bluuebunny/crossref_metadata_2025 huggingface dataset:
The N-Terminal Propeptide of Collagen Type III in Serum Reflects Activity and Degree of Fibrosis in Patients with Chronic Liver Disease†
Authors: Frei Alain , Zimmermann Arthur , Weigand Kurt
Published: August 2008

To evaluate the diagnostic significance of the collagen Type III (Col 1-3) JV-terminal propeptide of procollagen Type III, with respect to activity and degree of liver fibrosis, Col 1-3 serum concentrations were measured in 111 patients with chronic liver diseases and in 60 patients were correlated with liver histology and morphometry. Col 1-3 was measured by a specific radioimmunoassay. Biopsies were read without knowledge of diagnosis. Periportal and intralobular lesions were assessed semiquantitatively by allocating 1 of 4 severity grades to each parameter. All portal areas were measured morphometrically. Compared to 27 normal controls, Col 1-3 concentrations were significantly elevated in patients with untreated chronic active hepatitis, cirrhosis and primary biliary cirrhosis, but not in chronic persistent hepatitis or fatty liver. Morphometrically measured portal tract area significantly correlated with Col 1-3 plasma levels. Among the semiquantitatively measured periportal lesions, the number of fibroblasts exhibited the closest relationship with Col 1-3 levels; there was no relationship between Col 1-3 levels and intralobular lesions. These data suggest that Col 1-3 serum levels reliably reflect the activity and degree of liver fibrosis and are useful along with liver biopsy in follow-up of patients with chronic liver disease.
Long-Term Follow-Up of Serum N-Terminal Propeptide of Collagen Type III Levels in Patients with Chronic Liver Disease†
Authors: Weigand Kurt , Zaugg Pierre-Yves , Frei Alain , Zimmermann Arthur
Published: August 2008

To evaluate the diagnostic and prognostic significance of the N-terminal propeptide of collagen Type III (Col 1-3) in chronic liver disease, the peptide level was measured in the serum of 4 patients with primary biliary cirrhosis, 5 with chronic persistent hepatitis, 12 with chronic active hepatitis, and 1 with autoimmune hepatitis, for a period of 2 to 10 years and compared with liver function and histology. In primary biliary cirrhosis, Col 1-3 peptide levels were always elevated, regardless of medical therapy; however, after liver transplantation in one patient, the Col 1-3 peptide level decreased. In chronic persistent hepatitis, the peptide level fluctuated around the upper limit of normal. Among patients with chronic active hepatitis, the Col 1-3 peptide level normalized in 2 patients during remission, but was elevated in 7 patients who developed cirrhosis. Only in a patient with autoimmune hepatitis was the Col 1-3 peptide level normal, although the patient developed cirrhosis during prednisone therapy. When prednisone was withdrawn, the Col 1-3 peptide level increased. The data suggest that the serum Col 1-3 peptide may estimate the course of liver fibrosis in chronic liver disease and has prognostic value, particularly in chronic active hepatitis. Persistent elevation suggests ongoing fibrosis and development of cirrhosis; normalization suggests remission.
Acetaldehyde Stimulates Collagen and Noncollagen Protein Production by Human Fibroblasts†
Authors: Holt Kenneth , Bennett Michael , Chojkier Mario
Published: August 2008

The mechanisms responsible for the increased hepatic collagen deposition in alcoholic cirrhosis remain unknown. The question of whether ethanol or acetaldehyde has a direct effect on collagen and noncollagen protein production was investigated in human fibroblasts with no detectable activity of alcohol dehydrogenase to distinguish the effects of these metabolites. To eliminate environmental factors, protein production by confluent human skin, fetal and hepatic fibroblasts was studied after three passages. Cells were labeled with [5−3H]proline for 4 hr in the presence of 0.2 mM ascorbate alone or with addition of either ethanol (50 mM) or acetaldehyde (0 to 300 μM). Rates of protein production were calculated from the radioactivities of collagenase-sensitive and collagenase-resistant proteins. Skin fibroblasts from alcoholic individual either with cirrhosis or without liver disease have comparable rates of collagen and noncollagen protein production. Acetaldehyde, in a concentration found in the liver during ethanol abuse, significantly increased collagen production by human skin fibroblasts (up to 140%), fetal fibroblasts (up to 240%) and hepatic fibroblasts (up to 70%) but the addition of ethanol had no significant effect on basal collagen production. The effect of acetaldehyde was dose-related and affected noncollagen protein production in a similar manner. Acetaldehyde did not cause changes in either proline transport or the specific activity of the proline precursor pool. This newly recognized stimulation of collagen production by acetaldehyde may be a possible mechanism of fibrogenesis in alcoholic individuals.
Isolation, Culture and Characterization of Adult Human Hepatocytes from Surgical Liver Biopsies
Authors: Ballet Francois , Bouma Marie-Elisabeth , Wang Shu-Ren , Amit Norma , Marais Jacqueline , Infante Recaredo
Published: August 2008

A technique is described for isolation and culture of adult human hepatocytes from surgical liver biopsies. The mean cell yield was 1.75 ± 107 cells per gm liver and viability averaged 80%. Hepatocytes were maintained in primary culture for about 10 days. Cell morphology and histochemical characteristics were similar to hepatocytes in vivo. Bile canaliculi were observed by electron microscopy. Intracellular albumin was demonstrated up to the 7th day of culture; albumin secretion rate was maximal (0.6 ± 0.33 /tg per hr per 106 cells) 5 days after plating. These studies demonstrate that adult human hepatocytes can be isolated from surgical biopsies with high yield, and differentiated function can be maintained for several days.
Reduction of Hepatic Tetrahydrofolate and Inhibition of Exhalation of 14CO2 Formed from [Dimethylamino-14C]Aminopyrine in Nitrous Oxide-Treated Rats
Authors: Black Kurt A. , Virayotha Viravan , Tephly Thomas R.
Published: August 2008

