bluuebunny/crossref_metadata_2025 huggingface dataset:
Quantitative evaluation of parenchymal liver cell volume and total hepatocyte number in cirrhotic patients
Authors: Imamura Hiroshi , Kawasaki Seiji , Shiga Junji , Bandai Yasutsugu , Sanjo Kensho , Idezuki Yasuo
Published: March 2007

A simple morphometrical method was developed using a color image analyzing system, which allowed quantitative evaluation of parenchymal liver cell volume and total hepatocyte number in cirrhotic patients. With this method, we estimated these values in 29 cirrhotic patients who underwent hepatic resection (nine cases) or nonshunting operation (20 cases). Liver volume, calculated from computed tomographic images, was 976 ± 196 cm3 (range = 602 to 1,376 cm3); the parenchymal cell volume ratio, obtained based on liver histological appearance with silver stain, was 0.665 ± 0.092 (range = 0.510 to 0.881); and the parenchymal cell volume, calculated by multiplying the liver volume with the parenchymal cell volume ratio, was 645 ± 140 cm3 (range = 403 to 936 cm3). The total hepatocyte number obtained in a similar manner using hematoxylin-stained specimens was 1.72 ± 0.56 × 1011 (range = 0.74 to 2.94 × 1011). The validity and applicability of the method is discussed, and the data are compared with those reported in other studies.
The lack of relationship between hepatotoxicity and lithocholic-acid sulfation in biliary bile acids during chenodiol therapy in the national cooperative gallstone study
Authors: Fisher Rosemarie L. , Hofmann Alan F. , Converse James L. , Rossi Steven S. , Lan Shu-Ping
Published: March 2007

To test whether hepatotoxicity occurring in National Cooperative Gallstone Study patients was caused by a toxic effect of chenodiol per se or of lithocholate caused by defective sulfation, bile samples were analyzed using a new high-performance liquid chromatography method that measures the proportions of the four individual lithocholate amidates (sulfated and unsulfated lithocholylglycine and lithocholyltaurine) and all common bile acid amidates. Samples were obtained from National Cooperative Gallstone Study patients (n = 17) with abnormal light microscopic liver biopsy results or major aminotransferase elevations and from a matched control group of patients (n = 14) who received similar chenodiol doses but had no evidence of liver injury. Bile samples from 45 healthy subjects were also analyzed. The analytical method was validated by showing that the percentage of chenodiol and cholic and deoxycholic acid obtained by highperformance liquid chromatography correlated highly (r > 0.94) with previous gas-liquid chromatography analyses of these samples by the National Cooperative Gallstone Study Reference Laboratory. No significant differences were seen between gallstone patients with and without evidence of liver injury for percent total lithocholate amidates, percent sulfated or unsulfated lithocholate amidates or percent chenodiol amidates. Lithocholate was partially sulfated in all bile samples (52% ± 17% [mean ± S.D., n = 50]), but the extent of sulfation varied widely between and within patients during the course of therapy. Mean values of healthy subjects were similar and also showed a wide range in the extent of lithocholate sulfation. It is concluded that (a) liver injury caused by these doses of chenodiol could not be attributed to the accumulation of unsulfated lithocholate per se in circulating bile acids; (b) liver injury appeared to be, directly or indirectly, caused by enrichment in circulating bile acids with chenodiol or chenodiol together with lithocholate, suggesting that the hepatocytes of those patients with hepatotoxicity were injured by the change induced in bile-acid metabolism by the feeding of chenodiol; and (c) about half of lithocholate amidates in bile samples were sulfated, but the extent of sulfation was highly variable both in gallstone patients and healthy subjects.
Histological and morphometrical indicators for a biopsy diagnosis of well-differentiated hepatocellular carcinoma
Authors: Nagato Yoshinobu , Kondo Fukuo , Kondo Yoichiro , Ebara Masaaki , Ohto Masao
Published: March 2007

Among 597 patients with nodular hepatic lesions who underwent ultrasonically guided needle biopsy, 305 were histologically confirmed as having hepatocellular carcinoma, and 37 patients had borderline lesions. Histological reexamination was correlated with morphometrical analysis on selected cases of well-differentiated, microtrabecular hepatocellular carcinomas (n = 29), borderline lesion (n = 10), typical (mid-sized and macrotrabecular) hepatocellular carcinomas (n = 15) and cirrhotic liver tissue obtained from extranodular hepatic parenchyma of hepatocellular carcinoma patients (n = 47). Morphometrical analyses revealed that the mean cell size and nucleocytoplasmic ratio were most useful for distinguishing well-differentiated, microtrabecular hepatocellular carcinoma from cirrhosis. These two parameters were well correlated with nuclear density. The grade of nuclear density, therefore, seemed to be a convenient semiquantitative indicator for diagnosing well-differentiated hepatocellular carcinoma. A comparison between intranodular and extranodular hepatic tissues was particularly important for its assessment. It is concluded from the results that hepatic nodules presenting a nuclear density larger than two times that of controls could be classified into the overt hepatocellular carcinoma group. From the statistical aspect, the possibility of microtrabecular hepatocellular carcinoma should be considered when a nodule has a nuclear density exceeding 1.3 times that of the extranodular tissue.
Natural killer activity of human liver-derived lymphocytes in various liver diseases
Authors: Hata Kensaku , van Thiel David H. , Herberman Ronald B. , Whiteside Theresa L.
Published: March 2007

