bluuebunny/crossref_metadata_2025 huggingface dataset:
Inhibition of Rat Brain Na+,K+−ATPase Activity by Serum from Patients with Fulminant Hepatic Failure†
Authors: Seda Hosny W. M. , Hughes Robin D. , Gove Christopher D. , Williams Roger
Published: August 2008

Among the toxins accumulating in the circulation of patients with fulminant hepatic failure (FHF) are substances which inhibit leucocyte ouabain-sensitive sodium transport. A similar inhibition of brain Na+,K+−ATPase could lead to both coma and cerebral edema found in these patients which are associated with high mortality. In this study, we have investigated the effect of sera from FHF on normal rat brain Na+,K+−ATPase activity in vitro. Serum from patients with FHF significantly decreased the ouabain-sensitive Na+,K+−ATPase activity (13.58 ± S.D. 2.60 μmoles Pi mg protein−1 hr−1) in the rat brain membrane preparation in vitro as compared to normal serum (20.33 ± 3.24 jumoles Pi mg protein−1 hr−1, p < 0.001). A final serum dilution of 1 in 40 was required to abolish the inhibition of Na+,K+−ATPase activity. Cerebrospinal fluid obtained at postmortem from FHF patients also contained the inhibitory substances. Serum from patients in coma due to decompensated chronic liver disease inhibited the Na+,K+−ATPase activity (17.25 ± 1.37 μmoles Pi mg protein−1 hr−1), but this was less marked than with FHF serum. Hence, the inhibition of brain Na+,K+−ATPase by substances accumulating in the serum in FHF may be important in the pathogenesis of hepatic coma.
Asialoglycoprotein Receptor Expression in Murine Pregnancy and Development
Authors: Collins Janna C. , Stockert Richard J. , Morell Anatol G.
Published: August 2008

Hepatic asialoglycoprotein receptor activity of pregnant mice increased from the tenth day of gestation, reaching 3-fold the normal adult level just prior to parturition and returning to normal within 1 week postpartum. Increases in activity involved both intracellular and cell-surface associated receptors and were associated with enhanced endocytosis of asialoorosomucoid by isolated hepatocytes. Modulation of hepatic binding protein expression during murine pregnancy represents the first situation in which this receptor's concentration has been shown to exceed normal levels in response to a physiologic change. In the fetus, receptor activity was barely detectable throughout gestation, steadily increased in the neonatal period, and reached adult levels by the fifth postpartum day.
Ultrastructural Studies of Fibroblasts Transfected with Hepatitis B Virus DNA†
Authors: Aoki Naoto , Gerber Michael A. , Thung Swan N. , Chen Mei-Ling , Christman Judith K. , Price Peter M. , Flordellis Christos S. , Acs George
Published: August 2008

Cultured 3T3 mouse fibroblasts transfected with cloned hepatitis B virus genome and DNA coding for methotrexate-resistant dihydrofolate reductase, produce and secrete significant amounts of hepatitis B surface antigen (HBsAg). Ultrastructural morphometry revealed that fibroblasts transfected with hepatitis B virus DNA contained significantly more lysosomes than did fibroblasts transfected with the gene coding for methotrexate resistance or normal fibroblasts. Although abundant HBsAg was found in the cytoplasm of transfected fibroblasts by immunologic methods, HBsAg particles were not detected by electron microscopy. Immunoelectron microscopy localized HBsAg to the nuclear envelope, rough endoplasmic reticulum, and endoplasmic cisternae. These findings suggest that the transfected cells produce mainly nonparticulate HBsAg or that they have a defect in intracisternal packaging of HBsAg into particles.
The Effect of Bile Duct Obstruction on the Clearance of Circulating IgA Immune Complexes
Authors: Harmatz Paul R. , Kleinman Ronald E. , Bunnell Bruce W. , Mcclenathan Daniel T. , Walker Allan W. , Bloch Kurt J.
Published: August 2008

The clearance of circulating IgA immune complexes following acute bile duct obstruction was investigated in this study. IgA immune complexes were formed in vitro from MOPC-315, an IgA M-component with anti-dinitrophenyl (DNP) specificity, and 125I-DNP10 bovine serum albumin (BSA). Eighteen hours after laparotomy during which the common bile duct was either identified only or identified and ligated, the IgA immune complexes were injected intravenously. Groups of bile duct-ligated and bile duct-patent rats were also injected intravenously with IgG anti-DNP-125I-DNP 10BSA immune complexes and 125I-bovine liver β-glucuronidase to assess the hepatic clearance of materials not dependent on an intact biliary system. Clearance of IgA immune complexes was delayed after bile duct ligation. Although the clearance of IgA immune complexes was delayed, only 10% of these complexes remained in the circulation at 3 hr. The clearance of IgG immune complexes and β-glucuronidase was not affected by ligation. These experiments demonstrate the physiologic importance of a patent bile duct in the normal clearance of IgA immune complexes in the rat. The observation that clearance is delayed, but not completely inhibited by bile duct ligation suggests that alternate mechanisms exist for removing IgA immune complexes from the circulation.
The Nature of IgG Complexes in Alcoholic Liver Disease
Authors: Stoltenberg Phillip H. , Soltis Ronald D.
Published: August 2008

