bluuebunny/crossref_metadata_2025 huggingface dataset:
Detection of Cell-CAM 105 in the pericanalicular domain of the rat hepatocyte plasma membrane
Authors: Mowery Jeanette , Hixson Douglas C.
Published: March 2007

Cell-CAM 105 has been identified as a cell adhesion molecule based on the ability of anti-cell-CAM 105 monospecific Fab fragments to inhibit the reaggregation of rat hepatocytes. Because of its adhesive properties, it was expected that cell-CAM 105 would be present on the lateral cell surface where adhesive interactions predominate. Paradoxically, however, immunofluorescence analysis of frozen sections of rat liver using specific monoclonal antibodies indicated that cell-CAM 105 was present exclusively in the bile canalicular domain of the rat hepatocyte where there is no intercellular adhesion. To more precisely define the in situ localization of cell-CAM 105, immunoperoxidase labeling and electron microscopy were used to examine intact and mechanically dissociated liver tissue. Results showed that when accessibility was provided by mechanical dissociation of perfusion fixed liver tissue, cell-CAM 105 could be detected in the pericanalicular region of lateral membranes. In contrast, when hepatocytes were labeled after incubation in vitro under conditions used during adhesion assays to induce reaggregation, cell-CAM 105 rapidly redistributed to all areas of the plasma membrane. Immunofluorescence analysis of primary hepatocyte cultures further revealed that cell-CAM 105 and two other bile canalicular proteins relocalized to discrete domains reminiscent of bile canaliculi, whereas cell-CAM 105 was also present in areas of intercellular contact. Serial section electron microscopy analysis of well-defined, cross-sectional profiles of bile canaliculi suggested the presence of cell-CAM 105—positive membrane folds that extended along the length of the bile canalicular border. In sections from livers in which calcium-dependent adhesive contacts had been disrupted by treatment with ethylenediamine tetraacetate, intact bile canaliculi were found that remained attached only by these border folds. The implications of these results are discussed with regard to a possible role for cell-CAM 105 in bile canalicular formation. (Hepatology 1991; 13:47-56).
Hepatic extraction of organic anions in the rat depends on ligand hydrophobicity
Authors: Tokumo Hironori , Aoyama Nankei , Busch Norbert , Mancuso Daniel J. , Thomas Holzbach R.
Published: March 2007

Non-bile-salt cholephilic organic anions are efficiently taken up by the liver. Recent work from our group has suggested the possible importance of relative hydrophobicity among various organic anions in hepatic uptake. To further validate and clarify this, we studied hepatic extraction of five different cholephilic dyes using the isolated perfused rat liver in single-pass mode. Albumin binding affinities and capacities for each of the ligands were measured in vitro to permit evaluation of in vivo interactions for each of them over a spectrum of unbound ligand concentrations. As expected, a strong positive correlation was found between ligand hydrophobicity and the relative degree of albumin binding affinity and capacity. Using appropriate experimental conditions, we also found a strong positive correlation between hepatic extraction efficiency for a given ligand and both its hydrophobicity and its unbound concentration. These data indicate that where the unbound ligand concentration is significant, the greater the ligand hydrophobicity, the greater is its efficiency of hepatic extraction. We conclude that hepatic extraction efficiency for non-bile-salt cholephilic organic anions depends on a combination of ligand hydrophilic/hydrophobic balance and the availability of the unbound ligand for uptake. (Hepatology 1991;13:62-67).
Separation of periportal and perivenous rat hepatocytes by fluorescence-activated cell sorting: Confirmation with colloidal gold as an exogenous marker
Authors: Braakman Ineke , Keij Jan , Hardonk Machiel J. , Meijer Dirk K. F. , Groothuis Geny M. M.
Published: March 2007

Periportal and perivenous hepatocytes are known to display various functional differences. In this study we present a new method to separate periportal and perivenous cells: after selectively loading zone 1 or zone 3 with the fluorescent label acridine orange in an antegrade or retrograde perfusion, respectively, we separated the isolated hepatocytes on a fluorescence-activated cell sorter. The common way to check on proper separation is to estimate activities of enzymes known to exhibit a heterogeneous acinar distribution. Using enzyme histochemistry, however, we found that already on short collagenase perfusion, some enzymes displayed a more shallow gradient than in vivo, making enzyme activities less suitable as zonal markers. We therefore used colloidal gold granules (17 nm) injected intravenously (2.5 mg) into the rat 2 to 3 hr before cell isolation. The gold is taken up predominantly by perivenous hepatocytes, probably because of the efficient removal of gold granules in zone 1 by competing Kupffer cells. We compared acridine orange fluorescence, presence of gold particles and activities of six marker enzymes, three biochemically and three histochemically determined. Acridine orange and gold both pointed to a high enrichment of the fractions, whereas most enzyme activities were more randomly distributed among the cells as a result of the isolation procedure. Our separation procedure yielded fractions highly enriched in either viable periportal or perivenous cells, both from one liver. The use of colloidal gold as a marker to monitor separation is a valuable alternative to the more risky estimation of enzyme activities. (Hepatology 1991;13:73-82).
Selective dopamine DA1 stimulation with fenoldopam in cirrhotic patients with ascites: A systemic, splanchnic and renal hemodynamic study
Authors: Hadengue Antoine , Moreau Richard , Bacq Yannick , Gaudin Christophe , Braillon Alain , Lebrec Didier
Published: March 2007

