bluuebunny/crossref_metadata_2025 huggingface dataset:
A New Method to Measure Portal and Hepatic Blood Flow Using Taurocholate in the Rat
Authors: Corbic Michèle , Lebrec Didier , Le Dafniet Michele , Erlinger Serge
Published: August 2008

Hepatic artery and portal vein blood flows were assessed separately in anesthetized rats using radiolabeled taurocholate. The ratio of portal vein blood flow to total hepatic blood flow was calculated from measurements of taurocholate concentrations of portal vein, hepatic artery and hepatic vein, and of taurocholate hepatic extraction ratio. Animals were administered [14C]taurocholate intraperitoneally as a label of the taurocholate pool 24 hr before the experiments. In steady-state conditions, serum [14C]taurocholate activities were used as an index of taurocholate relative concentration and the hepatic extraction ratio of taurocholate was estimated using [3H]taurocholate infused into the systemic blood stream. Total hepatic blood flow was estimated by the Fick's principle using indocyanine green. The mean hepatic blood flow was 13.9 ml min−1 (S.E. 1.9), and the mean ratio of portal blood flow to hepatic blood flow was 72.2% (S.E. 6.7). It is concluded that taurocholate may be used to assess separately portal venous and hepatic arterial blood flow in the rat.
Renal Kallikrein Excretion in Cirrhotics with Ascites: Relationship to Renal Hemodynamics†
Authors: Pérez-Ayuso Rosa M. , Arroyo Vicente , Camps Jordi , Rimola Antoni , Costa Josep , Gaya Joan , Rivera Francisca , Rodes Joan
Published: August 2008

To investigate if the renal kallikrein-kinin system may be involved in the homeostasis of renal perfusion in cirrhosis, urinary kallikrein activity was measured in 11 normal subjects, 31 cirrhotics with ascites and preserved rena plasma flow (548.2 ± 32.2 ml per min) and glomerular filtration rate (85.8 ± 3.4 ml per min), and 18 cirrhotics with functional renal failure (renal plasma flow: 229.9 ± 23.4 ml per min; glomerular filtration rate: 34.9 ± 3.3 ml per min). Plasma renin activity, plasma norepinephrine concentration and the urinary excretion of prostaglandin E2 were also measured in these subjects.

Cirrhotics without renal failure showed a significantly higher renin (4.9 ± 1.1 ng per ml per hr), norepinephrine (458.2 ± 50.4 pg per ml), urinary kallikrein (15.4 ± 1.8 pkat per min) and urinary prostaglandin E2 (0.52 ± 0.08 ng per min) than did normal subjects (1.08 ± 0.1 ng per ml per hr, 218.1 ± 18.2 pg per ml; 8.4 ± 1.4 pkat per min and 0.24 ± 0.02 ng per min, respectively). Cirrhotics with renal failure showed a significantly higher renin (16.1 ± 3.4 ng per ml per hr) and norepinephrine (739.4 ± 79.2 pg per ml), and a significantly lower urinary kallikrein (5.2 ± 0.6 pkat per min) and urinary prostaglandin E2 (0.15 ± 0.02 ng per min) than did normal subjects and cirrhotics without renal failure. Glomerular filtration rate correlated (p < 0.001) with urinary kallikrein (r = 0.53), urinary prostaglandin E2 (r = 0.55), plasma renin (r = -0.41) and norepinephrine (r = -0.44).

These results suggest that renal kallikrein-kinin system may be involved in maintenance of renal blood flow in cirrhosis with ascites, and that functional renal failure in these patients could be a consequence of impaired renal production of kallikrein and prostaglandins in the setting of activation of the renin-angiotensin and sympathetic nervous systems.
Effect of Ethanol and Chlorpromazine on Transhepatic Transport and Biliary Secretion of Horseradish Peroxidase
Authors: Okanoue Takeshi , Kondo Isao , Ihrig Thomas J. , French Samuel W.
Published: August 2008

In order to demonstrate the effect to the acute administration of ethanol and chlorpromazine (CPZ) on bile flow and transhepatic transport of horseradish peroxidase (HRP) into bile, male rats were administered either 5 gm per kg ethanol intragastrically (E-rats) or 3 mg per kg CPZ intraperitoneally (CPZ rats). Control rats (C-rats) received saline. Two hours after ethanol feeding or 90 min after CPZ injection HRP was injected into the portal vein, and bile samples were collected at 1 0-min intervals for 2 hr. Tissue samples were removed at 1,10, 60, and 120 min to study HRP transport using electron microscopic cytochemical localization. Bile flow was reduced (p < 0.001) both in E- and CPZ-rats compared to C-rats. In E-rats HRP secretion was significantly decreased at 30 and 40 min post-HRP injection (p < 0.05) and the peak rate of HRP secretion was delayed by 10 min compared to C-rats. Uptake and transhepatic transport of HRP were similar to controls. These results suggest that bile secretion and flow were impaired by ethanol. CPZ inhibited secretion of HRP significantly (p < 0.001) during the first hr after HRP injection and by 25% after 2 hr. In CPZ rats studied cytochemically HRP reaction product decreased in the cytoplasm of hepatocytes 10 min after HRP injection (p < 0.01). These findings suggest that acute CPZ administration caused an inhibition of the uptake of HRP as well as secretion and bile flow.
Franco-Belgian Cooperative Study of Ursodeoxycholic Acid in the Medical Dissolution of Gallstones: A Double-Blind, Randomized, Dose-Response Study, and Comparison with Chenodeoxycholic Acid
Authors: Erlinger Serge , Le Go Alice , Husson Jean-Marc , Fevery Johan
Published: August 2008

