bluuebunny/crossref_metadata_2025 huggingface dataset:
α–Fetoprotein in the Woodchuck Model of Hepadnavirus Infection and Disease: Immunochemical Analysis of Woodchuck α–Fetoprotein and Measurement in Serum by Quantitative Monoclonal Radioimmunoassay
Authors: Cote Paul J. , Pohl Christoph , Boyd Juanell , Tennant Bud C. , Gerin John L.
Published: March 2007

Woodchuck hepatitis virus infection of the eastern woodchuck represents a useful model for the study of hepatitis B virus infection and disease in humans, including hepatocellular carcinoma. In man, hepatocellular carcinoma is frequently detected and monitored using assays for serum alpha–fetoprotein. To study the relationship between α–fetoprotein and woodchuck hepatitis virus–induced hepatocellular carcinoma in the woodchuck model, we produced a monoclonal antibody to woodchuck α–fetoprotein and used biophysical and immunochemical methods to demonstrate its specificity and affinity (7 × 108 L/mol) for woodchuck α–fetoprotein. A competition radioimmunoassay was then developed and standardized for measuring serum α–fetoprotein concentrations. In the radioimmunoassay system, woodchuck α–fetoprotein was detected between 20 ng/ml (20% to 25% inhibition) and 8,500 ng/ml (97% to 98% inhibition). Elevated serum α–fetoprotein concentrations (450 to 452,000 ng/ml) were measured in 21 of 23 woodchucks in the advanced stages of woodchuck hepatitis virus–induced hepatocellular carcinoma. Serum α–fetoprotein was elevated above normal ( ≥ 450 ng/ml) as early as 3 to 11 mo before terminal hepatocellular carcinoma in 11 of 16 of the woodchuck hepatitis virus–carrier woodchucks. In a pilot study, serum α–fetoprotein became markedly elevated above normal in woodchuck hepatitis virus–carrier woodchucks that developed hepatocellular carcinoma but not in serologically recovered or uninifected woodchucks (i. e., without hepatocellular carcinoma). Thus, α–fetoprotein may provide a useful nonivasive marker in the woodchuck model for detecting and monitoring woodchuck hepatitis virus–induced hepatocellular carcinoma from earlier stages.(HEPATOLOGY 1990; 11:824–833.)
Role of Hepatic γ–Glutamyltransferase in the Degradation of Circulating Glutathione: Studies in the Intact Guinea Pig Perfused Liver
Authors: Speisky Hernan , Shackel Nick , Varghese George , Wade Denis , Israel Yedy
Published: March 2007

The role of hepatic γ–glutamyltransferase in the breakdown of circulating glutathione was studied in the perfused guinea pig liver. Hepatic γ–glutamyltransferase activity in the guinea pig is sevenfold higher than in the rat and is comparable to its activity in man. Guinea pig livers were found to remove, in a single pass, 50% to 90% of glutathione ( 10 to 50 μmol/L) added to the portal perfusate. Removal of portal glutathione was totally dependent on the activity of γ–glutamyltransferase and led to the near quantitative appearance of cysteinyl–glycine and cysteine in the caval perfusate. Glutathione removal by the intact liver followed saturation with a Michaelis constant (Km) of 59 μmol/L for glutathione and a maximum velocity of 235 nmol glutathione/min/gm of liver weight. The capacity of the guinea pig liver to remove circulating glutathione was estimated to be sevenfold to 10–fold higher than its net rate of output of glutathione into the circulation. Inhibition of γ–glutamyltransferase activity in the perfused liver led to threefold to sixfold increases in the hepatic output of glutathione into the circulation, indicating that more than two thirds of glutathione transported extracellularly is broken down. Data obtained demonstrate a major role of hepatic γ–glutamyltransferase, both in the removal of portally carried glutathione and in the degradation of glutathione molecules released by the liver itself into the sinusoids. These findings suggest the existence of an intraorgan transport of glutathione in the liver, whereby periportal cells could provide glutathione precursors to pericentral cells.(HEPATOLOGY 1990; 11:843–849.)
Increased Densities of Peripheral–Type Benzodiazepine Receptors in Brain Autopsy Samples From Cirrhotic Patients With Hepatic Encephalopathy
Authors: Lavoie Joel , Layrargues Gilles Pomier , Butterworth Roger F.
Published: March 2007

Peripheral–type benzodiazepine receptors were evaluated using the specific ligand [3H]–PK 11195 in brain homogenates from nine cirrhotic patients who died in hepatic coma and from an equal number of age–matched control subjects. Histopathological studies showed evidence of severe Alzheimer type II astrocytosis in the brains of all cirrhotic patients. Saturation–binding assays revealed a single saturable binding site for [3H]–PK 11195 in brain, with affinities in the 2– to 3–nmol/L range. Diazepam was found to be a relatively potent inhibitor of 3H–PK 11195 binding (IC50 = 253 nmol/L), whereas the central benzodiazepine antagonist Ro 15–1788 displaced 3H–PK 11195 binding with low affinity (IC50 >40 μmol/L). Densities of [3H]–PK 11195 binding sites were found to be increased by 48% (p < 0.01) and 25% (p < 0.05) in frontal cortex and caudate nuclei, respectively, from cirrhotic patients. Densities of [3H]–PK 11195 binding sites in frontal cortex from two nonencephalopathic cirrhotic patients were not significantly different from control values. No concomitant changes of affinities of these binding sites were observed. Because it has been suggested that peripheral–type benzodiazepine receptors may be localized on mitochondrial membranes and may therefore be involved in cerebral oxidative metabolism, the alterations observed in this study could be of pathophysiological significance in hepatic encephalopathy.(HEPATOLOGY 1990; 11:874–878.)
Radioimmunolocation of Metastases From Adenocarcinoma of the Colon: Using A Hepatoma Antibody
Authors: Divgi Chaitanya R. , Cohen Alfred M.
Published: March 2007