We studied the effects of fenoldopam, a selective dopamine DA1 agonist on systemic and splanchnic hemodynamics, renal blood flow and sodium excretion in 12 patients with alcoholic cirrhosis and ascites. Hepatic, azygos and renal veins were catheterized before and after intravenous administration of fenoldopam, 0.05 μg/kg/min for 1 hr and increased to 0.1 μg/kg/min for another hour. Mean arterial pressure progressively decreased (from 83 ± 7 to a minimum of 77 ± 8 mm Hg 100 min after starting the infusion) but returned to baseline level at 120 min. Plasma norepinephrine and renin activity increased (respectively from 567 ± 297 to 919 ± 375 pg/ml, p < 0.05, and from 17 ± 14 to 23 ± 15 ng/ml/hr, p < 0.05). Renal blood flow, urine output or sodium excretion did not change. Sodium output decreased at 1 hr from 6.9 μmol/min to 4.0 μmol/min, p < 0.05. Both hepatic venous pressure gradient and azygos blood flow significantly increased by 21%. We conclude that the acute administration of fenoldopam did not improve renal hemodynamics or function in patients with cirrhosis and ascites. In addition, dopamine DA1 agonism caused further increases in norepinephrine concentration and plasma renin activity. Portal pressure also increased, probably because of an increase in mesenteric blood flow. These results question the renal benefit and raise concern about the use of dopamine agonists in patients with cirrhosis and ascites. (Hepatology 1991;13:111-116).
Significance of natural polymerized albumin and its receptor in hepatitis B infection of hepatocytes
Authors: Dash Srikant , Rao Kanury V. S. , Joshi Beenu , Nayak Nabeen C. , Panda Subrat K.
Published: March 2007

Lack of information regarding the presence of native albumin polymer in serum and its structural similarity to the one produced by glutaraldehyde treatment casts doubt on the postulate that hepatitis B virus attachment to hepatocytes is mediated through polymerized albumin. We used a sandwich enzyme-linked immunosorbent assay with murine monoclonal antibodies raised against glutaraldehyde-polymerized albumin to detect native albumin polymer in human serum and its cross-reactivity with other albumin polymers. Presence of polymerized albumin receptor on the HepG2 cell was studied by radioreceptor assay. Purified hepatitis B virus and synthetic peptide analogous to part of pre-S2 sequence (120-145) were used to study polymerized albumin-dependent attachment of the virus to HepG2 cells. Antibodies raised against pre-S2 peptide were used to inhibit the pre-S2 and hepatitis B virus attachment to HepG2 cells. Glutaraldehyde-treated polymerized albumin was found to be immunologically cross-reactive with native albumin polymer. Its levels were found to be significantly raised in sera of patients with liver diseases. Polymerized albumin has specific saturable receptor on HepG2 cells with two classes of binding sites of different equilibrium dissociation constant (Kd1) = (16 ± 9.6)pmol/L and Kd2 = (1,019 ± 172)pmol/L. Albumin monomer was unable to compete for the polymerized albumin receptor sites on HepG2 cells. Antipre-S2 antibodies inhibit hepatitis B virus and pre-S2 binding to hepatocyte by 40% and 70%, respectively. Added extraneous polymerized albumin and the antibody against it did not interfere with virus attachment to HepG2 cells. (Hepatology 1991;13:134-142).
Hepatic histological findings after transplantation for chronic hepatitis B virus infection, including a unique pattern of fibrosing cholestatic hepatitis
Authors: Davies Susan E. , Portmann Bernard C. , O′grady John G. , Aldis Peter M. , Chaggar Kanchan , Alexander Graeme J. M. , Williams Roger
Published: March 2007

Long-term follow-up of 27 patients with hepatitis B virus—related chronic liver disease treated by transplantation showed that 23 had hepatitis B virus recurrence. In 13 patients late changes in the grafts were similar to those described in other series: minor abnormalities in five cases, chronic active hepatitis in five cases and non-hepatitis B virus—related graft dysfunction in three cases. Three patients had incomplete histological follow-up. Analysis of the histological changes and viral antigen expression in six cases revealed a distinct and novel pattern termed fibrosing cholestatic hepatitis. Development of fibrosing cholestatic hepatitis was associated with rapidly progressive graft dysfunction. It is postulated that this pattern of fibrosing cholestatic hepatitis develops because of a high cytoplasmic expression of viral antigens, including HBsAg. The remaining case had some features of fibrosing cholestatic hepatitis. The main histological features of this unique syndrome include thin, perisinusoidal bands of fibrosis extending from portal tracts to surround plates of ductular-type epithelium; prominent cholestasis; ground-glass transformation; and ballooning of hepatocytes with cell loss and mild mixed inflammatory reaction. (Hepatology 1991;13:150-157).
New insights into canalicular organic anion secretion
Authors: Moseley Richard H.
Published: March 2007

The effect of partial and complete dissipation of the membrane potential and partial depletion of cellular ATP content on the efflux of dinitrophenyl-glutathione and oxidized glutathione was examined in hepatocytes isolated from normal and mutant (TR−) rats exhibiting defective organic anion transport. Whereas alterations in the membrane potential difference had no effect on the initial efflux rate of dinitrophenyl-glutathione and oxidized glutathione, depletion of cellular ATP inhibited dinitrophenyl-glutathione and oxidized glutathione efflux and a linear relationship between the cellular ATP content and the initial efflux rate of dinitrophenyl-glutathione was observed in normal isolated rat hepatocytes. In contrast, depletion of cellular ATP content had no significant effect on the slower rate of dinitrophenyl-glutathione efflux from TR− rat hepatocytes. These findings implicate an ATP-dependent hepatic transport system for oxidized glutathione and glutathione conjugates that is absent in TR− mutants.

