bluuebunny/crossref_metadata_2025 huggingface dataset:
Inappropriate expression of blood group antigens in hepatic allografts
Authors: Bloom Stuart , Fleming Ken , Chapman Roger , Neuberger James , Hubscher Stefan
Published: March 2007

We examined the expression of blood group antigens of the ABO, Lewis and Kell antigen systems using monoclonal antibodies and immunohistochemical study on 42 liver allograft specimens from 33 patients who underwent liver transplantation between 1986 and 1991 to learn whether altered blood group antigen expression might have a bearing on the immunopathogenesis of transplant rejection. Specimens were obtained at intervals of 0 days to 3 yr after transplant; they yielded the following histological diagnoses; time zero (n = 4), acute rejection (n = 4), pure cholestasis (n = 4), biliary obstruction (n = 4), early chronic rejection (n = 4), end-stage chronic rejection (n = 15) and miscellaneous late posttransplant biopsies (n = 7). Aberrant expression of blood group antigens was observed in 5 of 15 patients with chronic rejection. Two transplants into the same group O patient showed aberrant expression of AB antigens on hepatocytes, with a canalicular pattern, in group O-transplanted livers. In all three cases in which a group O liver was transplanted into a group A recipient and histological signs of chronic rejection were present, antibody staining showed acquisition of recipient blood phenotype by the donor liver bile ducts, endothelium or both. Aberrant expression of ABO antigens was seen only in chronic rejection. In seven cases we noted canalicular staining of periportal hepatocytes with the Lewis antibodies, normally confined to ducts and ductules. This was associated with severe cholestasis in six of the seven cases and may have represented early ductular metaplasia. These changes in carbohydrate cell surface phenotype may play a role in regulation of hepatic allograft susceptibility to immune-mediated damage. (Hepatology 1994;19:876-881.)
Short-term effects of simvastatin on bile acid synthesis and bile lipid secretion in human subjects
Authors: Loria Paola , Bertolotti Marco , Cassinadri Teresa M. , Dilengite Michele A. , Bozzoli Mara , Carubbi Francesca , Concari Mauro , Guicciardi Eugenia M. , Carulli Nicola
Published: March 2007

To test whether de novo synthesis of cholesterol is a limiting factor for bile acid synthesis, we studied the acute effect of simvastatin, an inhibitor of HMG-coenzyme A reductase (the limiting step of cholesterol synthesis) on bile acid synthesis and biliary lipid secretion in subjects with interrupted enterohepatic circulation. In these conditions bile acid synthesis is derepressed and is assumed to equal biliary bile acid secretion. Five cholecystectomized patients fitted with T-tubes were studied. All subjects were administered simvastatin (80 mg as a single dose) 5 days after surgery. Bile was collected in 3-hr intervals for 15 hr before and 30 hr after the administration of the drug. During the experiment we kept the enterohepatic circulation of bile acid interrupted by inflating an occludable balloon inserted, during cholecystectomy, in the common bile duct. Simvastatin induced significant decreases of plasma total and low density lipoprotein cholesterol concentrations, from 163 ± 29 mg/dl and 97 ± 24 mg/dl of the pretreatment value to 144 ± 30 mg/dl and 82 ± 22 mg/dl 18 hr after simvastatin administration, respectively. Bile flow tended to increase after simvastatin, and the mean values from the third to the 15th hour after simvastatin administration (22.1 ± 1.9 ml/hr) were significantly greater than the mean values of the pretreatment period (19.9 ± 2.8 ml/hr). Concomitantly biliary bile acid, cholesterol and phospholipid concentrations fell from basal values of 15.9 ± 5.1, 2.3 ± 0.3 and 5.5 ± 0.3 mmol/L to mean values, after treatment, of 9.0 ± 3.5, 1.9 ± 0.5 and 3.0 ± 0.9 mmol/L, respectively. Cholesterol saturation index increased from a mean value of 1.51 ± 0.31 in the pretreatment period to 1.98 ± 0.52 after simvastatin. Bile acid output decreased from a mean pretreatment value of 308.0 ± 79.1 μmol/hr to 191.9 ± 69.2 μmol/hr after simvastatin administration. Secretion rates of phospholipids decreased to a lesser extent, whereas cholesterol output remained constant. No correlation was found between bile acid output and bile flow, phospholipid secretion and cholesterol secretion. A significant correlation was present between phospholipid and cholesterol secretion. Our data show that, in conditions of derepressed bile acid synthesis, acute inhibition of HMG-coenzyme A reductase activity induces decreased bile acid synthesis and excretion. Our findings may suggest that the availability of newly synthesized cholesterol is a critical factor for bile acid synthesis and secretion but not for cholesterol secretion; alternatively HMG-coenzyme A reductase and cholesterol 7α-hydroxylase, the rate-limiting step of bile acid synthesis, may be coordinately regulated at the transcriptional level. (Hepatology 1994;19:882-888.)
α-Smooth muscle actin-positive perisinusoidal stromal cells in human hepatocellular carcinoma
Authors: Enzan Hideaki , Himeno Hiromi , Iwamura Shinichi , Onishi Saburo , Saibara Toshiji , Yamamoto Yasutake , Hara Hiroshi
Published: March 2007