We have established a large library of monoclonal antibodies against a human hepatoma cell line called FOCUS. One such monoclonal antibody (SF–25) detects a 125–kilodalton cell surface antigen found on FOCUS cells. As both the liver and the colon are of endodermal origin, we examined the possibility of expression in colon adenocarcinomas. This antigen was found in all 23 colon adenocarcinoma tissues surgically obtained but was absent in the adjacent normal mucosal counterpart as determined by a direct radioimmunohistologic technique. In the present study, we have established a model for human metastatic colon adenocarcinoma using the LS 180 cell line. Athymic mice were further immunosuppressed by intravenous injection of anti–NK cell antibodies (antiasialo GM1). After 24 h, mice were injected with LS 180 cells either via the tail vein or into the spleen followed by splenectomy. Macroscopic pulmonary and lymphatic metastasis developed within 2–3 wk after injection of cells and 9 of 10 mice died with advanced metastatic disease 2–3 wk later. In addition, macroscopic hepatic metastases were evident in 4 of 5 mice 3–4 wk after intrasplenic injection. Both hepatic as well as pulmonary and lymphatic tumor spread was localized by nuclear imaging with 125I–SF–25. Furthermore, micrometastases were detected by autoradiography 5–10 days later. Monoclonal antibody SF–25 is a potential candidate for tumor localization and the experimental metastatic colon cancer animal model may be useful for treatment evaluation of monoclonal antibody SF–25 either alone or in combination with other monoclonal antibodies when conjugated to radionucleotides and chemotherapeutic agents.
Blunt Hepatic and Splenic Trauma: Where Have All the Surgeons Gone?
Authors: Baker Christopher C.
Published: March 2007

To further define the computed tomographic (CT) criteria on which to guide the nonsurgical treatment of adult patients with blunt hepatic injury, the authors retrospectively reviewed abdominal CT scans obtained before surgery during a 35–month period. Blunt hepatic injury was diagnosed in 187 patients, and review revealed 37 patients in whom the liver was the site of sole or principal intraabdominal injury detected with the help of CT before surgery. A CT–based hepatic injury classification system partly derived from similar systems established with surgical assessment was devised to grade the severity of hepatic injury. CT–based injury scores ranging from grade 1 to 5 were compared with the clinical outcome in patients treated surgically and nonsurgically. Thirty–one patients (83.7%) were successfully treated without surgery, and four patients (10.8%) had findings at celiotomy that did not require further surgery. No patient who was initially treated without surgery required delayed celiotomy due to hepatic injury. The results indicate that even major hepatic injury up to and including grade 4 severity assessed with preoperative CT can usually be managed without surgery in hemodynamically stable patients.

Nonsurgical management of blunt splenic injury in children is a well–established method to salvage splenic function; however, nonsurgical management of adult blunt splenic trauma remains controversial. To assess the value of preoperative abdominal CT in predicting the outcome of blunt splenic injury in adults, a CT based injury–severity score consisting of four grades was devised and applied in 39 adult patients with blunt splenic injury as the sole or predominant intraperitoneal injury detected with preoperative CT. While patients with high grades of splenic injury generally required early surgery, eight (35%) of 23 patients with initial grade 3 or 4 injury were treated successfully without surgery, and four (29%) of 15 patients with grade 1 or 2 injury initially treated nonsurgically required delayed celiotomy (n = 3) or emergency rehospitalization. Results show that while CT remains an accurate method of identifying and quantifying initial splenic injury, as well as documenting progression or healing of critical injury, CT cannot reliably help predict the outcome of blunt splenic injury in adults. Treatment choices should therefore be based on the hemodynamic status of the patient and results of serial laboratory and bedside assessments.
Transient Induction of C–Jun During Hepatic Regeneration
Authors: Alcorn Joseph A. , Feitelberg Steven P. , Brenner David A.
Published: March 2007

The cellular oncogene c–jun is transiently expressed in cultured cells stimulated to proliferate but has not been identified in normal liver. Because partial hepatectomy results in coordinated cell proliferation in the remaining liver, we investigated c–jun expression after partial hepatectomy in mice. Northern analysis of whole liver mRNA demonstrated a transient increased expression of c–jun within half an hour of the operation. The related gene junB increased only 50%, whereas c–jun expression increased 13–fold compared with sham–operated controls. To determine the cell of origin of the c–jun transcript, both in situ hybridization and Northern analysis of mRNAs from parenchymal and nonparenchymal cell fractions were performed 2 hr after partial hepatectomy. C–jun expression was found in both cell populations. To investigate the mechanism of increased c–jun expression, cycloheximide was given to some animals preoperatively. C–jun induction occurred with cycloheximide alone, but partial hepatectomy further increased c–jun expression, indicating that new protein synthesis was not required for this effect. Furthermore, run–on transcriptional assay demonstrated a twofold increase in c–jun expression. Thus c–jun expression increase after hepatectomy by transcriptional and posttranscriptional mechanisms. Because the extracellular matrixdegrading enzyme transin, which bears the recognition site for jun/AP–1, showed sustained induction after hepatectomy, we speculate that an important function of c–jun expression could be the remodeling of extracellular matrices to accommodate cell proliferation.(HEPATOLOGY 1990;11:909–915.).
Acute Phase Responses After Acute Liver Injury by Parital Hepatectomy in Rats As Indicators of Cytokine Release
Authors: Cornell Robert P.
Published: March 2007

