in their plasma membrane, with an apparent molecular weight of 59 to 61 kD, specific for C-reactive protein and similar to the one previously isolated from macrophage cell lines. (HEPATOLOGY 1993;18:1216-1223).
Selectivity and sensitivity of changes in serum bile acids during induction of cirrhosis in ratsselectivity and sensitivity of changes in serum bile acids during induction of cirrhosis in rats
Authors: Azer Samy A. , Murray Michael , Farrell Geoffrey C. , Stacey Neill H.
Published: March 2007

Because some patients with cirrhosis have serum transaminase levels within the normal range, a prospective study was undertaken to determine whether the concentration of individual serum bile acids would be a sensitive indicator of development of cirrhosis. The choline-deficient rat has been used as a model for study of these changes. Using high-performance liquid chromatography, we measured the concentrations of individual serum bile acids at 3, 6, 10, 20 and 30 wk of dietary intake. Serum levels of total glycine- and taurine-conjugated bile acids were elevated at all stages tested as compared with levels in control groups (choline supplemented). Similarly, unconjugated bile acids and, particularly, cholic acid showed significantly higher levels at all stages except with the occurrence of cirrhosis at 30 wk, at which time there was a significantly lower level for unconjugated bile acids (0.48 ± 0.11 vs. 1.40 ± 0.36 in controls) and for cholic acid (0.17 ± 0.05 vs. 0.91 ± 0.39 in controls). The ratio of serum cholic acid to serum chenodeoxycholic acid changed in temporal relationship to progression in the histological lesions in livers of these rats. The ratio was at its highest at 78 ± 3 at 3 wk (no histological change) and decreased with increasing time and changes in histological appearance until 30 wk, at which time it was down to 1.6 ± 0.6. The routinely used markers of liver injury (serum ALT, alkaline phosphatase and bilirubin), however, did not match the progression of hepatic histological changes. The relationship of the increase in serum bile acids to the cirrhotic process is supported by qualitatively similar findings in a second rat model. Thus the hepatic pathological changes in cirrhotic rats were reflected by the changes in the individual serum bile acids together with the ratio of serum cholic acid to serum chenodeoxycholic acid, but not with other standard tests of liver function. (HEPATOLOGY 1993;18:1224-1231).
Regulation of human liver cytochromes P-450 in family 3A in primary and continuous culture of human hepatocytes
Authors: Schuetz Erin G. , Schuetz John D. , Strom Stephen C. , Thompson Melissa T. , Fisher Robert A. , Molowa David T. , Li Donna , Guzelian Philip S.
Published: March 2007

The cytochrome P-450 3A gene family comprises the dominant forms of cytochrome P-450 found in human liver. We examined as a possible useful system for studying the regulation of cytochrome P-450 3A under controlled conditions in vitro, primary monolayer cultures of human hepatocytes and compared the results with those obtained from the study of cytochrome P-450 3A in the human hepatoblastoma cell line HepG2 or in the human hepatocellular carcinoma cell line TONG/HCC. Using 3A antibodies, 3A cDNAs and 3A3, 3A4, 3A5 and 3A7 isozyme-specific oligonucleotides as probes, we determined that primary human hepatocyte cultures routinely expressed a 3A3/4* immunoreactive protein and 3A mRNA. These gene products were well maintained for many days and were induced by treatment of the cultures with dexamethasone, phenobarbital, macrolide antibiotics, the HMG CoA reductase inhibitor lovastatin or an antifungal agent, clotrimazole. Of six donor livers examined, only two contained mRNA or protein for 3A5, a form found in only a few adult human subjects. In cultures prepared from one of these two livers, 3A5 mRNA was detectable for several days. In cultures of hepatocytes from the remaining four human livers that did not contain 3A5 mRNA or protein, we detected neither spontaneous nor inducible 3A5 proteins or mRNAs. HepG2 cells contained only 3A7 protein, a form found in human fetal liver, even after treatment with inducers. Treatment of HepG2 cells with dexamethasone, macrolide antibiotics, phenobarbital and phenobarbital-like inducers or lovastatin produced dose-dependent induction of 3A7 mRNA and 3A7 immunoreactive protein. TONG/HCC cells contained 3A3, 3A4 and 3A5 mRNAs, but only 3A5 immunoreactive protein could be detected. Treatment of TONG/HCC cells with dexamethasone, macrolide antibiotics or phenobarbital induced 3A3 and 3A4 mRNAs, as observed in primary cultures, but failed to increase 3A5 mRNA or protein. We conclude that, in addition to primary cultures of human hepatocytes, replicating human hepatic lines HepG2 and TONG/HCC may be useful models for examining regulation of cytochrome P-450 3A genes polymorphically expressed in the living human being. (HEPATOLOGY 1993;18:1254-1262).
A new bile acid transporter?
Authors: Hardikar Winita , Suchy Frederick J.
Published: March 2007