A double-blind randomized, multicenter study was carried out to determine the efficacy and safety of ursodeoxycholic acid (UDCA) at 4 doses of 2.1 to 16.2 mg-kg−1 day−1, and chenodeoxycholic acid (CDCA) at the dose of 16.9 mg-kg−1 -day−1, in 197 patients treated for up to 1 year for radiolucent gallstones in functioning (opacified) gallbladders. There was confirmed complete dissolution in 5.9% of patients receiving UDCA at the dose of 2.1 mg-kg−1 day−1, 18.9% in those receiving 4.2, 28.9% in those receiving 8.4, 14.6% in those receiving 16.2, and 20.0% in patients receiving CDCA. Partial (over 50%) or complete dissolution occurred in 29.4% of patients receiving 2.1 mg-kg−1−day−1 of UDCA, 37.8% of those receiving 4.2, 55.2% in those receiving 8.4, 48.7% in those receiving 16.2, and 50.0% in patients receiving CDCA. Complete dissolution occurred significantly more frequently in small (less than 5 mm in diameter) than in large (5 to 15 and more than 15 mm) stones. There was no significant influence of treatment on serum cholesterol and triglycerides in any of the groups. Serum aminotransferases remained normal (or lower than twice the upper normal limit) in all patients treated with UDCA. Diarrhea leading to cessation of treatment occurred in 5% of patients receiving UDCA, but was significantly less frequent than in those receiving CDCA. These results confirm that, within a 1-year period, UDCA is equally effective and induces diarrhea less frequently than CDCA, with an optimal dose (8 mg-kg−1−day−1) approximately twice lower than that of CDCA.
Experimental Studies of Blood Brain Barrier Permeability in Acute Hepatic Failure
Authors: Zaki Ahmed E. O. , Ede Roland J. , Davis Michael , Williams Roger
Published: August 2008

Permeability of the blood brain barrier in relation to the development of hepatic encephalopathy was investigated in two animal models of acute hepatic failure, in one of which there was the potential for recovery (D-galactosamine-induced hepatitis). In both this and the hepatic devascularization model, there was an approximate 3-fold increase in the passive permeability of the blood brain barrier to inulin and sucrose. Transport of amino acids was also significantly affected, with approximate 30% increases in the brain uptake of phenylalanine, tyrosine and arginine and a 65% increase in uptake of leucine. These changes are attributed to the action of circulating toxic substances, some of which increase blood brain barrier permeability in normal animals.
Liver Ultrastructure in Mitochondrial Urea Cycle Enzyme Deficiencies and Comparison with Reye's Syndrome
Authors: Latham Patricia S. , Labrecque Douglas R. , Mcreynolds John W. , Klatskin Gerald
Published: August 2008

The liver ultrastructural findings in two girls with partial carbamyl phosphate synthetase I (CPS) deficiency and their heterozygote parents and two siblings with ornithine transcarbamylase (OTC) deficiency are described. Liver ultrastructure in the four patients with inherited deficiences of urea cycle enzymes showed minimal alterations with essentially normal mitochondria when biopsy was performed during periods of good control of their hyperammonemia. Mitochondrial ultrastructure was also essentially normal in the heterozygotes for carbamyl phosphate synthetase I deficiency. These findings are in contrast to the marked alterations in mitochondrial ultrastructure found in the study of two cases of Reye's syndrome in which severe depression of ornithine transcarbamylase and carbamyl phosphate synthetase I activities was noted.
Liver Disease in Cytomegalovirus Mononucleosis: A Light Microscopical and Immunoperoxidase Study of Six Cases
Authors: Snover Dale C. , Horwitz Charles A.
Published: August 2008

Six liver biopsies from previously healthy adult patients with cytomegalovirus (CMV) mononucleosis were studied by routine light microscopy and by the immunoperoxidase technique for CMV antigen. Light microscopical findings consisting of a mononuclear portal and sinusoidal infiltrate, increased hepatocellular mitotic activity and minimal hepatocellular necrosis were consistently found. Less common features were granuloma formation and bile duct epithelial damage. Typical CMV nuclear inclusions and CMV antigen were identified in only one case, a patient with marked leukopenia secondary to CMV who had received corticosteroid therapy. The other five cases contained no inclusions and CMV antigen could not be identified by immunoperoxidase staining. This data suggests that, as with hepatitis B, viral antigen is not identifiable in acute CMV hepatitis in the immunocompetent host, perhaps due to active destruction of infected cells. The immunoperoxidase technique for CMV appears to be of little value in the diagnosis of acute CMV hepatitis.
Mechanisms of the Impaired Diuretic and Natriuretic Responses to a Sustained and Moderate Saline Infusion in Rats with Experimental Cirrhosis
Authors: López-Novoa José M. , Santos Juan C. , Caramelo Carlos , Fernandez-Muñoz Dolores , Blanchart Alicia , Hernando Luis
Published: August 2008

Kidney function and tubular handling of water and sodium by superficial nephrons, packed cell volume, total plasma proteins and albumin distribution space were studied in control and cirrhotic rats before and after a moderate and sustained saline infusion (3% body weight per 30 min + reposition of urinary losses). Tubular fluid samples were obtained from late proximal, early distal and late distal convolutions of superficial nephrons using micropuncture. Protein distribution was assessed by intravenous injection of 0.5 μCi of (125I)-albumin. In basal conditions, both groups of rats showed similar glomerular filtration rate and renal plasma flow, but cirrhotic animals had lower sodium excretion (fractional excretion of sodium = 0.04 ± 0.01% vs. 0.22 ± 0.02%, p < 0.05) and urinary volume (4.31 ± 0.41 vs. 7.57 ± 0.53 μ1 per min; p < 0.05). After saline infusion, total plasma proteins decreased more in cirrhotic than in control rats (-−18.5 ± 2.7 vs. -12.9 ± 2.2%, p < 0.05). The opposite was observed for albumin distribution space (34.5 ± 6.1 vs. 22.1 ± 3.5%, p < 0.05). Fractional sodium excretion increased to 2.98 ± 0.15% in control rats but only to 0.61 ± 0.080% in cirrhotic rats. The ratio single nephron glomerular filtration rate/glomerular filtration rate increased from 19.6 ± 0.7 to 21.2 ± 1.0 (x10−6, p < 0.005) in control animals but did not change in cirrhotic rats. These animals were unable to decrease adequately fractional fluid reabsorption in the proximal tubule and the loop of Henle.