Fluorescence image analysis reveals normal secretion of a fluorescent bile acid fluorescein isothiocyanate glycocholate into the canalicular lumen of isolated normal and TR− mutant rat hepatocyte couplets, but negligible canalicular accumulation of a non-bileacid organic anion (carboxydichlorofluorescein diacetate) in TR− hepatocyte couplets. Canalicular membrane vesicles derived from normal rats exhibited saturable temperature- and ATP-dependent transport of sulfobromophthalein and sulfobromophthalein-glutathione that was absent in canalicular membrane vesicles from TR− rats. However, ATP-dependent daunomycin transport, reflecting transport mediated by the multidrug resistance gene product, p-glycoprotein, was present in canalicular membrane vesicles from both normal and TR− rats. Canalicular membrane vesicles from normal and TR− rats contained equal amounts of p-glycoprotein on immunoblots. These studies demonstrate that the conjugated hyperbilirubinemia in TR− mutant rats is the result of a functional absence of an ATP-dependent organic anion transport system on the canalicular membrane.
Responsiveness to phenobarbital in an adult with crigler-najjar disease associated with neurological involvement and skin hyperextensibility
Authors: Persico Marcello , Romano Marco , Muraca Maurizio
Published: March 2007

We present the case of a 23-yr-old man who had had since birth marked and sustained unconjugated nonhemolytic hyperbilirubinemia and who had had several attacks of grand mal seizures. Analysis of serum bilirubin by diazoreactive methods showed serum levels of unconjugated bilirubin as high as 445 μmol/L that were not affected by phenobarbital administration. However, analysis of serum bile pigments by high-pressure liquid chromatography demonstrated marked decrease of unconjugated bilirubin after phenobarbital treatment (from 432.4 μmol/L to 291.0 μmol/L) associated with slight increase of bilirubin monoconjugates and diconjugates (from 0.25 μmol/L to 0.42 μmol/L). Furthermore, in the past few years the patient had exhibited striking skin hyperextensibity and diaphragm eventration. This case confirms that alkaline methanolysis—high-pressure liquid chromatography is the most reliable method for assessment of serum fraction bilirubin levels; that clinical parameters such as neurological signs do not unequivocally discriminate between type I and II Crigler-Najjar disease and that response to phenobarbital treatment remains the main diagnostic tool. (Hepatology 1991;13:213-215).
Ultrastructural studies of hepatocyte cytoskeletons of phalloidin-treated rats by quick-freezing and deep-etching method
Authors: Naramoto Atsuhiko , Ohno Shinichi , Furuta Kiyoshi , Itoh Nobuo , Nakazawa Koh , Nakano Masayuki , Shigematsu Hidekazu
Published: March 2007

We observed hepatocyte cytoskeletons in phalloidintreated rats by the quick-freezing and deep-etching method in three dimensions and compared them with the ultrastructural findings on conventional ultrathin sections. The numbers of microvilli in dilated bile canaliculi were decreased in the rats treated with phalloidin for 1 wk. In hepatocytes the cytoplasm around bile canaliculi could be divided into three layers, increased microfilament layer, cell organelle layer of secretory system and increased smooth surface endoplasmic reticulum layer. In the rats treated with phalloidin for 4 wk, microfilaments were extended into the cytoplasm near the nucleus in addition to the increased number of large lysosomes and microtubules. In both groups, three-dimensional structures of microfilaments could be visualized around bile canaliculi and along cell borders by the quick-freezing and deep-etching method. The branching microfilaments with the diameters of 7 to 10 nm were directly attached to other filaments, cell organelles or cytoplasmic sides of cell membranes. Moreover, bundled intermediate filaments were increased around peribiliary microfilaments associated with long-term cholestasis. It is suggested that excessive accumulation of peribiliary microfilaments disturb the secretion of bile components into bile canaliculi. The cytoskeletal reorganizations of intermediate filaments seem to alter the arrangements of various cell organelles. (Hepatology 1991;222-229).
Metabolic response of isolated liver cells to in Vivo phagocytic challenge
Authors: Spolarics Zoltan , Bautista Abraham P. , Spitzer John J.
Published: March 2007

This study elucidates the in vivo metabolic response of different liver cells after a single phagocytic challenge. In vivo glucose uptake of different tissues and isolated liver cells was determined by a sequential double labeling version of the 2-deoxyglucose technique. After latex administration, glucose uptake more than doubled in the liver, increased by about 50% in the spleen and lung and was not changed in muscle and testis. Within 10 min after intravenous injection of latex beads, neutropenia developed, with no change in the number of lymphocytes. This was accompanied by a marked influx of polymorphonuclear leukocytes into the liver. Latex was found in 52%, 35%, and 14% of the isolated Kupffer cells, polymorphonuclear leukocytes and endothelial cells, respectively. In vivo glucose uptake increased by 111%, 142%, and 43% in these cells. Glucose uptake by the latex-free hepatocytes was also elevated, presumably by way of intercellular signals between parenchymal and non-parenchymal liver cells. Indomethacin pretreatment resulted in the delay of neutrophil immigration into tissues without any change in the glucose response of different liver cells. Thus phagocytic stimulation in vivo results in marked neutropenia, migration of neutrophils into the liver, increased glucose uptake by phagocytic cells of the liver and enhanced glucose metabolism by the nonphagocytic parenchymal cells. (Hepatology 1991;13:277—281).
Expression of laminin and its receptor LBP-32 in human and rat hepatoma cells
Authors: Rescan Pierre-Yves , Clement Bruno , Yamada Yoshihiko , Glaise Denise , Segui-Real Bartolome , Guguen-Guillouzo Christiane , Guillouzo André
Published: March 2007