The purpose of this study was to support the hypothesis that cytokines such as interleukin–1, tumor necrosis factor and interleukin–6 are released by macrophages or monocytes within 1 to 2 hr of phagocytosis of circulating, gut–derived backterial lipopolysaccharide translocated by acute liver injury. Time courses of fever, neutrophilia and low blood–zinc levels generally attributed to cytokines were quantified after partial (37%) hepatectomy of rats under ether anesthesia. These acute phase responses in hepatectomized rats were compared with those after intravenous injection of exogenous endotoxin and human natural interleukin–1. Fever commenced 30 min after interleukin–1 injection, 4 hr after exogenous lipopolysaccharide injection and 6 hr after 67% liver resection. Similarly, rectal temperatures were significantly elevated in recipient rats 30 min after intravenous administration of donor plasma from hepatectomized animals, indicating that cytokines, not lipopolysaccharide, elicited the febrile response. Neutrophilia was present 1, 2, and 4 hr after interleukin–1 injection, lipopolysaccharide injection and hepatectomy, respectively. Furthermore, the reduction in plasma zinc, which depends on cellular metallothionein synthesis, occurred 4 hr after interleukin–1 administration and 6 hr after lipopolysaccharide injection or partial hepatectomy. Donor plasma from hepatectomized rats also elicited neutrophilia at 1 hr and low blood–zinc levels 4 hr after injection in recipient animals. The timing of these responses, just as for the fever, implies that cytokines and not lipopolysaccharide in the donated plasma elicited the neutrophilia and hypozincemia. Evidence was reviewed that interleukin–1, tumor necrosis factor and interleukin–6 function as hepatotrophic factors and have been identified in the circulation of humans with liver damage. This complements the conclusion of this study that unspecified cytokines are released after partial hepatectomy of rats.(HEPATOLOGY 1990;11:923–931.).
Glucuronidation of Oxazepam Is Not Spared in Patients With Hepatic Encephalopathy
Authors: Sonne Jesper , Andreasen Per Buch , Loft Steffen , Døsing Martin , Andreasen Frederik
Published: March 2007

The disposition of oral oxazepam was investigated in seven patients with decompensated cirrhosis and encephalopathy and in nine healthy individuals to further examine the hypothesis of preservation of glucuronidation in liver disease. The patients showed a severe reduction in the quantitative liver function as assessed by estimation of the clearance of antipyrine; the median value was 9 ml. min-1 and the range was 6 to 12 ml. min-1. Apparent clearance of oxazepam in cirrhotic patients was 0.55 ml. min-1. kg-1, with a range of 0.46 to 1.24 ml. min-1. kg-1, compared with 1.19 ml. min-1. kg-1 and a range of 0.80 to 1.66 ml. min-1. kg-1 in the controls (p < 0.05). The unbound clearance of oxazepam in patients was 4.1 ml. min-1. kg-1, with a range of 3.4 to 5.5 ml. min-1. kg-1, compared with 25.4 ml. min-1. kg-1, and a range of 16.7 to 43.7 ml. min-1, kg-1, p < 0.001, in the controls. In patients with liver disease, the unbound clearance of oxazepam correlated significantly with antiyprine clearance (r = 0.88; p < 0.05). The results suggest a reduced capacity for glucuronidation in patients with decompensated liver disease and severe hepatic failure that corresponds to the general reduction in the quantitative liver function.(HEPATOLOGY 1990;11:951–956.).
Hepatocellular Carcinoma in Primary Biliary Cirrhosis: An Autopsy Study
Authors: Nakanuma Yasuni , Terada Tadashi , Doishita Kenji , Miwa Atsushi
Published: March 2007

A survey of Japanese autopsy cases of primary biliary cirrhosis disclosed that hepatocellular carcinoma is apparently becoming a better recognized complication of the advanced stage of primary biliary cirrhosis. Six autopsy cases (five women and one man) of primary biliary cirrhosis associated with hepatocellular carcinoma were obtained from several Japanese institutions and examined. All cases were in an established cirrhotic stage of primary biliary cirrhosis. Hepatocelluar carcinoma was incidentally found at autopsy in four cases and, in these, the carcinomas were small in size and number. The other two cases showed advanced hepatocellular carcinoma and one case showed extrahepatic metastasis. Histologically, all cases showed well–differentiated hepatocellular carcinomas. Fatty changes or bile plugs were frequently seen within the tumors. Mallory body clusters and focal deposition of copper–binding protein were consistently found in cirrhotic liver tissues and also in the carcinoma tissues of almost all cases. The presence of atypical adenomatous hyperplasia in the peripheries of some carcinomas suggested that hepatocellular carcinoma in primary biliary cirrhosis may evolve through multiple steps.(HEPATOLOGY 1990;11:1010–1016.).
Growth–Factor Independence of A New Differentiated Hepatitis B Virus Dna–Negative Human Hepatoma Cell Line
Authors: Bagnarelli Patrizia , Devescovi Giulia , Manzin Aldo , Bearzi Italo , Bonazzi Patrizia , Carloni Guido , Clementi Massimo
Published: March 2007

The establishment of a new, differentiated, hepatitis B virus DNA–negative, human hepatoma cell line (named PLC/AN/2) is described. Neoplastic liver tissue was obtained during hepatectomy in an HBsAg–negative man. The established cell line is negative for α–fetoprotein and carcinoembryonic antigen; it has retained in vitro some of the differentiated functions of normal hepatocytes. Additionally, it presents a distinctive rearrangement (translocation) at the long arm of chromosome 4. The high degree of independence from serum growth factor requirements appears to be a major in vitro characteristic of PLC/AN/2 cells, making them a suitable model system for the more precise definition of the human hepatocellular carcinoma phenotype, including mechanisms of growth control.(HEPATOLOGY 1990;11:1024–1032.).
Interaction Between Hepatitis B Surface Proteins and Monomeric Human Serum Albumin
Authors: Krone Bernd , Lenz Angela , Heermann Klaus-Hinrich , Seifer Maria , Xuangyong Lu , Gerlich Wolfram H.
Published: March 2007