Transport systems involved in uptake and biliary secretion of bile salts have been extensively studied in rat liver; however, little is known about these systems in the human liver. In this study, we investigated taurocholate (TC) transport in canalicular and basolateral plasma membrane vesicles isolated from 15 human livers (donor age 6-64 yr). ATP stimulated the uptake of TC into both canalicular and basolateral human liver plasma membrane vesicles (cLPM and bILPM, respectively). Considerable interindividual variations in the transport velocity were observed in the different membrane preparations used: 9.0 ± 1.3 (mean ± SEM, n = 17; range 1.6-18.0) and 9.3 ± 2.0 (range 1.1-29.8) pmol TC · protein−1 · min−1 at 1.0 μM TC for cLPM and bILPM, respectively. TC transport was temperature sensitive and showed saturation kinetics with a high affinity for TC (K
m 4.2 ± 0.7 μM and 3.7 ± 0.5 μM for cLPM and bILPM, respectively). Transport was dependent on the ATP concentration and saturable (K
m 0.25 ± 0.03 mM, n = 3). Neither nitrate, which reduces membrane potential, nor the protonophore FCCP strongly inhibited ATP-dependent TC transport, indicating that membrane potential and proton gradient are not involved in this process. TC transport was significantly inhibited by the classical anion transport inhibitor 4,4′-diisothiocyanostilbene-2,2′-disulfonate (250 μM) and the glutathione conjugate S-(2,4-dinitrophenyl)glutathione (100 μM). In conclusion, high affinity ATP-dependent TC transport is present in human liver at both the canalicular and the basolateral sides of the hepatocyte.
Rapid sequence variation of the hypervariable region of hepatitis C virus during the course of chronic infection
Authors: Kurosaki Masayuki , Enomoto Nobuyuki , Marumo Fumiaki , Sato Chifumi
Published: March 2007

To evaluate the clinical significance of sequence variations in the hypervariable region of hepatitis C virus during the course of chronic infection, we performed pairwise comparison of the predominant nucleotide sequences. Hepatitis C virus RNA was extracted from two plasma samples obtained from 12 chronically infected Japanese patients over approximately 1 yr. Complementary DNA containing the hypervariable region was amplified by means of reverse transcription-polymerase chain reaction and was directly sequenced for determination of predominant sequences. In all 12 individuals, the predominant sequence of the hypervariable region at the second time point differed from that at the first time point, and significant sequence variation was observed during this short period. A high proportion of these nucleotide substitutions (90%) resulted in changes of predicted amino acid sequences, indirect evidence of in vivo positive selection for these variants. There appeared to be an important difference in the rate of nucleotide sequence variation of the hypervariable region between four patients with flare-ups of their ALT levels (1.54 to 2.24 × 10-−1/genome site/yr) and eight patients with quiescent courses (0.13 to 1.21 × 10−1/genome site/yr). These results demonstrate that rapid sequence variations of the hypervariable region of predominant virus population take place during the natural course of chronic hepatitis C virus infection. These sequence variations seem to occur as an adaptive response of hepatitis C virus to evade host immunity and may play a major role in the establishment of persistent infection and in the occasional flare-up of hepatitis. (HEPATOLOGY 1993;18:1293-1299.)
Persistence of hepatitis B virus DNA in the liver after loss of HBsAg in chronic hepatitis B
Authors: Fong Tse-Ling , Di Bisceglie Adrian M. , Gerber Michael A. , Waggoner Jeanne G. , Hoofnagle Jay H.
Published: March 2007