Liver-derived lymphocytes were isolated from 40 human livers with end-stage liver disease that were removed at the time of orthotopic liver transplantation. In addition, 10 resection specimens or whole livers removed from patients with liver cancer and seven normal livers (unused donor organs) were studied as controls. Liver-derived lymphocytes were isolated from enzymatically digested tissue by gradient centrifugation and adherence to plastic. Their phenotypical characteristics were studied by two-color flow cytometry, and effector cell function was determined in 4-hr 51Cr-release assays against a natural killer-sensitive target, K562 (natural killer activity), natural killer-resistant Daudi line (lymphokine-activated killer activity) and by P815 line with or without phytohemagglutinin to assess lectin-dependent cellular cytotoxicity. Liver-derived lymphocytes isolated from normal liver contained equal proportions of T and natural killer lymphocytes (mean natural killer/T ratio = 0.7). CD3−CD56+ CD16− natural killer cells were the main natural killer subset present in liver-derived lymphocytes, in contrast to the predominant natural killer phenotype in the circulation (CD56+ CD16+). Control liver-derived lymphocytes had levels of cytotoxicity significantly greater than those of the normal peripheral-blood lymphocytes against all three targets. In contrast, liver-derived lymphocytes isolated from organs with advanced liver disease differed markedly in the natural killer/T cell ratio and levels of liver-derived lymphocyte cytotoxicity. Liver-derived lymphocytes obtained from hepatocellular carcinoma or rejecting allografts treated by immunosuppressive therapy had virtually no cytotoxicity. The highest levels of cytotoxicity, including spontaneous lymphokine-activated killer activity (which is not normally detectable in freshly isolated peripheral-blood lymphocytes) were observed in liver-derived lymphocytes obtained from patients with non-A, non-B hepatitis and metastatic liver cancer. These liver-derived lymphocytes also had the highest natural killer/T cell ratio of all liver-derived lymphocytes in patients with liver disease studied. Alterations in natural killer numbers and function of liver-derived lymphocytes, which are seen in various liver diseases, may reflect different pathological processes, disease origins or local activation states. Functionally active natural killer cells are present in the liver even in cases of end-stage disease and may play a pathophysiological role in viral and neoplastic liver disease.
Accurate prediction of death by serial determination of galactose elimination capacity in primary biliary cirrhosis: A comparison with the mayo model
Authors: Reichen Jürg , Widmer Thomas , Cotting Jacques
Published: March 2007

We retrospectively analyzed the predictive accuracy of serial determinations of galactose elimination capacity in 61 patients with primary biliary cirrhosis. Death was predicted from the time that the regression line describing the decline in galactose elimination capacity vs. time intersected a value of 4 mg · min−1· kg−1. Thirty-one patients exhibited decreasing galactose elimination capacity; in 11 patients it remained stable and in 19 patients only one value was available. Among those patients with decreasing galactose elimination capacity, 10 died and three underwent liver transplantation; prediction of death was accurate to 7 ± 19 mo. This criterion incorrectly predicted death in two patients with portal-vein thrombosis; otherwise, it did better than or as well as the Mayo clinic score. The latter was also tested on our patients and was found to adequately describe risk in yet another independent population of patients with primary biliary cirrhosis. Cox regression analysis selected only bilirubin and galactose elimination capacity, however, as independent predictors of death. We submit that serial determination of galactose elimination capacity in patients with primary biliary cirrhosis may be a useful adjunct to optimize the timing of liver transplantation and to evaluate new pharmacological treatment modalities of this disease.
Sodium retention and hepatic function after two-thirds hepatectomy in the rat
Authors: Ohno Toshiyuki , Sabra Ramzi , Branch Robert A.
Published: March 2007

Recently it was suggested that the onset of sodium retention in experimental cirrhosis in rats is related to a critical threshold of hepatic function, as assessed by the aminopyrine breath test. The aim of this study was to evaluate whether sodium retention occurred after two-thirds hepatectomy in rats and to investigate the relationship between sodium retention and changes in hepatic function associated with liver regeneration in this model. Sodium balance, creatinine clearance, serum sodium and the aminopyrine rate constant of elimination were evaluated daily for 4 days after surgery in partially hepatectomized (n = 6) and shamoperated rats (n = 6). All rats in the partial hepatectomy group exhibited sodium retention (sodium balance >0.7 mmol/day) 24 hr after surgery. This was associated with a 62% reduction of the aminopyrine rate constant of elimination. Spontaneous natriuresis, which occurred between 2 and 4 days after surgery, was associated with an increase in the aminopyrine rate constant of elimination (from 0.73 ± 0.02 × 10−2 min−1 on the last day of sodium retention to 0.95 ± 0.04 × 10−2 min−1 on the first day of natriuresis [p < 0.05]). In contrast, no change in creatinine clearance occurred over the same period. A negative curvilinear association was found between sodium balance and the aminopyrine rate constant of elimination in all animals (r = −0.72, p < 0.001). These observations indicate that natriuresis is related to the recovery of liver function, not to changes in creatinine clearance. In conclusion, the concept of a critical threshold of liver function below which sodium retention occurs has been substantiated in this model of hepatic dysfunction.
Effect of cyclosporine and steroids on canine bile flow
Authors: Brems John J. , Reese Jeffery , Kane Robert , Kaminski Donald L.
Published: March 2007

