bluuebunny/crossref_metadata_2025 huggingface dataset:
Decreased uptake of taurocholate and ouabain by hepatocytes isolated from cirrhotic rat liver
Authors: Reichen Jürg , Hoilien Catherine , Le Mysan , Jones Richard H.
Published: March 2007

To differentiate between the “intact” and “sick” cell hypothesis explaining decreased clearance of endo- and xenobiotics, we measured uptake of taurocholate and ouabain in hepatocytes isolated from cirrhotic rat liver. Cirrhosis was induced by chronic exposure of male Sprague-Dawley rats to phenobarbital and carbon tetrachloride. Uptake of [14C]taurocholate and [3H]ouabain was measured by a rapid filtration technique. Hepatocytes from cirrhotic liver were as viable as control hepatocytes—as judged by trypan blue exclusion and lactate dehydrogenase release—but consumed 28% less oxygen. Vmax of both taurocholate (3.16 ± 0.95 vs. 0.40 ± 0.35 nmoles·min−1·106 cells−1; p < 0.001) and ouabain (2.16 ± 0.78 vs. 0.83 ± 0.26 nmoles·min−1·106 cells−1; p < 0.005) was significantly reduced. These results are compatible with the “sick” cell hypothesis.
Metabolic state of the rat liver with ethanol: Comparison of in vivo 31phosphorus nuclear magnetic resonance spectroscopy with freeze clamp assessment
Authors: Helzberg John H. , Brown Mark S. , Smith Daniel J. , Gore John C. , Gordon Ellen R.
Published: March 2007

In vivo
31phosphorus nuclear magnetic resonance spectroscopy was used to measure the hepatic metabolic state in various groups of rats given ethanol, a control liquid diet or a solid chow diet. The use of selective presaturation pulses applied to the broad phosphorus resonances of immobile phospholipids permitted reliable determination of ATP/ADP ratios by quantitation of the ATP-β and ATP-γ peak areas. ATP/ADP ratios were depressed by both techniques in rats chronically ingesting ethanol compared to pair-fed animals consuming the control liquid diet. These differences were observed regardless of whether ethanol feeding was continued up to the time of investigation or whether it was discontinued for 24 hr prior to study. Acute alcohol administration in chow-fed rats, not previously ingesting ethanol, did not lower hepatic ATP/ADP ratios by either methodology. In all cases, liver ATP/ADP ratios assessed by 31phosphorus nuclear magnetic resonance spectroscopy were higher than those measured by high-performance liquid chromatography. However, parallel decreases in hepatic ATP/ADP ratios were observed with chronic ethanol consumption by both 31phosphorus nuclear magnetic resonance spectroscopy and the biochemical method, confirming the utility of in vivo
31phosphorus nuclear magnetic resonance spectroscopy for assessment of the hepatic bioenergetic status. The difference in absolute ATP/ADP ratios by the two methods may to some degree be explained by binding effects of ADP with proteins or mitochrondrial membranes, rendering it partially invisible to nuclear magnetic resonance or alternatively, by breakdown of high energy phosphate bonds with freeze clamp extraction.
Reactivities and clinical relevance of antimitochondrial antibodies to four mitochondrial inner membrane proteins in sera of patients with primary biliary cirrhosis
Authors: Ishii Hideo , Saifuku Koji , Namihisa Toshihiko
Published: March 2007

Antimitochondrial antibodies are characteristically detected in sera of patients with primary biliary cirrhosis. The antigens to which the antimitochondrial antibodies in primary biliary cirrhosis sera react have been located in the mitochondrial inner membrane. We have reported on four mitochondrial inner membrane proteins, extracted from beef heart, which reacted with antimitochondrial antibodies of primary biliary cirrhosis. These four proteins had molecular weights of 70, 54, 51 and 45 kd. Forty-six of the 50 PBC sera tested were positive with rat kidney and stomach specimens by the indirect immunofluorescent method. All sera of 50 primary biliary cirrhosis patients were positive with at least one and as many as four of the mitochondrial proteins extracted from beef heart using immunoblotting. Forty-seven primary biliary cirrhosis sera (94%) had IgG antibodies (antimitochondrial antibodies) to the antigen proteins, 43 patients (86%) IgM antimitochondrial antibodies and 38 patients (76%) IgA antimitochondrial antibodies. Seventy per cent of patients had antimitochondrial antibodies in all three immunoglobulin classes. In a few primary biliary cirrhosis sera, antimitochondrial antibodies were represented by only one class of immunoglobulin. The major antigenic proteins of mitochondrial inner membrane to which the antimitochondrial antibodies react were the 70 and 51 kd proteins. All primary biliary cirrhosis sera containing antibodies to the 54 and/or 45 kd proteins reacted with 70 kd as well. Patients with an elevation of serum bilirubin over 2 mg per dl were often found to produce antimitochondrial antibodies reactive with the 54 kd in addition to the 70 kd protein. No significant association was found between histological classifications and reactive patterns of antimitochondrial antibodies.
Opsonins in the hepatic clearance of pneumococci
Authors: Runyon B. A.
Published: March 2007