The exhalation of14CO2after the administration of [dimethylamino-14C]aminopyrine to an organism is assumed to reflect the demethylation of aminopyrine by hepatic mixed-function oxidase activity. The formaldehyde formed as a result of the demethylation of aminopyrine is then sequentially oxidized to formic acid and CO2. The last step in the pathway, i.e., formate oxidation, is dependent upon tetrahydrofolate; thus, factors which alter hepatic tetrahydrofolate potentially may modify14C-aminopyrine metabolism to14CO2in vivo.Exposure of rats to nitrous oxide (N2O) produces a significant reduction in hepatic tetrahydrofolate as a result of the inhibition of 5-methyltetrahydrofolate:homocysteine methyltransferase activity (E.C. 2.1.1.13). In the present study, exposure of rats to N2O/O2(1:1) for 4 hr prior to the administration of14C-aminopyrine (40 or 400 Mmoles per kg) produced a 60% reduction in the peak rate of14CO2exhalation and a 45% decrease in the total14CO2exhaled within 2 hr. In control experiments, exposure of rats to nitrogen/O2(1:1) produced no effect on14C-aminopyrine metabolism to14CO2. Administration of methionine (1.3 mmoles per kg) 30 min prior to14C-aminopyrine administration reversed the inhibition of14CO2exhalation and reduction in hepatic tetrahydrofolate observed in N2O-exposed animals. Aminopyrine (400 pmoles per kg) administration to air-breathing rats did not affect the level of urinary formate, but exposure to N2O produced a 40-fold increase. Aminopyrine administration to N2O-exposed rats produced a 75% increase in urinary formate as compared to rats treated with N2O alone. The first-order rate constant for aminopyrine elimination from the plasma, apparent volume of distribution, apparent plasma clearance and the level of hepatic cytochrome P-450 were not affected by N2O exposure. N2O exposure inhibited14CO2exhalation after [N-methyl-14C]morphine administration to a similar degree as that observed after aminopyrine administration. These results indicate that factors which alter the level of hepatic tetrahydrofolate, e.g., nutritional status, N20 exposure or certain drugs, may modify the metabolism of14C-amino-pyrine to14CO2in vivowithout affecting the demethylation of aminopyrine. Thus, such factors must be controlled when the exhalation of14CO2after14C-aminopyrine administration to an organism is used as anin vivomeasure of hepatic drug metabolism.
Serum α-L-Fucosidase: A New Marker for the Diagnosis of Primary Hepatic Carcinoma?
Authors: Deugnier Yves , David Véronique , Brissot Pierre , Mabo Philippe , Delamaire Damien , Messner Michel , Bourel Michel , Legall Jean-Yves
Published: August 2008

α-L-fucosidase, a lysosomal enzyme which catabolizes fucoproteins, was studied in sera from 30 controls, 32 patients with primary hepatic carcinomas, 24 patients with secondary metastatic liver carcinomas and 36 patients with cirrhosis.

Serum α-L-fucosidase was increased in primary hepatic carcinomas (145.5 ± 12 nkat per liter) with a high statistical significance versus controls (51.4 ± 4.5 - p & 10−7), secondary metastatic liver carcinomas (58.9 ± 6.4 - p < 10−5) and cirrhotics (71.3 ± 6 - p < 10−5). A level exceeding 110 was a useful marker for the diagnosis of primary hepatic carcinoma with 75% sensitivity and 90% specificity.
Serum Activity of Mitochondrial Aspartate Aminotransferase: A Sensitive Marker of Alcoholism With or Without Alcoholic Hepatitis
Authors: Nalpas Bertrand , Vassault Anne , Guillou Alain Le , Lesgourgues Bruno , Ferry Nicolas , Lacour Bernard , Berthelot Pierre
Published: August 2008

Serum activity of the mitochondrial isoenzyme of aspartate aminotransferase (mAST) was measured with an immunological method in 74 subjects. Fourty-six were chronic alcoholics with (30) or without (16) obvious alcoholic liver disease; 28 were nonalcoholic controls among whom 14 had acute or chronic viral hepatitis, the remaining 14 being healthy individuals. Mean mAST activity was much higher in all the alcoholic subjects, with or without liver disease, 10.4 and 1.95 units per liter, respectively, than in the healthy controls (0.43, p < 0.001). The mean mAST to total AST ratio was similar in the healthy controls and in the patients with viral hepatitis (2.98 and 3.19%, NS), whereas it was about 4 times higher in the alcoholics with a sensitivity which reached 93% in the patients with alcoholic liver disease and 100% in those without. Both -γ-glutamyl transpeptidase and glutamate dehydrogenase serum activities were far less sensitive and specific. As almost all chronic alcoholics had similar abnormal values of mAST/total AST ratio, this leads to question whether “normal” liver may really exist in any of such subjects.
Regulation of Low Density Lipoprotein Receptor Function in a Human Hepatoma Cell Line
Authors: Leichtner Alan M. , Krieger Monty , Schwartz Alan L.
Published: August 2008