Circulating immune complexes have been described in most liver diseases, including alcoholic liver disease, although their pathogenic significance remains unclear. Currently available immune complex assays do not distinguish immunoglobulin aggregates from antigen-antibody complexes. Immunoglobulin aggregate formation occursin vitroat 37°C in the presence of hypergammaglobulinemia and/or hypoalbuminemia, conditions common in liver disease. To determine if hypergammaglobulinemia and/or hypoalbuminemia could predispose to immunoglobulin aggregate formationin vivo, 25 patients with alcoholic liver disease were studied. Using sucrose density gradient fractionation followed by quantitation of IgG by radioimmunoassay, high molecular weight IgG complexes (>11S) were frequently present in alcoholic liver disease sera, and correlated with the degree of hypergammaglobulinemia and/or hypoalbuminemia, and with125I-Clq binding activity. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis of these complexes revealed only IgG and nonspecific trapping of several normal serum proteins. A specific complex-associated antigen could not be identified. While small, undetectable quantities of true antigen-antibody complexes might also be present, our data suggest that IgG complexes in alcoholic liver disease may represent immunoglobulin aggregate formationin vivo.
Effect of Ursodeoxycholic Acid on Biliary Bile Acid and Bile Lipid Composition in Gallstone Patients
Authors: Stiehl Adolf , Raedsch Richard , Rudolph Gerda , Walker Siegfried
Published: August 2008

In five patients with radiolucent gallstones, the effect of ursodeoxycholic acid (Urso) in doses of 250, 500, 750, 1,000, and 1,250 mg per day on biliary lipid and bile acid composition was studied. Biliary cholesterol decreased from 8.8 ± 0.8 mole% to 4.4 ± 0.2 mole% at 500 mg Urso per day (7.1 mg per kg) and to 4.2 ± 0.3 mole% at 750 mg Urso per day (10.7 mg per kg). Administration of 1,000 or 1,250 mg Urso per day produced no further decrease of biliary cholesterol. The biliary content of phospholipids and total bile acids remained unchanged.

During Urso treatment, the relative amounts of glyco-Urso and tauro-Urso in bile increased. Glyco-Urso reached a plateau at 49.1 ± 2.1% of total bile acids during treatment with 1,000 mg Urso per day, and tauro-Urso increased up to 4.3 ± 1.5% of total bile acids at 250 mg Urso per day. Simultaneously cholic acid, chenodeoxycholic acid, deoxycholic acid, and lithocholic acid decreased.

The data indicate that Urso treatment reduces biliary cholesterol efficiently already at a dose of 500 mg per day; biliary bile acid composition changes up to 1,000 mg Urso per day. Doses greater than 1,000 mg per day produced no additional alterations in bile composition.
Venocclusive Disease of the Liver after Bone Marrow Transplantation: Diagnosis, Incidence, and Predisposing Factors
Authors: Mcdonald George B. , Sharma Pankaj , Matthews David E. , Shulman Howard M. , Thomas Donnall E.
Published: August 2008

Venocclusive disease (VOD) of the liver, a fibrous obliteration of small hepatic venules, can be caused by chemoradiation therapy. We reviewed 255 consecutive patients undergoing bone marrow transplantation for malignancy during 1978 to 1980 in order to determine the incidence of VOD and the predisposing factors. Fifty-three of 255 patients met our criteria for VOD, for an incidence of 21%. Multivariate analysis showed that the most significant risk factors for VOD were age over 15, an underlying malignancy other than acute lymphocytic leukemia and hepatitis prior to transplantation. Patients with hepatitis had a 3. 4-fold risk of developing VOD, as compared to patients with normal SGOT values (p = 0.0004). Hepatitis in this setting is probably of non-A, non-B viral etiology and represents a relative contraindication to marrow transplantation because of enhanced toxicity from conditioning chemoradiotherapy.
Hepatic Graft Versus Host Disease: A Study of the Predictive Value of Liver Biopsy in Diagnosis
Authors: Snover Dale C. , Weisdorf Sally A. , Ramsay Norma K. , Mcglave Philip , Kersey John H.
Published: August 2008

Ninety-six liver biopsies [32 bone marrow transplant (BMT), 7 pre-BMT, and 57 non-BMT] are reviewed for histological evidence of graft versus host disease (GVHD), based on bile duct atypia and related inflammatory changes. In addition, the presence of cholestasis, piecemeal necrosis, and attachment of lymphocytes to vascular endothelium (endothelialitis) are evaluated. The 57 non-BMT biopsies include examples of viral hepatitis (acute and chronic), nonviral chronic hepatitis, extrahepatic biliary obstruction, cytomegalovirus hepatitis, primary biliary cirrhosis, and orthotopic liver transplant rejection. Although the sensitivity of bile duct damage as an indicator of GVHD appears high (only one probable false negative was noted), there is considerable overlap between the changes of GVHD and occasional cases of acute and chronic hepatitis and extrahepatic biliary obstruction. Nine of the 57 non-BMT biopsies (15%) were felt to be consistent with GVHD and represent “false positives”. Despite this relative lack of specificity, analysis of several features in combination provided clues to improve accuracy of diagnosis. The findings of extensive bile duct damage with minimal inflammatory changes is characteristic of GVHD. Possibly more predictive is the presence of endothelialitis of portal or central veins, which was seen in only three non-BMT biopsies, being present in eight cases of GVHD.
Biochemical and Morphological Alterations of Baboon Hepatic Mitochondria After Chronic Ethanol Consumption
Authors: Arai Masao , Leo Maria A. , Nakano Masayuki , Gordon Ellen R. , Lieber Charles S.
Published: August 2008