The results suggest that the lack of adequate diuresis and natriuresis after saline infusion in cirrhotic rats depends on lower plasma volume expansion by extravasation of high protein content fluid, lack of intrarenal glomerular filtration rate redistribution, and inadequate decrease in proximal and Henle's loop fluid reabsorption.
Prognostic Value of Child-Turcotte Criteria in Medically Treated Cirrhosis
Authors: Christensen Erik , Schlichting Poul , Fauerholdt Lis , Gluud Christian , Andersen Per Kragh , Juhl Erik , Poulsen Hemming , Tygstrup Niels
Published: August 2008

The Child-Turcotte criteria (CTC) (based on serum bilirubin and albumin, ascites, neurological disorder and nutrition) are established prognostic factors in patients with cirrhosis having portacaval shunt surgery. The objective of this study was to evaluate the prognostic value of CTC in conservatively treated cirrhosis. Patients (n = 245) with histologically verified cirrhosis from a control group of a controlled clinical trial were studied. Data at entry into the trial were used to classify patients according to CTC. Survival curves for up to 16 years were made, and survival rates were compared using the log-rank test. Survival decreased significantly with increasing degree of abnormality (A ± B ± C) of albumin (p < 0.001), ascites (p < 0.001), bilirubin (p = 0.02) and nutritional status (p = 0.03). Survival was insignificantly influenced by neurological status (p = 0.11) probably because none of the patients had hepatic coma at entry into the trial. The five variables in CTC were combined to a score. With increasing score, the median survival time decreased from 6.4 years (score 5) to 2 months (scores 12 or more). Furthermore, the mortality from hepatic failure, gastrointestinal bleeding or hepatocellular carcinoma increased significantly with increasing score. CTC provide valuable and easily obtainable prognostic information in cirrhosis. However, CTC are inferior to a prognostic index based on multivariate analysis of prognostic factors.
Comparison of Intravenous Somatostatin and Vasopressin Infusions in Treatment of Acute Variceal Hemorrhage†
Authors: Kravetz David , Bosch Jaime , Terés Josep , Bruix Jordi , Rimola Antoni , Rodés Juan
Published: August 2008

The present trial compared the effectiveness and complications of intravenous somatostatin and vasopressin in treatment of variceal bleeding. Sixty-one cirrhotic patients with endoscopically proven active variceal bleeding were included. Both drugs were given as continuous intravenous infusions for 48 hr. Thirty patients received somatostatin (250 #ig per hr after a bolus of 50 ng) and 31 vasopressin (0.4 units per min).

Initial control of bleeding was achieved in 26 (87%) patients receiving somatostatin and in 23 (74%) of those treated with vasopressin.

However, 10 patients [not significant statistically] in the somatostatin group and 5 in the vasopressin group rebled during treatment, after a mean of 15 and 20 hr, respectively. Therefore, complete control of bleeding during the 48 hr of therapy was achieved in 16 (53%) patients treated with somatostatin and in 18 (58%) of those receiving vasopressin. Mortality during hospitalization was similar in both groups (somatostatin 47%, vasopressin 45%).

Differences were observed in complications associated with each therapy. Vasopressin produced major complications in 8 patients (left ventricular failure in 4 and severe abdominal pain requiring drug withdrawal in 4), and minor complications in 14; somatostatin infusion produced minor complications in 3 patients (p < 0.01). In addition, the serum sodium concentration was significantly reduced by vasopressin (from 134.3 ± 1.6 to 128.3 ± 1.4 mEq per liter, p < 0.001) but not by somatostatin (134.6 ± 1.1 vs. 133.2 ± 1.1 mEq per liter).

This study shows that somatostatin is as effective as vasopressin in controlling variceal hemorrhage, but has a much lower rate of complications.
Effect of Albumin Binding on Extraction of Sulfobromophthalein by Perfused Elasmobranch Liver: Evidence for Dissociation-Limited Uptake
Authors: Weisiger Richard A. , Zacks Charles M. , Smith Neil D. , Boyer James L.
Published: August 2008

We investigated the role of serum albumin in the hepatic uptake of organic anions by determining the effect of added bovine albumin on sulfobromophthalein (BSP) uptake in skates, an animal which naturally lacks this protein. A single-pass perfused liver model was used to determine steady-state net uptake rates as the BSP or BSP and albumin concentrations were systematically varied. Results indicated that two different steps in the uptake process are capable of limiting the uptake rate depending on the albumin and BSP concentrations selected. At higher rates of uptake corresponding to higher BSP concentrations (up to 112 μM), saturation kinetics were observed as the BSP concentration was varied, with apparent Kra and Vmax values which were independent of the albumin concentration (0.05 to 0.75%). These data suggest that under these conditions uptake is limited by a saturable step intrinsic to the liver. In contrast, for lower BSP concentrations and albumin concentrations below about 0.5%, a different kinetic pattern was seen which suggested that the rate-limiting step in uptake was transfer of BSP from albumin to the liver. The latter data were found to be consistent with a model in which the limiting step in the transfer process is spontaneous dissociation of the BSP from binding sites on albumin within the sinusoid.