Low density lipoprotein (LDL) processing was investigated in a human hepatoma-derived cell line, Hep G2. Hep G2 cells bound, internalized and degraded LDL via a saturable, high affinity (Kd — 2 ± 10−8
M) pathway similar to that present in other mammalian cells. Although 80% of the uptake and degradation of 125I-LDL was inhibited by 40-fold excess native LDL, the same concentration of methylated LDL, which cannot bind to LDL receptors, had virtually no effect on processing. When added at low concentrations, the lysosomotropic agent, chloroquine, inhibited degradation (I50 — 15 μM) without affecting the rate of lipoprotein internalization. Receptor activity was decreased 60% by preincubation of the cells in medium containing a source of cholesterol (LDL or unesterified cholesterol) and increased 1.7-fold by preincubation with compactin, a competitive inhibitor of 3-hydroxy-3-methylglutaryl coenzyme A reductase. The Hep G2 cell line may prove a useful system both for the further study of hepatic lipoprotein metabolism and for the evaluation of new antihypercholesterolemic agents.
Urea Synthesis After Protein Feeding Reflects Hepatic Mass in Rats† ‡
Authors: Brewer Thomas G. , Berry William R. , Harmon John W. , Walker Strothers H. , Dunn Michael A.
Published: August 2008

Urea synthesis is an exclusive biosynthetic function of the liver. Since the exact relationship between urea synthesis in vivo and functional liver mass remains unclear, we established an animal model using oral protein loading and measurement of resultant urea synthesis in rats. We studied rats subjected to sham operation, 40% hepatectomy, 66% hepatectomy, portacaval shunt and CC14-induced cirrhosis. Urea synthesis was calculated as the sum of urinary urea excretion and accumulation of urea in body water during the 6-hr period after oral administration of a casein protein load equivalent to 20 gm per kg body weight.

Peak urea synthesis rate in the sham-operated group of rats was 142 ± 11 μmoles per hr per gm wet liver weight (mean ± 1 S.D.), 473 ± 34 μmoles per hr per gm liver protein and 80 ± 5 μmoles per hr per mg liver DNA. This rate closely matched those of the other groups for each type of liver mass measurement. Marked reduction (p < 0.01) of urea synthesis on a DNA basis was noted only in the CCl4-cirrhotic livers, related to the significantly higher (p < 0.01) DNA content of the cirrhotic livers. Similarly, increased (p < 0.05) liver protein content of the sham-operated rats when compared with the other groups was reflected in slightly lower urea synthesis rates expressed on the basis of liver protein (p < 0.05) when compared to that of all other groups. The slopes of the trend lines which define the relationship of urea synthesis rates in each group to total liver mass (liver weight, protein content and DNA content) were not significantly different.

We conclude that urea synthesis rates after oral administration of a protein load reflect liver mass in terms of wet weight, liver protein content and liver DNA content. This relationship remains valid in the presence of changes in liver parenchymal mass, portacaval shunting and CC14-cirrhosis.
UDP-Glucuronyltransferase-Catalyzed Deconjugation of Bilirubin Monoglucuronide
Authors: Cuypers Theo H.M. , Haar El Terlen M. , Jansen Peter L. M.
Published: August 2008

Bilirubin monoglucuronide is rapidly deconjugated when incubated with UDP and rat liver microsomal preparations at pH 5.1. The following evidence was found that this reaction is catalyzed by UDP-glucuronyltransferase: (i) unconjugated bilirubin and UDP-glucuronic acid were identified as the reaction products; (ii) Gunn rat microsomal preparations lack bilirubin UDP-glucuronyltransferase deficiency and do not catalyze the deconjugation reaction, and (iii) neither saccharo-1,4-lactone, a β-glucuronidase inhibitor, nor butylated hydroxy toluene, an inhibitor of spontaneous isomerisation, affect the rate of the deconjugation reaction. Deconjugation appears to be the reverse of UDP-glucuronyltransferase-catalyzed glucuronidation.

The conditions for the reverse reaction differ in the following aspects from those of the forward reaction: (i) nucleotide triphosphates stimulate the reverse reaction probably allosterically; (ii) UDP-N-acetylglucosamine stimulates the forward reaction but has no effect on the reverse reaction; (iii) the optimal pH for the reverse reaction is pH 5.1 and for the forward reaction is pH 7.8, and (iv) Mg++ ion is not required for the reverse reaction but stimulates the forward reaction. Detergents stimulate both reactions. Stimulation of the reverse reaction by nucleotide triphosphates and detergents is mutually independent and additive which suggests different mechanisms of action.

Deconjugation reactions may become important during parenchymatous liver disease when, as a result of anaerobic glycolysis, intracellular pH decreases. Elevated levels of unconjugated bilirubin in the serum of patients with parenchymatous liver disease may be a sign of sick liver cells rather than decreased UDP-glucuronyltransferase activity.
The Comparative Effect of Administration of Substances Via the Hepatic Artery or Portal Vein on Hepatic Arterial Resistance, Liver Blood Volume and Hepatic Extraction in Cats
Authors: Lautt Wayne W. , Legare Dallas J. , Daniels Timothy R.
Published: August 2008

Compounds reaching the liver do so via either the hepatic artery or the portal vein. This paper reports on the effectiveness of administration of compounds into these alternate routes for their effects on the hepatic parenchymal cells, the hepatic arterial resistance vessels (blood flow) and hepatic capacitance (blood volume responses). All tests were done on cats under pentobarbital anesthesia. Perfusion of the parenchymal cell mass was assessed by comparing the hepatic elimination of indocyanine green (ICG) administered via the two vascular routes. The ICG uptake was assessed by measuring relative areas under the hepatic venous outflow curve obtained following bolus injections of ICG into the artery and portal vein. In a separate series, using different methods, the hepatic venous levels reached early (2 min) and later (5 min) during a constant infusion were compared during administration via the two routes and found to be equal. Parenchymal cell functions (ICG extraction, bile salt stimulation of bile flow) indicate that blood from the artery and portal vein supplies the hepatic parenchymal cells equally well. This suggests a well-mixed blood supply prior to exposure of either blood stream to parenchymal cells. Substances being processed by the liver are thus equally well handled if reaching the liver via either the arterial or portal blood stream. This has significance in validating the use of some isolated liver perfusion methods that perfuse only via the portal vein.