An in situ isolated, perfused rat liver system was used to evaluate various opsonins for hepatic trapping and killing of encapsulated, virulent, type 3 pneumococci. Pneumococci were rapidly trapped in the liver in the presence of all potential opsonins including Hanks balanced salt solution with added colloid. However, with some of the potential opsonins the organisms remained viable and could be recovered from the liver. With others there was killing of pneumococci. The combination of rat antibody and complement was effective as an opsonin and produced about the same degree of killing as the same human components. When tested separately killing was not promoted by rat complement, human complement, or human antibody. However, when ethylene glycol-bis(β-aminoethyl ether)-N,N,N′,N′-tetraacetic acid-Mg2+ was added to block the classical pathway and preserve the alternative pathway there was significant pneumococcal killing which may have been complement mediated or due to an adverse effect of ethylene glycol-bis(β-aminoethyl ether)-N,N,N′,N′-tetraacetic acid-Mg2+ on the organisms. Fibronectin and C-reactive protein reduced the number of surviving microorganisms, although the change was not significant. Immunoglobulin G (IgG), IgM, IgG, or IgM with complement, and antibody + the classical complement pathway, significantly reduced survival of the organisms. The best killing occurred with antibody plus complement plus fibronectin, which reduced mean survival to 24% during the 1-h perfusion.
Isolated rat hepatocyte couplets in short-term culture: Structural characteristics and plasma membrane reorganization
Authors: Gautam Anil , Ng Oi-Cheng , Boyer James L.
Published: March 2007

Studies of canalicular bile secretion have been limited due to lack of direct access to the canalicular lumen. Isolated rat hepatocyte couplets, consisting of two hepatocytes enclosing a canalicular space, have been proposed as a primary secretory unit that may be useful for direct studies of unmodified canalicular bile secretion. The present study examines their structural characteristics and plasma membrane reorganization. The canalicular space of freshly isolated hepatocyte couplets is freely permeable to ruthenium red, but within 4 hr the junctional borders reseal in most couplets which then exclude ruthenium red from the luminal area. These resealed spaces expand in 61.8 ± 10% of couplets as secretion is elaborated and after 4 hr in monolayer culture, 12.7 ± 4.7% of the canalicular spaces are dilated to diameters greater than 3 μ. Normal-appearing microvilli line these canalicular membranes in the majority of dilated spaces as assessed by electron microscopy. Immediately after isolation, Mg++-ATPase, a histochemical marker for canalicular membranes, is located as a discrete band corresponding to the normal in vivo circumferential distribution of the canalicular membrane domain. However, this pattern of Mg++-ATP-ase staining rapidly diminishes and reorganizes at the remaining canalicular pole within several hours. This membrane reorganization is a microfilament-dependent process, since it is inhibited by cytochalasin D but not by colchicine, an inhibitor of microtubular function or cycloheximide, an inhibitor of new protein synthesis.

Thus, 3 to 4 hr after isolation, the isolated hepatocyte couplet has reorganized its canalicular membrane, retaining polarity of its excretory domain only at the remaining junctional contacts between adjacent hepatocytes. Secretion is elaborated into the ruthenium red impermeant canalicular lumen permitting direct studies of canalicular bile secretion by micropuncture, electrophysiologic and quantitative microscopic techniques.
Comparative effects of deoxycholate and 7-methyl-deoxycholate in the hamster
Authors: Kuroki Syoji , Mosbach Erwin H. , Stenger Richard J. , Cohen Bertram I. , McSherry Charles K.
Published: March 2007

The metabolism and effect on biliary lipids of a new bile acid analog, 7-methyl-deoxycholic acid, were studied and compared with those of deoxycholic acid in the hamster. 14C-Labeled 7-methyl-deoxycholic acid and deoxycholic acid were administered intravenously or intraduodenally to bile fistula hamsters at 1.0 or 4.0 μmoles per min·kg, and hepatic bile was analyzed for radioactive metabolites and biliary lipid outputs. Deoxycholic acid and 7-methyl-deoxycholic acid were efficiently absorbed from the intestine, extracted by the liver and excreted into bile as taurine and glycine conjugates. Twenty per cent of deoxycholic acid was 7α-hydroxylated to cholic acid while 7-methyl-deoxycholic acid did not undergo hydroxylation. During deoxycholic acid infusion, the biliary secretion of phospholipid did not increase, and the bile became more lithogenic. In contrast, 7-methyl-deoxycholic acid stimulated phospholipid secretion, and bile became less lithogenic. Although pathologic changes in the liver were inconstant and mostly mild, both bile acids were toxic in the hamster; hemolysis and death due to respiratory distress were observed.
Ultracentrifugal isolation of vesicular carriers of biliary cholesterol in native human and rat bile
Authors: Ulloa Natalia , Garrido Jorge , Nervi Flavio
Published: March 2007