Baboons fed ethanol (50% of total calories) chronically develop ultrastructural alterations of hepatic mitochondria. To determine whether mitochondrial functions are also altered, mitochondria were isolated from nine baboons fed ethanol chronically and their pair-fed controls. At the fatty liver stage, ADP-stimulated respiration was depressed in ethanol-fed baboons by 59.4% with glutamate, 43.2% with acetaldehyde, 45.1% with succinate and 51.1% with ascorbate as substrates. A similar decrease was noted in the ADP/O ratio (14 to 28%) and respiratory control ratio (20 to 44%) with all substrates. Similar alterations of mitochondrial functions were observed in baboons with more advanced stages of liver disease, namely fibrosis. These changes after ethanol treatment were associated with decreases in the enzyme activities of mitochondrial respiratory chain: glutamate, NADH and succinate dehydrogenase (42, 24 and 28%, respectively), glutamate-, NADH-or succinate-cytochrome c reductase (42, 27 and 32%, respectively) and cytochrome oxidase (59.6%). The content of all cytochromes was also decreased in ethanol-fed baboons, especially aa3 (57%). Moreover, [14C] leucine incorporation into mitochondrial membranes was depressed by 21% after ethanol treatment. On the other hand, glutamate dehydrogenase activities of serum and cytosol in ethanol-fed baboons were significantly higher than those in pair-fed controls. Morphologically, mitochondria of ethanol-fed baboons were larger than those of pair-fed controls. However, the mitochondrial protein content per mitochondrial DNA was unchanged. From these results, we conclude that, morphologically and functionally, hepatic mitochondria in baboons are altered by chronic ethanol consumption; it is noteworthy that these changes are fully developed already at the fatty liver stage, and that morphological alteration appears to reflect the damage of mitochondrial membranes rather than an adaptive hypertrophy.
A Nonhuman Primate Model of Gilbert's Syndrome
Authors: Portman Oscar W. , Chowdhury Jayanta Roy , Chowdhury Namita Roy , Alexander Manfred , Cornelius Charles E. , Arias Irwin M.
Published: August 2008

A Bolivian population of squirrel monkeys, Saimiri sciureus, exhibits several features of Gilbert's syndrome in man, and is proposed as a nonhuman primate model of the condition. The Bolivian population was found to have higher fasting (40.6 ± 2.7 μM; mean ± S.E.) and postcibal (9.9 ± 0.9 μM) plasma unconjugated bilirubin concentrations (p < 0.001) than a closely related Brazilian population (fasting 5.5 ± 0.7 μM); postcibal (2.4 ± 0.7 μM). After intravenous administration of [3H]bilirubin as a tracer dose or at 3.4 μmoles per kg body weight, there was delayed plasma clearance in the Bolivian monkeys. Hepatic UDP-glucuronyl transferase activity for bilirubin (164 ± 25 nmoles per 30 min per gm liver) and biliary bilirubin diglucuronide to monoglucuronide ratios (2.9 ± 0.2) were lower in Bolivian monkeys than in Brazilians (421 ± 36 nmoles per 30 min per gm liver-p < 0.01 and 4.1 ± 0.1-p < 0.02, respectively). Hepatic cytosol glutathione-S-transferase B activity (ligandin) levels were similar for the two populations. After phenobarbital therapy, fasting (11.1 ± 0.9 μM) and postcibal (5.3 ± 1 μM) plasma bilirubin concentrations in Bolivian monkeys were significantly reduced (p < 0.001). Sulfobromophthalein clearance was slightly slower in the Bolivian than in the Brazilian monkeys. SGOT, lactate dehydrogenase, -γ-glutamyl transpeptidase and alkaline phosphatase activities were not increased in Bolivians. Fasting serum conjugated bile salt concentrations in Bolivian monkeys were lower than that in Brazilian monkeys (p < 0.01). Erythrocyte survival and hematological measurements were comparable in both populations.
Identification of an Acceptor System for γ-Aminobutyric Acid on Isolated Rat Hepatocytes†
Authors: Minuk Gerald Y. , Vergalla John , Ferenci Peter , Jones Anthony E.
Published: August 2008

γ-Aminobutyric acid (GABA) is a potent inhibitory neurotransmitter which is synthesized by the enteric bacterial flora and delivered into portal venous blood. To determine whether the liver is likely to play an important role in regulating serum GABA levels, the uptake and metabolism of [3H]GABA by three populations of cells isolated from rat liver were studied. GABA was specifically taken up by hepatocytes but not by endothelial or Kupffer cells. Uptake by hepatocytes was saturable, as well as time and sodium dependent. At 0.5°C, a temperature at which binding of GABA to the cell surface is considered to be the predominant component of the uptake process, the apparent affinity constant (Km) was 0.82 μM and a minimum value for binding velocity (Vmax) was 0.13 μM per min per 5 ± 105 cells. Uptake of [3H]GABA by hepatocytes was markedly inhibited by excess unlabeled GABA (95%), a-aminoisobutyric acid (66%) and bicuculline (58%), but was inhibited much less by alanine (16%) and leucine (29%). These findings suggest that GABA binds specifically to the high affinity acceptor of the A amino acid transport system of rat hepatocytes. Impaired function of this transport system in liver failure could contribute to increased circulating levels of GABA.
Changes in the Status of Neurotransmitter Receptors in a Rabbit Model of Hepatic Encephalopathy
Authors: Ferenci Peter , Pappas Chris S. , Munson Peter J. , Henson Ken , Jones Anthony E.
Published: August 2008

It has previously been shown in an animal model of hepatic encephalopathy (HE) that the number of receptors for the inhibitory neurotransmitter, γ-aminobutyric acid (GABA), increases and that the number of receptors for the excitatory neurotransmitter, glutamate, decreases. To determine the functional status of other neurotransmitter systems in HE, measurements were made of the specific binding of other neurotransmitters to synaptic membranes prepared from the brains of normal rabbits and rabbits in HE due to galactosamine-induced acute liver failure. The development of HE was associated with: (i) a decrease in the density (Bmax) of receptors for the two excitatory amino acid neurotransmitters, aspartate and kainic acid; (ii) an increase in the Bmaxof both the low and high affinity binding site for strychnine, a marker for the inhibitory neurotransmitter glycine; (iii) a decrease in the affinity (Kd) of receptors for dopamine, and (iv) no appreciable change in either the specific binding of [3H]D-ala2-methionine enkephalinamide or [3H]naloxone, markers for opiate receptors, or in the Bmaxor the Kdof receptors for acetylcholine. If it is assumed that the sensitivity of the brain to neurotransmitters varies directly with the density of neurotransmitter receptors, HE may be associated with increased sensitivity to inhibitory amino acid neurotransmitters and decreased sensitivity to excitatory amino acid neurotransmitters. Thus, the observed changes in neurotransmitter receptors in HE afford a feasible pathophysiological basis for the mediation of the neural inhibition of HE.
The Specificity of Human Liver Membrane Lipoprotein: Studies with Monoclonal Antibodies
Authors: Murakami Hiroshi , Kuriki Junsuke , Kakumu Shinichi , Fukui Kazuhiko , Sakamoto Nobuo
Published: August 2008