Access of vasoactive compounds in the two blood streams to hepatic arterial resistance vessels was assessed using electromagnetic flow probes. The infusion of adenosine and angiotensin produced dilation and constriction respectively of the hepatic artery with the effect being about 3 times as large at a given dose when infused via the artery. The data show that vasoactive substances in the portal blood can affect the hepatic arterial blood flow by “transvascular” route and that portal blood has quite a high access to hepatic arterial resistance vessels. It does indicate, however, that the arterial resistance sites are not being perfused with mixed portal and arterial blood.

The hepatic blood volume changes in responses to intraportal and intraarterial infusions of angiotensin and noradrenaline were measured using an in vivo plethysmograph assembled around the intact liver. Constriction of hepatic blood volume was equal with administration via the two routes indicating that the capacitance vessels are perfused by well-mixed blood and are, therefore, likely to consist mostly of sinusoidal and postsinusoidal vessels with presinusoidal roles being minor or absent.
Pneumococcal Vaccination: The Response of Patients with Alcoholic Liver Cirrhosis
Authors: Pirovino Mauro , Lydick Eva , Grob Peter J. , Arrenbrecht Stephan , Altorfer Josef , Schmid Martin
Published: August 2008

Patients with cirrhosis have an increased risk of dying from pneumococcal pneumonia. Their immunological response to pneumococcal vaccination is not known. We compared the antibody response to challenge with a 14-valent purified pneumococcal polysaccharide vaccine in 15 patients with biopsy-proven alcoholic liver cirrhosis, 10 healthy volunteers and 10 patients with chronic obstructive pulmonary disease. Preimmunization levels for Danish type 1 pneumococcal polysaccharide were lower among cirrhotics than patients with chronic obstructive pulmonary disease. Four and 12 weeks after immunization, no significant differences were found in antibody concentrations between the three groups. We conclude that patients with alcoholic cirrhosis can produce an adequate antibody response to pneumococcal polysaccharide antigen.
Spinal Cord Paralysis Following Sclerotherapy for Esophageal Varices
Authors: Seidman Ernest , Weber Andrée M. , Morin Claude L. , Ethier Roméo , Lamarche Jacques B. , Guerguérian Ara J. , Geoffroy Guy , Roy Claude C.
Published: August 2008

A child with cryptogenic cirrhosis underwent a third session of elective sclerotherapy. Endoscopic therapy consisted of intravascular injection of ethanolamine oleate in varices newly developed at the midesophagus level. Irreversible paraplegia was documented within 8 hr postoperatively. Two years later she eventually died from gastrointestinal bleeding. Autopsy findings were compatible with an infarct of the spinal cord secondary to an occlusion of the anterior spinal artery. Various hypotheses which might explain the passage of the sclerosing material from the esophagus to the anterior spinal artery include: arterial occlusion secondary to venous thrombosis and spinal cord necrosis, accidental injection in an intercostal artery or azygous vein through the esophageal wall, the presence of a congenital arteriovenous fistula or the opening of arteriovenous shunt. Paravasal injection of dilute sclerosing agent might protect against this unusual but dramatic complication.
Pit Cell-Hepatocyte Contact in Autoimmune Hepatitis
Authors: Kaneda Kenji , Kurioka Narito , Seki Shuichi , Wake Kenjiro , Yamamoto Sukeo
Published: August 2008

This is the first report to document pit cells in the human liver. These cells were identified by characteristic electron-dense granules and rod-cored vesicles. The granules of human pit cells were smaller in number and size than those of the rat. In the present case of autoimmune hepatitis, pit cells and conventional agranular lymphocytes migrated into hepatic parenchyma and contacted degenerating and immature hepatocytes. Pit cells show natural killer activity (Kaneda et al., 1983), and may participate in development of lesions and cellular damage.
HLA-DR Antigens in HBsAg-Positive Chronic Active Liver Disease with and without Associated Delta Infection
Authors: Forzani Barbara , Actis Giovanni C. , Verme Giorgio , Amoroso Antonio , Borelli Iolanda , Curtoni Emilio S. , Rumi Maria Grazia , Picciotto Antonio , Marinucci Giovanni , Freni Maria Antonietta , Rizzetto Mario
Published: August 2008

The A, B, C and DR locus specificities of the human leukocyte antigens system (HLA) were determined in 45 delta-positive and 44 delta-negative Italian patients, all with HBsAg-positive chronic active liver disease; controls were 526 healthy Italian blood donors matched for age, sex and geographical origin. HLA-A, B, C gene frequencies were not significantly changed. In delta-positive patients, the frequencies of the DR locus specificities were: DR2, 37.8%; DR3, 20%; DR4, 11.1%. In the delta-negative patients, the frequencies were: DR2, 13.6%; DR3, 36.4%; DR4, 0%. Control frequencies were: DR2, 19.4%; DR3, 17.1%; DR4, 18.5%. The corrected p values of the differences between controls and delta-positive patients were: DR2, pc = 0.046; DR3, pc = NS (not significant); DR4, pc = NS. The corrected p values of the differences between controls and delta-negative patients were: DR2, pc = NS; DR3, pc = 0.03; DR4, pc = 0.002. These findings show that: (a) DR3, a genetic marker of autoimmunity, might assist the establishment of chronic HBsAg liver disease in the absence of delta superinfection; (b) DR2 is linked with failure to clear the delta agent, and (c) DR4 may protect from virus B persistence. Identification of adventitious factors such as delta may help uncover a subgroup of HBsAg carriers who are genetically predisposed to develop chronic liver disease.
Hepatitis B Markers in United States Drug Addicts with Special Emphasis on the Delta Hepatitis Virus
Authors: Ponzetto Antonio , Seeff Leonard B. , Buskell-Bales Zelma , Ishak Kamal G. , Hoofnagle Jay H. , Zimmerman Hyman J. , Purcell Robert H. , Gerin John L.
Published: August 2008