We have utilized ultracentrifugation of native bile-Metrizamide density gradients to isolate a vesicular transport system or biliary lipids in both man and rat. We identified vesicular structures by electron microscopy. Fresh bile specimens were obtained from bile fistula rats (unsaturated bile) and from patients 1 week after bile duct surgery (supersaturated bile). Metrizamide was dissolved in bile (33% w/v), and continuous density gradients were performed with undiluted bile (density limits = 1.020 to 1.300 gm per ml). The relative distribution of biliary cholesterol, phospholipid and bile salt was studied as a function of the density of the fractions. Approximately 50% of total rat biliary cholesterol and between 61 and 90% of human biliary cholesterol was concentrated in the lightest fractions of the gradients (density < 1.060 gm per ml). In contrast, less than 20% of bile salts was present in fractions with densities lower than 1.060 gm per ml. The highest amounts of bile salts and phospholipids of the bile-Metrizamide density gradients were found in the density range of 1.075 to 1.100 gm per ml in both human and rat bile. More than 80% of biliary proteins was found in fractions with densities < 1.075 gm per ml, and only 2% was found in the cholesterol-rich fraction with density < 1.060 gm per ml in both species. When bile salt concentration was raised in rat bile from 38 to 97 mM by adding taurocholate, the low density cholesterol-rich fraction almost disappeared. Electron microscopy of negatively stained preparations of the fractions with density < 1.060 gm per ml showed 40 to 120 nm vesicles, which were not apparent in the other fractions. Similar vesicles were demonstrated also in fresh rat bile and within the canaliculi after acute depletion of the bile salt pool (biliary bile salt concentration of 3.45 mM; total biliary lipid concentration of 0.25 gm%). The structure of these vesicles was shown in thin sections of liver specimens. They appeared as internal cavities surrounded by a single, continuous 6-nm-thick bilayer. These studies demonstrate that a high proportion of biliary cholesterol is transported in vesicles in human supersaturated native bile and that vesicular carriers are also responsible for the transport of a significant amount of biliary cholesterol in unsaturated rat bile. The presence of vesicles in unsaturated hepatic bile strongly supports the thesis that biliary lipids may be secreted as vesicles from the hepatocyte into the canaliculi.
Liver levels of vitamin a and cellular retinol-binding protein for patients wth biliary atresia
Authors: Ong David E. , Amédée-Manesme Olivier
Published: March 2007

We have examined whether the amount of cellular retinol-binding protein in human liver is related to the amount of vitamin A stored in the liver. Levels of vitamin A, as retinol and retinol esters, and of cellular retinol-binding protein have been determined in liver samples from 6 normal adults and 11 children with biliary atresia, with and without vitamin A treatment. The level of cellular retinol-binding protein in the liver was not related to the liver vitamin A concentration examined over a 300-fold range of vitamin A levels. Also, biliary atresia did not appear to interfere with storage of vitamin A, and the level of cellular retinol-binding protein was comparable to that observed in the liver of normal adults. The demonstration of proper vitamin A storage in treated children as well as normal levels of cellular retinol-binding protein suggest the vitamin A deficiency frequently observed in children with biliary atresia may be due primarily to faulty absorption rather than a combination of poor absorption and impaired hepatic vitamin A metabolism.
Influence of branched-chain amino acids and branched-chain keto acids on protein synthesis in isolated hepatocytes
Authors: Base Wolfgang , Barsigian Carl , Schaeffer Alisa , Shaw Ellen , Martinez Jose , Maddrey Willis C.
Published: March 2007

We investigated the effects of the branched-chain amino acids—valine, leucine and isoleucine—or their keto analogs, the branched-chain keto acids—α-ketoisovaleric acid, α-ketoisocaproic acid and α-keto-β-methylvaleric acid—on protein synthesis and secretion by monolayers of rabbit hepatocytes incubated with [35S] methionine in pulse-chase and steady-state experiments. The branched-chain amino acids (2.0 mM or 1.0 mM), in the presence or absence of insulin (2 × 10−4 IU per dish) and in both types of experiments, reduced the trichloroacetic acid-precipitable 35S-protein secreted into the medium. The branched-chain keto acids (2.0 mM or 1.0 mM) had a stimulatory effect on secreted trichloroacetic acid-precipitable 35S-protein which was observed only by the pulse-chase technique in the presence of insulin. Immunoaffinity chromatography of medium demonstrated a slight inhibition by branched-chain amino acids and a slight stimulation by branched-chain keto acids on secretion of 35S-albumin and no effect of either treatment on secretion of 35S-fibrinogen. ELISA analysis of total (i.e., 35S-labeled and unlabeled) secreted albumin revealed an inhibitory effect of the branched-chain amino acids in both pulse-chase and steady-state experiments, and a small stimulatory effect, in steady-state experiments, of the branched-chain keto acids; both effects were insulin-dependent. Total secreted fibrinogen, under steady-state conditions, was increased by the branched-chain keto acids in the presence of insulin, while transferrin production was unaffected by any treatment. Intracellular trichloroacetic acid-precipitable 35S-protein was increased by the branched-chain keto acids in the presence of insulin and was reduced by the branched-chain amino acids in the presence or absence of insulin, suggesting that the effects of these agents may have been related to synthesis and not to effects on secretion.
Cytophotometric measurements of nuclear DNA content in hepatocellular carcinomas
Authors: Kuo Sow-Hsong , Sheu Jin-Chuan , Chen Ding-Shinn , Sung Juei-Low , Lin Chi-Chung , Hsu Hey-Chi
Published: March 2007

The nuclear DNA contents of 54 hepatocellular carcinomas were measured on cytologic smears using a scanning microdensitometer at a wavelength of 550 nm. The cellular materials were aspirated under ultrasonographic guidance and stained with modified Feulgen method. The DNA modal values, which could be defined in 51 cases, ranged from 2.2 to 9.8 ploidy, with 46.3% (25/54) in hypertetraploidy. The hepatocellular carcinomas in patients with higher serum levels of α-fetoprotein contained more hyperploid cells. The DNA modal values did not correlate with the age, HBsAg status, tumor size and histological grading of hepatocellular carcinoma. The S-phase fraction values, which could be estimated on the DNA histograms in 43 cases (79.6%), ranged from 1 to 35 (mean ± S.D. = 10.6 ± 8.1). The S-phase fraction values were higher in high grade hepatocellular carcinomas (13.4 ± 8.5) than low grade hepatocellular carcinomas (6.1 ± 4.4), p < 0.01. The mean S-phase fraction values were also significantly higher in hepatocellular carcinomas with α-fetoprotein levels of more than 100 ng per ml than those of less than 100 ng per ml (12.9 ± 8.0 and 7.8 ± 5.6, respectively). The S-phase fraction values also did not correlate with the age, HBsAg status and tumor size.
Portacaval shunt and the reacquisition of γ-glutamyl transpeptidase activity
Authors: Colombo J. P.
Published: March 2007