Cyclosporine and hydrocortisone are the main immunosuppressants used in transplant surgery. The purpose of this study was to determine the effect of intravenous administration of cyclosporine and hydrocortisone on bile flow in dogs. Cyclosporine in doses of 0.5, 1.0 and 1.5 mg · kg−1 · hr−1 and hydrocortisone in doses of 1.25, 2.5 and 5 mg · kg−1 · hr−1 were administered along with 18 μmol/min intravenous sodium taurocholate to dogs with chronic biliary and gastric fistulas. Bile volume and bile chloride concentration and output were increased by cyclosporine in a dose-related manner, whereas bile salt concentration decreased and bile salt output was unchanged. Hydrocortisone produced small but significant increases in bile flow only at the highest dose of hydrocortisone administered. Subsequently, experiments were performed when sodium taurocholate was administered in progressively increasing doses (9, 18 and 36 μmol/min), with the dose changed every hour. Bile volume, [14C]erythritol clearance in bile and bile salt concentrations were measured with and without cyclosporine and hydrocortisone administration. Cyclosporine increased the bile salt-independent fraction of canalicular bile flow and ductular bile flow. Experiments evaluating the role of the cyclosporine carrier polyoxyethylated castor oil (Cremophor EL) demonstrated that this substance had no independent choleretic activity, whereas cyclosporine dissolved in ethanol and administered without Cremophor EL significantly increased bile flow. The results of this study indicate that cyclosporine stimulates chloride-rich choleresis independent of bile salt secretion.
Inhibition of glutathione synthesis in the liver leads to S -adenosyl-L-methionine synthetase reduction
Authors: Corrales Fernando , Ochoa Pilar , Rivas Carmen , Martin-Lomas Manuel , Mato José M. , Pajares Maria A.
Published: March 2007

The hepatic levels of glutathione in rats treated with buthionine sulfoximine (4 mmol/kg), an inhibitor of glutathione synthesis, were 72.5% ± 4.9% of those determined in control animals. This decrease in glutathione concentration was prevented by the administration of glutathione monoethyl ester (7.5 mmol/kg). S-Adenosyl-L-methionine-synthetase activity in the liver of rats treated with buthionine sulfoximine was 39.4% ± 6.5% of that determined in control animals. Again, glutathione monoethyl ester prevented the effect of buthionine sulfoximine on S-adenosyl-L-methionine-synthetase activity. There was a close correlation (r = 0.936) between the hepatic levels of glutathione and S-adenosyl-L-methionine-synthetase activity. The hepatic concentration of S-adenosyl-L-methionine in buthionine sulfoximine-treated animals was 59.7% ± 3.7% of that measured in control rats. Contrasting with the protective effects mentioned above, glutathione monoester had no preventive action on buthionine sulfoximine-induced S-adenosyl-L-methionine depletion. Electron microscopic examination of liver samples of rats after buthionine sulfoximine administration showed evidence of liver degeneration, which was attenuated by glutathione monoethyl ester treatment. Glutathione (7.5 mmol/kg) treatment was less effective than glutathione monoethyl ester in attenuating buthionine sulfoximine effects on hepatic S-adenosyl-L-methionine metabolism and morphology. The reduction of S-adenosyl-L-methionine-synthetase activity observed after treatment with buthionine sulfoximine and its prevention by glutathione monoethyl ester, as well as the correlation between the activity of this enzyme and glutathione levels, indicate that glutathione plays an important role in maintaining S-adenosyl-L-methionine-synthetase activity in the liver.
Vanadate inhibits glucose output from isolated perfused rat liver
Authors: Bruck Rafael , Prigozin Haia , Krepel Zipora , Rotenberg Paul , Shechter Yoram , Bar-Meir Simon
Published: March 2007

Previous studies have demonstrated that vanadate ions mimic many of the actions of insulin in in vitro systems. Also, vanadate administered to diabetic hyperglycemic rats lowers their blood glucose levels to normal values. In this study we demonstrate that vanadate inhibits glucose output in the isolated perfused rat liver. Glucose production was suppressed maximally (about 50% to 60%), on addition of extremely low vanadate ion concentrations (0.5 to 1 μmol/L). This concentration is about two log units lower than the vanadate ion concentrations that are required to activate hexose uptake and glucose metabolism in vitro and is within the range of endogenous intracellular vanadium concentration. Insulin had little or no effect in inhibiting hepatic glucose output in this experimental system. The effect of vanadate ions is rapid in onset and is not accompanied by any signs of liver toxicity as assessed by various criteria. In conclusion, the study indicates that (a) vanadate ions inhibits hepatic glucose output, maximally and at extremely low, nontoxic concentrations (ID50 = 0.7 ± 0.1 μmol/L). (b) The modulation action of the ion is fast and probably occurs at point(s) distal to the insulin receptor itself. (c) The liver participates in the process of maintaining euglycemia in diabetic rats receiving optimal doses of vanadate orally.
Is intrapulmonary arteriovenous shunting and hypoxemia a contraindication for liver transplantation?
Authors: Eriksson Siw L.
Published: March 2007

A 12-year-old boy with Wilson’s disease developed exertional dyspnea, cyanosis, and finger clubbing 10 months after diagnosis. The hypoxemia was caused by arteriovenous shunting, demonstrated by radionuclide scanning and pulmonary arteriography. Orthotopic liver transplantation was performed after the development of severe hypoxemia. There was no apparent reversal of the intrapulmonary arteriovenous shunting and he died 10 days posttransplantation of multiple organ failure secondary to hypoxemia. Monitoring arterial oxygen saturation in children with cirrhosis is warranted since the presence of significant arteriovenous shunting may influence prognosis and decisions regarding liver transplantation.
Antibodies to hepatitis B virus and hepatitis C virus in alcoholic hepatitis and cirrhosis: Their prevalence and clinical relevance
Authors: Mendenhall Charles L. , Seeff Leonard , Diehl Anna Mae , Ghosn Saad J. , French Samuel W. , Gartside Peter S. , Rouster Susan D. , Buskell-Bales Zelma , Grossman Charles J. , Roselle Gary A. , Weesner Robert E. , Garcia-Pont Pedro , Goldberg Stephen J. , Kiernan Thomas W. , Tamburro Carlo H. , Zetterman Rowen , Chedid Antonio , Chen Thomas , Rabin Lionel
Published: March 2007