Hepatitis B virus and hepatitis delta virus co-infection in drug addicts has been well described in Europe, the latter agent appearing to have been introduced there in the mid-1970’s. Currently, similar data are scanty among United States addicts. We therefore reevaluated 99 drug addicts from three different geographic locations in the United States who had participated in a Veterans Administration Cooperative Study between 1972 and 1975. Almost all were asymptomatic, and all had been subjected to liver biopsy because of prolonged aminotransferase abnormalities. Stored sera were tested for antibody to hepatitis delta antigen (anti-HD) by radioimmunoassay and available liver biopsies examined for hepatitis delta antigen (HDAg) by immunofluorescence.

Overall, 19.2% were HBsAg positive, 9.1% HBeAg positive, 90% anti-HBc positive and 10.1%, positive for anti-HD. Anti-HD was identified in 42.1% of addicts who were HBsAg positive and in 3.3% who were anti-HBs positive. No correlation was found between HBeAg and anti-HD, but anti-HD was present significantly more frequently in those with chronic active hepatitis than in those with chronic persistent hepatitis. We conclude that hepatitis delta virus infection is common in HBsAg-positive drug addicts in the United States dating back to at least 1972 and probably earlier.
HBsAg-Serum Protein Complexes Stimulate Immune T Lymphocytes More Efficiently Than Do Pure HBsAg
Authors: Celis Esteban , Chang Tse Wen
Published: August 2008

HBsAg from plasma of chronic hepatitis B carriers was purified by affinity chomatography using a mouse monoclonal antibody specific for HBsAg. Elution with buffer at two different pH values separated HBsAg into two fractions: one contained high amounts of immune complexes associated with HBsAg; the other contained larger quantities of the HBsAg polypeptides P24 and GP27 and only small amounts of immunoglobulin. When compared for effects on stimulating the proliferative response of freshly isolated lymphocytes and an HBsAg-specific T cell clone, the HBsAg fraction containing a high proportion of immunoglobulin was much more potent than HBsAg with low amounts of immunoglobulins or pure HBsAg, which was isolated from the culture supernatant of the human hepatoma cell line (PLC/PRF/5). The plasma-derived HBsAg with low amounts of complexed immunoglobulins became more immunogenic in the presence of an anti-HBsAg monoclonal IgG. The present results, combined with earlier findings, suggest that HBsAg associated with immune complexes is a more potent stimulator of T cells than purer HBsAg preparations due to an increase in the efficiency of monocytes to capture the antigen through binding to immune complexes for subsequent processing and presentation of the antigen. These observations could be of relevance for the preparation of effective hepatitis B vaccines from recombinant DNA and peptide synthesis technologies.
Liver Orcein Stain and Viral DNA in Duck Hepatitis B Virus Infection in Chinese Ducks and Experimentally Infected Japanese Ducklings
Authors: Mori Junko , Omata Masao , Yokosuka Osamu , Imazeki Fumio , Ito Yoshimi , Uchiumi Katsuo , Matsuyama Yasuhisa , Weifa Ye , Okuda Kunio
Published: August 2008

Liver sections were stained with orcein, and duck hepatitis B virus was identified in sera and livers by the hybridization technique in 106 ducks (44 Chinese ducks, 15 Japanese ducks and 47 Japanese ducklings). Orcein-positive hepatocytes were found in 18 of 38 (47%) duck hepatitis B virus DNA seropositive ducks, and only in 3 of 68 (4%) seronegative ducks. The three ducks were all from a heavily infected flock in southern China.

Serial analyses of viral DNA by Southern blot and spot hybridizations in experimentally infected Japanese ducklings revealed a dissociation or a time gap between the amount of viral DNA in serum and the emergence of orcein positive hepatocytes. Orcein-positive hepatocytes were generally associated with prolonged presence of viral infection for at least 4 to 6 months. These findings support the clinical hypothesis that the presence of orcein-positive hepatocytes indicates persistent rather than acute infection.

Since orcein-positive hepatocytes have been seen in infection with hepatitis B, woodchuck hepatitis, ground squirrel and duck hepatitis B viruses, accumulation of orcein-positive material in liver cells may be one of the common properties these viruses share. This stain may be utilized for screening new hepatitis B virus-like viruses.
Detection of Anti-HBc IgM Following Prednisone Treatment in Patients with Chronic Active Hepatitis B Virus Infection
Authors: Nowicki Marek J. , Tong Myron J. , Nair Prem V. , Stevenson Douglas
Published: August 2008