Portacaval shunt (PCS) operations were performed on male inbred SD rats. The activity of liver γ-glutamyltranspeptidase (GGT) increased a few days after the operation and remained high after several months. However, the activity was only present in periportal areas of liver lobules and was mainly restricted to the endothelial lining cells of periportal blood vessels. Phenobarbital sodium (CAS: 57-03-7) administration did not change the distribution of GGT. The activity of liver glucose-6-phosphatase disappeared in the centrilobular area a few days after the operation but was present in the periportal areas. The distribution of this activity returned to normal after several months. The capacity of liver cells to store glycogen was significantly decreased following the PCS operation. Morphologic changes in hepatocytes observed after the PCS operation did not show similarities to those seen in preneoplastic livers. The nuclei of the small compact hepatocytes found in the animals with PCS were irregular in shape and stained intensively with hematoxylin and eosin. The large pale-staining cells in the periphery of liver lobes were GGT negative and their nuclei had a normal morphology. Neither morphologic nor histochemical changes consistent with preneoplastic and/or neoplastic stage were observed after 10 months in the liver when the PCS operation was performed on rats having received an ip dose (10 mg/kg) of diethylnitrosamine (CAS: 55-18-5) 2 weeks prior to the operation.
Diuresis in Portuguese
Authors: Conn Harold O.
Published: March 2007

Standardization of a therapeutic approach for ascites due to chronic liver disease was prospectively tried in 100 patients. The four progressive steps were: I—relative bed rest associated to with-drawal of toxic agents, and restriction of salt (40 mEq) and water (1.000ml per day) for 5 to 7 days. In twenty two patients a good diuretic response was observed. Step II, initiated only for patients who did not respond to step I, corresponded to the introduction of espironolactone, 150 mg/day, for at least 6 days. A good response was defined as an increased diuresis and weight loss between 300 and 900g/day. 57% of the patients included in step II, showed such a good diuretic response. In step III the dosage of espironolactone was increased to 300 mg/d and 56% of the patients included in this step presented good diuretic response. Out of 100 patients who began treatment only 12 needed a IV step treatment which consisted of the association with furosemide in the dosage of 40 to 80mg/d. Apart from two patients who did not respond, developing spontaneous bacterial peritonitis during treatment and dying, the remaining 10 cases had a good diuretic response. General mortality due to chronic liver disease occurred in 10% of the patients during treatment for ascitis. Hepatic encephalopathy developed in 22 cases but in only four it was due to the diuretic therapy and reversed. Hyperpotassemia in 16 cases and hyperazotemia in 24 cases were generally transient.
The effects of galactosamine-induced hepatic failure upon blood-brain barrier permeability
Authors: Lo Warren D. , Ennis Steven R. , Goldstein Gary W. , McNeely David L. , Betz Lorris A.
Published: March 2007

The role of changes in blood-brain barrier permeability in the pathogenesis of hepatic encephalopathy remains uncertain. To test the hypothesis that brain microvessel permeability is nonselectively increased in hepatic encephalopathy we measured the blood-brain barrier permeability-surface area product in rats with acute liver failure induced by intraperitoneal injection of galactosamine. The permeability-surface area products to the diffusion-limited tracers, sucrose and methylaminoisobutyric acid, were determined as a measure of blood-brain barrier permeability. Animals were examined 24, 36 and 42 hr after injection, at times when they were stuporous, but not comatose. No significant elevations of the permeability-surface area products for either compound were detected in clinically affected experimental animals when compared to controls. Our results indicate there is no generalized increase in brain vascular permeability during hepatic insufficiency in precomatose animals.
The evolution of changes in quantitative liver function tests in a rat model of biliary cirrhosis: Correlation with morphometric measurement of hepatocyte mass
Authors: Gross John B. , Reichen Juerg , Zeltner Thomas B. , Zimmermann Arthur
Published: March 2007

The aim of this study was to determine the prognostic significance of functional changes in the liver during progression of cirrhosis. Liver function was quantitated weekly by the aminopyrine breath test (measuring microsomal function) and the galactose breath test (measuring cytosolic function) in rats made cirrhotic by bile duct ligation (n = 14) and in sham-surgery controls (n = 9). Nine rats died spontaneously of cirrhosis. Both the aminopyrine breath test and galactose breath test were sensitive (89%) predictors of death within 1 week, but the galactose breath test was more specific (83%). Morphometric measurements of livers from surviving cirrhotic animals and controls (n = 5 each) showed that mean hepatocyte mass was maintained in the cirrhotic livers [cirrhosis (17.0 ± 2.0) vs. controls (13.9 ± 0.9 gm)]. The galactose breath test was also maintained, whereas the aminopyrine breath test was significantly decreased in the surviving cirrhotics. The galactose breath test, but not the aminopyrine breath test, correlated with hepatocyte mass (r = 0.67). The aminopyrine breath test correlated with microsomal aminopyrineN-demethylase activity (r = 0.78). Serial use of quantitative liver tests allows prediction of time of death from cirrhosis in this model.
The aminopyrine breath test does not correlate with histologic disease severity in patients with cholestasis
Authors: Baker Alfred L. , Krager Patricia S. , Kotake Alvin N. , Schoeller Dale A.
Published: March 2007