Patients with overt alcoholic liver disease who had participated in a multicenter therapeutic trial and subgroups of controls (i.e., alcoholic patients without liver disease and patients with neither alcoholism nor liver disease) were tested for hepatitis B virus and hepatitis C virus antibodies to determine the prevalence of these antibodies and any clinical association in the progression and outcome of alcoholic liver disease. Antibodies to hepatitis B (anti-HBs and/or anti-HBc) were found in 29.2% of patients with alcoholic liver disease, in 26.1% of hospitalized alcoholic patients without liver disease and in 24.2% of hospitalized nonalcoholic patients without liver disease; frequencies were not significantly different from one another. HBsAg was not evaluated because HBsAg+ patients had been excluded from the original trial. The presence of these antibody markers correlated with ethnic origin of and immunoglobulin levels in the patients. In contrast, antibody to hepatitis C, as detected by enzyme immunoassay, was positive in 27.1%, 4.8% and 3.0% of the three groups, respectively, the first differing significantly from the other two. Antibody to hepatitis C virus positivity correlated significantly with clinical severity of the disease and with the presence of histological features that imply chronic viral infection (periportal inflammation, cirrhosis), despite the fact that the supplementary assay for antibody to hepatitis C virus, using recombinant immunoblot assay, reduced the positive rate by 79%. Although the presence of hepatitis B antibodies did not correlate with patient survival, some categories of patients with antibody to hepatitis C virus did; the survival of those with antibody to hepatitis C virus and recombinant immunoblot assay reactivity was 67%; for those with antibody to hepatitis C virus positivity but recombinant immunoblot assay negativity the survival was 29% (p < 0.01). The reason for the poor prognosis for those with antibody to hepatitis C virus positivity and recombinant immunoblot assay negativity is not clear, although some theories are offered. (Hepatology 1991;14:581-589.)
Molecular forms and biological activity of atrial natriuretic factor in patients with cirrhosis and ascites
Authors: Jiménez Wladimiro , Gutkowska Jolanta , Ginés Pere , Arroyo Vicente , Rivera Francisca , Rodés Joan
Published: March 2007

Patients with cirrhosis and ascites show sodium retention and normal or increased plasma levels of atrial natriuretic factor, a peptide with powerful natriuretic activity. To investigate whether this paradoxical observation could be related to a dysregulation in the process of synthesis and maturation of atrial natriuretic factor leading to abnormal molecular forms lacking biological activity, the chromatographic patterns of atrial natriuretic factor contained in plasma extracts from 10 patients with cirrhosis and ascites and 6 healthy subjects were compared. Atrial natriuretic factor from cirrhotic patients was also tested in two different radioreceptor assays, which detect the biologically active form(s) of this peptide. Patients with cirrhosis and ascites had higher plasma levels of atrial natriuretic factor (81.3 ± 8.5 pg/ml, p < 0.001) than control subjects (29.8 ± 3.2 pg/ml). High-performance liquid chromatography analysis of atrial natriuretic factor showed an identical chromatographic pattern in cirrhotic patients and control subjects. Three peaks related to the atrial natriuretic factor prohormone were observed in cirrhotic patients and control subjects, accounting for 64%, 23% and 11% of the total atrial natriuretic factor in cirrhotic patients and 63%, 18% and 8% of the total atrial natriuretic factor in control subjects. The main peak eluted at the same position of synthetic human atrial natriuretic factor (Ser 99-Tyr 126), which represents the major active form of the circulating hormone. Cirrhotic atrial natriuretic factor displayed the same ability to inhibit the binding of 125I-atrial natriuretic factor to rat glomerular and bovine adrenal membrane receptors as synthetic human atrial natriuretic factor. In conclusion this study demonstrates that atrial natriuretic factor of patients with cirrhosis and ascites has an equipotent binding activity to its receptor as to that of synthetic human atrial natriuretic factor and possesses the same molecular weight and biologically active forms as atrial natriuretic factor of normal subjects. These data indicate that in cirrhosis there is no dysregulation in the atrial natriuretic factor maturation process. (Hepatology 1991;14:601-607.)
Estrogen and progesterone receptors in the gallbladders from patients with gallstones
Authors: Ranelletti Franco O. , Piantelli Mauro , Zanella Everardo , Capelli Arnaldo , Farinon Attilio M.
Published: March 2007

Cytosolic receptors for estrogen and progesterone were assayed in 50 gallbladders from 29 women and 21 men who had cholecystectomies because of choleithiasis. High-affinity (equilibrium dissociation constant, KD = 0.46 ± 0.23 nmol/L of 24 cases) estrogen receptors were detected in 20 of 29 gallbladders from women (range = 1.6 to 32 fmol/mg protein; mean ± S.D. = 10.9 ± 8.1), whereas in men only 4 of 21 specimens contained detectable estrogen receptors. High-affinity (KD = 0.45 ± 0.17 nmol/L; mean ± S.D. of 41 cases) progesterone receptors were found in 25 of 29 gallbladders of women (range = 2 to 62 fmol/mg protein; mean ± S.D. = 19.2 ± 14.4) and in 16 of 21 gallbladders of men (range = 4 to 36 fmol/mg protein; mean ± S.D. = 12.5 ± 8.5). There is a statistically significant difference between men and women in the proportion of estrogen receptor-positive gallbladders, 19% and 69% for men and women, respectively. Progesterone receptors are present in similar proportion in the gallbladders of men (72.2%) and women (86.2%). In women a positive correlation between estrogen and progesterone receptors was found. Even in the absence of estrogen receptors, the gallbladders of men express progesterone receptors at levels similar to those observed in women. At least in cholelithiasic gallbladders, this suggests that a sex difference exists in the coexpression of estrogen and progesterone receptors. Such a difference could be related with the higher gallstone incidence in women than in men. (Hepatology 1991;14:608-612.)
Extracellular ATP, intracellular calcium and canalicular contraction in rat hepatocyte doublets
Authors: Kitamura Tsuneo , Brauneis Ulrike , Gatmaitan Zenaida , Arias Irwin M.
Published: March 2007