Dramatic cellular changes that occur during hepatocarcinogenesis are associated with major alterations in extracellular matrix formation and in the relationships between cells and their microenvironment. We have studied the expression of laminin, the major noncollagenous glycoprotein of basement membrane, and the laminin receptor 32 kD laminin-binding protein in two rat (Faza 967 and HTC) and two human (HepG2 and HBGC2) hepatoma cell lines that express a variety of liver-specific functions. Laminin was found in the rough endoplasmic reticulum of these cells when the indirect immunoperoxidase method and electron microscopic examination were used. Radiolabeled laminin, immunoprecipitated from both media and cell extracts, was resolved by electrophoresis on sodium dodecyl sulfate gel in two major polypeptides that comigrated with the A and B subunits from Engelbreth-Holm-Swarm tumor laminin. Immunoblot analysis showed that the Mr = 400,000 polypeptide did not correspond to the A subunit of laminin. Northern blot analyses demonstrated large amounts of B1 and B2 mRNAs but no A chain mRNA. We conclude that the tumor cells produce the laminin B chains only. In contrast, normal adult hepatocytes from either man or rat lacked laminin mRNAs, whereas in 1-day primary culture, B chain mRNAs became detectable. The steady-state level of 32 kD laminin-binding protein mRNA was 10-fold and threefold higher in rat hepatoma cells than in freshly isolated and 1-day cultured normal rat hepatocytes, respectively. In human hepatocytes, the steady-state levels of 32 kD laminin-binding protein mRNAs varied depending on the donor and never reached the level of the human hepatoma cells. Subsequently, Faza, HTC and HepG2 cells, which were selected for their ability to grow on soft agar, were injected into the flank of nude mice and formed solid tumors with no formation of secondary tumors. The tumors contained laminin in sparse deposits between adjacent cells and accumulated around vessels. None of the hepatoma cell lines studied was found to penetrate a reconstituted basement membrane using a chemoinvasion assay with a Boyden chamber. Our data show that hepatoma cells produce the B chains of laminin and express a high level of receptor 32 kD laminin-binding protein mRNA. However these properties appear to be insufficient for these cells to form metastases in vivo. (Hepatology 1991;13:289-296).
High doses of recombinant α-interferon or γ-interferon for chronic hepatitis C: A randomized, controlled trial
Authors: Sáez-royuela Federico , Porres Juan Carlos , Moreno Alberto , Castillo Inmaculada , Martinez Gracia , Galiana Fernando , Carreño Vicente
Published: March 2007

Chronic hepatitis C is often a progressive liver disease for which there is no satisfactory treatment. We studied the efficacy of recombinant α-interferon or γ-interferon in the treatment of this disease in comparison with a control group. Thirty patients were randomly assigned to three groups. Ten patients received 7.5 MU α-interferon/m2 body surface three times weekly for 3 mo, then 5 MU/m2 for 3 mo and 2.5 MU/m2 for 6 mo. Ten patients were treated with γ-interferon at a dose of 2 MU/m2 for 6 mo and the other 10 served as controls without treatment.

The mean serum ALT levels and liver histological findings improved significantly only in the patients treated with α-interferon. No changes were observed in patients treated with γ-interferon or in controls. Five of 10 patients treated with α-interferon had complete responses (mean ALT normal during therapy). After treatment ALT returned to pretreatment levels in two of 5 patients. The long-term response rate after α-interferon therapy was 30% at 18 mo. We conclude that α-interferon is effective in controlling disease activity in a portion of patients with chronic hepatitis C. High doses of α-interferon do not appear to add further benefit in the response rate or relapse rate. γ-Interferon therapy is ineffective. (Hepatology 1991;13:327-331).
Effects of hepatitis B virus, alcohol drinking, cigarette smoking and familial tendency on hepatocellular carcinoma
Authors: Chen Chien-Jen , Liang Kung-Yee , Chang Ah-Seng , Chang Yo-Chi , Lu Sheng-Nan , Liaw Yun-Fan , Chang Wen-Yu , Sheen Maw-Chang , Lin Tong-Ming
Published: March 2007