HBsAg is known to bind to human serum albumin polymerized by glutaraldehyde, human serum albumin has been found in preparations of HBsAg by several investigators. However, it is not yet known whether natural human serum albumin binds to hepatitis B virus under physiological conditions. We studied the binding between natural or recombinant HBsAg and monomeric human serum albumin by immunological, biochemical and biophysical methods. The binding capacity of 20–nm HBs spheres was variable but ranged up to six molecules HSA/sphere. A reversible binding site for human serum albumin was exclusively localized in the preS2 domain, whereas the S domain was inactive in vitro. Human serum albumin copurified with HBsAg of human origin during gel chromatography or sucrose–gradient centrifugation. This human serum albumin was monomeric in sodium dodecyl sulfate–polyacrylamide gel electrophoresis. The preS2 bound part of the human serum albumin could be removed from HBsAg by high–salt, such as CsCl centrifugation, but another part could only be removed by treatment with a disulfide cleaving reagent. Most of this covalently bound human serum albumin was retained at the HBsAg particle after complete cleavage of medium–sized HBs protein with trypsin. This indicates a second way in which albumin binds irreversible to cysteine(s) of the small HBs protein (SHBs, P24 and GP27)(HEPATOLOGY 1990;11:1050–1056.).
Radiologic Ablation of the Gallbladder: An Alternative to Cholecystectomy in the Twenty–First Century
Authors: Martin E. C.
Published: March 2007

Twenty pigs underwent (a) cystic duct occlusion by means of fluoroscopically guided transcatheter endoluminal bipolar radio–frequency (RF) electrocoagulation and (b) gallbladder sclerotherapy with one of two different regimens of ethanol and sodium tetradecylsulfate (STS). Serum ethanol levels and hepatic enzyme tests showed no acute toxicity. Postmortem histologic studies showed that the bile ducts beyond the occlusion site remained entirely unaffected in all animals. In three of four animals followed up for 2 weeks, the sclerosants induced necrosis of the gallbladder mucosa, but the adjacent liver, serosa, and blood vessels remained intact. In 13 of 16 animals followed up for 8 weeks, the gallbladder lumen was obliterated by fibrous scar tissue. In the animals treated with 95% ethanol and 3% STS, the gallbladder mucosa was necrotic in all areas after 2 weeks (two of two animals) and eradicated completely after 8 weeks (six of eight animals); the other regimen (70% ethanol plus 1% STS) was somewhat less effective. In this study, the combination of RF–mediated cystic duct occlusion and gallbladder sclerotherapy with ethanol and STS permitted gallbladder ablation in swine without toxic side effects.
Pathogenesis of Hepatocellular Carcinoma in Hereditary Hemochromatosis: Occurrence in Noncirrhotic Patients
Authors: Kew Michael D.
Published: March 2007

Previous reports have emphasized the association of primary hepatocellular carcinoma in patients with idiopathic hemochromatosis with cirrhosis. In contrast, patients with idiopathic hemochromatosis without cirrhosis have no increased risk of hepatocellular carcinoma. Phlebotomy therapy, by preventing the accumulation of parenchymal iron and subsequent cirrhosis, is believed to prevent hepatocellular carcinoma in the precirrhotic stage of the disease. We report the case of a 67–yr–old man with a 32–yr history of idiopathic hemochromatosis compliacated by cirrhosis, who had reversal of cirrhosis with phlebotomy theray, yet developed hepatocellular carcinoma. There was no serologic or tissue evidence of hepatitis B infection.
Propranolol–Induced Reduction in Recurrent Variceal Hemorrhage in Schistosomiasis
Authors: Conn Harold O.
Published: March 2007

Fifty patients with non–cirrhotic portal fibrosis who were admitted to hospital because of upper gastrointestinal bleeding were randomly assigned to treatment with either oral propranolol given in does that reduced the resting pulse rate by 25% (25 patients) or with a placebo (25 patients). One year after the start of the study 20 patients in the propranolol group and five patients in the placebo group were free from recurrent gastrointestinal bleeding (p < 0.0001).

Giving continuous oral propranolol treatment is therefore effective in preventing recurrent upper gastriontestinal bledding in patients with noncirrhotic portal fibrosis.
Improved Survival After Prophylactic Portal Nondecompression Surgery for Esophageal Varices: A Randomized Clinical Trial
Authors: Inokuchi Kiyoshi
Published: March 2007

To evaluate prophylactic surgery for esophageal varices, a prospective randomized controlled trial was begun in 1980 by the Japanese Research Society for Portal Hypertension. Methods of operation included selective shunts and nonshunting interruption procedures. One hundred and twelve Japanese patients, in whom endoscopic findings suggested risk of bleeding but who had no bleeding episode, were randomly allocated to the operated group of 60 patients or nonoperated group of 52 patients. Nine patients with idiopathic portal hypertension, histologically proven noncirrhotic disease, which all fell in the operated group, were excluded from the study and the remaining 103 patients (51 operated and 52 nonoperated) were analyzed. Long–term follow–up of patients for a median of 49 mo with a maximum of 73 mo showed a total of 11 (22%) deaths, including 2 operative deaths, in the operated group compared with 23 (49%) deaths in the nonoperated group. The cumulative survival rate at 5 yr in the operated group was 72%, which was significantly higher than the 45% of the nonoperated group (p < 0.05). The cumulative variceal bleeding rate at 5 yr was 7% in the operated group, which was significantly lower than that of the nonoperated group of 46% (p < 0.001). It was concluded that portal nondecompression surgery was effective in preventing the variceal bleeding and in improving survival. (Hepatology 1990; 12:1-6).
Computerized Measurement of Iron in Liver Biopsies: A Comparison With Biochemical Iron Measurement
Authors: Olynyk John , Hall Pauline , Sallie Richard , Reed William , Shilkin Keith , Mackinnon Malcolm
Published: March 2007