Hybrid cell lines which secreted antibodies to liver-specific membrane lipoprotein (LSP) were obtained by immunizing SMA and BALB/c mice with human LSP and fusing their splenocytes with the myeloma cell line P 3-NSI/l-Ag 4-l. The secretion of antibody to LSP (anti-LSP) was monitored by binding to a human hepatocellular carcinoma cell line, SK-Hep-1, which possesses surface membrane LSP, and to 125I-antimouse F(ab')2 antibody in radiobinding assay, and by reacting with 125I-LSP in double-antibody radioimmunoassay. From four separate cell fusions, seven secreting hybrids were cloned by dilutional techniques. Of these, four cell lines produced antibodies reacting with a wide variety of cells. The culture supernatants of the remaining three (6D6, 6G3 and 8F10) demonstrated the strongest binding activities against SK-Hep-1 among the various kinds of cell lines tested. However, binding with other cell lines, including renal cancer cells (SK-RC-6) and myeloid cell (HL-60) also occurred. Absorption test of ascitic fluids derived from 6D6 showed that ascitic fluids lost their capacity to bind to target SK-Hep-1 cells when absorbed with SK-Hep-1. Similarly absorption by SK-RC-6 and HL-60 removed almost all of the binding activity of ascitic fluids. Moreover, the binding activities of the ascitic fluids to SK-RC-6 and HL-60 were eliminated when absorbed with SK-RC-6, HL-60 and SK-Hep-1. The present study indicates that our LSP preparation contains nonspecific organ antigens, and although LSP exists on liver cell membrane, It is also found on the cell memberane of other albeit in less quantity.
Liver Membrane Antibodies Detected by Immunoradiometric Assay in Acute and Chronic Virus-Induced and Autoimmune Liver Disease
Authors: Wiedmann K. H. , Bartholemew T. C. , Brown D. J. C. , Thomas Howard C.
Published: August 2008

In this study, we describe a radioimmunoassay to detect liver membrane binding antibodies. The assay was designed to exclude binding of aggregated IgG or immune complexes to Fc 7 receptors of hepatocytes. When this assay was applied to sera from 142 patients, antibodies were found in highest titer in patients with autoimmune chronic active hepatitis, rarely in patients with hepatitis B virus-induced chronic active liver disease, and in 32% of patients with primary biliary cirrhosis. IgM antibodies were found in 100% of patients with acute Type A but not B or non-A, non-B hepatitis. IgA class antibodies were found in the sera of 57% of patients with alcohol-induced hepatitis. All patient groups showing significant titers of liver membrane antibodies display the lesion of piecemeal necrosis except those with alcohol-induced hepatitis. Further studies are needed to determine whether this antibody is the cause of the lesion.
Induction of Taurocholate Release from Isolated Rat Hepatocytes in Suspension by α-Adrenergie Agents and Vasopressin: Implications for Control of Bile Salt Secretion
Authors: Gewirtz David A. , Randolph Joyce K. , Goldman David I.
Published: August 2008

Hepatocytes incubated with 25μM.[3H]taurocholate rapidly deplete the extracellular medium of [3H]taurocholate and achieve a steady-state level of intracellular bile salt within 15 min. Exposure of cells at steady state ith extracellular taurocholate to the catecholamines norepinephrine or epinephrine results in release of3H from the cells into the incubation medium; the3H released represents almost exclusively unmetabolized [3H]taurocholate. The hierarchy of effectiveness of the catecholamines, norepinephrine ≈ epinephrine > phenylephrine ≫ isoproterenol, is indicative of an α-adrenergic mechanism. Induction of [3H]taurocholate release by norepinephrine is inhibited by the β-antagonists phenoxybenzamine and phentolamine and by chlorpromazine, but is not affected by the β-antagonist propranolol, further supporting an α-adrenergic basis for this phenomenon. Arginine vasopressin, at concentrations of 1 ± 10−9M and greater, also induces bile salt release. Classicalα- and β-antagonists have minimal effects on vasopressin induced bile salt release. While the peptide hormones angiotensin and oxytocin are, alone, relatively ineffective inducers of bile salt release, oxytocin potentiates the induction of bile salt release by vasopressin, suggesting complex interactions with membrane receptor function. Further studies assessing the interaction of sympathetic neurotransmitters and peptide hormones with bile salt transport and release in the hepatocyte may provide insight into the regulation of hepatic secretory function in the intact animal.
Importance of Local Proliferation in the Expanding Kupffer Cell Population of Rat Liver After Zymosan Stimulation and Partial Hepatectomy
Authors: Bouwens Luc , Baekeland Marijke , Wisse Eddie
Published: August 2008

Partial hepatectomy and a single intravenous injection of zymosan were used to provoke expansion of the Kupffer cell population in rat liver. The number of Kupffer cells per microscopic field increased exponentially for 4 to 5 days after either stimulation. During this exponential growth phase, high mitotic activity of Kupffer cells was observed.