The purpose of this study is to clarify the morphological characteristics and functional significance of the perisinusoidal stromal cells in hepatocellular carcinoma. The liver specimens surgically resected from 24 patients with hepatocellular carcinoma were studied by electron microscopy and immunohistochemistry using monoclonal antibodies against α-smooth muscle actin, vimentin and desmin. In the tissue space between endothelial cells and trabeculae of cancer cells, the stromal cells were frequently found. They were strongly positive for α-smooth muscle actin, weakly and less frequently positive for vimentin but negative for desmin. They varied in shape, size and distribution, stretching cytoplasmic processes and occasionally surrounding the trabeculae of cancer cells. They contained considerable amounts of microfilaments that were positive for α-smooth muscle actin and condensed in cell periphery. Along the cell membrane, the short dense areas and pinocytotic vesicles were seen. The external lamina incompletely invested the stromal cells. They were always surrounded by amorphous material. In the granulation tissue and fibrotic areas around necrotic cancer tissue, they were increased in size and number. On the other hand, immunohistochemically and ultrastructurally, they closely resembled the Ito cells in the piecemeal necrosis that showed myofibroblastic transformation. These results suggest that the perisinusoidal stromal cells in nonnecrotic cancer tissue produce the extracellular matrix in the tissue space and maintain the cancerous trabecular structure. After necrosis of cancer tissue, they may become activated and actively participate in the fibrosis. (Hepatology 1994;19:895-903.)
Cyclosporine withdrawal for nephrotoxicity in liver transplant recipients does not result in sustained improvement in kidney function and causes cellular and ductopenic rejection
Authors: Sandborn William J. , Hay Eileen J. , Porayko Michael K. , Gores Gregory J. , Steers Jeffery L. , Krom Ruud A. F. , Wiesner Russell H.
Published: March 2007

Twelve consecutive liver transplant recipients with stable allograft function and cyclosporine nephrotoxicity were subjected to cyclosporine withdrawal in an attempt to halt and possibly reverse renal damage. Only patients who met the following criteria were included: (a) triple immunosuppression with cyclosporine, azathioprine and prednisone; (b) stable graft function for at least 1 yr without rejection; and (c) serum creatinine greater than 2.1 mg/dl or renal clearance less than 35 ml/min. Cyclosporine was reduced by 50 mg every 3 wk until discontinuation, prednisone was temporarily increased from 10 to 20 mg/day and azathioprine was maintained at 2 mg/kg/day. Sustained improvement in kidney function in the 12 patients was minimal, with the mean serum creatinine level decreasing from 2.5 ± 0.5 mg/dl (mean ± S.D.) at study entry to 2.4 ± 1.2 mg/dl after a mean follow-up of 18 ± 6 mo. In six patients, histologically confirmed cellular rejection developed after a mean of 5 ± 6 mo from the time that cyclosporine withdrawal was begun. Two of six patients with rejection responded to bolus steroid therapy and are in stable condition at this writing with low-dose cyclosporine (2.8 and 3.2 mg/kg/day). Two patients initially responded to bolus steroids but later exhibited ductopenic rejection; one responded to treatment with FK 506 and the other died of sepsis. The two remaining patients were steroid unresponsive. One responded to treatment with OKT3 and is now stable on low-dose cyclosporine (2.3 mg/kg/day), but in the other ductopenic rejection developed and the patient died of sepsis during rescue therapy with FK 506. By comparison, in none of 12 matched control patients did rejection develop during a similar follow-up period. We conclude that cyclosporine withdrawal for nephrotoxicity in liver transplant recipients does not result in sustained improvement in kidney function and causes a high incidence of cellular rejection, which can progress to ductopenic rejection or death from infection. (Hepatology 1994;19:925-932.)
Hepatic lidocaine metabolism and liver histology in patients with chronic hepatitis and cirrhosis
Authors: Shiffman Mitchell L. , Luketic Velimir A. , Sanyal Arun J. , Duckworth Frederick P. , Purdum Preston P. , Contos Melissa J. , Mills Scott A. , Edinboro Leslie E. , Poklis Alphonse
Published: March 2007

Recent advances in the medical and surgical treatment of chronic hepatitis and cirrhosis have made it increasingly important to develop noninvasive tests of liver function. Our study has evaluated the hepatic conversion of lidocaine to its primary metabolite monoethylglycinexylodide and compared this with liver histological findings in 225 patients with chronic hepatitis (161 with hepatitis C, 23 with hepatitis B, 21 with autoimmune hepatitis and 20 with cryptogenic hepatitis). One hundred seven (47.7%) patients had cirrhosis at the time of evaluation. A decline in monoethylglycinexylodide production was observed with worsening liver histological conditions from a mean of 81.5 ± 7.0 ng/ml in patients with chronic persistent hepatitis to 61.2 ± 5.5 ng/ml for chronic active hepatitis and 20.9 ± 1.5 ng/ml in patients with cirrhosis (p < 0.05). A further stepwise decline in monoethylglycine xylodide production was observed with worsening Child class: from 25.5 ± 2.2 ng/ml for class A patients to 8.9 ± 1.4 ng/ml for patients with Child class C disease (p < 0.05). All patients with monoethylglycinexylodide production less than 20 ng/ml had cirrhosis confirmed on histological examination. In contrast, no relationship was observed between liver histological status and serum transaminases (AST or ALT), bilirubin, albumin and prothrombin time. Thirty-five patients underwent repeat histological evaluation and monoethylglycinexylodide testing after receiving at least 6 mo treatment for chronic hepatitis (interferon for hepatitis B and C and corticosteroids for autoimmune hepatitis). The change in monoethylglycinexylodide production observed in these patients was a linear function of the change in Knodell histological index (r = 0.73, p < 0.005). We conclude that hepatic monoethylglycinexylodide production parallels liver histological condition in patients with chronic hepatitis and cirrhosis. Monoethylglycinexylodide testing may therefore prove to be a useful noninvasive test by which to monitor hepatic function and assess response to drug treatment in patients with chronic hepatitis. (Hepatology 1994;19:933-940.)
Effects of different bile salts on steady-state mRNA levels and transcriptional activity of cholesterol 7α-hydroxylase
Authors: Pandak William M. , Vlahcevic Reno Z. , Heuman Douglas M. , Redford Kaye S. , Chiang John Y. L. , Hylemon Philip B.
Published: March 2007