The measurement of stainable hepatic iron using a microcomputer image analysis system was compared with standard biochemical measurements of liver iron content in 103 liver biopsy specimens-29 of idiopathic hemochromatosis, 51 of alcoholic liver disease and 23 of various nonalcoholic liver diseases. Sections were stained using Perls' method for iron; the mean area staining positively for iron was measured and expressed as a percentage of the area of biopsy measured. Biochemical (biochemical hepatic iron [μmol/gm dry wt]/age) and morphometrical (morphometrical hepatic iron [%]/age × 100) hepatic iron indices were calculated. Patients in the idiopathic hemochromatosis group had significantly higher biochemical hepatic iron concentrations (p < 0.001) compared with the alcoholic liver disease and nonalcoholic liver disease groups: 284 (range = 119 to 631), 21 (range = 2 to 65) and 15 (range = 3 to 31) μmol/gm dry wt, respectively. The biochemical hepatic iron index was also significantly higher (p < 0.001) in the hemochromatosis group compared with the alcoholic liver disease and nonalcoholic liver disease groups: 5.8 (range = 2.1 to 13.7), 0.4 (range = 0 to 1.6) and 0.4 (range = 0 to 1.1), respectively. Computerized measurements were significantly higher in the hemochromatosis group (p < 0.001) compared with the alcoholic liver disease and nonalcoholic liver disease groups: 9.72% (range = 1.50% to 29.26%), 0.13% (range = 0% to 1.20%) and 0.03% (range = 0% to 0.40%), respectively. The morphometrical hepatic iron index was also significantly higher (p < 0.001) in the hemochromatosis group compared with the alcoholic liver disease and nonalcoholic liver disease groups: 19.32 (range = 2.60 to 75.10), 0.21 (range = 0 to 1.90) and 0.09 (range = 0 to 1.10), respectively. There was no overlap between the hemochromatosis and the alcoholic liver disease and nonalcoholic liver disease groups. Significant linear relationships were demonstrated between morphometrical and biochemical measurements of hepatic iron (p < 0.001) and morphometrical and biochemical hepatic iron indices (p < 0.001).

We conclude that computerized measurements of stainable hepatic iron show a significant linear relationship with biochemical iron measurements. Also, the morphometrical hepatic iron index, like the biochemical hepatic iron index, may help differentiate idiopathic hemochromatosis from other disorders with increased hepatic iron levles. (Hepatology 1990;12:26-30).
Immunoelectron Microscopic Observation of Intrahepatic Hbeag in Patients With Chronic Hepatitis B
Authors: Yamada Gotaro , Takaguchi Kouichi , Matsueda Kazuhiro , Nishimoto Hiroshi , Takahashi Michiko , Fujiki Shigeatsu , Mizuno Motowo , Kinoyama Shingo , Tsuji Takao
Published: March 2007

Immune light and electron microscopic studies using monoclonal antibodies have been applied to localize HBeAg in liver biopsy specimens of 19 patients with chronic hepatitis B. Under the light microscope, HBeAg was demonstrated in nuclei, cytoplasm and on the cell surface of hepatocytes. The number of HBeAg–positive hepatocytes correlated well with the serum levels of HBeAg (enzyme immunoassay) and DNA–polymerase. Of 11 patients in whom high numbers of HBeAg–positive hepatocytes were found at the light microscopic level, HBeAg was also studied in hepatocytes at the electron microscopic level. The HBeAg in nuclei was either found as aggregates or dispersed diffusely. In the aggregates of HBeAg, the 27–nm core particles were frequently found. In addition, the antigen was found in the cytosol of some hepatocytes as amorphous mass and in some hepatocytes in the cisternae of perinuclear space, endoplasmic reticulum and Golgi saccules. Occasionally the antigen was found on the membranes of the cell organelles and on the plasma membranes that faced the intercellular and the Disse spaces.

These findings suggest that cytoplasmic HBeAg in hepatocytes may be ultrastructurally classified into two different patterns by its distribution, mainly in endoplasmic reticulum or in cytosol. The ratio varied between hepatocytes with these two types of patterns. The titer of serum HBeAg tended to be higher when the corresponding liver biopsy specimens contained more hepatocytes with HBeAg in endoplasmic reticulum than those with HBeAg in the cytosol. (Hepatology 1990;12:133-140).
Binding of the Major and Large Hbsag to Human Hepatocytes and Liver Plasma Membranes: Putative External and Internal Receptors for Infection and Secretion of Hepatitis B Virus
Authors: Leenders William P.J. , Glansbeek Harrie L. , De Bruin Wieke C.C. , Yap Sing-Hiem
Published: March 2007

A likely mechanism of the strong hepatotrophism of the hepatitis B virus is the presence of specific receptors for the surface antigen of hepatitis B virus on hepatocyte membranes. To examine this hypothesis, we have performed binding studies using recombinant large (preS1 + preS2 + S) and major (S) proteins with adult human hepatocytes, rat hepatocytes, human fibroblasts, human peripheral blood mononuclear cells and plasma membranes derived from these cell types. We found that major HBsAg was able to bind specifically to human hepatocytes, human fibroblasts and human blood mononuclear cells. This binding could be inhibited by recombinant middle (preS2 + S) protein but not by the recombinant large protein. No binding could be demonstrated between large HBsAg and human hepatocytes. However, large protein bound specifically to plasma membranes derived from human liver tissue, human fibroblasts and HepG2A16 cells. This binding could be partially inhibited by the major protein and by a synthetic preS1 peptide but not by a synthetic preS2 peptide. These results support the assumption that hepatitis B virus absorption and penetration into human hepatocytes is mediated by specific receptors recognizing an amino acid sequence in the S–region. This recognition site is not displayed by the recombinant large protein. However, the large protein is recognized by its preS1 region and by a second binding site in the S–region by a receptor molecule, located on the inner surface of the plasma membranes or intracellular membranes of human hepatocytes and of some other cell types derived from human tissue. (Hepatology 1990;12:141-147).
Recombinant Human γ–Interferon in Patients With Chronic Active Hepatitis B: Pharmacokinetics, Tolerance and Biological Effects
Authors: Marcellin Patrick , Loriot Marie-Anne , Boyer Nathalie , Martinot-Peignoux Michéle , Degott Claude , Degos Françoise , Brandely Maud , Lenfant Bernard , Benhamou Jean-Pierre
Published: March 2007