Bile-canaliculus contraction in rat hepatocyte doublets is postulated to involve activation of an actinmyosin system. We examined this hypothesis by determining the relationship between canalicular contraction and cystolic free Ca2+ ([Ca2+]i) concentration after extracellular addition of ATP or microdialysis of myosin light chain kinase or its Ca2+-independent fragment, which retains catalytic activity. After incubation of doublets with 200 μmol/L ATP in the absence of extracellular Ca2+, [Ca2+]i peaked at 40 sec and 71% of canaliculi contracted within 4 min. Decreasing effects were observed with equimolar ADP, AMP and nonhydrolyzable ATP, but no effect was observed with adenosine. The effect of extracellular ATP on [Ca2+]i and canalicular contraction was dose dependent. Addition of extracellular Ca2+ and ATP resulted in a plateau level of [Ca2+]i. Cytochalasin D, which depolymerizes actin filaments, inhibited ATP-induced canalicular contraction, but not the increase in [Ca2+ Microdialysis of myosin light chain kinase and its Ca2+-independent fragment (but not the heatdenatured fragment, albumin, trypsin plus soybean inhibitor or buffer) into one hepatocyte of a doublet resulted in canalicular contraction in 86% of doublets. Injection of myosin light chain kinase or its Ca2+-independent fragment did not increase [Ca2+]i within 5 min. These results indicate that (a) the basolateral plasma membrane of hepatocytes has a P2Y-class purinoceptor, (b) increased [Ca2+]i after incubation with ATP is initially due to mobilization from internal sites and (c) canalicular contraction is directly related to [Ca2+]i and activation of an actin-myosin system. The physiological role of extracellular ATP in canalicular contraction is uncertain. (Hepatology 1991;14:640-647.)
Role of extracellular zinc and copper on metallothionein regulation in cultured rat hepatocytes
Authors: Hidalgo Juan , Dingman Allen , Garvey Justine S.
Published: March 2007

Cellular and extracellular metallothionein contents of rat hepatocytes cultured in the presence of albumin (30 μmol/L) with zinc (1, 10, 50 and 100 μmol/L), copper (1, 10 and 50 μmol/L), zinc and copper (1, 10 and 50 μmol/L of each metal) or no metals in the culture medium have been measured by radioimmunoassay. Cellular metallothionein levels increased steadily with culture time regardless of the metal treatment and showed little dependence (only a twofold increase) on extracellular zinc or copper at all metal concentrations and times (up to 3 days) studied. In contrast, the presence of both metals simultaneously in the culture medium strongly increased cellular metallothionein contents, acting synergistically in some cases. Significant extracellular metallothionein was observed when copper or zinc and copper were present in the culture medium, most of which is likely a consequence of cell leakage because no evidence of physiological secretion was observed. Total metallothionein production (cellular and extracellular metallothionein levels) indicated that copper was a better metallothionein inducer than zinc in these experimental conditions. These results indicate that metallothionein regulation in the hepatocyte is different depending on the extracellular metal levels and composition and that attention must be given to metallothionein release from the hepatocyte. (Hepatology 1991;14:648-654.)
ATP-dependent taurocholate transport by rat liver canalicular membrane vesicles
Authors: Adachi Yukihiko , Kobayashi Hiroaki , Kurumi Yoshiaki , Shouji Mika , Kitano Motokazu , Yamamoto Toshio
Published: March 2007

We conducted an experimental study to examine the possibility that ATP is involved in the mechanism by which bile acids are excreted through the liver canalicular membrane in opposing the concentration gradient. Canalicular membrane vesicles were purified from the livers of Sprague-Dawley rats, and the uptake of tritiated sodium taurocholate into canalicular membrane vesicles was determined by rapid filtration technique. Vesicle-associated sodium taurocholate was increased in the presence of ATP and ATP-regenerating system. This was also observed at a voltage-clamped condition. ATP-dependent uptake into the osmotically reactive intravesicular space was saturated with increasing concentrations of sodium taurocholate (Km = 47 μmol/L, Vmax = 270 pmoles/mg protein · 20s). ATP-dependent uptake increased to the point of saturation when the sodium taurocholate concentration was 50 μmol/L and the ATP concentration was increased from 0 to 1 mmol/L (Km = 64 μmol/L). Among the several nucleotides used, ATP was a potent stimulator of transport, whereas a nonhydrolyzable analogue (i.e., adenosine 5′-[β,γ-imino]triphosphate) showed no effect. In addition, ATP-dependent transport was inhibited by vanadate in a dose-dependent manner. From these results it was concluded that the primary active transport of sodium taurocholate is present in hepatocellular canalicular membranes. This transport is directly dependent on ATP, and hydrolysis of γ-phosphate of ATP is required. (Hepatology 1991;14:655-659.)
Dietary fish oil effects on biliary lipid secretion and cholesterol gallstone formation in the African green monkey
Authors: Scobey Martin W. , Johnson Fred L. , Parks John S. , Rudel Lawrence L.
Published: March 2007