Cholesterol 7α-hydroxylase, the rate-limiting enzyme in the bile acid synthesis pathway, is downregulated by taurocholate by way of negative feedback control at the level of gene transcription. The molecular basis of regulation of cholesterol 7α-hydroxylase by other hydrophobic bile salts and under more physiological conditions is not known. The aim of this study was to investigate the molecular basis of regulation of cholesterol 7α-hydroxylase by several naturally occurring bile salts in rats with intact enterohepatic circulation. Male Sprague-Dawley rats were pair-fed for 14 days normal chow (control), cholestyramine (5% of diet), cholic acid (1%), chenodeoxycholic acid (1%) or deoxycholic acid (0.25%). When rats were killed, livers were harvested and HMG-CoA reductase specific activity and cholesterol 7α-hydroxylase specific activities, steady-state mRNA levels and transcriptional activity were determined Nuovo Cimento with those of control rats fed normal chow. Compared with results in paired controls, cholestyramine feeding led to an approximate threefold increase in HMG-CoA reductase specific activity. Feeding of hydrophobic bile salts profoundly decreased the specific activity of HMG-CoA reductase. Cholestyramine led to a three-fold increase in cholesterol 7α-hydroxylase specific activity, steady-state mRNA levels and gene transcriptional activity. The feeding of cholic (1%), chenodeoxycholic (1%) and deoxycholic acid (0.25%) led to significant decreases in cholesterol 7α-hydroxylase specific activities (62%, 84% and 97%, respectively), steady-state mRNA levels (72%, 29% and 61%, respectively) and transcriptional activities (44%, 43% and 54%, respectively). Down-regulation of cholesterol 7α-hydroxylase specific activity was in order of increasing hydrophobicity of bile salts (cholic < chenodeoxycholic < deoxycholic acid). No such clear correlation was observed between bile salt hydrophobicity and steadystate mRNA levels or gene transcriptional activity. We conclude that down-regulation of cholesterol 7α-hydroxylase activity by cholic, chenodeoxycholic and deoxycholic acids occurred as the result of decreased transcriptional activity of the cholesterol 7α-hydroxylase gene. Because chenodeoxycholic and deoxycholic acids led to greater fractional suppression of cholesterol 7α-hydroxylase specific activity than of gene transcriptional activity, we postulate the existence of posttranscriptional regulation of cholesterol 7α-hydroxylase by these two hydrophobic bile salts. (Hepatology 1994;19:941-947.)
Hepatocyte growth factor in transgenic mice: Effects on hepatocyte growth, liver regeneration and gene expression
Authors: Shiota Goshi , Wang Timothy C. , Nakamura Toshikazu , Schmidt Emmett V.
Published: March 2007

Attention has recently been focused on hepatocyte growth factor as a major candidate factor in liver regeneration because it is the most potent known mitogen for hepatocytes in vitro. However, hepatocyte growth factor also displays diverse activities in vitro as scatter factor, as an epithelial morphogen, as a pluripotent mitogen and as a growth inhibitor. Consequently, we developed transgenic mice that expressed hepatocyte growth factor under the control of albumin regulatory sequences to examine its in vivo role in hepatocyte growth. Hepatocytes of these mice expressed increased levels of hepatocyte growth factor as an autocrine growth factor. Hepatocyte growth factor was a potent stimulus for liver repair; the livers of hepatocyte growth factor-transgenic mice recovered completely in half the time needed for their normal siblings after partial hepatectomy. This transgenic model also enabled us to study the chronic effects of hepatocyte growth factor expression. During several months of observation, the labeling index of hepatocytes in albumin-hepatocyte growth factor mice was doubled, and liver DNA content was increased compared with that in wild-type mice. To identify intermediate signaling pathways for hepatocyte growth factor that might regulate this increased growth response, we examined transgenic mice for changes in expression of genes that are known to be regulated during liver regeneration. We found that levels of c-myc and c-jun mRNA were increased in the hepatocyte growth factor-transgenic mice. In additional experiments the increased c-myc expression was the consequence of increased transcription rates as seen in nuclear run-on and myc-CAT reporter gene experiments. We conclude that hepatocyte growth factor increases growth and repair processes when expressed for long periods in the liver and that c-myc and c-jun may be important intermediaries in the hepatocyte growth response caused by hepatocyte growth factor. (Hepatology 1994;19:962-972.)
Contribution of complement-stimulated hepatic macrophages and neutrophils to endotoxin-induced liver injury in rats
Authors: Jaeschke Hartmut , Farhood Anwar , Smith Wayne C.
Published: March 2007