The effect of a short course of prednisone therapy on serum IgM type antibody to the hepatitis B core antigen (anti-HBc IgM) was studied in 14 male patients with chronic active type B hepatitis. Eleven patients (78.5%) became positive for serum anti-HBc IgM either during or shortly after prednisone withdrawal. Detection of anti-HBc IgM correlated with an increase in hepatitis B virus specific DNA-polymerase activity and was followed by a rise in serum transaminase levels. Six patients with histologic evidence of cirrhosis developed anti-HBc IgM which lasted six or more months after prednisone therapy and had a rapid onset of hepatic decompensation manifested by encephalopathy with ascites and/or variceal bleeding. In 17 untreated chronic active type B hepatitis patients who served as controls, anti-HBc IgM was detected at low levels in only a single serum sample from each of two patients during the same observation period.
Protective Effect of 4-(3,7,11,15-Tetramethyl-6,10,14-Hexadecatrienoyl)Morpholine on Liver Injury Induced by Hepatotoxins in Rats
Authors: Fujiwara Kenji , Ohta Yasuhiko , Ogata Itsuro , Sato Yuzuru , Oka Yuji , Hayashi Shigeki , Takatsuki Katsuyoshi , Oka Hiroshi
Published: August 2008

The effect of 4-(3,7,ll,15-tetramethyl-6,10,14-hexadecatrienoyl)morpholine (E-0712), a new synthetic compound, on liver injury induced by two hepatotoxins in rats was studied. Oral doses of E-0712 four times at 0, 6, 12 and 18 hr significantly attenuated elevated SGPT values and prolonged prothrombin time (PT) at 24, 36, 48, 60 and 72 hr in rats with a single s.c. dose of D-galactosamine in which SGPT values and PT reached the peak within 48 hr. In cases in which a dose of carbon tetrachloride (i.p.) produced peak SGPT values and PT within 36 hr attenuation occurred likewise at 24, 36 and 60 hr. In the control and E-0712-treated rats dosed with D-galactosamine or carbon tetrachloride, SGPT values were positively correlated with PT at 24 hr. In addition, morphological degrees of acidophilic body formation or centrizonal necrosis were significantly reduced by E-0712. These results suggest that E-0712 protects against liver injury, particularly hepatocellular necrosis, induced by D-galactosamine and carbon tetrachloride in rats.
The Hepatotoxicity of Valproic Acid and Its Metabolites in Rats. I. Toxicologic, Biochemical and Histopathologic Studies
Authors: Kesterson James W. , Granneman Richard G. , Machinist Joseph M.
Published: August 2008

Valproic acid (VPA), its unsaturated metabolites and pent-4-enoate (4-PA) were studied for potential hepatotoxicity in rats. 4-PA, 4-en-VPA and 2,4-dien-VPA were potent inducers of microvesicular steatosis in young rats. Microvesicular steatosis induced by the 4-en-VPA was accompanied by ultrastructural changes characterized by myeloid bodies, lipid vacuoles and mitochondrial abnormalities. Myeloid bodies and lipid vacuoles were seen to a lesser extent in 2,4-dien-VPA and 4-PA-treated rats. VPA failed to induce discernible liver lesions in young rats even at near lethal doses of 700 mg per kg per day. The drug did, however, induce hepatic lipid accumulation in mature rats and in young rats dosed concomitantly with phenobarbital. β-oxidation inhibition and several other biochemical alterations were observed in rats dosed with VPA, its unsaturated metabolites and 4-PA. It was suggested that β-oxidation inhibition observed in both VPA and en-metabolite-treated rats occurred by different mechanisms. VPA inhibits by a transient sequestering of CoA while the CoA esters of some en-V,PA-metabolites, particularly 4-en-VPA, inhibit specific enzyme(s) in the β-oxidation sequences.
The Hepatotoxicity of Valproic Acid and Its Metabolites in Rats. II. Intermediary and Valproic Acid Metabolism
Authors: Granneman Richard G. , Wang Shyh-Ing , Kesterson James W. , Machinist Joseph M.
Published: August 2008

The role of metabolites in valproic acid (VPA)-associated hepatotoxicity was studied in rats. The most steatogenic mono-unsaturated metabolite, 4-en-VPA, caused the greatest changes in indicators of β-oxidation inhibition (dicarboxylic aciduria, β-hydroxybutyrate reduction); however, the biochemical effects were much less pronounced than those reported for hypoglycin. Steatosis in VPA-treated rats occurred only at nearly lethal doses. Phenobarbital induction was confirmed as a predisposing factor; however, it appeared not to greatly enhance production of 4-en-VPA or its recognized metabolites, which collectively comprised only 0.5% of the dose. Elevated oxo-VPA metabolites in serum and 2-propylglutarate in liver were associated with toxicity. Among the newly discovered minor metabolites with possible biologic effects were diols (suggesting epoxide precursors) and a series of dienes and trienes. The rarity of severe human hepatotoxicity indicates that, normally, β-oxidation inhibition is compensated, and cellular defense mechanisms prevail over reactive metabolites. This requires adequate nutrition; on the other hand, severe glycogen depletion may promote toxicity by compromising glucuronidation, the major clearance route. Other literature comments are also supported: (i) caution is indicated for patients with various unusual congenital disorders (e.g., organic acidurias or other mitochondrial defects), and (ii) monotherapy obviates both the predisposition to toxicity and the requirement of large doses to produce therapeutic levels.
Basement Membrane Production by Hepatocytes in Chronic Liver Disease
Authors: Bianchi Francesco B. , Biagini Graziella , Ballardini Giorgio , Cenacchi Giovanna , Faccani Augusto , Pisi Emilio , Laschi Renzo , Liotta Lance , Garbisa Spiridione
Published: August 2008