To determine whether the aminopyrine breath test can be used to document the presence of cirrhosis in patients with cholestatic liver disease, 19 patients (13 primary biliary cirrhosis, 4 sclerosing cholangitis and 2 chronic extrahepatic bile duct obstruction) underwent clinical and biochemical evaluations, liver biopsies and an aminopyrine breath test. Results were compared with those in 10 patients with biopsy-proven chronic active hepatitis with bridging and/or cirrhosis and in 22 healthy subjects. The aminopyrine breath test results in the 10 cholestatic patients with cirrhosis were not significantly different from the results in precirrhotic cholestatic patients (mean ± S.D., 11.2 ± 5.0 vs. 11.6 ± 2.8 %dose per 2 hr, p < 0.05) or healthy subjects (11.5 ± 2.9 %dose per 2 hr). In contrast, the results in the patients with chronic hepatitis were markedly depressed (3.2 ± 1.9 %dose per 2 hr, p < 0.05). The aminopyrine breath test results did not correlate with results of conventional liver function tests in the cholestatic patients. These results demonstrate that the aminopyrine breath test is not clinically useful in identifying the presence of cirrhosis in patients with cholestatic liver disease, and provide further evidence that decreased microsomal enzyme function is a late feature of cholestatic liver disease.
Orthotopic liver transplantation for acute and subacute hepatic failure in adults
Authors: Peleman Rene R. , Gavaler Judith S. , Van Thiel David H. , Esquivel Carlos , Gordon Robert , Iwatsuki Shunzaburo , Starzl Thomas E.
Published: March 2007

The role of liver transplantation in 29 patients with fulminant and subacute hepatic failure due to a variety of different causes was examined by comparing the outcome and a variety of “hospitalization” variables. Transplanted patients (n = 13) were more likely to survive (p < 0.05), were younger (p < 0.05) and spent more time in the hospital (p < 0.025) than did those who were not transplanted (n = 16). Despite spending a much longer time in the hospital, transplanted patients spent less time in the intensive care unit (p < 0.05) in coma (p < 0.01) and on a respirator (p < 0.01) than did those not transplanted. Most importantly, the survival rate for transplanted patients was significantly improved (p < 0.05) as compared to those not transplanted. We conclude that liver transplantation can be applied successfully to the difficult clinical problem of fulminant and subacute hepatic failure.
Oleic acid-induced cholelithiasis in the rabbit: Conversion of dietary oleic acid to cholestanol as a cause of calcium-bile salt gallstones
Authors: Lee Sum P. , Tasman-Jones Cliff , Carlisle Valerie F.
Published: March 2007

Rabbits fed a diet rich in oleic acid develop gallstones consisting of calcium salts of (5α)-glyco-allodeoxycholic acid. To study the metabolic pathway of oleic acid, we followed the changes in plasma, hepatic and biliary lip-ids in this animal model. In addition, to also determine the role played by intestinal microflora on biliary lipid metabolism, we added kanamycin to the oleic acid diet. Oleic acid-fed rabbits rapidly developed hypercholes-terolemia. This was associated with an increase in liver 3-hydroxy-3-methylglutaryl coenzyme A reductase activity, accumulation of cholesterol as well as cholestanol in the liver and progressive saturation of cholesterol in bile. [14C]oleic acid fed orally to rabbits was recovered in liver extracts as both cholesterol and cholestanol. With oleic acid feeding, there was a progressive increase in glyco-allodeoxycholic acid culminating in the formation of gallstones. Kanamycin supplement to the oleic acid diet resulted in the same changes in plasma and hepatic sterol metabolism compared with oleic acid-fed rabbits. There was, however, a striking difference in the biliary bile acid profile. Kanamycin supplementation dramatically reduced the proportion of 5α-dihydroxy bile acids, increased the proportion of 5β-trihydroxy bile acids and completely abolished gallstone formation. We postulate that, in the rabbit, oleic acid is used as a carbon source for cholesterol synthesis, and a high oleic acid diet increases hepatic cholesterogenesis. Hepatic cholesterol is then metabolized to form cholestanol, followed by (5α)-glyco-allocholic acid which is secreted into bile and transformed by gut bacteria to form (5α)-allodeoxycholic acid. Kanamycin abolished gallstone formation by inhibiting intestinal bacterial dehydroxylation.
Establishment and characterization of a human combined hepatocholangiocarcinoma cell line and its heterologous transplantation in nude mice
Authors: Murakami Tastuo , Yano Hirohisa , Maruiwa Masafumi , Sugihara Shigetaka , Kojiro Masamichi
Published: March 2007

A human cell line (KMCH-1) derived from a surgical specimen of combined hepatocellular and cholangiocarcinoma has been established. The original tumor consisted of both hepatocellular carcinoma of the trabecular type and cholangiocellular carcinoma. This cell line has been maintained for 26 months through 75 passages. KMCH-1 cells show characteristics of adenocarcinoma on light and electron microscopy. They proliferate in culture in a pave stone arrangement. The doubling time of these cells at the 24th passage was 39 hr. Chromosome analysis revealed a chromosome number ranging from 60 to 98, with a modal number of 74. KMCH-1 cells produced tumors several months after subcutaneous and intraperitoneal transplantations into athymic nude mice. Histologically, the subcutaneous tumors were poorly differentiated adrenocarcinoma, while intraperitoneal tumors were moderately to well-differentiated adenocarcinoma. Hepatocellular carcinoma components were not observed. Thus, KMCH-1 cells demonstrate the feature of cholangiocellular carcinoma in vitro and in vivo. This KMCH-1 cell line is the first established combined hepatocellular and cholangiocarcinoma cell line and may contribute to further investigation of combined hepatocellular and cholangiocarcinoma.
Spot-blot hybridization assay for the detection of hepatitis b virus dna in serum: Factors determining its sensitivity and specificity
Authors: Walter Eike , Blum Hubert E. , Offensperger Wolf-Bernhard , Zeschnigk Christine , Offensperger Silke , Gerok Wolfgang
Published: March 2007