The role of complement as potential activator for tissue macrophages and neutrophils was investigated in an experimental model of endotoxin-induced liver injury in male Fischer rats. Injection of Salmonella enteritidis endotoxin (1 mg/kg) into Corynebacterium parvum-pretreated animals (7 mg/kg; single dose 6 days before endotoxin) resulted in severe oxidant stress, as indicated by a 37-fold increase of plasma levels of glutathione disulfide (basal concentration, 0.36 ± 14 μmol/L), accumulation of neutrophils in the liver (600 ± 31 neutrophils/50 high-power fields) and liver injury (plasma ALT, 1184 ± 185 U/l; necrosis; 19% ± 3%) 10 hr after endotoxin. The oxidant stress induced by 1 mg/kg endotoxin in the C. parvum-treated animals was always significantly higher than that in control animals receiving the same dose of endotoxin. Inhibition of complement activation with the soluble complement receptor type 1 attenuated the oxidant stress and liver injury by 50% to 65% but had no effect on hepatic neutrophil accumulation or plasma tumor necrosis factor-α levels. Treatment with a monoclonal antibody directed against the α-chain of CD11b/CD18 adhesion proteins (clone 17), which was highly effective in attenuating ischemia-reperfusion injury in the liver by reducing the number of neutrophils and functionally inactivating these cells, neither protected against parenchymal cell injury nor affected hepatic neutrophil infiltration in the C. parvum model. We conclude that reactive oxygen derived from complement-stimulated macrophages is critical for the development of liver injury in the C. parvum/endotoxin model. (Hepatology 1994; 19:973-979.)
The role of major histocompatibility complex and non-major histocompatibility complex encoded antigens in generation of bile duct lesions during hepatic graft-vs.-host responses mediated by helper or cytotoxic T cells
Authors: Williams Fred H. , Thiele Dwain L.
Published: March 2007

In these studies, we examined the role of discrete classes of alloantigen differences in generating nonsuppurative cholangitis during graft-vs.-host disease. Transfer of C57BL/6J (B6) splenocytes to class I major histocompatibility complex-disparate bm1 × B6 F1, class II major histocompatibility complex-disparate B6 × bm12 F1, or multiple non-major histocompatibility complex antigen-disparate Balb,B × B6 F1 mice led to the development of periportal inflammatory infiltrates and lymphocyte invasion of bile duct walls. However, frank destruction of bile duct walls was observed only in strain combinations with class I majorhistocompatibility complex or multiple non-major histocompatibility complex-encoded disparities. The concomitant presence of class II major histocompatibility complex differences and class I major histocompatibility complex or multiple non-major histocompatibility complex differences did not increase and in some cases was associated with less severe bile duct disease than was observed in strain combinations with discrete histocompatibility antigen differences. Depletion of l-leucyl-l-leucine methyl ester-sensitive cytotoxic T lymphocytes from donor inocula reduced the incidence of destructive bile duct lesions observed early in the course of graft-vs.-host disease in B6→Balb.B × B6 F1 or B6→bm1 × B6 F1 mice. However, transfer of CD8-negative, L-leucyl-L-leucine methyl ester-resistant T helper cells alone was sufficient to generate destructive cholangitis in class I + II major histocompatibility complex-disparate or multiple non-major histocompatibility complex antigen-disparate strain combinations. These findings demonstrate that histologically similar patterns of destructive nonsuppurative cholangitis can be generated by diverse T effector mechanisms directed at class I major histocompatibility complex or non-major histocompatibility complex-encoded antigen differences. However, in this murine graft-vs.-host disease model, the presence of class II major histocompatibility complex-encoded alloantigen differences provided neither a direct nor an additive stimulus for generation of destructive duct lesions. (Hepatology 1994;19:980-988.)
Tauro α-muricholate is as effective as tauro β-muricholate and tauroursodeoxycholate in preventing taurochenodeoxycholate-induced liver damage in the rat
Authors: Kitani Kenichi , Kanai Setsuko , Sato Yuko , Ohta Minoru
Published: March 2007

Male Wistar rats were infused intravenously with taurochenodeoxycholate (0.4 μmol/min/100 gm) alone (group A) or with one of the three bile salts (tauroursodeoxycholate [group B], tauro β-muricholate [group C] or tauro α-muricholate [group D]) at a rate of 0.2 μmol/min/100/gm for 1 hr. One-hour bile flow and bile salt excretion rates were significantly lower in group A than in the other three coinfused (B, C, D) groups. Biliary 1-hr outputs of lactate dehydrogenase and albumin in the bile, on the other hand, were significantly higher in group A than in the other groups. Plasma concentrations of lactate dehydrogenase at the time of killing (1 hr) were two to three times higher in group A than in the other groups. Although tauro α-muricholate does not possess a 7β-hydroxy group, the 6β-hydroxy group that tauro α-muricholate possesses thus appears to be as effective as a 7β-hydroxy group in reducing the liver damage caused by toxic bile salts such as taurochenodeoxycholate. The so-called hepatoprotective effects of tauroursodeoxycholate and tauro β-muricholate found in previous studies may require explanation(s) other than the presence of a 7β-hydroxy group in their molecular structures. (Hepatology 1994;19:1007-1012.)
Comparison of peptidoglycan-polysaccharide and lipopolysaccharide stimulation of Kupffer cells to produce tumor necrosis factor and interleukin-1
Authors: Lichtman Steven N. , Wang Jian , Schwab John H. , Lemasters John J.
Published: March 2007