Pharmacokinetics, tolerance and biological effects of human recombinant γ–interferon were studied in 12 patients with chronic active hepatitis B. Serum concentrations of γ–interferon were measured by radioimmunoassay in four patients after a subcutaneous injection of 10 million U (0.5 mg); the peak serum concentration of γ–interferon (29 ± 7 U/ml) was reached after 5 to 8 hr and γ–interferon remained detectable for 24 to 36 hr. Twelve patients received recombinant γ–interferon, 2.5 to 10 million U daily, for 4 mo. All suffered from a dose–dependent, flulike syndrome similar to that induced by α–interferon. Recombinant γ–interferon induced a marked increase of serum ALT and a significant decrease of serum hepatitis B virus–DNA. Serum hepatitis B virus–DNA disappeared in one patient during administration of recombinant γ–interferon. Serum hepatitis B virus–DNA disappeared in four additional patients, and HBeAg disappeared in two patients during the 12 mo after administration of recombinant γ–interferon. These results indicate that subcutaneous injection is suitable for administration of recombinant γ–interferon and that recombinant γ–interferon has an antiviral effect in patients with chronic active hepatitis B. (Hepatology 1990;12:155-158).
Familial Polyposis Coli and Hepatocellular Neoplasms
Authors: Bussey H. J.R.
Published: March 2007

Two cases of hepatocellular carcinoma in patients with familial polyposis coli (FPC) have recently been reported in the literature. Hepatoblastoma was also reported in five children with maternal ancestors with FPC. We report a case of hepatoblastoma in a female infant whose father has FPC as further evidence of an association between hepatoblastoma and FPC.
Detection of Antibodies Against the Polymerase Gene Product in Hepatitis B Virus Infection
Authors: Yuki Nobukazu , Hayashi Norio , Kasahara Akinori , Katayama Kazuhiro , Ueda Keiji , Fusamoto Hideyuki , Kamada Takenobu
Published: March 2007

We have studied antibodies (anti–pol antibody) against the polymerase gene product of hepatitis B virus by solid–phase enzyme immunoassay using synthetic peptides coded for by this gene. Sera from six patients with acute hepatitis B, 112 chronic hepatitis B virus carriers and six healthy individuals with naturally acquired immunity to hepatitis B virus were tested for anti–pol antibody. In acute hepatitis B virus infection, anti–pol antibody was detected in three of six patients. In chronic hepatitis B virus infection, antipol antibody was detected in 17 of 29 (59%), in 23 of 33 (70%) of cirrhotic patients and in 18 of 24 (75%) patients with cirrhosis complicated by hepatocellular carcinoma, compared with 4 of 19 (21%) asymptomatic carriers and 2 of 7 (29%) patients with chronic persistent hepatitis. Titers of anti–pol antibody were higher in cirrhotic patients with and without hepatocellular carcinoma than in patients with chronic active hepatitis. The presence of anti–pol antibody, however, had no relationship with hepatitis B virus–associated DNA polymerase activities and other viral replicative markers. As for sera from six healthy individuals with naturally acquired immunity to hepatitis B virus, two (33%) were positive for anti–pol antibody. These results indicate that the immune response toward the polymerase gene product is induced during acute and chronic hepatitis B virus infection. In chronic hepatitis B virus infection, anti–pol antibody may serve as a new marker indicative of a long period of hepatitis B virus–induced hepatitis. (Hepatology 1990;12:193-198).
Abnormal Hepatic Uptake of Low Doses of Sulfobromophthalein in Gilbert's Syndrome: the Role of Reduced Affinity of the Plasma Membrane Carrier of Organic Anions
Authors: Gentile Sandro , Persico Marcello , Tiribelli Claudio
Published: March 2007

The plasma disappearance rate of sulfobromophthalein (VBSP; μmol/kg/min) was measured in 15 Gilbert's syndrome patients and 12 control subjects after intravenous injection of two different doses (0.59 and 5.90 μmol/kg) of the dye. Plasma disappearance rate was significantly reduced in Gilbert's syndrome patients after administration of 0.59 μmol sulfobromophthalein/kg (0.119 ± 0.016 vs. 0.146 ± 0.018 μmol/kg/min; mean ± S.D.; p < 0.001), whereas no difference was found with the higher dose (0.754 ± 0.040 vs. 0.767 ± 0.072 μmol/kg/min). Significant reduction was also found after administration to four Gilbert's syndrome patients and four control subjects of 0.29 and 2.95 μmol sulfobromophthalein (0.060 ± 0.005 μmol/kg/min vs. 0.077 ± 0.07 μmol/kg/min and 0.480 ± 0.012 μmol/kg/min vs. 0.591 ± 0.015 μmol/kg/min, respectively; p < 0.01). Competition studies with combined administration of sulfobromophthalein (0.59 μmol/kg) and different doses of rifamycin SV (0.59, 1.47 and 2.95 μmol/kg) showed a significant (p < 0.001) reduction in plasma disappearance rate in Gilbert's syndrome patients but not in controls. The rifamycin SV dose at which a 50% inhibition in plasma disappearance rate was observed was 0.8 μmol/kg. The apparent affinity (Km) of the hepatic transport was higher in Gilbert's syndrome patients than in control subjects (3.61 ± 0.37 μmol sulfobromophthalein/kg vs. 2.76 ± 0.29 μmol sulfobromophthalein/kg, mean ± S.D.; p < 0.01), whereas no difference was found in Vmax (0.95 ± 0.11 μmol sulfobromophthalein/kg vs. 0.93 ± 0.10 μmol sulfobromophthalein/kg/min, mean ± S.D.; N.S.). We conclude that a defective sulfobromophthalein hepatic transport is present in Gilbert's syndrome, and it may be revealed by lowering the does of the dye. The data sugest that the defect in Gilbert's syndrome is an impaired affinity for sulfobromophthalein of one or more of the putative transport proteins in the basolateral plasma membrane of the hepatocyte. (Hepatology 1990;12:213-217).
Class I and Class Ii Major Histocompatibility Complex Antigen Expression on Hepatocytes: A Study in Children With Liver Disease
Authors: Lobo-Yeo Ava , Senaldi Giorgio , Portmann Bernard , Mowat Alex P. , Mieli-Vergani Giorgina , Vergani Diego
Published: March 2007