Independent and interactive effects related to the development of hepatocellular carcinoma were assessed using a community-based case-control study for hepatitis B virus, habitual alcohol drinking, cigarette smoking, peanut consumption and history of hepatocellular carcinoma among the immediate family. All 200 male newly diagnosed hepatocellular carcinoma patients were recruited consecutively through the period of study as the case group from two teaching medical centers in northern and southern Taiwan. Healthy community residents matched one-to-one with cases on age, sex, ethnic group and residential area were selected as the control group. The carrier status of HBsAg and HBeAg was determined by blind radioimmunoassays, and other risk factors were obtained through standardized interviews according to a structured questionnaire. Conditional logistic regression analysis showed a significant association between hepatocellular carcinoma and the carrier status of HBsAg and HBeAg with an odds ratio of 16.7 and 56.5, respectively, for carriers of HBsAg alone and for carriers of both HBsAg and HBeAg. There was a dose-response relationship between cigarette smoking and hepatocellular carcinoma with an odds ratio of 1.1, 1.5 and 2.6, respectively, for those who smoked 1 to 10, 11 to 20 and more than 20 cigarettes a day. A significant association with hepatocellular carcinoma was also observed for the habitual alcohol consumer with an odds ratio of 3.4. Those whose immediate family had a history of hepatocellular carcinoma were more likely to have the disease develop, with an odds ratio of 4.6. However, the frequency of peanut consumption was not significantly associated with hepatocellular carcinoma. Whereas the modification of age was not statistically significant, hepatocellular carcinomaassociated risk factors were multiplicatively independent under the conditional logistic model. (Hepatology 1991;13:398-406.)
Hepatitis B virus replication modulates pathogenesis of hepatitis D virus in chronic hepatitis D
Authors: Smedile Antonina , Rosina Floriano , Saracco Giorgio , Chiaberge Elisabetta , Lattore Valter , Fabino Antonina , Brunetto Maurizia R. , Verme Giorgio , Rizzetto Mario , Bonino Ferruccio
Published: March 2007

Hepatitis D virus and hepatitis B virus nucleic acids were detected by Northern and Southern blot hybridlzation in the sera and livers of 85 chronic carriers of HBsAg and anti-hepatitis D followed up for a mean of 10 yr. We identified three subsets of patients: 13 with hepatitis D virus and hepatitis B virus viremia, 53 with serum hepatitis D virus RNA, but without hepatitis B virus DNA and 19 negative for both nucleic acids.

Genomic and subgenomic forms of hepatitis D virus RNA were detected only in patients with hepatitis D virus and hepatitis B virus viremia. Histological findings and disease activity at admission were comparable in the three groups of patients, but the outcome was significantly worse in patients with active replication of both viruses; two of them died of terminal liver failure and hepatocellular carcinoma developed in two; the remaining patients had an uneventful course. These results suggest that active hepatitis B virus replication represents an important previously unrecognized determinant of severe liver damage in patients with chronic hepatitis D virus infection. Since hepatitis B virus provides the means for hepatitis D virus secretion and release from infected cells, active hepatitis B virus multiplication favoring the spread of hepatitis D virus from cell to cell may increase the pathogenetic potential of the defective agent. (Hepatology 1991;13:413-416.)
Development and application of an in vitro model for screening anti-hepatitis B virus therapeutics
Authors: Lampertico Pietro , Malter James S. , Gerber Michael A.
Published: March 2007

The development of effective anti-hepatitis B virus agents has been hampered by the lack of reliable in vitro systems for the screening of new therapeutics. In an effort to circumvent this problem, we have developed an in vitro system for screening antihepatitis B virus drugs using hepatitis B virus DNAtransfected Hep G2 cells. The cell line designated 2.2.15 produces replicative viral DNA intermediates, mature Dane particles and high levels of viral antigens. Subconfluent 2.2.15 cells were treated with a variety of commonly used anti-hepatitis B virus therapeutics, and their efficacy was determined by analyzing changes in the replicative cellular or extracellular hepatitis B virus DNA content by Southern blotting or slot-blot hybridization. The slot-blot method was sensitive, reproducible and rapid and correlated well with Southern blotting. Analysis of the media for hepatitis B virus DNA was indicative of changes in intracellular, replicative hepatitis B virus DNA, permitting sampling of the media. Therefore 2.2.15 cells may provide a valuable method for identifying and monitoring effective anti-hepatitis B virus therapeutics. Using this system to test various agents, we confirm that 2′-deoxyguanosine strongly inhibited viral replication, whereas others tested were less effective. Correlation with in vivo systems is now needed. (Hepatology 1991;13:422-426.)
In micronodular cirrhosis, hepatocytes retain a normal C-25 hydroxylation capacity toward vitamin D3: A study using the rat carbon tetrachloride-induced cirrhotic model
Authors: Dubé Catherine , Vallières Sylvie , Éthier Chantal , Benbrahim Nawel , Tremblay Chantale , Gascon-Barré Marielle
Published: March 2007