To determine whether patients with chronic hepatitis B who lose hepatitis B virus DNA and HBsAg from the serum completely resolve the hepatitis and virus infection, we evaluated serum and liver tissue from 11 patients who had lost HBsAg. These patients were evaluated for clinical, histological and serological features and for hepatitis B virus DNA by use of hybridization and polymerase chain reaction techniques. Liver biochemical test results were normal in all except two patients who had mild aminotransferase elevations. All sera were negative for hepatitis B virus DNA by direct hybridization, and only one was positive transiently by polymerase chain reaction. Liver histology was abnormal in all patients, but the changes were mild and markedly improved compared with biopsy specimens taken before loss of HBsAg. Liver tissue from 10 patients was positive for hepatitis B virus DNA by polymerase chain reaction but not by direct hybridization. These findings indicate that loss of HBsAg is associated with marked improvements in clinical and serum biochemical features of chronic hepatitis B, yet mild degrees of hepatitis and low levels of viral DNA may persist in the liver (HEPATOLOGY 1993;18:1313-1318.
Quantitation and typing of serum hepatitis C virus RNA in patients with chronic hepatitis C treated with interferon-β
Authors: Kobayashi Yoshinao , Watanabe Shozo , Konishi Masayoshi , Yokoi Masato , Kakehashi Ryuichi , Kaito Masahiko , Kondo Masahiro , Hayashi Yuji , Jomori Takahito , Suzuki Shiro
Published: March 2007

We quantified serum hepatitis C virus RNA titers and determined hepatitis C virus subtypes in chronic hepatitis C patients treated with interferon-β to investigate relationships among serum ALT response, serum hepatitis C virus titer and hepatitis C virus subtype. Of 146 chronic hepatitis C patients who received interferon-β therapy, 24 patients with sustained serum ALT normalization (complete responders) and 26 patients without serum ALT normalization (nonresponders) were randomly selected. Detection, typing and quantitation of hepatitis C virus were performed by means of the “single-tube” polymerase chain reaction method. Of the 24 complete responders, 21 (87.5%) became negative for hepatitis C virus RNA, whereas 21 (80.8%) of the 26 nonresponders remained positive. Hepatitis C virus infections with types I, II, III, IV, II + III and III + IV occurred in 0 (0%), 22 (51.2%), 10 (23.3%), 1 (2.3%), 7 (16.5%) and 3 (7.9%) patients, respectively. The mean pretreatment hepatitis C virus RNA titer of complete responders (0.4 ± 2.0 × 104 CID50/ml) was significantly lower than that of nonresponders (3.8 ± 4.5 × 104 CID50/ml) (p < 0.01). Regardless of HCV subtype, patients with more than 104 CID50/ml of HCV did not show serum ALT normalization, whereas complete serum ALT response was seen in most cases with less than 102 CID50/ml HCV. These results show that mixed infections with different hepatitis C virus subtypes appear to be more common than previously reported and that the pretreatment serum level of hepatitis C virus RNA is a more important predictor of outcome of interferon therapy than is virus genotype. (HEPATOLOGY 1993;18:1319-1325.)
Viral pathogenesis of hepatocellular carcinoma in the United States
Authors: Liang Jake T. , Jeffers Lennox J. , Reddy Rajender K. , De Medina Maria , Parker Talley I. , Cheinquer Hugo , Idrovo Victor , Rabassa Alfredo , Schiff Eugene R.
Published: March 2007