Controversy exists regarding major histocompatibility complex antigen expression on hepatocytes. In this study, hepatocyte expression of class I and II major histocompatibility complex antigens was investigated in diseased and normal livers, using indirect immunofluorescent staining of mechanically isolated, viable hepatocytes. Hepatocytes were obtained from 76 children: 10 with autoimmune chronic active hepatitis, nine with primary sclerosing cholangitis, nine with chronic hepatitis B virus infection, five after liver transplantation, 19 with extrahepatic biliary atresia, 11 with α1–antitrypsin deficiency, four with idiopathic neonatal hepatitis and nine with histologically normal liver. Immunohistochemistry was performed in all cases; flow cytofluorimetry was performed for class I antigens in 38 cases and performed for class II antigens in 18 cases. From three children with autoimmune chronic active hepatitis and two with chronic hepatitis B virus infection, isolated hepatocytes were also incubated with γ–interferon before staining and analysis. By fluorescence microscopy, class I antigens were detected on hepatocytes from all children, the highest percentage (100%) of positive cells and the most intense staining were observed in untreated patients with autoimmune chronic active hepatitis or primary sclerosing cholangitis and in those with acute rejection of a liver transplant. Reduced class I antigen expression occurred in chronic hepatitis B virus infection. Class II antigens were only detected on hepatocytes from eight patients: three with autoimmune chronic active hepatitis and five with primary sclerosingcholangitis, all untreated. Flow cytofluorimetric analysis confirmed the results obtained by fluorescence microscopy, but it also demonstrated a weak class II antigen expression during liver allograft rejection. After culture with γ–interferon, the hepatocytes from all five children showed an increase in staining intensity for class I and the acquisition of staining for class II antigens. This study shows that, whereas class I antigens are invariably expressed on liver cells, class II antigens are only found on hepatocytes from untreated patients with immune–mediated liver disorders in whom class I antigen display is also enhanced. (Hepatology 1990;12:224-232).
Isolated Perfused Cirrhotic Human Liver Obtained From Liver Transplant Patients: A Feasibility Study
Authors: Villeneuve Jean-Pierre , Huet P.-Michel , Gariepy Louise , Fenyves Daphna , Willems Bernard , Côté Jean , Lapointe Réal , Marleau Denis
Published: March 2007

Cirrhotic livers obtained from eight patients who underwent orthotopic liver transplantation were perfused through the portal vein and hepatic artery in a closed recycling system for periods ranging from 2 to 7 hr. An average perfusion flow of 451 ml/min was used, with about 80% coming from the portal vein and 20% from the hepatic artery. The livers appeared to remain viable as assessed by gross appearance, stable portal vein and hepatic artery pressures, oxygen consumption and bile prodúction. The extraction ratio of indocyanine green by the perfused livers averaged 0.098 (range = 0.023 to 0.168); that of propranolol averaged 0.445 (range = 0.126 to 0.813). Using the multipleindicator dilution–curve method, shunts >15 μm in diameter were demonstrated between the portal and hepatic veins in six of eight cases, whereas shunts from the hepatic artery to the hepatic veins were absent. Perfusion of human livers obtained during hepatic transplantation is a fairly simple procedure that will allow researchers to gain new insights into cirrhosis in humans. (Hepatology 1990;12:257-263).
Changes in Brain Metabolism During Hyperammonemia and Acute Liver Failure: Results of A Comparative 1H–Nmr Spectroscopy and Biochemical Investigation
Authors: Bosman Diederik K. , Deutz Nicolaas E.P. , De Graaf Albert A. , Vd Hulst Rene W.N. , Van Eijk Hans M.H. , Bovée Wim M.M.J. , Maas Martinus A.W. , Jörning George G.A. , Chamuleau Robert A.F.M.
Published: March 2007

The effects of hyperammonemia on brain function have been studied in three different experimental models in the rat: acute liver ischemia, urease–treated animals and methionine sulfoximine-treated animals. To quantify the development of encephalopathy, clinical grading and electroencephalographic spectral analysis were used as indicators.

In all three experimental models brain ammonia concentrations increased remarkably associated with comparable increases in severity of encephalopathy. Furthermore, in vivo
1H–nuclear magnetic resonance spectroscopy of a localized cerebral cortex region showed a decrease in glutamate concentration in each of the aforementioned experimental models. This decreased cerebral cortex glutamate concentration was confirmed by biochemical analysis of cerebral cortex tissue post mortem. Furthermore, an increase in cerebral cortex glutamine and lactate concentration was observed in urease–treated rats and acute liver ischemia rats. As expected, no increase in cerebral cortex glutamine was observed in methionine sulfoximine-treated rats.