These results suggest that skate liver clears BSP from albumin solutions by a two-step mechanism in which dissociation from albumin is followed by a saturable process consistent with carrier-mediated transport. The albumin concentration and uptake velocity are important factors in determining which of these steps limits the uptake rate. Clearance is efficient and irreversible even though elasmobranch liver lacks high-affinity cytosolic binding proteins for BSP, and elasmobranch plasma does not normally contain albumin.
Acute Hepatitis A Infection in Hepatitis B Chimpanzee Carriers†
Authors: Tsiquaye Kwesi N. , Harrison Tim J. , Portmann Bernard , Hu Shanlian , Zuckerman Arie J.
Published: August 2008

Two hepatitis B virus carrier chimpanzees which were superinfected with hepatitis A virus developed acute hepatitis followed by the production of antibodies to hepatitis A virus. The Southern blot technique employed to monitor liver hepatitis B virus DNA revealed that the amount of viral DNA in both animals was significantly reduced during the acute phase of hepatitis A infection. The levels of plasma hepatitis B DNA polymerase activity were also reduced in one chimpanzee. The high titers of HBsAg in the circulation remained unchanged throughout the study, and antibodies to the surface antigen and to e antigen were not detected. The morphological lesions in the liver were severe in one chimpanzee from whom one specimen showed both periportal focal necrosis and zonal parenchymal necrosis.
Lack of Glucagon Receptors in Morris Hepatoma 7800
Authors: Mourelle Marisabel , Rojkind Marcos
Published: August 2008

When compared to normal liver membranes, purified plasma membranes of regenerating liver and Morris hepatomas contain low but variable capacities to bind glucagon. This property is inversely related to the capacity of the isolated hepatocytes to bind to heterologous biomatrix glycoproteins. Since these parameters are characteristic of the proliferative state of the cells, it was important to further study the glucagon receptor protein and stimulation of adenylate cyclase activity. Our results show that 125I-iodinated plasma membranes obtained from normal liver contain three molecular species (117000, 98000, 86000 molecular weight) that can be eluted specifically with glucagon from a sepharose-glucagon affinity column. These proteins contain the putative glucagon receptor since binding of 125I-iodoglucagon is increased 150-fold as compared to unfractionated membranes. Plasma membranes obtained from Morris hepatoma (7800) and liver of chemically hepatectomized rats do not bind glucagon and lack these proteins. After inactivation with N-ethylmaleimide of the adenylate cyclase activity of the normal plasma membranes, they were fused with membrane of the hepatoma. The hybrid membranes showed 60% recovery of glucagon-stimulated cyclase activity. These results suggest that the plasma membranes of the proliferating liver cells do not contain receptor protein but have intact regulatory and catalytic subunits of the adenylate cyclase system.
Quantitative Microscopy Comparison of Peroxisome Proliferation by the Lipid-Regulating Agent Gemfibrozil in Several Species
Authors: Gray Robert H. , La De Iglesia Felix A.
Published: August 2008

Peroxisome proliferation, a well-documented subcellular reaction which follows the administration of hypolipidemic agents, has been well studied in rodents. However, quantitative studies of this phenomenon in other species of laboratory animals are not readily available even though these species are commonly used as predictors of tolerance or safety in humans. The quantitative stereologic studies reported here compared the effects of the new hypolipidemic agent gemfibrozil on hepatic peroxisomes of monkeys, dogs, hamsters and rats of both sexes under several treatment schedules. Gemfibrozil was administered to rats at 300 mg per kg per day for 1 year in the diet; to hamsters at 400 mg per kg per day for 2 weeks by diet admixture; to dogs at 300 mg per kg per day in gelatin capsules for 1 year; and to monkeys at 300 mg per kg day for 3 months by gavage. These dose levels were selected on the basis of tolerance from preliminary studies in each species. At the end of each experimental interval, liver samples were processed for quantitative microscopy. Peroxisomes from male rats were enlarged and the number of peroxisomes per cell were increased 7-fold over controls, resulting in a 20-fold increased peroxisome volume per cell. Statistically, significant increases also occurred in female rats and the difference between treated and controls was 3-fold for both number and volume of peroxisomes per cytoplasmic unit volume. In hamsters, peroxisomes were proliferated and were of significantly smaller size to the extent that the volume of cytoplasm occupied by peroxisomes was not significantly changed. In dogs, the number of peroxisomes per cell was increased and the volume fraction was significantly increased in females only. The number of peroxisomes in young monkeys did not change after treatment, and the peroxisome volume was decreased in males and increased in females. Aged monkeys had increased number of peroxisomes per hepatocyte with increased volume fraction. These results indicate significant differences in the magnitude and direction of peroxisome changes, reflecting species-dependent organelle response to hypolipidemic agents. The order of susceptibility of peroxisome proliferation in laboratory animals is dog < monkey < hamster < rat.
Phthalate Esters and Their Effect on the Liver
Authors: Ganning Agneta E. , Brunk Ulf , Dallner Gustav
Published: August 2008

The advantageous properties of polyvinyl chloride plastics make these materials indispensable at all levels of human life, and there is no reason to believe that the upward trend of their use will change. One third of most polyvinyl chloride plastics consists of plasticizers, mostly phthalate esters, which may migrate out contaminating our environment and consequently our bodies. Experimental results indicate that chronic exposure to phthalic esters may be responsible for a number of adverse health effects. Phthalates change the structure and function of the liver in a profound manner by inducing peroxisomes, mitochondria and enzymes which participate in fatty acid transport and β-oxidation. Prolonged administration of phthalate esters, in doses comparable to those occurring in human exposures, seems to have an accumulative effect on the liver. Liver biopsies taken from dialysis patients show peroxisome proliferation which again warn of the possibility that human health may be influenced by plasticizers.
Plasma Lipoprotein and Apolipoprotein Profile in Alcoholic Patients with and without Liver Disease: On the Relative Roles of Alcohol and Liver Injury
Authors: Duhamel Gilles , Nalpas Bertrand , Goldstein Sonia , Laplaud Michel P. , Berthelot Pierre , Chapman John M.
Published: August 2008