Endotoxin (lipopolysaccharide) is a cell wall polymer from gram-negative bacteria that stimulates Kupffer cell release of cytokines such as tumor necrosis factor-α and interleukin-1. Another bacterial cell wall polymer in both gram-negative and gram-positive organisms is peptidoglycan-polysaccharide. Lipopolysaccharide and peptidoglycan-polysaccharide exist together in the intestinal lumen and can cross the intestinal mucosa, enter the portal vein and activate Kupffer cells. The purpose of this study was to compare the effects of lipopolysaccharide stimulation and peptidoglycan-polysaccharide stimulation of Kupffer cells on release of tumor necrosis factor-α and interleukin-1. Both bacterial polymers caused maximum tumor necrosis factor-α release from Kupffer cells after incubation for 4 to 8 hr. Maximum tumor necrosis factor-α release induced by 400 ng/ml lipopolysaccharide was 704 ± 258 pg/ml, compared with 329 ± 91 pg/ml tumor necrosis factor-α after 100 μg/ml peptidoglycan-polysaccharide (p < 0.001). Polymyxin B blocked lipopolysaccharide stimulation of tumor necrosis factor-α by 95% ± 5% but blocked peptidoglycan-polysaccharide-stimulated tumor necrosis factor-α by 30% ± 14% (p < 0.001). Repeat incubation of Kupffer cells with lipopolysaccharide after prior lipopolysaccharide incubation induced low tumor necrosis factor-α release (tolerance). Repeat incubation with peptidoglycan-polysaccharide induced no tolerance to tumor necrosis factor-α release. Incubation of lipopolysaccharide plus peptidoglycan-polysaccharide released less tumor necrosis factor-α than did each polymer used alone, but this inhibition was prevented by indomethacin. Dibutyryl cyclic AMP, prostaglandin E1, prostaglandin E2 and the adenosine A2-receptor agonist N-ethylcarboxyamideadenosine inhibited lipopolysaccharide-stimulated tumor necrosis factor-α release by 83%, 97%, 90% and 94%, respectively, but inhibited peptidoglycan-polysaccharide-stimulated tumor necrosis factor-α release by 52%, 60%, 45% and 51%, respectively (p < 0.001 for each). This indicates that intracellular signaling pathways differ for lipopolysaccharide-stimulated and peptidoglycan-polysaccharide-stimulated tumor necrosis factor-α release. After incubation for 8 and 24 hr, 100 μg/ml peptidoglycan-polysaccharide had induced significantly more interleukin-1 release from cultured Kupffer cells than had 400 ng/ml lipopolysaccharide (p < 0.001). Lipopolysaccharide induced tolerance to interleukin-1 release after repeat incubation, but peptidoglycan-polysaccharide caused no tolerance. These studies show that peptidoglycan-polysaccharide, a ubiquitous bacterial cell wall polymer, shares several proinflammatory properties with lipopolysaccharide but that there are differences that may have pathophysiological significance. (Hepatology 1994;19:1013-1022.)
Enzyme inhibitory autoantibodies to pyruvate dehydrogenase complex in primary biliary cirrhosis differ for mammalian, yeast and bacterial enzymes: Implications for molecular mimicry
Authors: Teoh Khay-Lin , Mackay Ian R. , Rowley Merrill J. , Fussey Shelley P. M.
Published: March 2007

Primary biliary cirrhosis is a chronic autoimmune disease in which serum autoantibodies against the mitochondrial 2-oxo acid dehydrogenase enzyme complexes (M2 antibodies) are regularly present. Molecular mimicry of host proteins by bacterial counterparts is a suggested explanation for the origin of these autoantibodies. We tested this hypothesis by measuring the functional reactivity of serum autoantibodies by means of an enzyme inhibition assay against pyruvate dehydrogenase complex from different sources: mammalian, Saccharomyces cerevisiae and Escherichia coli. The 10 primary biliary cirrhosis sera all reacted on immunofluorescence study for M2 antibodies and on immunoblotting with the pyruvate dehydrogenase complex E2 subunit from each of the three enzymes, but there were strikingly different inhibitory capacities. The primary biliary cirrhosis sera were highly inhibitory for mammalian pyruvate dehydrogenase complex (10 of 10 inhibitory; mean level of inhibition, 99%), moderately inhibitory for yeast pyruvate dehydrogenase complex (10 of 10 inhibitory; mean level, 70%) and weakly inhibitory for Escherichia coli pyruvate dehydrogenase complex (4 of 10 inhibitory; mean level, 26%). Thus, with a functional assay that depends on epitope recognition of primary biliary cirrhosis sera, cross-reactivity between mammalian and bacterial pyruvate dehydrogenase complex enzymes is low and molecular mimicry, at least at the B-lymphocyte level, is not supported. (Hepatology 1994; 19:1029-1033.)
Role of Na,K-ATPase in regulating acidification of early rat liver endocytic vesicles
Authors: Anbari Maan , Root Karen V. , Van Dyke Rebecca W.
Published: March 2007

Endocytic vesicles are acidified by an electrogenic proton pump and a parallel chloride conductance; however, acidification might be decreased if electrogenic transporters, such as Na,K-ATPase, that increase vesicle interior-positive membrane potential were also present. We examined this issue in early rat liver endosomes using ion substitution and inhibitors to alter Na,K-ATPase activity. These early endosomes, labeled for 2 min with the fluorescent fluid-phase marker fluorescein isothiocyanate-dextran, consistently acidified faster than endosomes similarily labeled for a 10-min period. In chloride-free media initial rates of acidification of early endosomes were faster in K+ media than in Na+ medium, although addition of K+ to Na+ or Na+ to K+ media to allow Na,K-ATPase to function did not decrease the rate of acidification. In chloride-containing media, rates were the same regardless of cation composition. The Na,K-ATPase inhibitor vanadate was prepared from orthovanadate by several methods, all of which inhibited liver ATPase activity. Two hundred μmol/L vanadate, prepared Cl−-free, tended to decrease rates of acidification in all media tested and these effects achieved statistical significance in Cl−-free media containing 150 mmol/L K+ or mixtures of Na+ and K+ and in 145 mmol/L KCl/5 mmol/L NaCl medium. Vanadate stocks pH-adjusted with hydrogen chloride increased rates of acidification in sodium gluconate buffers, probably as a result of the effects of the included Cl−. Five mmol/L ouabain (loaded into vesicles by endocytosis) and the membrane-permeable analog strophanthidin (2 mmol/L) both markedly inhibited endosome acidification, regardless of buffer ion composition. Collectively, these results suggest that Na,K-ATPase does not regulate acidification of rat liver early endocytic vesicles, that vanadate may modestly inhibit endosome acidification and that ouabain at high concentrations may inhibit acidification from the vesicle interior face. (Hepatology 1994;19:1034-1043.)
Where does the gene for hemochromatosis lie in relation to HLA-A?
Authors: Jazwinska Elizabeth C. , Halliday June W. , Powell Lawrie W.
Published: March 2007