We studied two groups of adult male African green monkeys to assess the effects of dietary fish oil on biliary lipid secretion and cholesterol gallstone formation. One group was fed 0.8 mg cholesterol/kcal and 42% of calories as fat with half of the fat calories derived from lard; the other group was fed a similar diet except for the isocaloric substitution of menhaden oil for lard. After 21/2 to 3 yr, necropsies were performed and the presence of gallstones was determined. Gallbladder bile specimens were analyzed for cholesterol saturation index, as well as for bile acid species and biliary phospholipid fatty acid composition. Our results showed that 67% of animals fed the lard diet had cholesterol gallstones compared with only 22% of animals in the group fed the fish oil diet (p = 0.08). The cholesterol saturation index of gallbladder bile also tended to be higher in the lard-fed group (1.15 ± 0.11) compared with the fish oil-fed group (0.86 ± 0.09, p = 0.06). No differences between the two dietary groups were noted in the percentages of the various types of bile acids. However, a greater percentage of ω-3 fatty acids and a lesser percentage of 18:1, 18:2 and 20:4 fatty acids were found in the biliary phospholipids from the fish oil-fed group compared with the lard-fed group. Biliary lipid secretion rates were then measured during isolated recirculating liver perfusion performed with a constant sodium taurocholate infusion. A significantly greater biliary phospholipid secretion rate was noted in the fish oil-fed animals compared with the lard-fed animals, whereas bile acid and cholesterol secretion rates were similar between the diet groups. We conclude that the increased biliary phospholipid secretion observed in the fish oil-fed group produced a less lithogenic gallbladder bile, which in turn resulted in a less frequent occurrence of gallstones in that group. In addition the increased phospholipid secretion was probably favored by the enrichment of biliary phospholipids with relatively less hydrophobic ω-3 fatty acids. (Hepatology 1991;14:679-684.)
Hepatitis C: Improving the diagnostic armamentarium
Authors: Gumucio Jorge J. , Martin Paul
Published: March 2007

A new four-antigen recombinant immunoblot assay (4-RIBA) for confirmation of hepatitis C virus (HCV) C-100 enzyme-linked immunosorbent assay (ELISA) reactivity was tested in stored serum samples (1984-86) of blood donors and recipients and compared with results from polymerase chain reaction (PCR) analysis of fresh (1990) plasma samples in donors and recipients from the original study. Of 37 HCV C-100 ELISA-positive blood products, 8 were 4-RIBA positive, of which 7 were implicated in post-transfusion non-A, non-B hepatitis (PT-NANBH) and/or PCR confirmed recipient HCV infection. Of 9 recipients with PTNANBH, 8 were reactive in 4-RIBA (6 positive and 2 indeterminate). With fresh plasma samples, 3 donors and 6 recipients who were 4-RIBA positive were also PCR positive. 4 4-RIBA indeterminate and 78 4-RIBA negative samples of donors and recipients were PCR negative. Of 6 4-RIBA positive recipients, 5 were PCR positive four to six years later. 1.6% of the 383 recipients became chronically infected with HCV. The new 4-RIBA represents a candidate confirmation test to discriminate between infective and non-infective HCV C-100 ELISA-positive blood donors.
Transforming growth factor B and hepatic fibrosis: Cause or effect?
Authors: Brenner David A.
Published: March 2007

Background.

Cirrhosis is a diffuse process of hepatic fibrosis and regenerative nodule formation of unknown pathogenesis. Transforming growth factor (TGF) β1 induces the production of extracellular matrix proteins by liver cells and has been implicated in the pathogenesis of hepatic fibrosis in laboratory animals. TGFα is a hepatocyte mitogen that participates in liver regeneration.

Methods.

Using Northern blot analysis, we studied the expression of TGFβ1 messenger RNA (mRNA) in liver specimens from 42 patients with chronic hepatitis and cirrhosis and 12 subjects with either normal or fatty livers. The results were correlated with measurements of procollagen Type I mRNA in liver tissue, procollagen Type III peptide in serum, and the degree of histologic injury. We also investigated whether TGFα mRNA would be detectable in biopsy specimens of livers with proliferative activity.

Results.

TGFβ1 mRNA expression correlated closely with the expression of procollagen Type I mRNA (r = 0.94) and serum procollagen Type III peptide (r = 0.89) and with the histologic activity index (r = 0.73). All patients with increased fibrogenic activity (serum procollagen Type III peptide level, > 11.9 μg per liter) had increased levels of TGFβ1 mRNA (2 to 14 times the levels in the control group or in patients with normal fibrogenic activity), and both TGFα and H3 histone (a marker of DNA synthesis) mRNAs were detectable in patients with regenerative nodules. Six of eight patients with hepatitis C treated with interferon alfa for one year had sustained clinical responses with normalization of serum procollagen Type III peptide and aminotransferase activity. All these patients had normal levels of TGFβ1 mRNA in liver specimens obtained at the end of the year.

Conclusions.