The rate of mitosis, based on counts of cells arrested in metaphase by vinblastin, was compared with the increase in total cell population. During the first 3 days after partial hepatectomy, local proliferation was sufficient to explain the increase in population since the mean potential doubling time was 1.3 days compared to an observed doubling time of 3.7 days. During the first 3 days after zymosan stimulation, observed doubling time was 4.3 days compared with the mean potential doubling time of 5.2 days which led to the conclusion that growth is largely due to local proliferation of Kupffer cells. In both experimental situations, extrahepatic recruitment of “resident-type” macrophages was required during the last 2 days of the growth phase, since mitotic activity became too low to explain the observed growth. Proliferating Kupffer cells had characteristics of mature resident macrophages and were, therefore, considered to be different from elicited “exudate-type” cells, such as monocytes.
An Antibody Which Precipitates Dane Particles in Acute Hepatitis Type B: Relation to Receptor Sites Which Bind Polymerized Human Serum Albumin on Virus Particles
Authors: Alberti A. , Pontisso P. , Schiavon E. , Realdi G.
Published: August 2008

An antibody, which is distinct from the HBsAg, and reacts with antigenic sites on Dane particles HBcAg and HBeAg, was studied by radioimmunoprecipitation of radioactive intact hepatitis B virions in sera obtained early in the course of acute hepatitis type B. The antibody, previously termed anti-Dane particle (anti-DP) antibody, was reactive with Dane particles and HBsAg particles obtained from HBeAg-positive sera but not with HBsAg particles from anti-HBe containing sera. The expression on virus particles of the evoking antigen correlated with levels of binding sites for polymerized human serum albumin (pHSA) as detected by solid-phase radioimmunoassay. In acute hepatitis B sera, levels of anti-DP antibody activity showed inverse correlation with expression of pHSA receptors on circulating virus particles, although the two reactivities were not mutually exclusive. In inhibition experiments, pHSA blocked precipitation of Dane particles by anti-DP positive sera, while native human albumin and polymerized bovine albumin had no effect. The inhibition by pHSA of the anti-DP reaction appeared specific since identical concentrations of pHSA did not interfere with precipitation of virus particles by anti-HBs. Affinity chromatography studies with anti-DP insolubilized on Sepharose 4B columns showed selective binding to the gel of radioactive Dane particles; 125I-HBsAg was not reactive. The binding of Dane particles to anti-DP columns was completely inhibited when virus particles were applied to the gel in pHSA; pretreat-ment of the column with pHSA did not affect the reaction. These results and the demonstration that anti-DP positive sera were free of anti-pHSA antibodies indicate that anti-DP reacts with antigenic determinants on Dane particles that are involved in or by pHSA binding sites. Anti-DP positive sera and IgG fractions inhibited expression of pHSA receptors on purified virus particles. Our findings support suggestions that the anti-DP response may be relevant to the phase of virus neutralization, on the basis of the hypothesis that the virus receptor for pHSA could play a role in mediating virus access to hepatocytes which may have similar albumin receptors on their membrane.
Biosynthesis of Abnormally Glycosylated α-Antitrypsin by a Human Hepatoma Cell Line†
Authors: Carlson Joyce , Eriksson Sten , Alm Ragnar , Kjellstrom Thomas
Published: August 2008

The human hepatoma cell line PLC/PRF/5 synthesized and secreted a functioal α-antitrypsin (α-AT) glycoprotein with normal molecular size but retarded electrophoretic mobility. The total process of translation, glycosylation and export required about 40 min and followed the same synthetic pattern as seen in rat hepatocytes, i.e., a signal peptide is cleaved cotranslationally; a core-glycosylated protein in the high-mannose form is formed in the rough endoplasmic reticulum, trimmed, and a stable complex-glycosylated α 1-AT is found intracellularly prior to export. α 1-AT export to medium was delayed by tunicamycin, inhibited by cycloheximide but unaffected by colchicine. After addition of exogenous α 1-AT to culture medium, neither negative nor positive feedback induction of synthesis could be demonstrated.

Electrophoretic techniques indicated the presence of atypical, highly branched but incompletely sialylated carbohydrate chains in the hepatoma cell-derived α 1-AT. The accumulation of intracellular α 1-AT inclusions seen in the endoplasmic reticulum may reflect an imbalance between a high rate of polypeptide synthesis and terminal glycosylation.
Intestinal Calcium Absorption and Vitamin D Status in Chronic Cholestatic Liver Disease
Authors: Bengoa Jose M. , Sitrin Michael D. , Meredith Stephen , Kelly Susan E. , Shah Nikunj , Baker Alfred L. , Rosenberg Irwin H.
Published: August 2008

Metabolic bone disease is common in patients with cholestatic liver disease. The importance of vitamin D status and calcium malabsorption in the pathogenesis of bone disease in these patients remains undefined. We have measured intestinal calcium absorption in relation to vitamin D status in 14 patients with chronic cholestatic liver disease including 11 with primary biliary cirrhosis. Fractional calcium absorption was determined from radioactive counts in the right forearm after separate oral and intravenous doses of 47CaCl2 in the fasting state. Eight of 14 patients (57%) had a decreased calcium absorption compared to controls. A significant correlation was observed between serum 2 5-hydroxyvitamin D levels and fractional calcium absorption (r = 0.623, p < 0.02). Treatment with oral 2 5-hydroxyvitamin D3 in three patients with low serum 2 5-hydroxyvitamin D levels resulted in correction of serum 2 5-hydroxyvitamin D levels and improvement in fractional calcium absorption. No correlation was found between serum 1,2 5-dihydroxyvitamin D levels and fractional calcium absorption (r = 0.221). Calcium malabsorption was common in this series of patients, and serum 25-hydroxyvitamin D levels were useful in predicting fractional calcium absorption. Treatment with oral 25-hydroxyvitamin D3 was accompanied by improved calcium absorption.
Metabolic Bone Disease in Alcoholic Cirrhosis: A Comparison of the Effect of Vitamin D2 25-Hydroxyvitamin D, or Supportive Treatment
Authors: Mobarhan Sohrab A. , Russell Robert M. , Recker Robert R. , Posner David B. , Iber Frank L. , Miller Pamela
Published: August 2008