To test further the competence of the cirrhotic liver to metabolize vitamin D3 at C-25, hepatocytes were isolated from controls and from CCl4-induced cirrhotic rat livers, as well as from partially hepatectomized rats. The transformation of D3 into 25-hydroxyvitamin D3 was studied in the presence of 107 hepatocytes at D3 concentrations of 20 nmol/L to 15.4 μmol/L. Histologically, micronodular cirrhosis was present in all CCl4-treated rats, whereas controls had normal livers; portal venous pressure (p < 0.008) and intrahepatic collagen content (p < 0.0001) were significantly increased in CCl4-treated rats, whereas no difference was found between the two groups in the total and ionized serum calcium, D3 metabolites, ALT, AST and alkaline phosphatase. Cytochrome P-450 was 0.27 ± 0.02 and 0.25 ± 0.02 nmol/106 hepatocytes in controls and cirrhotic rats (N.S.), and it significantly increased in both groups after phenobarbital or 3-methylcholanthrene administration (p < 0.0001). 25-Hydroxyvitamin D3 formation was best described by power law equations and varied between 0.02 ± 0.0004 and 29.57 ± 2.8 in controls, and 0.024 ± 0.0004 and 32.0 ± 7.0 pmol ± hr−1 ± 106 hepatocytes−1 in cirrhotic rats. No statistically significant difference was found in the slopes of the 25-hydroxyvitamin D3 formation, but the y-axis intercept was found to be lower in cirrhotic rats under basal resting conditions (p < 0.005). Inducers of the mixed function oxidases significantly increased 25-hydroxyvitamin D3 formation in controls as well as in cirrhotic rats (p < 0.005). Moreover, both groups were found to respond similarly to the addition of modulators of the enzyme such as the calcium ionophore A23187 and parathyroid hormone. Partial hepatectomy was also without effect on the activation of D3. Furthermore, the cell sequestration of D3 was also found to be unperturbed in hepatocytes obtained from either cirrhotic or partially hepatectomized livers. The data indicate that in well-compensated micronodular cirrhosis, the C-25 hydroxylation of D3 is generally intrinsically normal at the cellular level and that it also remains fully responsive to in vivo and in vitro modulators of its activity. (Hepatology 1991;13:489-499.)
Hypothermic preservation of hepatocytes. III. Effects of resuspension media on viability after up to 7 days of storage
Authors: Marsh Diane C. , Hjelmhaug Julie A. , Vreugdenhil Paul K. , Kerr Julie A. , Rice Mark J. , Belzer Folkert O. , Southard James H.
Published: March 2007

Hepatocyte suspensions provide a rapid method to determine how hypothermic storage affects liver cell metabolism and viability. Using these studies, improved methods of hypothermic liver preservation for transplantation may be developed. In this study, rat hepatocytes were cold-stored for up to 7 days in University of Wisconsin liver preservation solution. At the end of each day of storage hepatocytes were resuspended in Krebs-Henseleit buffer or tissueculture medium (Liebovitz-15; Fischer's; modified Fischer's, which was similar to Fischer's but with glycine and cysteine added; or Waymouth's medium). Hepatocyte viability was assessed by rewarming and oxygenating the suspensions and measuring the percentage of leakage of lactate dehydrogenase from the cells, the cellular concentration of potassium and the stimulation of respiration by succinate, all measures of plasma membrane integrity. Additionally, concentrations of ATP and glutathione after rewarming and reoxygenation in the various resuspension media were measured. Hepatocyte permeability to lactate dehydrogenase did not increase during cold storage of 1 to 7 days (7.2% ± 2% leakage), indicating that most of the hepatocytes remained viable during cold storage. However, when rewarmed, loss of viability (leakage of lactate dehydrogenase) was dependent on the composition of the resuspension media. In Krebs-Henseleit buffer, viability was reduced after 2 and 3 days of storage (lactate dehydrogenase leakage on rewarming = 70% to 90%). Leakage of lactate dehydrogenase was reduced significantly after resuspension in tissue-culture media. After 6 days of storage, lactate dehydrogenase leakage from hepatocytes stored in Liebovitz-15 or modified Fischer's was only about 30%. Other measures of membrane permeability-loss of potassium and succinate stimulation of respiration - correlated with lactate dehydrogenase release. Our results show that cold-stored hepatocytes are sensitized to reperfusion injury, which can be suppressed when an appropriate resuspension medium is used for rewarming the cells. Resuspension in an appropriate medium showed that hepatocyte viability can be maintained for up to 6 days in cold storage. (Hepatology 1991;13:500-508.)
Laparoscopic cholecystectomy: What is really known about it?
Authors: Gumucio Jorge J. , Malet Peter F.
Published: March 2007

Endoscopic intra abdominal surgery yet routinely used for gynecologic operations can be extended for performing cholecystectomy in uncomplicated cholelithiasis.

With the available but not yet completely satisfactory endoscopic instruments, 330 patients underwent coelioscopic cholecystectomy with few and only benign complications. The main advantages are the cosmetic aspect and strength of scars, moderation of post-operative pain and short hospital stay.
Orthotopic liver transplantation for patients with hepatitis B virus-related liver disease
Authors: Todo Satoru , Demetris Anthony J. , van Thiel David , Teperman Lewis , Fung John J. , Starzl Thomas E.
Published: March 2007

Fifty-nine patients with prior hepatitis B virus infection underwent orthotopic liver transplantation. During the first 2 mo, mortality was not significantly different in the hepatitis B virus-infected group (25.5%) vs. a hepatitis B virus-immune control group (21%). Beyond 2 mo, the mortality, rate of graft loss, need for retransplantation and incidence of abnormal liver function were significantly higher in the hepatitis B virus-infected group. Treatment of the hepatitis B virus infection was attempted with passive immunization, combined active and passive immunization, α-interferon or nothing. The clinical outcome was not significantly influenced by any of these therapies. However, of the patients who lived more than 60 days, 6 of 22 treated with active plus passive immunization were cleared of HBsAg, something achieved once in 16 patients treated with α-interferon, never in 3 patients with passive immunization only and once in 4 patients with no therapy. In patients with recurrent hepatitis B virus infection, the pace of hepatitis development in the graft appeared to be accelerated, and this was particularly striking in patients who underwent multiple retransplantations at progressively shorter intervals. None of the patients who became HBsAg-negative had HBeAg preoperatively. (Hepatology 1991;13:619-626.)
Determination of hepatitis B virus DNA in serum using the polymerase chain reaction: Clinical significance and correlation with serological and biochemical markers
Authors: Baker Bennie L. , Di Bisceglie Adrian M. , Kaneko Shuichi , Miller Roger , Feinstone Stephen M. , Waggoner Jeanne G. , Hoofnagle Jay H.
Published: March 2007

Sera from 98 patients with various stages of chronic hepatitis B virus infection were studied to determine the clinical significance of hepatitis B virus DNA in serum detected by the polymerase chain reaction. Patients were divided into three groups according to their HBsAg and HBeAg status. Group I (n = 31) had detectable HBsAg and HBeAg, group II (n = 46) had HBsAg but not HBeAg and group III (n = 21) consisted of patients who were once chronic hepatitis B virus carriers but had lost HBsAg during follow-up.