The immunohistologic distribution of fibronectin, laminin, type IV collagen and whole basement membrane was evaluated in liver biopsies from patients with chronic active liver disease. Fibronectin was consistently increased in the areas of piecemeal necrosis, portal tracts and fibrous septa. Laminin was not detected in normal liver parenchyma. In contrast, laminin positive linear basement membrane structures were prevalent in portal tracts, fibrous septa and the peripheral sinusoids of cirrhotic nodules. In areas of piecemeal necrosis, the hepatocytes, single or assembled in “rosettes”, were frequently underlined by linear deposits of laminin and type IV collagen. This immunoreactivity was often polarized, being confined to the stromal side of liver cells, while the parenchymal side was negative for both proteins. Electron microscopy revealed a typical basement membrane in corresponding areas. Hepatocytes normally do not produce a basement membrane, but do so following chronic injury. We suggest that the polarized basement membrane accumulation by hepatocytes is a hallmark of hepatocyte regeneration following damage.
The Fate of Polymeric and Secretory Immunoglobulin A After Retrograde Infusion into the Common Bile Duct in Rats
Authors: Jones Albert L. , Hradek Gary T. , Schmucker Douglas L. , Underdown Brian J.
Published: August 2008

In the rat, plasma IgA is rapidly endocytosed by hepatocytes and translocated to the bile via a receptor-mediated vesicular transport system which appears to remain intact even during cholestasis. During the latter phenomenon, there is an accumulation of secretory IgA (sIgA) in plasma. These data suggest that biliary IgA can be regurgitated into the plasma compartment. The present study was designed to determine the location and mechanism(s) by which this might occur. 125I-labeled human polymeric IgA (plgA) or slgA was infused retrograde into the rat common bile duct at a flow rate of 20 μ1 per min (5 to 10 μg per min pig A; 7 μg per min sIgA) over 1 hr. Blood and liver samples were collected 10, 30 and 60 min, and radioactivity determined. Radioactive label appeared in the blood by 10 min and increased linearly with time. By 30 min, however, the liver had reached saturation. All of the label found in the blood was intact starting material, i.e., plgA or slgA. Electron microscopic autoradiography analysis clearly demonstrated the presence of grains in vesicles in hepatocyte pericanalicular cytoplasm, as well as in vesicles near the sinusoidal plasma membrane. No grains were observed associated with bile duct or ductule epithelium at any time period. Further, there was no grain accumulation near the parenchymal cell intercellular spaces indicating that paracellular flow plays little or no role in large protein regurgitation. In addition, by 60 min, there were grains associated with Kupffer cells. These data provide the first evidence that hepatocytes, during times of elevated biliary pressure can readily transport macro-molecules from bile to plasma via nonreceptor-mediated membrane-limited vesicles.
Immunohistochemical Distribution of Glucagon, Substance P and Vasoactive Intestinal Polypeptide in Hepatic Vasculature of the Rat
Authors: Sasaki Yutaka , Kamada Takenobu , Hayashi Norio , Sato Nobuhiro , Kasahara Akinori , Fusamoto Hideyuki , Shiosaka Sadao , Tohyama Masaya , Shiotani Yahe
Published: August 2008

The distribution of immunoreactive glucagon, substance P (SP) and vasoactive intestinal polypeptide (VIP)-like structures was investigated in the rat liver, with special reference to the hepatic vasculature by means of the indirect immunofluorescence method. Immunoreactive structures of glucagon were seen in the walls of the portal vein, hepatic artery and hepatic vein, but not in the central vein. Immunoreactive glucagon was localized in the smooth muscle cells of these blood vessels. SP and VIP-like immunoreactive (SPI and VIPI) structures were seen in the neuronal elements. In the porta hepatis, thick, compact SPI and VIPI fibers, which were dissociated from their fiber bundles, reached the tunica adventitia where they were distributed. No SPI and VIPI structures were seen in the tunica media or the tunica interna. No SPI- and VIPI-containing cell bodies could be detected in the liver. These observations suggest that these peptides may have an important role in the neural regulation of hepatic hemodynamics.
Activity and Regulation of Low Density Lipoprotein Receptors in a Human Hepatoblastoma Cell Line†
Authors: Wu George Y. , Wu Catherine H. , Rifici Vincent A. , Stockert Richard J.
Published: August 2008

The responsiveness of low density lipoprotein (LDL) binding and uptake was measured in a human hepatoblastoma cell line, Hep G2. These cells exhibited both saturable, high-affinity and nonsaturable low-affinity components similar to that described for LDL binding in other mammalian cells. In addition, receptor-mediated uptake of [125I]LDL was dependent on the presence of Ca++ and required intact lysine residues in LDL as evidenced by abolition of binding following reductive methylation of LDL. The receptor activity was rapidly responsive to changes made in LDL concentrations in the medium, up-regulating by 200% in the absence of LDL, and down-regulating by 56% in the presence of LDL, both changes occuring within 2 hr. Cholesterol synthesis was estimated by measuring 3-hydroxymethylglutaryl coenzyme A reductase activity. The activity of this enzyme was also found to increase rapidly in the absence of LDL and decrease rapidly in the presence of LDL in the media. The human cell line Hep G2 possesses an active and responsive receptor for LDL and provides a useful model for the study of liproprotein uptake and metabolism in intact human liver.
Regulation of Bile Salt Sulfotransferase Isoenzymes by Gonadal Hormones†
Authors: Kane Robert E. , Chen Lee J. , Thaler Michael M.
Published: August 2008