In a study of 56 alcoholics with liver cirrhosis, 18 (32%) had decreased bone density and low levels of serum 25-hydroxyvitamin D (25-OH-D) (<20 ng per ml). To compare the efficacy of vitamin D2 and 25-OH-D treatment in correcting the metabolic bone disease in alcoholic cirrhosis, the 18 patients were randomized in the following manner, in groups of six patients each: Group 1, control (received no supplemental vitamin D treatment); Group 2, given vitamin D2 (50,000 IU p.o.) two to three times weekly, and Group 3, treated with 25-OH-D (20 to 50 mg p.o.) daily as required to attain normal serum 25-OH-D levels. The study period lasted 6 to 12 months (mean, 10.7 months). Initial histomorphometric study of transiliac bone biopsy with double tetracycline labeling in nine patients in whom biopsy was feasible showed only osteoporosis without evidence of osteomalacia. By the end of the study, serum 25-OH-D levels in the control group (Group 1) raised slightly while showing marked improvement in Groups 2 and 3. Bone density results remained unchanged in control patients but demonstrated a significant increase in both treatment groups. Vitamin D2 and 25-OH-D were equally effective in increasing bone density measurements. Posttreatment biopsies were performed in three patients of Group 2 and two patients of Group 3. While the histomorphometric results in Group 3 were not conclusive, in Group 2 improvement in static measures of bone remodeling was noted. Osteoporosis is the usual form of bone disease in alcoholic cirrhosis and a response to either vitamin D2 or 25-OH-D treatment is suggested.
Reversal of Intrahepatic Cholestasis of Pregnancy in Women after High Dose S-Adenosyl-L-Methionine Administration†
Authors: Frezza Mario , Pozzato Gabriele , Chiesa Lucio , Stramentinoli Giorgio , Di Padova Carlo
Published: August 2008

Previous investigations have indicated that S-adenosyl-L-methionine (SAMe) leads to reversal of estrogen-induced bile flow impairment in rats. This randomized, single-blind clinical trial was performed to determine whether SAMe reverses intrahepatic cholestasis of pregnancy (ICP) which occurs in hypersensitive women associated with increased estrogen levels in late pregnancy. Eighteen women with ICP were randomly divided into three groups of six and treated for 20 days as follows: Group I received 200 mg per day of i.v. SAMe; Group II received 800 mg per day of i.v. SAMe; Group III served as control. At the beginning of the study, clinical and biochemical parameters were similar among groups. After 10 and 20 days of treatment with the higher dose of SAMe, the mean values of serum transaminases, conjugated bilirubin and total bile acids fell significantly in respect to initial levels; opposite results were found in the other two treatment groups. The final values of these selected parameters were lower in the group of subjects treated with 800 mg per day SAMe than in the other two groups of women. Pruritus graded on a 0 to 4+ scale significantly was reduced only in patients treated with the higher dose of SAMe. These results indicate a trend toward remission of ICP in women treated with 800 mg per day SAMe and suggest that SAMe administration may be a new therapeutic modality for ICP.
Effect of the Oral Administration of Branched Chain Amino Acids on Hepatic Encephalopathy in the Rat
Authors: Beaubernard Christian , Delorme Marie-Louise , Opolon Pierre , Boschat Mlchele , Morin Joelle , Oryszcyn Marie-Plerre , Franco Dominique
Published: August 2008

The aim of this study was to investigate the effect of oral administration of branched-chain amino acid (BCAA)-enriched diets after portacaval shunt (PCS) in rats. Fifty-one Sprague-Dawley male rats (200 gm) underwent PCS and 55 a sham operation. Half of the animals received BCAA 142 mg per day through a gastric tube; the other half underwent a sham procedure. Sleep disturbances were evaluated at 7, 14, and 28 days postoperatively by measurement of the excitability of the reticular brain-stem formation during the slow-wave sleep and paradoxical sleep. Animals were killed at the same intervals and liver/body weight ratio, plasma, and brain amino acids, brain norepinephrine, brain serotonin, 5-hydroxyindolacetic acid and histamine were measured. Each group of animals was matched with a similar group of sham-operated rats, i.e., receiving or not receiving BCAA. After PCS (as compared to sham-operated animals) a significant hyper-excitability of the reticular brain-stem formation was found during the slow-wave sleep. The liver/ body weight ratio was significantly lower. Tryptophan (free tryptophan in the plasma), phenylalanine, tyrosine, and histidine increased in the plasma and the brain. Leucine and isoleucine were decreased in the plasma. After PCS, an initial decrease at 7 days postoperatively of brain norepinephrine and blockade of the intracerebral metabolism of tryptophan were observed. These changes were transient and progressively disappeared at 14 and 28 days postoperatively. Brain histamine remained at a very high level through the experiment. A good correlation was demonstrated between modification of the sleep disturbance and tryptophan (or 5-hydroxyindolacetic acid) and histamine brain levels. In the PCS animals, BCAA restored the excitability of the reticular brainstem formation toward normal. The oral administration of BCAA was followed by decreased levels and a more rapid turnover of the intracerebral tryptophan. The decrease of brain serotonin and increase of histamine in the brain were corrected only transiently.
Effects of Chenodeoxycholic and Ursodeoxycholic Acids on Lipid Metabolism and Gallstone Formation in the Prairie Dog
Authors: Cohen Bertram I. , Singhal Anil K. , Stenger Richard J. , May-Donath Patricia , Finver-Sadowsky Judith , Mcsherry Charles K. , Mosbach Erwin H.
Published: August 2008