In the present study, we report on alterations in plasma lipid, lipoprotein and apolipoprotein patterns in three separate populations of alcoholic patients, one without liver damage (Group I), a second presenting steatosis or mild alcoholic hepatitis or both (Group II) and a third with alcoholic cirrhosis (Group III), using a healthy, normolipidemic, nonalcoholic group as controls (Group C). Total plasma cholesterol levels were elevated in Groups II and III when compared with Groups I and C, while the ratio of esterified to free cholesterol was considerably lower in Group III than in the other groups. Plasma apo-AI levels were higher in Groups I and II than in Group C, but varied over a wide range in Group III. Apo-AII was present at higher concentrations in Groups I and II than in both Groups III and C. In contrast, no significant differences were detected in total apo-B levels, irrespective of the group. Modifications in the chemical composition of plasma lipoproteins primarily concerned a reduction in the cholesteryl ester content of low-density lipoproteins (LDL) and high-density lipoprotein (HDL) in Group III, this being compensated by a reciprocal increase in triglyceride. In addition, Group III lipoproteins, with the exception of HDL3 (density 1.100 to 1.140 gm per ml), exhibited a greater content of phospholipids than those of corresponding density from patients in Groups I and II. No significant differences were found in very low-density lipoprotein concentrations, while LDL levels increased in parallel with the severity of liver injury. In Groups I and II, HDL2 concentrations were elevated relative to Group C, while HDL3 decreased in parallel with the degree of impairment of liver function and thus from Group C to Group III. Such opposing tendencies led to an HDL2:HDL3 ratio which was more than 5-fold higher than normal in Group III.

The distribution of apoprotein B in the ultracentrifugal subfractions revealed no significant modification between the different groups. By contrast, in Groups I and II, apo-AI and apo-AII levels increased consequent to higher concentrations of HDL2. Our data suggest that abnormal HDL particles preferentially enriched in apo-AII and possibly apo-AI were present in Group I, while the lowest levels of both apo-AI and apo-AII were seen in Group III.

No simple association is evident between a well-defined plasma lipid, lipoprotein and/or apolipoprotein profile and chronic alcoholism in the presence or absence of liver injury. Nonetheless, our findings suggest that measurement of the absolute concentrations and ratio of HDL2 and HDL3, as well as of the levels and ratio of apo-AI and apo-AII in these subclasses, may permit differentiation of certain of the subpopulations of chronic alcoholics described herein.
Hepatitis B Virus Replication in Southern African Blacks with HBsAg-Positive Hepatocellular Carcinoma
Authors: Song Ernest , Dusheiko Geoffrey M. , Bowyer Sheila , Kew Michael C.
Published: August 2008

Sera from 106 southern African blacks with hepatocellular carcinoma and hepatitis B surface antigenemia (HBsAg) were tested for hepatitis B viral DNA (HBV-DNA) activity, HBV-DNA polymerase concentrations, and HBV e antigen (HBeAg) and antibody (anti-HBe) to investigate the state of viral replication in these patients. HBeAg and anti-HBe were detected by radioimmunoassay, HBV-DNA by molecular hybridization using a 32p-labeled HBV-DNA probe, and HBV-DNA polymerase was measured by incorporation of 3H-labeled thymidine triphosphate into double-stranded HBV-DNA. HBeAg was present in 30.2% (32/106) of the patients, almost always in low titer; 63.8% of the patients were anti-HBe positive. Circulating HBV-DNA was detected in 18.8% (20/106) of patients, including 14 of 32 (43.7%) who were HBeAg positive and 6 of 74 (8.1%) who were anti-HBe positive. In most patients, only trace amounts of HBV-DNA were evident. Raised HBV-DNA polymerase activity was found in 5.6% (6/106) of the patients, all of whom were HBeAg positive and 4 of whom had detectable amounts of circulating HBV-DNA. The HBV-DNA polymerase activity was relatively low in these patients. HBV replication thus appears to be present in only a minority of southern African Blacks with HBV-related hepatocellular carcinoma, and when
IgM Antibody to Hepatitis B Core Antigen in Korean Patients with Hepatocellular Carcinoma† ‡
Authors: Sjogren Maria H. , Lemon Stanley M. , Chung Whan K. , Sun Hee S. , Hoofnagle Jay H.
Published: August 2008

Primary hepatocellular carc1inoma (PHC) has been linked etiologically to persistent hepatitis B virus (HBV) infections by epidemiologic, serologic and molecular lines of evidence. To evaluate the frequency of IgM antibody to the viral core antigen (IgM anti-HBc) detected by a highly sensitive radioimmunoassay, we compared 110 Korean patients with PHC to a group of 63 age-and sex-matched control patients with other tumors. Results were correlated with those of commercially available HBV assays. IgM anti-HBc was found in 74 of 110 PHC patients (67%), but only 1 of 63 (1.6%) control patients. Although HBsAg was found in a larger percentage of PHC patients (81%), it was also present in more control patients (14%). Thus, IgM anti-HBc was more specifically associated with PHC than was the presence of HBsAg. IgM anti-HBc was found in 91% of PHC patients with detectable HBeAg and 74% of PHC patients with positive anti-HBe tests (p < 0.04). The frequency of IgM anti-HBc was similar among HBsAg-positive PHC patients with and without anti-HBs, or those with low or high levels of serum a-fetoprotein. In 18 patients with PHC, IgM anti-HBc was further characterized by rate-zonal centrifugation of sera, all were found to have 19S IgM anti-HBc although 6 also had greater or equal IgM anti-HBc reactivity in the low molecular weight region. The presence of IgM anti-HBc in adult Korean HBsAg carriers may indicate an especially high risk for the development of PHC, and this should be evaluated in prospective studies.
Evidence for Valine Intolerance in Patients with Cirrhosis
Authors: Schauder Peter , SchrÖDer Karsten , Herbertz Lothar , Langer Klaus , Langenbeck Ulrich
Published: August 2008