TGFβ1 may have an important role in the pathogenesis of fibrosis in patients with chronic liver disease, and TGFα expression may be associated with liver regeneration in these patients.
Focusing in on P53 in hepatocellular carcinoma
Authors: Carbone David
Published: March 2007

Human hepatocellular carcinomas (HCC) from patients in Qidong, an area of high incidence in China, in which both hepatitis B virus and aflatoxin B1 are risk factors, were analysed for mutations in p53, a putative tumour-suppressor gene. Eight of the 16 HCC had a point mutation at the third base position of codon 249. The G→T transversion in seven HCC DNA samples and the G→C transversion in the other HCC are consistent with mutations caused by aflatoxin B1 in mutagenesis experiments. No mutations were found in exons 5, 6, 8 or the remainder of exon 7. These results contrast with p53 mutations previously reported in carcinomas and sarcomas of human lung, colon, oesophagus and breast; these are primarily scattered over four of the five evolutionarily conserved domains, which include codon 249. We suggest that the mutant p53 protein may be responsible for a selective clonal expansion of hepatocytes during carcinogenesis.

Bressac B, Kew M, Wands J, Ozturk M. Selective G to T mutations of p53 gene in hepatocellular carcinoma from southern Africa. Nature 1991;350:429-431.

Hepatocellular carcinoma (HCC) is a prevalent cancer in sub-Saharan Africa and eastern Asia. Hepatitis B virus and aflatoxins are risk factors for HCC, but the molecular mechanism of human hepatocellular carcinogenesis is largely unknown. Abnormalities in the structure and expression of the tumour-suppressor gene p53 are frequent in HCC cell lines, and allelic losses from chromosome 17p have been found in HCCs from China and Japan. Here we report on allelic deletions from chromosome 17p and mutations of the p53 gene found in 50% of primary HCCs from southern Africa. Four of five mutations detected were G→T substitutions, with clustering at codon 249. This mutation specificity could reflect exposure to a specific carcinogen, one candidate being aflatoxin B1, a food contaminant in Africa, which is both a mutagen that induces G to T substitution and a liver-specific carcinogen.
Failure to detect hepatitis C virus genome in human secretions with the polymerase chain reaction
Authors: Hsu Henry H. , Wright Teresa L. , Luba Daniel , Martin Mary , Feinstone Stephen M. , Garcia Gabriel , Greenberg Harry B.
Published: March 2007

Although hepatitis C infection has been clearly demonstrated to be transmitted through blood products or blood contamination, most cases of sporadic hepatitis C infection are unassociated with parenteral risk factors, and it is unclear how infection might be acquired by nonparenteral means. One potential mode of nonparenteral transmission is through body secretions. We used a highly sensitive and specific polymerase chain reaction assay to determine whether hepatitis C viral genomic RNA could be detected in secretions obtained from nineteen individuals with chronic hepatitis C virus infection. Although hepatitis C genomic RNA was found in all 19 sera, hepatitis C virus RNA was not detected in any samples of saliva, semen, urine, stool or vaginal secretions from these patients. Viral titers in serum ranged from 102 to 107 polymerase chain reaction units/ml. The sensitivity of our polymerase chain reaction assay indicates that, if hepatitis C virus were in secretions, it would be present in amounts less than 1 to 4 polymerase chain reaction units/ml. This contrasts with hepatitis B virus infection, in which serum titers frequently are in excess of 109 copies of hepatitis B genomes/ml. Body secretions have been found to contain up to 106 copies of hepatitis B genomes/ml. Our findings support seroepidemiological studies indicating that nonparenteral transmission of hepatitis C through secretions is uncommon and probably much less efficient than hepatitis B virus infection. (Hepatology 1991;14:763-767).
Gallstones in cystic fibrosis: A critical reappraisal
Authors: Angelico Mario , Gandin Claudia , Canuzzi Pietro , Bertasi Serenella , Cantafora Alfredo , de Santis Adriano , Quattrucci Serena , Antonelli Mariano
Published: March 2007

Radiolucent gallstones are common in young adults with cystic fibrosis. In the mid-1970s, it was suggested that gallstones are made of cholesterol, but this hypothesis has never been tested. Several recent studies have shown that the detection of cholesterol monohydrate crystals in bile has high sensitivity and specificity for the diagnosis of cholesterol gallstones. We therefore used this approach to study 17 young adults with cystic fibrosis, 10 of whom had radiolucent gallstones. The two groups of patients were comparable in age and gender (all patients but one were male). Duodenal bile was obtained after gallbladder contraction with intravenous cerulein; it was used for lipid and protein chemistry studies and for polarizing microscopy. The latter was performed both in whole bile and in the postultracentrifugation (100,000 g) sediment. Bile cholesterol saturation did not significantly differ between patients with (1.21 ± 0.28) or without gallstones (0.99 ± 0.54). Slight cholesterol supersaturation was found in 7 of 10 gallstone and three of seven nongallstone patients. At no time were cholesterol crystals detected in either the group, even after bile ultracentrifugation. Two more cystic fibrosis patients with gallstones died of severe bronchopneumopathy, and small pigment gallstones were obtained at autopsy. At stone analysis, cholesterol content was 44% and 28% of dry weight, respectively. Infrared spectroscopy of stone powder was compatible with the presence of calcium bilirubinate and proteins as major components. We conclude that radiolucent gallstones of cystic fibrosis are not of the conventional cholesterol type. (Hepatology 1991;14:768-775).
Reduced retinoid content in hepatocellular carcinoma with special reference to alcohol consumption
Authors: Adachi Sadashi , Moriwaki Hisataka , Muto Yasutoshi , Yamada Yasuhiro , Fukutomi Yasushi , Shimazaki Makoto , Okuno Masataka , Ninomiya Mitsuo
Published: March 2007