These data support the hypothesis that ammonia is of key importance in the pathogenesis of acute hepatic encephalopathy. Decreased availability of cerebral cortex glutamate for neurotransmission might be a contributing factor to the pathogenesis of hyperammonemic encephalopathy. A surprising new finding revealed by 1H–nuclear magnetic resonance spectroscopy was a decrease of cerebral cortex phosphocholine compounds in all three experimental models. The significance of this finding, however, remains speculative. (Hepatology 1990;12:281-290).
Analysis of Hepatic T Lymphocyte and Immunoglobulin Deposits in Patients With Primary Biliary Cirrhosis
Authors: Krams Sheri M. , van de Water Judy , Coppel Ross L. , Esquivel Carlos , Roberts John , Ansari Aftab , Gershwin Eric M.
Published: March 2007

The histological findings in patients with primary biiary cirrhosis have been well–defined and are often used in the clinical staging of disease. However, it has only been with the development of reagents that phenotypically characterize the lymphoid infiltrate that attempts have been made to correlate pathophysiology with immune effector populations. Indeed, the inflammatory hepatic lesions in primary biliary cirrhosis have been described as containing CD4–positive and CD8–positive T cells. Less clear, however, have been the T cell receptors in these lesions. Further, the data on immunoglobulin deposits in hepatic lesions have been less well–defined; this deficit may be a result of the quality of polyspecific sera and difficulties in background. To address these issues, we have used a battery of well–defined monospecific and polyspecific reagents to phenotypically define the occurrence of lymphoid cells in the livers of patients undergoing transplantation. Furthermore, we have defined these same markers on T cell lines derived from liver, regional lymph node and peripheral blood. The predominant cell type in the mononuclear infiltrate is the CD3+, CD4+ T lymphocyte bearing the T cell receptor αβ. T cell lines from the same patients demonstrate similar findings. Of special importance, however, was the detection of CD20+ B cells and Ig+ cells in the lymphoid infiltrate. Indeed, we also readily demonstrated the presence of immunoglobulin on the surface of biliary epithelium. These data suggest that mechanisms involved in the pathophysiology of primary biliary cirrhosis may include both T cell and antibody mechanisms. The results also underscore the need to develop a functional, and not just a phenotypical, assay of the inflammatory infiltrate. (Hepatology 1990;12:306-313).
Comprehensive Study of the Biliary Bile Acid Composition of Patients With Cystic Fibrosis and Associated Liver Disease Before and After Udca Administration
Authors: Nakagawa Michiko , Colombo Carla , Setchell Kenneth D.R.
Published: March 2007

The biliary bile acid composition was determined for patients with cystic fibrosis and associated liver disease before and after the administration of ursodeoxycholic acid (10 to 15 mg/kg body wt/day). Bile acids were analyzed by fast atom bombardment ionizationmass spectrometry, high performance liquid chromatography and gas chromatography–mass spectrometry after individual bile acids were separated according to their mode of conjugation using the lipophilic anion exchanger, diethylaminohydroxypropyl Sephadex LH–20. More than 50 individual bile acids were identified in the bile of cystic fibrosis patients and these acids were predominantly secreted as glycine and taurine conjugates. Small proportions (<8% of the total) of unconjugated and sulfate conjugates were present. Of interest was the identification of two side–chain-elongated (C25) bile acids, homocholic and homochenodeoxycholic acids. After ursodeoxycholic acid was administered, duodenal bile became enriched with the conjugated species of ursodeoxycholic acid (accounting for 11.9% to 32.5% of the total biliary bile acids), but to a lesser extent than reported previously for patients with other liver diseases or gallstones who received comparable doses of ursodeoxycholic acid, and this presumably occurs because of bile acid malabsorption that is a feature of cystic fibrosis. The mean glycine/taurine ratio increased from 2.4 before ursodeoxycholic acid administration to 5 after ursodeoxycholic acid administration even though these patients also received taurine. Despite the relatively low enrichment of the bile by ursodeoxycholic acid, biochemical indices of liver function all improved in these patients after ursodeoxycholic acid administration. (Hepatology 1990;12:322-334).
Demonstration of “Naturally Occurring Mitochondrial Antibodies” in Family Members of Patients With Primary Biliary Cirrhosis
Authors: Klein Reinhild , Berg Peter A.
Published: March 2007

Sera from 81 healthy family members (including husbands) of 13 patients with primary biliary cirrhosis were tested by Western blotting against the antigen fractions M9 and M2 derived respectively from rat liver and beef heart mitochondria. Fifty–eight (70%) were positive and recognized four major (molecular weights 98 kD, 65 kD, 61 kD, 58 kD) and eight minor determinants (85 kD, 81 kD, 78 kD, 54 kD, 48 kD, 45 kD, 40 kD, 30 kD) labeled α–my. Each of the 58 sera recognized at least one of the four major polypeptides. In contrast, only 6% of 80 primary biliary cirrhosis patients had antibodies against one of these epitopes. Sera from 25 patients with different infectious disorders previously shown to react with submitochondrial particles by enzyme–linked immunosorbent assay also recognized at least one of the four major determinants.

From these findings it was concluded that these antibodies may belong to the family of natural autoantibodies. Since they reacted with submitochondrial fractions, they were defined as “naturally occurring mitochondrial antibodies.”

The high incidence of “naturally occurring mitochondrial antibodies” in primary biliary cirrhosis contact persons may be taken as indirect evidence for a contagious immunogenic agent circulating in the blood of primary biliary cirrhosis-patients. In contrast, the absence of “naturally occurring mitochondrial antibodies” in primary biliary cirrhosis-patients themselves implies that an underlying B–cell defect is responsible for this lack of antibody production. Considering the protective role of naturally occurring antibodies in general, this postulated B–cell defect could be a major factor in the etiopathogenesis of primary biliary cirrhosis. (Hepatology 1990; 12:335-341).