Hereditary Hemochromatosis (HFE) is one of the most common inherited disorders with an estimated frequency of homozygous patients of 0.002-0.0045. The disease is characterized by increased intestinal iron absorption and progressive iron overload. Affected subjects show clinical symptoms of parenchymal organ damage after the third-fourth decade of life and have a 200-fold increased risk of developing hepatocellular carcinoma. Early diagnosis and treatment prevent complications and may normalize life expectancy of patients. The biochemical and genetic defects leading to progressive iron accumulation are still unknown, but the HFE gene is tightly linked to HLA complex on the short arm of chromosome 6. Utilizing HLA serotypes and the study of several polymorphic markers of 6p21, a linkage analysis of the disease locus was performed in a series of Italian hemochromatosis families. The data obtained by linkage analysis and the study of a family with a double recombinant allowed us to better define the HFE gene location with respect to HLA-class I A and F loci.
Hold that needle: Octreotide for acute variceal hemorrhage
Authors: Katkov William N.
Published: March 2007

To compare octreotide with injection sclerotherapy in the treatment of acute variceal hemorrhage, patients admitted with gastrointestinal bleeding and oesophageal varices confirmed by endoscopy were randomised to receive either emergency sclerotherapy with 3% sodium tetradecyl sulphate or octreotide (50 μg intravenous bolus plus 50 μg per hr intravenous infusion for 48 hr). At the end of the study period (48 hr), the octreotide group also had sclerotherapy to obliterate the varices.

One hundred patients were recruited. Demographic features including the aetiology of portal hypertension and the Child-Pugh’s grading of the two groups were similar. Bleeding was initially controlled in 90% of patients by emergency sclerotherapy and in 84% by octreotide infusion (95% confidence interval 0-19.5, p = 0.55). There were no significant differences between the two groups in early (within 48 hr of randomisation) rebleeding (16% vs 14%), blood transfusion (3 units vs 3.51, hospital stay (5 days vs 6 days), or hospital mortality (27% vs 20%). No notable side-effects were associated with octreotide.

We conclude that octreotide infusion and emergency sclerotherapy are equally effective in controlling variceal haemorrhage.
A multicenter study on the prognosis of fulminant viral hepatitis: Early prediction for liver transplantation
Authors: Takahashi Yoshiyata , Kumada Hiromitsu , Shimizu Masaru , Tanikawa Kyuichi , Kumashiro Ryukichi , Omata Masao , Ehata Toshiki , Tsuji Takao , Ukida Minoru , Yasunaga Mitsuru , Okita Kiwamu , Sato Shunichi , Takeuchi Toshihiko , Tsukada Katsuhiko , Obata Hiroshi , Hashimoto Etsuko , Ohta Yasuyuki , Tada Kouji , Kosaka Yoshitane , Takase Koujiro , Yoshiba Makoto , Sekiyama Kazuhiko , Kano Takashi , Mizoguchi Yasuhiro
Published: March 2007

To determine the risk of death at an early stage of fulminant viral hepatitis, we created severity indexes drawn from clinical data on the day of development of encephalopathy in 128 patients with fulminant hepatitis B and 103 with fulminant hepatitis non-A, non-B. In fulminant hepatitis B, the risk score was 2.75×BL+2.75×BR+2.7×AG+2.3×WB+1.67×CD+1.56×AL−0.098×PR−0.88, where BL is 1 if total bilirubin is higher than 20 mg/dl, BR is 1 if the ratio of total to direct bilirubin exceeds 2.2, AG is 1 if age is above 40 yr, WB is 1 if white blood cell count is less than 4,000 cells/mm3 or more than 18,000 cells/mm3, CD is 1 if a hazardous disease coexists and AL is 1 if ALT is less than 100 times the upper limit of normal (otherwise all are 0), and PR is prothrombin time (percentage of normal value). Using a cutoff score of 0, we found the positive predictive value, negative predictive value and predictive accuracy to be 0.90, 0.86 and 0.89, respectively. Sensitivity and specificity were 0.94 and 0.77, respectively. In fulminant non-A, non-B hepatitis, the risk score was 2.66×BR+2.25×BL+2.24×DI+2.05×AL ± 1.38×AG+0.00021×WB−6.33. BR is 1 if the ratio of total to direct bilirubin is more than 1.5, BL is 1 if total bilirubin is higher than 15 mg/dl, DI is 1 if the duration of illness before encephalopathy is less than 4 days or more than 12 days, AL is 1 if the ALT level exceeds 40 times the upper limit of normal and AG is 1 if age exceeds 50; otherwise all variables are 0. WB is WBC count per cubic millimeter. Positive predictive value, negative predictive value and predictive accuracy were 0.92, 0.80 and 0.89, respectively. Sensitivity and specificity were 0.95 and 0.70, respectively. These prediction equations were validated prospectively on a population of testing samples comprising 52 patients with fulminant hepatitis B and 47 with fulminant non-A, non-B hepatitis. Positive predictive values were 0.92 and 0.92, negative predictive values were 0.71 and 0.78 and predictive accuracies were 0.87 and 0.89 for patients with fulminant hepatitis B and fulminant non-A, non-B hepatitis, respectively. They may be used for selecting candidates for liver transplantation and for evaluating efficacy of a new therapy. (Hepatology 1994;19:1065-1071.)
Nuclear ploidy of normal and neoplastic hepatocytes from woodchuck hepatitis virus-infected and uninfected woodchucks
Authors: Cullen John M. , Linzey David W. , Gebhard Douglas H.
Published: March 2007