Factors determining the sensitivity and specificity of the spot-blot hybridization technique for the detection of hepatitis B virus DNA in serum were systematically investigated. Methods for pretreatment of serum samples, mode of application of the samples to the transfer membranes, blot treatment and hybridization conditions were all found to affect the sensitivity of the assay. The optimum hybridization procedure was found to be incubation of serum samples with salt, NaOH, formaldehyde and detergent, followed by spot application of the samples. This method specifically detected hepatitis B virus DNA in serum with a sensitivity 5 to 15 times higher than the presently used assay procedures.
IgM anti-HBc in anti-HBe positive chronic type B hepatitis with acute exacerbations
Authors: Koike Kazuhiko , Iino Shiro , Kurai Kiyohiko , Mitamura Keiji , Endo Yasuo , Oka Hiroshi
Published: March 2007

IgM anti-HBc was measured by radioimmunoassay in serially collected serum samples during 20 acute exacerbations which developed in 14 patients with anti-HBe positive chronic type B hepatitis. IgM anti-HBc became positive in 12 of the 14 (86%) patients and in 18 of the 20 (90%) exacerbations, and elevation of IgM anti-HBc which was believed to be significant was also observed in the remaining two patients. In most of the patients who also had determinations of serum hepatitis B virus DNA, the hepatitis B virus DNA became positive. These results suggest that ALT elevation in anti-HBe positive chronic type B hepatitis is associated with active replication of hepatitis B virus. It is considered to be useful to measure IgM anti-HBc for the purpose of identifying the causes of ALT elevation in anti-HBe positive chronic type B hepatitis.
Interferon-γ production by peripheral blood mononuclear cells of patients with chronic liver disease
Authors: Fuji Akihiko , Kakumu Shinichi , Ohtani Yoshiyuki , Murase Kenichi , Hirofuji Hideo , Tahara Hirofumi
Published: March 2007

We investigated the role of the interferon system in the pathogenesis of chronic liver disease. Interferon-γ production by peripheral blood mononuclear cells was measured with an ELISA. While concanavalin A-stimulated and recombinant interleukin 2-stimulated production of interferon-γ in patients with chronic active hepatitis and liver cirrhosis was significantly decreased when compared with that of controls (518 ± 189 and 729 ± 195 units per ml, mean ± S.D.), there was also a lot of overlap. Addition of indomethacin to the cultures partially restored interferon-γ production in patients with chronic active hepatitis and liver cirrhosis, indicating that suppressor function of monocytes was, in part, responsible for the diminished interferon-γ production. Serial studies showed that interferon-γ production rose during acute deterioration of illness, during treatment with interleukin 2 and with the improvement of clinical course. Interferon-γ production was not different among hepatitis B e antigen or antibody positive, and non-A, non-B patients with chronic active hepatitis and liver cirrhosis. Our findings suggest that diminished interferon-γ production is associated with disease severity in chronic liver disease, irrespective of the hepatitis B virus carrier state. It would be interesting to compare the efficacy of treatment with interferon-γ or interferon-γ inducers such as interleukin 2 in chronic hepatitis B patients with and without decreased in vitro interferon-γ production.
Liver transplantation across abo blood groups
Authors: Bove Joseph R. , Conn Harold O.
Published: March 2007

Six hundred seventy-one first, second and third orthotopic liver allografts in 520 patients were reviewed to determine the effect of donor-recipient mismatches or incompatibilities for the ABO blood groups on graft survival. A significant advantage for ABO donor-recipient identity was found, especially in adults for first grafts. However, a surprisingly large number of ABO incompatible grafts were successful. We recommend that nonidentical or incompatible grafts be limited to patients such as small children for whom the supply of available donors is severely limited for patients in urgent need of transplantation or retransplantation.
The jaundiced heart
Authors: Deckelbaum Lawrence
Published: March 2007

Patients with obstructive jaundice are susceptible to postoperative shock and kidney failure. The cause of these potentially fatal complications has not been fully clarified. The present study was designed to assess the role of myocardial dysfunction in the hemodynamic disturbance of obstructive jaundice. We studied the effect of isolated cholemia on left ventricular performance in five conscious dogs before and 2 weeks after choledochocaval anastomosis by using measurements of systolic time intervals (STIs) and maximal dp/dt. Mean left ventricular ejection time (LVET) decreased after cholemia from 159 ± 2.8 msec to 139 ± 2.6 msec (p < 0.005), while mean preejection period (PEP) and mean PEP/LVET were increased from 41 ± 8.5 msec to 87 ± 14 msec (p < 0.05) and from 0.39 ± 0.06 to 0.62 ± 0.1 (p < 0.01), respectively. During cholemia, STIs were unchanged after intravenous administration of ouabain, whereas in the control period, there was shortening of mean PEP from 71 ± 8.8 msec to 58 ± 7.6 msec (p < 0.05) and of Q-S2 from 257 ± 12 msec to 235 ± 14 msec (p < 0.005) in response to ouabain. Maximal dp/dt decreased after choledochocaval anastomosis from 4543 ± 593 mm Hg/sec to 3666 ± 648 Hg/sec (p < 0.025). We conclude that cholemia in the dog is clearly associated with impaired left ventricular performance. The present data also support a previously published in vitro study from our laboratory showing that cholemia blunts the myocardial contractile response to sympathomimetic agents. The cardiodepressor effect of cholemia may explain the increased tendency of patients with obstructive jaundice to postoperative shock and renal failure.
Endoscopic measurement of pressure in esophageal varices: Variations and complexities
Authors: Polio John
Published: March 2007