Valine (62.5 mg per kg), leucine (70 mg per kg) and equal amounts of the calcium salts of the corresponding keto acids, i.e., α1-ketoisovaleric acid (KIVA) and α1-ketoisocaproic acid (KICA) were orally administered to patients with cirrhosis and to control subjects.Valine or leucine ingestion increased serum valine and leucine levels and the corresponding keto acids, KIVA and KICA, in cirrhotics and controls. KIVA or KICA ingestion increased serum KIVA and KICA concentrations within a few minutes associated with a rise in valine and leucine.

In cirrhotics, administration of valine or KIVA resulted in significantly higher serum valine or KIVA concentrations than in control subjects. The clearance of valine and KIVA from blood was also delayed in cirrhotic patients. No such differences were observed after leucine or KICA ingestion.

It is suggested that cirrhotics have a diminished tolerance for valine. Since the tolerance for KIVA, but not KICA, is also impaired, it appears that cirrhotics have a derangement in one or more metabolic steps distal to the branched-chain keto acid dehydrogenase.
Immunocytochemical Detection of Desmin in Fat-Storing Cells (Ito Cells)
Authors: Yokoi Yukio , Namihisa Toshihiko , Kuroda Hlroyuki , Komatsu Ikutoshi , Miyazaki Akihisa , Watanabe Sumio , Usui Koh
Published: August 2008

Desmin is a smooth muscle protein consisting of intermediate filaments. It is also present in other cell types. We obtained desmin antibodies from New Zealand white rabbits which were immunized with desmin following its extraction from chicken gizzard smooth muscle. By immunocytochemical methods, the antibody stained the Z-line of rat skeletal myofibrils and produced only one band with chicken gizzard desmin by immunoblotting. The antibody was specific for desmin, but was not tissue-specific. Wistar rat liver was studied by indirect immunoperoxidase and immunofluo-rescent methods using the antibody. Cells which reacted were diffusely distributed in the hepatic lobule, had stellate shape, bore elongated cytoplasmic extensions and contained few fat droplets. The fat droplets increased in size and amount after administration of vitamin A, and contained vitamin A as determined by ultraviolet exposure and gold impregnation. The fat-containing cells did not phagocytose carbon particles after injection of colloidal carbon and were, therefore, classified as fat-storing cells (Ito cells).
Studies on the Secretion of Serum Proteins from Rat Hepatoma Cells
Authors: Mccracken Ardythe A. , Emmett Merrill , Crowle Alfred J. , Brown Jerry L.
Published: August 2008

We have used crossed Immunoelectrophoresis to identify and establish the relative amounts of serum proteins secreted by a differentiated cell line (Fao) derived from a Reuber H35 rat hepatoma. Our results show that these cells secrete at least 15 plasma proteins. Ten of these: albumin, α1-antitrypsin, α1-lipoprotein, α1-macroglobulin, α1-antichymotrypsin, GC-globulin (transcalciferin), fibronectin, hemopexin, transferrin and the C3 component of complement have been identified. To examine the feasibility of using the Fao cell line as a model for studies on the regulation of hepatic protein secretion, we measured the relative amounts of 10 serum proteins secreted into the growth medium after exposure of these cells to dibutyryl cyclic AMP, hydrocortisone and a combination of both compounds. We also examined the effects of growth temperature (33.5°, 37° and 39°C) and the removal of fetal calf serum from the growth medium on the relative amounts of these proteins secreted. We found that the rates of secretion of most of the serum proteins were altered by one or more of the treatments used in these experiments. In addition, detectable levels of secretion of three serum proteins, fibronectin and two unidentified, occurred only under certain of the experimental conditions. These results demonstrate that the pattern of proteins secreted from Fao cells can be experimentally altered and indicate that this cell line may be a useful model for studies on the control of hepatic protein secretion.
An Enzyme-Linked Immunosorbant Assay (ELISA) for Detecting Antimitochondrial Antibody
Authors: Kaplan Marshall M. , Gandolfo John V. , Quaroni Elaine G.
Published: August 2008

We have developed an enzyme linked immunosorbant assay (ELISA) for antimitochondrial antibody. Polyvinyl microtiter plate wells are coated with partially purified rat kidney mitochondria, and excess protein binding sites are blocked with bovine serum albumin. Human serum, diluted 1:1,000, is incubated for 1 hr. Then β-galactosidase-goat-anti-human IgG (H+L) is added followed by the substrate, p-nitrophenyl-β-D-galactopyranoside. The plates are then read at 404 nM in a microelisa autoreader. A positive result was defined as optical density ± 0.100, more than 5 standard deviations above the mean of 36 normal individuals. With this techique, 56 of 60 patients with primary biliary cirrhosis were positive for antimitochondrial antibody (93%), mean O.D., 0.456 ± 0.031 S.E. Seventeen of 17 patients with extrahepatic bile duct obstruction and 14 or 14 patients with alcoholic cirrhosis were negative. Only 1 of 29 patients with chronic active liver disease was positive (4%). Antinuclear antibody and antimicrosomal antibody do not bind in this assay, and activity is absorbed from sera by preincubation with suspensions of rat kidney mitochondria. The ELISA is approximately 20 times more sensitive than a quantitive microtiter complement fixation technique and more convenient than radioimmunoassay. It is rapid, quantitative and uses stable reagents. In contrast to immunofluorescence techniques, it is not affected by observer interpretation.
Clinical Significance of Enhanced Detection of HBsAg by a Monoclonal Radioimmunoassay†
Authors: Ben-Porath Edna , Wands Jack , Gruia Mignon , Isselbacher Kurt
Published: August 2008