Flow cytometric analysis of the ploidy of normal and neoplastic hepatocyte nuclei obtained from adult woodchucks, a model of human hepadnavirus-induced hepatocellular carcinoma, was performed. All 36 samples of nuclei from non-neoplastic liver from woodchuck hepatitis virus-infected or uninfected liver were diploid, indicating that age-related nuclear polyploidization does not occur in this species, unlike other rodents. Individual or multiple hepatic neoplasms were obtained from each of 14 woodchuck hepatitis virus-infected woodchucks. Nineteen samples of hepatocellular carcinoma and eight adenomas were examined. Aneuploid nuclei were detected in 10 of the hepatocellular carcinomas and three of the adenoma samples. Similar DNA indexes, ranging from 1.11 to 1.22, were found in 7 of the 10 aneuploid HCCs and all 3 aneuploid adenomas. Nine of the 19 hepatocellular carcinoma samples and 5 of the 8 adenomas were diploid. Four of the diploid hepatocellular carcinomas had increased proportions of tetraploid nuclei. The presence of aneuploid nuclei was not related to histological appearance of the neoplasms or serum γ-glutamyltranspeptidase levels. Because none of the hepatocellular carcinomas metastasized, the presence of aneuploidy could not be related to biological behavior. We determined the proportion of uninucleate and binucleate hepatocytes in hepatocellular carcinoma and nonneoplastic liver. Approximately 7% of hepatocytes were binucleate in nonneoplastic liver from woodchuck hepatitis virus-infected and uninfected liver. Only 2% of malignant hepatocytes were binucleate. The results of this study indicate that aneuploidy is a common change in hepatic neoplasms from woodchuck hepatitis virus-infected woodchucks. (Hepatology 1994;19:1072-1078.)
M13 bacteriophage DNA inhibits duck hepatitis B virus during acute infection
Authors: Iizuka Akira , Watanabe Tatsuro , Kubo Toshiyuki , Yamamoto Masahiro , Ogawa Kazuo , Ohkuma Tetsuo , Kaji Akira
Published: March 2007

We investigated effects of various DNAs on duck hepatitis B virus replication in vivo. One-day-old ducks were infected intravenously with DHBV. Various DNAs were then injected intravenously, and duck hepatitis B virus levels were followed for up to 20 days after the inoculation. When M13 bacteriophage DNA (M13 DNA), heat-denatured Escherichia coli DNA or φX 174 phage DNA was injected intravenously at a dose of 2.45 mg/kg body wt daily for 10 days, a significant decrease of serum duck hepatitis B virus DNA was detected within 10 days. The efficacy was twice that reported with antisense DNA on a weight basis and far more than that reported on a molar basis. M13 DNA was superior, on the basis of effective dose, to acyclovir as an anti-duck hepatitis B virus agent. On treatment with M13 DNA, serum 2-5 A synthetase level was increased five to six times, suggesting that the antiviral effect of M13 DNA is at least partly due to induction of endogenous interferon, which in turn induces 2-5 A synthetase. No significant inhibitory effect on replication of duck hepatitis B virus was demonstrated by DNAs obtained from herring testes, herring sperm, salmon testes, human placenta or calf thymus. On discontinuation of M13 DNA injection on day 10, duck hepatitis B virus reappeared in the serum at later time points. Digestion of M13 DNA with S1 nuclease resulted in marked reduction of antiviral activity. These results show that M13 DNA, not its digested product, has potent antiviral activity. (Hepatology 1994;19:1079-1087.)
Factors predictive of response to interferon-α therapy in hepatitis C virus infection
Authors: Tsubota Akihito , Chayama Kazuaki , Ikeda Kenji , Yasuji Arase , Koida Isao , Saitoh Satoshi , Hashimoto Michiyo , Iwasaki Satomi , Kobayashi Mariko , Hiromitsu Kumada
Published: March 2007

To determine the patient-dependent and virus-related factors that may predict sustained response to interferon-α therapy, we prospectively evaluated 60 consecutive patients with chronic hepatitis C who received a standardized treatment schedule of interferon-α. Twenty-eight patients achieved a long-term sustained remission, 14 patients had temporary responses during treatment but relapsed after completing it and 14 patients did not respond. Four patients dropped out because of severe side effects and were excluded from evaluation of efficacy. Twenty-one variables were chosen as possible predictors of sustained response and were analyzed by means of multi-variate analysis. Variables related to the hepatitis C virus included genotype and concentration in serum before treatment. The former was determined by means of the polymerase chain reaction with genotype-specific primers for genotypes PT, K1, K2a and K2b, which were deduced from nonstructural region 5 of the hepatitis C virus genome. The latter was measured with a competitive polymerase chain reaction technique. Three variables were statistically significant (p<0.05) on univariate analysis: viral genotype, pretreatment level of viremia and Knodell’s fibrosis score. In multivariate analysis viral genotype and Knodell’s fibrosis score were correlated independently with a sustained response (p<0.05 for both). Incidence of sustained response was 40% and 91% in patients with genotypes K1 and K2a, respectively. We conclude that with the treatment schedule used, viral genotype was the most useful predictor of a sustained response to treatment with interferon-α. (Hepatology 1994;19:1088-1094.)
Effects of low-sodium diet and spironolactone on portal pressure in patients with compensated cirrhosis†
Authors: García-Pagán Juan Carlos , Salmerón Juan Manuel , Feu Faust , Luca Angelo , Ginés Pere , Pizcueta Pilar , Claria Juan , Piera Carlos , Arroyo Vicente , Bosch Jaume , Rodés Juan
Published: March 2007