An endoscopic pressure sensor has been evaluated in the measurement of oesophageal variceal pressure and its response to drug ingestion. The variceal pressure showed a highly significant correlation with the splenic pulp pressure (r = 0.97) in six patients with liver disease of diverse aetiology and with hepatic venous wedge pressure (r = 0.92) in eight alcoholic cirrhotic men. Intraduodenal infusion of isosorbide dinitrate in the cirrhotics produced no change in wedge pressure or endoscopic variceal pressure despite profound falls in arterial systolic pressure. Thus isosorbide dinitrate appears to be of no value in treating portal hypertension. This study establishes the endoscopic pressure sensor as a valuable tool in screening drugs in this condition.
Endoscopic sclerotherapy with portal-systemic shunt rescue
Authors: Conn Harold O.
Published: March 2007

This paper reports the preliminary results of a prospective randomized trial comparing endoscopic variceal sclerosis and distal splenorenal shunt (DSRS) in the management of patients with cirrhosis and variceal bleeding. Seventy-one patients have been entered; 36 have received sclerosis and 35 DSRS. Randomization of the study population was stratified on Child's A/B (56%) and Child's C (44%). Sixty-one per cent had alcoholic and 39% nonalcoholic cirrhosis. No patients have been lost to follow-up, which currently stands at a median of 26 months. Rebleeding occurred significantly (p < 0.05) more frequently in patients in the sclerosis group (19 of 36: 53%) compared to DSRS (1 of 35: 3%), but only 11 of 36 (31%) were not controlled by further sclerosis and failed that therapy. Patients in whom sclerosis failed underwent surgery. Survival was significantly (p < 0.01) improved in the sclerosis group (+ surgery in 31%), with an 84% 2-year survival compared to a 59% 2-year survival in the DSRS group. Portal perfusion was significantly (p < 0.05) better maintained in the sclerosis (95%) compared to the DSRS (53%) group. Galactose elimination capacity improved significantly (p < 0.05) in 21 patients successfully managed by sclerosis at 1 year and was significantly (p < 0.01) better maintained in the sclerosis compared to DSRS group. The authors conclude that endoscopic sclerosis: (1) has a higher rebleeding rate than DSRS, with one third of patients failing therapy from rebleeding; (2) allows significant improvement in liver function when successful; and (3) gives significantly improved survival in the management of variceal bleeding when backed up by surgical therapy for patients with uncontrolled rebleeding.

The authors report the results of a randomized comparison of emergency portacaval anastomosis (PCA) (32 patients) with endoscopic sclerotherapy (EST) (32 patients). Short-term survival was similar in the two groups, respectively (44% vs. 50%). The amount of blood transfusions required and the length of the hospital admissions were significantly less in the EST group (p < 0.001). Recurrences of variceal hemorrhage, the number of transfusions required to treat them and the duration of hospital readmissions were all greater in the EST group (p < 0.001). Almost half of the EST patients who were discharged from the hospital alive subsequently rebled and required surgical treatment. The authors conclude that EST is as effective as PCA in the acute treatment of bleeding, but that when EST fails, PCA may be effective management.
The synthesis of proteoglycans in fat-storing cells of rat liver
Authors: Schäfer Stephan , Zerbe Ottwin , Gressner Axel M.
Published: March 2007

Fat-storing cells (perisinusoidal stellate cells) were isolated by enzymatic digestion of rat liver and purified by a single-step Nycodenz ® gradient to yield 11.4·106 cells per liver, with a purity of 74% and a viability of 76%. Monolayer cultures of fat-storing cells incorporated both [35S]sulfate and [3H]glucosamine into glycosaminoglycans; the rate of incorporation increased with culture time (3-fold between the third and eighth days in culture). About 80% of newly formed glycosaminoglycans were secreted into the medium. Analysis of the types of glycosaminoglycans revealed a different pattern for cells and medium, respectively, which is subject to culture time. Heparan sulfate remains primarily cell-bound and, therefore, has a low fractional secretion rate. Chondroitin sulfate and even more dermatan sulfate are the main types of glycosaminoglycans in the medium. Dermatan sulfate represents about 60% of total medium glycosaminoglycans. In advanced cultures (eighth day), this type becomes the predominant one in the cell layer. The reduction of the molecular weight of native medium-sulfated molecules by papain digestion and β-elimination and the puromycin-induced inhibition of their synthesis by more than 75% suggest the formation of glycosaminoglycans as complex proteoglycans. It is concluded that fat-storing cells are a major cellular source of dermatan sulfate and chondroitin sulfate in liver connective tissue. Since the pattern of proteoglycans of fat-storing cells closely resembles that found in the fibrotic liver matrix, this cell type might be of pathogenetic significance for the accumulation of chondroitin sulfate and dermatan sulfate in cirrhotic connective tissue.
The effect of recombinant α-interferon treatment on serum levels of hepatitis B virus-encoded proteins in man
Authors: Hess Georg , Gerlich Wolfram , Gerken Guido , Manns Michael , Hütteroth Thomas H. , Büschenfelde Karl-Hermann Meyer Zum
Published: March 2007