We assessed the significance of the enhanced detection by monoclonal radioimmunoassay (M-RIA) of HBsAg in serum of patients with hepatitis B virus (HBV) infection. In acute HBV infection, the M-RIA detected HBsAg in the blood for a far longer period of time than previously recognized. In some patients, the “window phase” of HBV infection (defined as the presence of anti-HBc and the lack of detectable HBsAg and anti-HBs) was shortened or completely eliminated. Furthermore, 26% of individuals with anti-HBc as the only serologic marker of recent or past HBV infection were reactive only in the M-RIA. Analysis of patients tested for HBsAg because of suspected underlying liver disease revealed additional HBsAg-positive sera, unrecognized by polyvalent RIA, thus adding important information in the immunodiagnosis of HBV infection. The enhanced performance of the monoclonal RIA compared to conventional polyvalent RIA was due in part to the increased sensitivity of the assay and the observation that M-RIA may detect “hidden” HBV antigens in immune complexes.
Long-Term Sequelae of Non-A, Non-B Hepatitis in Experimentally Infected Chimpanzees
Authors: Burk Kenneth H. , Dreesman Gordon R. , Cabral Guy A. , Peters Robert L.
Published: August 2008

We have observed the development of long-term sequelae in four cases of experimentally induced non-A, non-B (NANB) hepatitis in chimpanzees. These sequelae were characterized by the following manifestations: nonprotection against challenge with autologous infectious plasma following acute disease and subtle histopathological alterations typical of long-lasting viral hepatitis. These manifestations were observed in animals infected with either of two human inocula. Whether or not these inocula represent sources of single or multiple etiologic agents is not known. However, our studies suggest that these inocula share at least one common etiologic agent. Further, these results may represent an atypical chronology of convalescence from viral hepatitis infection. For example, the convalescent stage of a type B hepatitis infection may be expected to occur within 6 to 8 months following exposure, whereas true convalescence in NANB hepatitis may be protracted over several months to several years. Thus, future efforts to identify the causative agent(s) of NANB hepatitis, and efforts to define the immune response in NANB, must take into consideration these studies.
Sclerotherapy of Esophageal Varices: An Endoscopic and Portographic Study
Authors: Söderlund Claes , Backman Lars , Erwald Rolf , Forsgren Lennart , Marions Olle , Wiechel Karl-Ludvig
Published: August 2008

A prospective series of 26 patients with portal hypertension and recent bleeding from esophageal varices was investigated with percutaneous transhepatic selective portography (PTP). PTP was performed immediately prior to and, in 23 patients, just after the initial endoscopic injection sclerotherapy (ST) session to study the acute effects of ST on the mediastinal portal-systemic collaterals. Late follow-up PTP was performed after a median of 8 months in 21 of 26 patients considered endoscopically to be free from esophageal varices after a median of 6 ST sessions. Five patients rebled from esophageal or gastric varices during the follow-up period of 15 months, but there were no fatalities due to variceal hemorrhage. In all patients, the initial PTP showed portal-systemic mediastinal collaterals. Immediately after ST, it was not possible to opacify esophageal varices at all (10 patients) or only partially (7 patients). Five patients died prior to late follow-up PTP. Endoscopic judgment of complete eradication of esophageal varices after repeated ST was in agreement with the late PTP results in 18 of 21 patients. In one patient, PTP showed residual esophageal varices subsequently confirmed by endoscopy. The results were uncertain in two patients for technical reasons. This study supports the opinion that submucosal esophageal varicose veins, as visualized by PTP, can be efficiently eradicated by serial ST, leaving the other mediastinal collaterals unaffected.
Rat Liver Regeneration After 90% Partial Hepatectomy
Authors: Gaub Johannes , Iversen Johan
Published: August 2008

In previous studies, 90% partial hepatectomy in the rat was invariably accompanied by 100% mortality within 40 hr. This paper describes a technique by which 90% of the liver mass can be removed with only 14% mortality, provided that rats have free access to glucose-containing drinking water. Measurements of total liver DNA, [3H]thymidine labeling index and mitotic index suggest rapid cell proliferation, commensurate with a powerful regenerative stimulus. In the presence of normoglycemia, peripheral insulin and glucagon concentrations were elevated to levels normally observed in the portal vein.
Does Z-Protein Have a Role in Transport of Bilirubin and Bromosulfophthalein by Isolated Perfused Rat Liver?
Authors: Theilmann Lorenz , Stollman Yacov R. , Arias Irwin M. , Wolkoff Allan W.
Published: August 2008

Bilirubin and other organic anions are transported in serum avidly bound to albumin from which they are extracted and transferred into the hepatocyte where they bind to cytosolic proteins. Two abundant organic anion binding proteins, ligandin and Z-protein, were previously purified from liver cytosol and characterized. Other studies in isolated perfused rat liver revealed that selectively increased cytosolic ligandin concentration, following phenobarbital treatment or thyroidectomy, directly correlated with net bilirubin uptake which resulted from reduced bilirubin efflux. To clarify the role of Z-protein in hepatic organic anion transport, we have now determined the kinetics of bilirubin and bromosulfophthalein (BSP) uptake in isolated perfused liver of normal rats and compared results to rats in which Z-protein, but not ligandin, was selectively increased following treatment with clofibrate (ethylchlorophenoxy-isobutyrate). These studies revealed that despite a 147% induction of Z-protein in treated animals, there was no effect on influx or efflux of tracer doses of bilirubin or BSP. Addition of albumin to the protein-free 10% fluorocarbon perfusate reduced influx of 3H-bilirubin (p < 0.03) and tended to reduce influx of BSP. In this situation, there was still no influence of Z-protein concentration on efflux. These studies indicate that Z-protein does not appear to play a role in the hepatic uptake of bilirubin and BSP.
The Persistence of Hepatitis A IgM Antibody After Acute Clinical Hepatitis A†
Authors: Kao Henry W. , Ashcavai Mary , Redeker Allan G.
Published: August 2008