We studied the regulation of hepatic bile salt sulfotransferase activity by gonadal hormones and the effect of gonadal hormones on two bile salt sulfotransferase isoenzymes. Bile salt sulfotransferase enzyme activity was three times greater in the female than in the male rats. Oophorectomy significantly decreased bile salt sulfotransferase activity in the female, but orchidectomy had no effect on bile salt sulfotransferase activity in the male. Estrogen treatment of intact as well as orchidectomized males markedly stimulated the enzyme activity, while testosterone treatment of intact or oophorectomized females did not effect bile salt sulfotransferase activity. We concluded that the 3-fold greater activity in female rats is due to the striking stimulatory effect of estrogen on bile salt sulfotransferase activity, and the testosterone has little or no role in the sexually related differences in bile salt sulfotransferase activity in mature rats. These sex-related differences in bile salt sulfotransferase activity were investigated further using DEAE-Sephadex A50 ion-exchange chromatography of rat hepatic cytosol. Two bile salt sulfotransferase isoenzymes were identified both with an approximate molecular weight of 130,000. Bile salt sulfotransferase I eluted with 0.05 M NaCl, had an isoelectric point at pH 6.8, was stimulated by estrogen, and was responsible for 90% of total bile salt sulfotransferase activity in the mature female. Bile salt sulfotransferase II eluted with 0.14 Af NaCl, had an isoelectric point at pH 5.3, was unresponsive to estrogen, and accounted for 75 to 80% of bile salt sulfotransferase activity in the mature male. Bile salt sulfotransferase I was 3.8-fold more active toward 3/3-hydroxy-5-cholenoate as glycolith-ocholate, and its activity for glycolithocholate was completely inhibited by 3-ketolithocholate. Conversely, bile salt sulfotransferase II had very little detectable activity towards 3β-hydroxy-5-cholenoate, and its activity toward glycolithocholate was only marginally (13%) inhibited by 3-ketolithocholate. The activity of both bile salt sulfotransferase isoenzymes with various bile acids was: monohydroxylated > dihydroxylated > trihydroxylated. Hepatic bile salt sulfation in the mature rat is catalyzed by two isoenzymes that had the same approximate molecular weight, but differ in sex distribution, substrate activities, regulation by gonadal hormones and inhibition of activity for glycolithocholate by keto bile acids.
Effects of Propranolol on Azygos Venous Blood Flow and Hepatic and Systemic Hemodynamics in Cirrhosis
Authors: Bosch Jaime , Masti Ricardo , Kravetz David , Bruix Jordi , Gaya Joan , Rigau Joaquin , Rodes Joan
Published: August 2008

The effects of propranolol on blood flow through gastroesophageal collaterals and on systemic and hepatic hemodynamies were investigated in 23 cirrhotic patients with portal hypertension. Gastroesophageal collateral blood flow was evaluated by the measurement of azygos venous blood flow by continuous thermal dilution. Azygos venous blood flow was markedly increased in these patients (544 ± 48 ml per min, as compared with 132 ± 18 ml per min in subjects without portal hypertension (p < 0.001).

Propranolol at doses achieving effective β-blockage (83 ± 5 mg) (mean ± S.E.M.) markedly reduced azygos venous blood flow (to 354 ± 34 ml per min, p < 0.001). Reduction of azygos venous blood flow (-34.2 ± 3.6%) was significantly greater (p < 0.01) than reductions in cardiac output (-22.6 ± 1.9%), hepatic venous pressure gradient (-11.5 ± 2.4%) and hepatic blood flow (-13.4 ± 7.4%). The hemodynamic effects of propranolol were not related to plasma norepinephrine levels.

Reduction of gastroesophageal collateral blood flow may be the mechanism by which oral propranolol therapy reduces the risk of repeated episodes of variceal bleeding in cirrhotic patients with portal hypertension.
The Nicotinic Acid Test in Constitutional Conjugated Hyperbilirubinemias and Effects of Corticosteroid
Authors: Ohkubo Hideki , Okuda Kunio
Published: August 2008

The nicotinic acid test was performed in 5 patients with Dubin-Johnson syndrome, 5 with Rotor syndrome and 13 with Gilbert’s syndrome. The increment in serum bilirubin concentration and bilirubin retention 5 hr later differed; the proportion of conjugated bilirubin in the increment of total bilirubin in Dubin-Johnson and Rotor syndromes was greater than in Gilbert’s syndrome. These observations suggest that the nicotinic acid test reflects, in part, impaired biliary excretion of conjugated bilirubin. The results did not differentiate the two conjugated hyperbilirubinemias. The nicotinic acid test was also performed before and after corticosteroid treatment in four patients with Dubin-Johnson or Rotor syndrome. Although serum total and unconjugated bilirubin concentrations were reduced by corticosteroids, no significant change occurred in the parameters of the nicotinic acid test, suggesting that corticosteroids may enhance uptake of bilirubin without significantly altering biliary excretion of conjugated bilirubin.
Morphologic Features of the Liver in Abetalipoproteinemia
Authors: Avigan Mark I. , Ishak Kamal G. , Gregg Richard E. , Hoofnagle Jay H.
Published: August 2008

Liver tissue of a newly diagnosed 30-year-old patient with abetalipoproteinemia was studied by light and electron microscopy. Despite accumulation of large quantities of lipid droplets in hepatocytes, the lobular architecture remained intact, and there was no fibrosis. Acanthocytes were readily identified in sinusoids by scanning electron microscopy. Profiles of rough endoplasmic reticulum, smooth endoplasmic reticulum and Golgi apparatus appeared normal by transmission electron microscopy. Lipid droplets were not bound to membranes and were not associated with the endoplasmic reticulum or Golgi apparatus. In abetalipoproteinemia, these morphological features are consistent with defective apolipoprotein B synthesis. This prevents assembly of lipoproteins that can be secreted through the normal intracellular communicating tubular network and, consequently, fat accumulates in the cytoplasm of the hepatocytes.