The prevention of cholesterol cholelithiasis by dietary chenodeoxycholic and ursodeoxycholic acids was studied in the male prairie dog (Cynotttys ludovicianus). Gallstones were induced by administration of a semisynthetic diet containing 0.4% cholesterol for a period of 8 weeks. Groups of 5 or 6 animals received the lithogenic diet with added chenodeoxycholic or ursodeoxycholic acid (0.03% “low dose” or 0.06% “high dose”). Under the conditions used, the incidence of gallstones was reduced with the high dose of chenodeoxycholic acid and the low dose of ursodeoxycholic acid, but cholesterol crystals were detected in the biles of 20 of the 22 animals fed these bile acids. A control group maintained on a low (0.08%) cholesterol semisynthetic diet exhibited neither crystals nor stones and was the only group with a lithogenic index below 1.0. The administered bile acids tended to reduce the accumulation of cholesterol in liver and plasma. The activity of hepatic HMG-CoA reductase was significantly inhibited with all cholesterol-supplemented diets. Cholesterol 7α-hydroxylase activity was elevated 83% in prairie dogs fed 0.4% cholesterol, but tended to return to normal levels when bile acids were added to this diet. Histologically, the livers of all animals on the semisynthetic (cholesterol-supplemented) diet exhibited bile duct proliferation, as well as portal fibrosis and inflammatory infiltration. These morphologic alterations were ameliorated by low dose supplementation with either chenodeoxycholic or ursodeoxycholic acid, but high dose bile acid supplementation failed to reduce these pathologic changes.
Frequency of Papillary Dysfunction Among Cholecystectomized Patients
Authors: Bar-Meir Simon , Halpern Zamir , Bardan Eithan , Gilat Tuvia
Published: August 2008

Four hundred and fifty-four consecutive patients who had had their gallbladder removed were interviewed to determine the presence of upper abdominal pain, increased serum alkaline phosphatase and/or serum amylase activity. Patients with unexplained upper abdominal pain and/or enzyme abnormalities were offered endoscopic retrograde cholangiopancreatography (ERCP) and manometric evaluations. Dysfunction of the sphincter of Oddi diagnosed by ERCP manometry may account for the abdominal pain seen in 14% of the patients with postcholecystectomy syndrome. It may rarely be the cause of an elevated serum alkaline phosphatase and/or amylase when abdominal pain is not present. Papillary dysfunction is seen in less than 1% of the patients who have had their gallbladders removed. ERCP manometry is recommended in cholecystectomized patients with unexplained abdominal pain suggesting pancreaticobiliary origin.
(+)-Cyanidanol-3 in the Treatment of Acute Viral Hepatitis: A Randomized Controlled Trial
Authors: Schomerus Hans , Wiedmann Karl H. , Dölle Wolfgang , Peerenboom Herrmann , Strohmeyer Georg , Balzer Klaus , Goebell Harald , Dürr Hans K. , Bode Christian , Blum André L. , Frösner Gert , Gerlich Wolfram , Berg Peter A. , Dietz Klaus
Published: August 2008

One-hundred sixty patients (81 cyanidanol, 79 placebo) were included in a double-blind placebo-controlled clinical trial to evaluate the effect of 3 gm (+)-cyanidanol-3 per day for 8 weeks on the clinical course of acute viral hepatitis and HBsAg elimination. Quantitative determination of HBsAg was performed at frequent intervals.

The mean time for serum bilirubin to decrease to 1.3 mg per dl was 30.8 ± 3.5 days in the treated group and 52.2 ± 9.8 days in the control group, (p < 0.025). The time for SGOT to decrease to 100 IU per liter was 17.98 ± 1.82 in the treated group and 26.53 ± 3.7 in the control group (p < 0.025). No significant difference in the evolution of other laboratory values or symptoms was found. The elimination rate of HBsAg was identical in both groups. Treatment did not alter the incidence of chronicity.
A Randomized Controlled Study of Propranolol for Prevention of Recurrent Gastrointestinal Bleeding in Patients with Cirrhosis: A Final Report
Authors: Lebrec Didier , Poynard Thierry , Bernuau Jacques , Bercoff Eric , Nouel Olivier , Capron Jean-Pierre , Poupon Raoul , Bouvry Michel , Rueff Bernard , Benhamou Jean-Pierre
Published: August 2008

We have previously reported the results of a controlled trial showing that continuous oral administration of propranolol reduced the risk of recurrent gastrointestinal bleeding in patients with cirrhosis; only part of our patients had been followed for 1 year. This controlled trial was continued for an additional year; accordingly, all of our patients have now been followed for at least 2 years. The purpose of the present study is to determine whether prolonged administration enhances the efficacy of this therapy.

Seventy-four patients with cirrhosis, admitted for an episode of gastrointestinal bleeding, were included in this study; ascites, jaundice and encephalopathy were absent or mild and transient. The patients were randomly assigned to two groups; one group of 38 patients received propranolol twice daily at doses that reduced the resting heart rate by 25%, the other group of 36 patients received a placebo twice daily. The cumulative percentages of patients free of recurrent gastrointestinal bleeding 1 and 2 years after inclusion were 87 and 79% in the propranolol group, and 42 and 32% in the placebo group; both differences were highly significant (p < 0.0001). Furthermore, the cumulative percentages of surviving patients 1 and 2 years after inclusion were 94 and 90% in the propranolol group, and 84 and 57% in the placebo group; the difference between the two groups was not significant at 1 year, but was statistically significant at 2 years (p < 0.02). We conclude that, in patients with cirrhosis in good condition, propranolol reduced the risk of recurrent gastrointestinal bleeding and the mortality rate during the 2-year period of continuous oral administration of the drug.
Hepatitis B Viral Nucleotide Sequences in Non-A, Non-B or Hepatitis B Virus-Related Chronic Liver Disease†
Authors: Figus Annalena , Blum Hubert E. , Vyas Girish N. , De Virgilis Stefano , Cao Antonio , Lippi Marco , Lai Eliana , Balestrieri Angelo
Published: August 2008