Hepatitis A IgM antibody (IgM anti-HAV), detected by commercially available solid-phase radioimmunoassay, is an accepted marker of acute viral hepatitis A infection. However, persistence of this serological marker far beyond the acute illness and immediate convalescent period has been reported. To determine the persistence of IgM anti-HAV following clinically manifest acute hepatitis A infection, 59 patients with this diagnosis were followed prospectively until this marker disappeared or persisted for greater than 60 days. Timed from the onset of jaundice, IgM anti-HAV persisted for less than 30 to greater than 420 days; most patients became seronegative by 120 days. These findings suggest that some patients may become seronegative early in the disease course while others (13.5%) remain positive for prolonged periods greater than 200 days. Awareness of this marked variability is important in the interpretation of IgM anti-HAV as a serologic marker of recent hepatitis A infection.
Ultrastructural Liver Pathology in Patients with Minimal Liver Disease and α1-Antitrypsin Deficiency: A Comparison Between Heterozygous and Homozygous Patients
Authors: Hultcrantz Rolf , Mengarelli Silva
Published: August 2008

A light and electron microscopic investigation of liver tissue from three homozygous (Pi-type ZZ) and three heterozygous (Pi-type MZ) α1-antitrypsin deficiency patients is presented. All had slightly elevated levels of serum amino transferases as the only signs of liver damage. Light microscopical investigation showed minor periportal fibrosis and periodic acid-Schiff-positive globules in the homozygous patients, but no specific findings besides fatty changes were seen in the heterozygous patients.

Electron microscopical investigation was performed on periportal and centrilobular areas separately. In the three homozygous patients, dilatation of endoplasmic reticulum was found in many periportal liver parenchymal cells. In these cells, a marked increase in peroxisomes was also noted and in some instances they showed signs of cell necrosis. Evidence of Rupffer cell heterophagy of such cells was also found, thus indicating that they had died. Only in one liver parenchymal cell from one of the three heterozygous patients was evidence of a dilated endoplasmic reticulum found. No increase in peroxisomes was found and no apparent signs of liver cell necrosis were noted. An increased amount of lipofuscin-like material in the lysosomes was encountered in liver cells both from patients with homozygous and heterozygous ai-antitrypsin deficiency.

Based on these findings, it is suggested that cell death occurs in cells with a dilatation of the endoplasmic reticulum. If these findings are important for the pathogenesis in liver disease in α1-antitrypsin deficiency, the present study suggests that the risk to develop liver disease is less in heterozygous patients since, with one exception, alterations of the same nature were not found in their hepatocytes.
Close Association Between Basal Cell Layer Antibodies and Hepatitis B Virus-Associated Chronic Delta Infection
Authors: Zauli Daniela , Fusconi Marco , Crespi Cristina , Bianchi Francesco B. , Craxi Antonio , Pisi Emilio
Published: August 2008

Antibodies reacting in immunofluorescence with the basal cell layer of rat forestomach (BCLA) have been detected in 36 of 121 (30%) hepatitis B virus (HBV)-mediated chronic liver disease (CLD), in 1 of 30 (3%) HBV-negative CLD, in 3 of 36 (8%) alcoholic liver disease (with no correlation with serum HBV markers), in 1 of 25 (4%) primary biliary cirrhosis, in none of 19 HBV-related HBsAg-negative CLD and 60 healthy blood donors. Of 352 hospitalized patients with miscellaneous diseases (including immunological conditions), the antibodies were found in four (1%).

In the 36 BCLA positive cases from HBV-mediated CLD, evidence of chronic delta infection was found in 34. The overall prevalence of BCLA in 68 delta cases was 50% (58% in chronic active hepatitis, 46% in cirrhosis), and in 28 delta negative cases was 4% (p < 0.00002).

BCLA of delta cases were mainly of the IgG class (38 of 41 sera), and high titers (up to 40,960) were found in the majority (66% >1:640).

The high titer BCLA has to be considered a marker of chronic delta infection in HBV cases.
Detection of Reovirus Type 3 in the Porta Hepatis of an Infant with Extrahepatic Biliary Atresia: Ultrastructural and Immunocytochemical Study
Authors: Morecki Rachel , Glaser Joy H. , Johnson Anne B. , Kress Yvonne
Published: August 2008

This report describes immunocytochemical and ultrastructural methods which led to the identification of Reovirus type 3 (Reo-3) in the porta hepatis of a patient with extrahepatic biliary atresia. The study indicates that Reo-3 antigenic sites are demonstrable by the avidin-biotinylated complex peroxidase method following formalin fixation and paraffin embedding, but are destroyed by freezing and thawing prior to fixation. Deparaffinization of the block and subsequent rembed-ding in epon-araldite did not alter immunoperoxidase staining. This procedure offered the advantage of higher light microscopic resolution of semithin (1 jtm) sections and assisted in the selection of specific areas for ultrastructural studies.

Localization of Reo-3 in extrahepatic biliary atresia was confined to a biliary remnant in which there were acutely inflammed, partially necrotic microscopic ducts. Electron microscopic examination of the immunoreactive sites revealed virus-like particles similar in appearance to Reo-3 particles in infected tissue culture cells. The observations presented here support previously reported serologic data which have shown an association between Reo-3 infection and extrahepatic biliary atresia.