Chronic hepatitis B virus infection is closely associated with the development of hepatocellular carcinoma, which is a major cause of cancer death worldwide. Recent studies have implicated hepatitis C virus infection as a major pathogenic agent of HBsAg-negative hepatocellular carcinoma. The significance of hepatitis C virus and hepatitis B virus infections in the occurrence of HBsAg-negative hepatocellular carcinoma has not been well established in the United States. We studied 91 HBsAg-negative American patients with hepatocellular carcinoma for evidence of hepatitis C virus or hepatitis B virus infection. These patients had no other predisposing factors to hepatocellular carcinoma. A sensitive polymerase chain reaction was employed to detect hepatitis C virus RNA and hepatitis B virus DNA in serum and liver. Three sets of hepatitis C virus and hepatitis B virus primers were used to optimize the detection of viral genomes. Hepatitis C virus antibodies were measured with second-generation immunoassays. Twenty-six (29%) of these patients carried low levels of hepatitis B virus DNA in either serum, liver/tumor tissue or both. On the basis of the results from serological and polymerase chain reaction analyses of serum and liver, we found that 53 of 91 patients (58%) exhibited evidence of hepatitis C virus infection. When data were combined, 14 patients (15%) had evidence of hepatitis B virus/hepatitis C virus coinfection, whereas 12 (13%) were infected with hepatitis B virus alone and 39 (43%) had hepatitis C virus only. Twenty-six (29%) had no markers of hepatitis B virus or hepatitis C virus infection. All patients with identifiable viral markers had coexisting chronic liver disease. Our study suggests that hepatitis C virus and occult hepatitis B virus infections account for most (71%) hepatocellular carcinoma cases of unknown pathogenesis in the United States. However, in some patients with hepatocellular carcinoma no defined pathogenesis is associated with development of disease. (HEPATOLOGY 1993;18:1326-1333.)
Human leukocyte antigen A1-B8-DR3-DQ2-DPB1*0401 extended haplotype in autoimmune hepatitis
Authors: Manabe Koji , Donaldson Peter T. , Underhill James A. , Doherty Derek G. , Mieli-Vergani Giorgina , McFarlane Ian G. , Eddleston Adrian L. W. F. , Williams Roger
Published: March 2007

Genetic susceptibility to autoimmune hepatitis is associated with the human leukocyte antigen haplotype A1-B8-DR3 and DR4. To date, only one study in Japan has considered the human leukocyte antigen DP locus in this disease, and no studies have been reported in whites. In this study we used a series of sequencespecific oligonucleotide probes to determine human leukocyte antigen DPB1 genotypes in 101 unrelated white northern European patients and 105 racially and geographically matched controls. The aims of the study were twofold: first, to determine the degree of DPBencoded susceptibility to autoimmune hepatitis, and, second, to establish whether susceptibility can be extended to include human leukocyte antigen DPB. None of 17 DPB1 alleles was significantly associated with the susceptibility to autoimmune hepatitis. Although one particular seven-locus haplotype AlB8-DRB3*010-DRB1*0301-DQA1*0501-DQB1*02001-DPB1*0401 was significantly associated with the disease (27% vs. 7%, relative risk = 5.14, p < 0.0005), the association with this haplotype was weaker than that for the six-locus haplotype excluding DPB (40% vs. 11%, RR = 5.52, p < 0.0005). When the patients first seen at ages younger than 16 yr (pediatric patients) were considered separately, the greatest relative risk was for the seven-locus haplotype (41% vs. 7%; relative risk = 9.60, p < 0.0005). The results of this study further confirm that major histocompatibility complex-encoded susceptibility to autoimmune hepatitis is located at or close to the human leukocyte antigen DR locus; however, the A1-B8-DR3-DQ2-DPB1*0401 extended haplotype may be important in determining the age of onset and severity of disease. (HEPATOLOGY 1993;18:1334-1337.)