The effect of α-interferon treatment on serum levels of hepatitis B virus-encoded proteins was analyzed in eight patients with chronic type B hepatitis who participated in a pilot study of interferon therapy. Three individuals became HBsAg-negative, 4 lost HBeAg but remained HBsAg-positive and 1 remained positive for both HBsAg and HBeAg. Initiation of interferon treatment was rapidly followed by reduction or loss of hepatitis B virus DNA in the serum but by little immediate change in hepatitis B virus antigen levels. Changes in hepatitis B virus antigens were usually delayed. Loss of HBsAg from the serum was preceded by the sequential disappearance of pre-S-encoded proteins (pre-S1 and polymerized human serum albumin) and HBeAg. In patients who lost HBeAg but remained HBsAg-positive, serum levels of pre-S1 and polymerized human serum albumin usually, but did not always, decrease. The individual who remained HBsAg- and HBeAg-positive had unchanged serum levels of pre-S1, polymerized human serum albumin and HBsAg. These results suggest that α-interferon inhibits hepatitis B virus DNA replication but has little direct effect on synthesis of hepatitis B virus gene products.
A multicenter, prospective study of posttransfusion hepatitis in milan
Authors: Colombo Massimo , Oldani Silvia , Donato Maria F. , Borzio Mauro , Santese Raffaele , Roffi Luigi , Viganó Paolo , Cargnel Antonietta
Published: March 2007

We studied the risk of posttransfusion hepatitis in recipients of blood collected from volunteer donors who tested negative for HBsAg and had serum ALT levels less than 1.5 times the upper limit of the normal range. Between October, 1983 and September, 1984, 676 consecutive patients who needed blood or plasma transfusions during or after elective surgery, who had no history of liver disease and had never received blood previously, were studied. The patients were given a total of 4,813 (mean = 7) units. Ninety-six patients developed posttransfusion hepatitis, which yielded a hepatic incidence of 20 cases per 1,000 units of transfused blood. Ninety-two patients had non-A, non-B hepatitis, 3 had hepatitis B and 1 had cytomegalovirus infection. The incubation periods for non-A, non-B hepatitis ranged from 2 to 26 (mean = 9.5 ± 4) weeks. In 68 (73%) patients, the hepatitis was completely asymptomatic; only 24 (27%) patients developed symptoms, including jaundice and hepatomegaly. There were no cases of fulminant hepatitis. Sixty per cent of the patients still had elevated serum ALT levels 1 year after the onset of hepatitis. The 96 patients with hepatitis had received a mean of 9.6 blood units, as compared to a mean of 6.7 units for the unaffected patients (p < 0.001). This study demonstrated that non-A, non-B hepatitis remains a common and important complication of blood transfusion despite screening of blood donors for HBsAg and elevated serum ALT levels.
Spontaneous reactivation of hepatitis B in Chinese patients with Hbsag-positive chronic active hepatitis
Authors: Tong Myron J. , Sampliner Richard E. , Govindarajan Sugantha , Co Ruth L.
Published: March 2007

Eleven patients of Chinese origin experienced spontaneous reactivation of chronic active hepatitis B. Eight HBsAg-positive patients were followed for an average of 15 months prior to, while three others presented during reactivation. Fatigue, hepatomegaly and jaundice were frequent findings. Elevation of both serum ALT (average = 1,212 units per liter) and hepatitis B virus DNA levels were noted in all patients, and reactivation lasted an average of 4.4 months. During resolution, clinical symptoms abated, serum ALT levels reverted toward normal, and in nine patients, the hepatitis B virus DNA values became undetectable. All patients lacked evidence for acute hepatitis A, Epstein-Barr Virus, cytomegalovirus or hepatitis delta virus infection

Histologic findings of liver tissue from eight patients showed piecemeal necrosis and fibrosis. Within the parenchyma, varying degrees of hepatocytolysis with cuffing, perivenular necrosis and acidophilic bodies were noted. Ground-glass cells and regenerative changes also were observed. Cirrhosis was not present in any of the liver biopsies

These findings suggest that spontaneous reactivation of hepatitis B occurs in heterosexual patients with chronic active hepatitis B and contributes to chronic inflammation and to the progression of their liver disease.
Evidence for the cell-surface localization of antigens cross-reacting with the “mitochondrial antibodies” of primary biliary cirrhosis
Authors: Ghadiminejad Iraj , Baum Harold
Published: March 2007

Studies with subfractions of Saccharomyces cerevisiae obtained by differential centrifugation showed only two primary biliary cirrhosis-specific antigens. These antigenic species were shown, using preabsorption studies, to have determinants cross-reactive with their mammalian counterpart

Distribution profiles of marker enzymes and primary biliary cirrhosis antigens between sucrose density gradient subcellular fractions of yeast showed that a relatively high concentration of primary biliary cirrhosis-specific antigens was associated with fractions containing plasma membranes, as well as those containing mitochondria. The possible cell-surface localization of the primary biliary cirrhosis antigens was further investigated using an indirect immunofluorescent technique on a number of different mammalian cells. Rat hepatoma cells, isolated rat hepatocytes and human polymorphonuclear leukocytes and lymphocytes stained positively with primary biliary cirrhosis sera, but not with normal sera or primary biliary cirrhosis sera preabsorbed with beef heart mitochondria. However, blood cells from primary biliary cirrhotic patients gave positive immunofluorescence in all tests, which is compatible with prior binding of the patients' own antimi-tochondrial antibodies to the surface of the cells.