Although alcohol is known to enhance hepatocarcinogenesis, the mechanism of this action remains to be explained. To test the hypothesis that ethanol depletes the liver of antitumor promoters such as retinoid, we measured the retinoid concentration in hepatocellular carcinoma tissues and noncancerous surrounding liver tissues in humans known to have a history of alcohol consumption. By high-performance liquid chromatography, the retinoid contents of 29 surgically resected hepatocellular carcinoma specimens and their noncancerous surrounding tissues were measured. Retinoid contents were decreased in both the cancerous and the surrounding noncancerous liver tissues of patients with a high intake of alcohol. The levels correlated inversely with the estimated cumulative lifetime ethanol consumption. The decrease in the retinoid content of hepatic parenchymal cells paralleled that in stellate cells. When compared with the surrounding liver tissues, the cancerous liver tissues were in the state of retinoid deficiency. In summary, alcohol abuse may help promote the hepatocarcinogenesis in man by depleting the liver of the antitumor promoter, retinoid. (Hepatology 1991;14:776-780).
Detection of proliferating liver cells in various diseases by a monoclonal antibody against DNA polymerase-α: With special reference to the relationship between hepatocytes and sinusoidal cells
Authors: Seki Shuichi , Sakaguchi Hiroki , Kawakita Nobuyoshi , Yanai Atsushi , Kuroki Tetsuo , Mizoguchi Yasuhiro , Kobayashi Kenzo , Monna Takeyuki
Published: March 2007

Proliferating cells in liver specimens from patients with various diseases were detected by use of a monoclonal antibody against human DNA polymerase-α, which is present in the nuclei of cells in the G1, S, M and G2 phases of the mitotic cell cycle and absent in the G0 phase, to clarify the kinetics and morphological characteristics of these cells. This monoclonal antibody was supernatant derived from clone CL222-42B, and the peroxidase antiperoxidase method was used.

Not only epithelial cells (hepatocytes, biliary epithelial cells and hepatocellular carcinoma cells) but also nonepithelial cells (Kupffer cells and other macrophages, endothelial cells, fat-storing cells, lymphocytes and fibroblasts) were stained for DNA polymerase-α. In acute viral hepatitis with confluent necrosis, small hepatocytes with basophilic cytoplasm next to the necrosis accounted for most of the proliferating cells. In these areas, Kupffer cells and other macrophages and lymphocytes had often proliferated. Hepatocellular carcinoma cells were frequently stained for DNA polymerase-α, in addition to endothelial cells, macrophages and lymphocytes. These nonepithelial cells were stained more frequently in specimens with many stained carcinoma cells than in those with only a few cells stained. In fibrotic areas, fibroblasts were often stained for this enzyme. In proliferating bile ducts, both small epithelial cells and large mature cells were stained. The differences between stained and nonstained cells that were not hepatocytes could not be defined by their ultrastructural characteristics. From these findings, it seemed possible that sinusoidal cells, especially Kupffer cells and other macrophages, might be much involved in hepatocytic proliferation during regeneration of the liver and also in the occurrence of malignant tumors. (Hepatology 1991;14:781-788).
In situ immunophenotyping study of endothelial cells of the human hepatic sinusoid: Results and functional implications
Authors: Scoazec Jean-Yves , Feldmann Gérard
Published: March 2007

Hepatic sinusoids are highly specialized capillary vessels characterized by the presence of resident macrophages adhering to the endothelial lining. Although it is likely that sinusoidal endothelial cells have specific adaptations, little is known about the roles that they actually play in vivo. We therefore designed an in situ immunophenotyping study of sinusoidal endothelial cells in normal human liver to compare this population with those of the other vascular compartments of the liver and to determine whether it expresses molecules involved in the following physiological processes: scavenging and nonspecific immune functions, antigen presentation and endothelialleukocyte adhesion. Our study showed that sinusoidal endothelial cells displayed a highly distinctive immunophenotype characterized by the expression of several molecules not found in the other vascular compartments of the liver. A first group of molecules restricted to sinusoidal endothelial cells were: the receptors II and III for the Fc fragment of IgG; the CD14 molecule, serving as a receptor for the lipopolysaccharide-binding protein; and aminopeptidase N. The presence of those molecules suggests that sinusoidal endothelial cells contribute to the scavenger and nonspecific immune functions of hepatic sinusoids. No direct evidence for an antigenpresenting function of this cell population was obtained. Another group of proteins restricted to sinusoidal endothelial cells comprised the leukocyte adhesion molecules CD4 and intercellular adhesion molecule-1, which may be involved in the adhesion of Kupffer cells to the sinusoidal wall. Therefore our immunophenotyping study makes it possible to provide an in situ demonstration of the phenotypical and functional diversity of endothelial cells in normal human liver. The distinctive phenotype of sinusoidal endothelial cells is likely to correlate with specific functional adaptations to their particular microenvironments. (Hepatology 1991;14:789-797).
Azathioprine hepatotoxicity after liver transplantation
Authors: Sterneck Martina , Wiesner Russell , Ascher Nancy , Roberts John , Ferrell Linda , Ludwig Jurgen , Lake John
Published: March 2007

We report the first two cases of apparent azathioprine hepatotoxicity occurring after liver transplantation. The two patients exhibited jaundice, elevated serum transaminase activities and histopathological features of sinusoidal congestion and centrilobular hepatocellular degeneration 17 and 61 days after transplantation. After withdrawal of azathioprine, liver test results improved immediately in both patients. Recurrence of liver injury after another challenge with azathioprine was seen in the first case. Previously, fatal venoocclusive disease occurring after kidney transplantation had been attributed to azathioprine. Irreversible venoocclusive disease and the reversible hepatotoxicity described in this report (i.e., sinusoidal congestion with centrizonal necrosis) most likely represent different stages of hepatic endotheliitis caused by azathioprine. Thus early diagnosis of azathioprine hepatotoxicity is of great importance. (Hepatology 1991;14:806-810).