The presence of serological markers of hepatitis B virus (HBV) infection and of hepatocellular HBV DNA were investigated in 19 HBsAg-negative patients with clinically and histologically significant chronic liver disease. Four cases negative for antibodies to HBsAg (anti-HBs), to the core antigen (anti-HBc), and to the e antigen (anti-HBe) were classified as non-A, non-B hepatitis. The remainder, positive for one or more of the three antibodies, were classified as hepatitis B. Histologic diagnosis was chronic active hepatitis in five, chronic persistent hepatitis in 11, micron-odular cirrhosis in two, and fatty liver in one patient. The DNA extracted from limited amounts of liver biopsies, without cleavage by restriction endonucleases, was analyzed by the Southern blot technique for the presence of episomal HBV DNA. Autoradiographs showed a single band of less than 4.0 kilobase (kb) corresponding to the monomeric form of HBV DNA in five patients, several bands of larger forms (4.0 to 18.0 kb) in three patients, both the monomeric and the larger forms in eight patients, and no HBV DNA in three patients. While HBV DNA was detected in the hepatocellular DNA of six patients who underwent splenectomy, hybridization was negative with the DNA extracted from their spleens. The episomal viral DNA larger than 4.0 kb may represent concatemeric forms or free oligomers which could not be distinguished from rearranged and/or integrated viral DNA in the limited analyses of the hepatocellular DNA hydrolyzed with HindIII or EcoRI. Our observations suggest the presence of HBV-like agents in the liver of serologically HBsAg-negative patients with chronic liver disease.
Etiology of Fulminant Viral Hepatitis in Greece
Authors: Papaevangelou George , Tassopoulos Nicolaos , Roumeliotou-Karayannis Anastasia , Richardson Clive
Published: August 2008

The etiology of fulminant viral hepatitis was determined in 65 consecutive cases among 1,814 (3.58%) adults with acute viral hepatitis who were admitted to the Infectious Diseases Hospital of Athens from May, 1981 to August, 1983. Radioimmunoassays were used to detect hepatitis B virus, hepatitis A virus and δ-agent markers. Enzyme immunoassay was used to detect anti-HBc IgM. Hepatitis B virus was responsible for 48 (73.9%), non-A, non-B for 16 (24.6%) and hepatitis A virus for one case (1.5%). The use of anti-HBc IgM identified 10 cases of fulminant type B hepatitis among HBsAg negative, anti-HBc and anti-HBs positive patients. Fulminant non-A, non-B hepatitis was superimposed on 10 asymptomatic HBsAg carriers; δ-agent co-infection was diagnosed in two fulminant type B hepatitis cases. These data show that hepatitis B virus is the major cause of fulminant hepatitis in Greece. HBsAg carriers are at high risk for fulminant non-A, non-B hepatitis. α-agent does not seem to be as important as in Italy and other countries.
Long-Term Co-Cultures of Adult Human Hepatocytes with Rat Liver Epithelial Cells: Modulation of Albumin Secretion and Accumulation of Extracellular Material
Authors: Clement Bruno , Guguen-Guillouzo Christiane , Campion Jean-Pierre , Glaise Denise , Bourel Michel , Guillouzo Andre
Published: August 2008

High yields of viable human hepatocytes were obtained by enzymatic perfusion of the left hepatic lobe of kidney donors and cultured alone or with an epithelial cell line derived from rat liver. In conventional cultures, human hepatocytes did not survive more than 2 to 3 weeks and by Day 8 decreased their ability to secrete albumin. When co-cultured, they survived for more than 2 months and secreted high levels of albumin even in a serum-free medium. This long-term survival appeared to correlate with production of an extracellular material which is rich in Type III collagen.In vitrophenotypic alterations of parenchymal cells were reversed by addition of rat liver cells and were characterized by recovery of cuboidal morphology, increased albumin secretion and a shift from Type I to Type III collagen deposition. Rat liver epithelial cells could not be replaced by nonhepatic epithelial cells. These observations suggest that when adult human hepatocytes are maintained in a culture which closely resembles theirin vivoenvironment, they are capable of continuing to actively express specific cell functions.
Alterations in Endothelial Fenestrations in Liver Sinusoids of Baboons Fed Alcohol: A Scanning Electron Microscopic Study
Authors: Mak Ki M. , Lieber Charles S.
Published: August 2008

The effects of chronic alcohol consumption on the ultrastructure of endothelial fenestrations in liver sinusoids were studied by scanning electron microscopy in surgical liver biopsies of 16 baboons pair-fed with nutritionally adequate diets containing alcohol or isocaloric carbohydrate for up to 112 months. Alcohol consumption for 4 to 24 months resulted in a decreased number of fenestrations (1.4 per μm2 of the endothelial surface vs. 3.3 in pair-fed controls; p < 0.01) and an increase in their geometric mean diameter (115.6 vs. 82.3 nm in controls; p < 0.001). After 61 to 112 months of alcohol feeding, the number of fenestrations was 1.9 (vs. 4.6 in controls; p < 0.005) and the fenestration diameter was 91.8 nm (vs. 76.7 nm in controls; p < 0.01). The fractional areas occupied by the fenestrations on the endothelial surface of the sinusoids in baboons fed alcohol for 4 to 24 and 61 to 112 months were calculated to be 84 and 58% of their respective controls. The alterations in the sinusoidal endothelium revealed in this study are most likely associated with a disturbance in the exchanges between the sinusoidal blood stream and the liver parenchyma and may thereby contribute to alcohol-induced liver injury.