Group I patients usually had significant liver disease (raised serum aminotransferases), had higher titers of HBsAg and had been infected with hepatitis B virus for a shorter period than patients in the other two groups. All patients in group I had hepatitis B virus DNA detectable by polymerase chain reaction and 94% had sufficient hepatitis B virus DNA present for detection by dot-blot hybridization. Group II patients had lower mean serum aminotransferase activities and titers of HBsAg than those in group I. Serum hepatitis B virus DNA was detectable by polymerase chain reaction in 78% but in only 30% of group II patients by dot-blot hybridization. Group II patients who did not have hepatitis B virus DNA detectable by polymerase chain reaction had mean serum aminotransferase levels within the normal range and had a younger mean age than those with hepatitis B virus DNA. Group III patients generally had no evidence of active liver disease. Of five patients in this group seropositive for hepatitis B virus DNA by polymerase chain reaction on initial testing, all eventually became negative for hepatitis B virus DNA on subsequent testing. There was a tendency for those patients in group III with hepatitis B virus DNA detectable by polymerase chain reaction to have elevated serum aminotransferase levels (40% vs. 25%); however, these differences were not statistically significant.

Taken together, these data show that the presence of hepatitis B virus DNA in serum as detected by polymerase chain reaction appears to be a good marker of the level of viremia, can be correlated with aminotransferase levels and parallels the presence of HBsAg in serum. (Hepatology 1991;13:632-636.)
Urinary excretion of bile acid glucosides and glucuronides in extrahepatic cholestasis
Authors: Wietholtz Hubertus , Marschall Hanns-Ulrich , Reuschenbach Regina , Matern Heidrun , Matern Siegfried
Published: March 2007

Recently the formation of bile acid glucosides has been described as a novel conjugation mechanism in vitro and in vivo. In 10 patients with extrahepatic cholestasis caused by carcinoma of the head of the pancreas we investigated excretion rates and profiles of urinary bile acid glucosides. Urinary bile acid glucosides and, for comparison, bile acid glucuronides were extracted and characterized according to established methods. In controls total urinary bile acid glucoside excretion was 0.22 ± 0.03 μmol/24 hr (mean ± S.E.M.)-in the range of bile acid glucuronide excretion (0.41 ± 0.06 μmol/24 hr; mean ± S.E.M.). A gas chromatography-mass spectrometry-characterized trihydroxy bile acid glucoside of stillunknown hydroxyl positions accounted for 65% of total urinary bile acid glucosides. In extrahepatic cholestasis total urinary bile acid glucoside excretion was 0.52 ± 0.13 μmol/24 hr (mean ± SEM), yet significantly lower than bile acid glucuronide excretion (1.53 ± 0.13 μmol/24 hr; mean ± SEM; p <a 0.001). In cholestasis the primary bile acid derivatives cholic and chenodeoxycholic acid glucosides amounted to 90%, whereas the trihydroxy bile acid glucoside had decreased to 5% of total bile acid glucoside excretion, indicating its alteration during enterohepatic circulation. The data establish the composition and quantity of urinary bile acid glucosides in healthy controls and cholestasis and constitute a quantitative comparison with another glycosidic conjugation reaction, bile acid glucuronidation. (Hepatology 1991;13:656-662.)
Hepatocellular carcinoma presenting with pyrexia and leukocytosis: Report of five cases
Authors: Okuda Kunio , Kondo Yoichiro , Nakano Masayuki , Kage Masayoshi , Arakawa Masahiro , Kojiro Masamichi , Ohtsuki Toshio , Shimokawa Yutaka , Hirata Masao , Kohno Kunihiko
Published: March 2007

In the past 26 years we have encountered five patients with primary liver malignancy clinically characterized by high remittent fever and leukocytosis mimicking liver abscess. Two patients underwent exploratory laparotomy, and drainage was carried out in another. The clinical courses went rapidly downhill. The liver was cirrhotic in two patients. The interior of the main mass was almost totally necrotic in four cases. Histologically, the malignant cells in the main portion resembled sarcoma, but in some areas cells appeared epithelial with eosinophilic cytoplasm and were in a trabecular arrangement, except for one case not subjected to autopsy in which histological study was inadequate because of extensive necrosis. It seems that these neoplasms were very poorly differentiated hepatocellular carcinomas rather than combinations of sarcoma and hepatocellular carcinoma. These patients, therefore, may represent a distinct clinicopathological type of hepatocellular carcinoma that is very rare in Japan but perhaps more common in South Africa, where similar cases have been clinically described in larger numbers. (Hepatology 1991;13:695-700.)
Extrahepatic bile duct growth in mice repeatedly injected with human normal serum, IgA-deficient serum or purified secretory IgA
Authors: Solbreux Pierre M. E. , Maldague Paul , Vaerman Jean Pierre
Published: March 2007

Another group of researchers has reported that seven intraperitoneal injections into mice of purified human serum IgA or normal human serum-but not IgA-deficient serum, mouse serum or saline-induced considerable growth of the extrahepatic bile duct epithelium. They stated that heterologous IgA was principally responsible for this effect. We have quantitated this growth by another method: enumeration of total and radiolabeled nuclei of the lumenal and glandular epithelium in transverse duct sections after tritiated thymidine injection and autoradiography. Our data show maximal incorporation between 18 and 24 hr after the last injection, with the highest labeling index in the lumenal epithelium after only three injections and before any duct enlargement. After four to seven injections, thymidine incorporation continued in juxtalumenal glands, together with massive glandular proliferation into the thickening wall and obvious inflammation. These changes were more pronounced in the liver-proximal part of the duct without affecting intrahepatic duct and gallbladder epithelia. Subcutaneous serum injections were less active. We confirm the inactivity of buffered saline, but, unlike previous authors, demonstrate strong activity in purified human milk secretory IgA and in IgA-deficient serum. We suggest that this different approach of quantitating epithelial proliferation will allow comparison of the bile duct growth effects of different, well-characterized human and animal IgA preparations. (Hepatology 1991;13:735-742.)
Localization of tumor necrosis factor-stimulated DNA synthesis in the liver
Authors: Feingold Kenneth R. , Barker Mary E. , Jones Albert L. , Grunfeld Carl
Published: March 2007

Tumor necrosis factor and other cytokines mediate the body's response to infection and inflammation. Long-term administration of tumor necrosis factor causes liver hypertrophy, and our laboratory has shown that tumor necrosis factor acutely increases hepatic DNA synthesis. The purpose of this study was to determine which specific cell types in the liver undergo DNA synthesis in response to tumor necrosis factor. Light microscopic autoradiographs of 3H-thymidine incorporation demonstrate that tumor necrosis factor stimulates DNA synthesis in nonparenchymal cells, whereas the low level of DNA synthesis normally found in hepatocytes remains unaffected. Tumor necrosis factor increased labeling in sinusoidal cells that appeared to be Kupffer cells and in cells localized to the portal tracts. With electron microscopy and histochemical staining for endogenous peroxidase, the sinusoidal cells were clearly identified as Kupffer cells. The exact identity of the cells in the portal tract is less certain, but some of the cells shared the characteristics of bile duct precursors. With long-term administration of tumor necrosis factor, a striking proliferation of bile ducts was seen. The functional role of the proliferation of bile ducts in response to tumor necrosis factor is unclear, and it is not known whether the increase is a beneficial or deleterious response. The increased DNA synthesis in Kupffer cells could result in a potentially beneficial response to infection by increasing an animal's ability to phagocytize and clear microorganisms. However, under some circumstances, an expansion of the number of Kupffer cells could be deleterious by increasing an animal's ability to produce tumor necrosis factor and other cytokines that in large quantities can have toxic effects. (Hepatology 1991;13:773-779.)
More on glucose transporters: The acinar organization for hepatic glucose transport
Authors: Gumucio Jorge J. , Bilir Bahri , Gumucio Jorge J.
Published: March 2007

The “erythroid/brain” glucose transporter (GT) isoform is expressed only in a subset of hepatocytes, those forming the first row around the terminal hepatic venules, while the “liver” GT is expressed in all hepatocytes. After 3 d of starvation, a three- to fourfold elevation of expression of the erythroid/brain GT mRNA and protein is detected in the liver as a whole; this correlates with the expression of this GT in more hepatocytes, those forming the first three to four rows around the hepatic venules. Starvation-dependent expression of the erythroid/brain GT on the plasma membrane of these additional hepatocytes is lost within 3 h of glucose refeeding; however, by immunoblotting we show that the protein is still present. Its loss from the surface is possibly explained by internalization.
Transcellular transport of organic anions in the isolated perfused rat liver: The differential effects of monensin and colchicine
Authors: Aoyama Nankei , Ohya Toshihide , Chandler Kimberly , Gresky Susan , Holzbach Thomas R.
Published: March 2007

Nonbile salt cholephiles and bile salts are two classes of organic anions that are efficiently taken up and excreted by the liver. Recent evidence suggests that a microtubular system-dependent, colchicine-sensitive transcellular pathway may transport both classes of these ligands. The relationship of this pathway to flux rates, however, remains unclear. Some structural evidence suggests an important role for a Golgi-associated vesicular system. Monensin, like colchicine, is a perturbing agent that is believed to target primarily Golgi and related organelles. The effects of a minimal effective dose of both colchicine (0.06 mg to 0.12 mg/100 gm body wt) and monensin (0.6 mg/100 gm body wt) were examined in the isolated perfused rat liver in a single-pass mode. The nonbile salt cholephile, phenol red, was studied at two doses: 1 nmol and 5 μmol. Sodium taurocholate was studied at three doses: 2 nmol, 1 μmol and 5 μmol. Colchicine affected the transcellular transport for both classes of organic anions equally. Partially inhibitory effects on both anions occurred only at high ligand flux rates. In contrast, monensin greatly impaired the transport of nonbile salt cholephiles but had no influence on transcellular bile salt flux. We conclude that the monensin effect appears to define a distinct transcellular transport pathway for each of the two classes of organic anions. (HEPATOLOGY 1991;14:1-9.)