The aim of this study was to investigate the hemodynamic effects of spironolactone associated with a low-sodium diet (n=14) or a low-sodium diet alone (n=9) in patients with compensated cirrhosis and portal hypertension. Spironolactone significantly reduced the plasma volume. This effect was associated with a significant reduction in the hepatic venous pressure gradient, from 17.6 ± 3.6 mm Hg to 15.3 ± 3.5 mm Hg (−13% ± 13%; p<0.01). Azygos blood flow (−20% ± 20%), cardiac output (−16.2% ± 10.5%) and mean arterial pressure (−9% ± 9%) also decreased significantly. However, there were no significant changes in hepatic blood flow. Patients receiving low-sodium diet alone experienced a mild but significant reduction in hepatic venous pressure gradient (−6.3% ± 6%) and in mean arterial pressure (−4% ± 5%). There were no significant changes in cardiac output and in hepatic or azygos blood flows. This study indicates that low-sodium diet plus administration of spironolactone reduces portal pressure and azygos blood flow in patients with compensated cirrhosis. Low-sodium diet alone only produces mild effects that are likely to be clinically irrelevant. (Hepatology 1994;19:1095-1099.)
Selective uptake of high-density lipoprotein-associated cholesteryl esters by human hepatocytes in primary culture
Authors: Rinninger Franz , Brundert May , Jäckle Stefan , Galle Peter R. , Busch Christoph , Izbicki Jakob R. , Rogiers Xavier , Henne-Bruns Doris , Kremer Bernd , Broelsch Christoph E. , Greten Heiner
Published: March 2007

High-density lipoprotein cholesteryl esters are taken up by many cells without simultaneous uptake of high-density lipoprotein apolipoproteins. This selective uptake was investigated in human hepatocytes in primary culture. Human high-density lipoprotein-3 (density, 1.125 to 1.21 gm/ml) was radiolabeled in both its apolipoprotein and in its cholesteryl ester moiety; uptake of these high-density lipoprotein3 tracers by hepatocytes was investigated. Apparent high-density lipoprotein3 particle uptake as measured with the cholesteryl ester tracer was in excess of that from the apolipoprotein tracer, indicating selective uptake of high-density lipoprotein3 cholesteryl esters by hepatocytes. This selective uptake is a regulated pathway in hepatocytes, as demonstrated by an inverse relationship between cell cholesterol and the rate of selective uptake. Studies on the mechanism of selective uptake have used inhibitors such as monensin, chloroquine, heparin, and a monoclonal antibody directed against low-density lipoprotein receptors. These experiments provide no evidence for a role of cell-secreted apolipoprotein E, endocytosis or retroendocytosis in selective uptake. The intracellular fate of high-density lipoprotein3-associated cholesteryl esters was investigated with [3H]cholesteryl oleatelabeled high-density lipoprotein3. Hepatocytes hydrolyzed [3H]cholesteryl oleate internalized from labeled high-density lipoprotein3; this catabolism was not inhibited by the presence of chloroquine. In parallel hepatocytes were incubated with [3H]cholesteryl oleate-labeled low-density lipoprotein. Cells hydrolyzed [3H]cholesteryl oleate taken up with low-density lipoprotein; however, this hydrolysis was inhibited by chloroquine, indicating lysosomal low-density lipoprotein cholesteryl ester catabolism. These experiments show that high-density lipoprotein3 cholesteryl esters selectively taken up by hepatocytes are hydrolyzed independently from the classical lysosomal catabolic pathway. The question was addressed if selective uptake mediates a net mass uptake of cholesterol rather than an isotope exchange phenomenon. Incubation of hepatocytes with high-density lipoprotein-3 suppressed endogenous sterol synthesis from sodium [14C]acetate. Hepatocytes were incubated in the presence of high-density lipoprotein3; medium cholesteryl esters decreased as a result of incubation with hepatocytes. These results show a net mass delivery of high-density lipoprotein cholesteryl esters to hepatocytes. In conclusion, the pathway for selective uptake of high-density lipoprotein cholesteryl esters could be demonstrated in human hepatocytes in primary culture. A role for selective uptake in high-density lipoprotein-mediated cholesterol delivery to the liver in human beings in vivo is proposed. (Hepatology 1994;19:1100-1114.)
The distribution of calcium salt precipitates in the core, periphery and shell of cholesterol, black pigment and brown pigment gallstones
Authors: Kaufman Howard S. , Magnuson Thomas H. , Pitt Henry A. , Frasca Peter , Lillemoe Keith D.
Published: March 2007

Calcium bilirubinate, palmitate, carbonate and phosphate have been identified in the cores of cholesterol and pigment gallstones, suggesting a role for precipitated calcium salts in the early events of gallstone formation. Previous studies that compared the calcium salt contents of cholesterol and pigment stones required destruction of gallstone structure. We have used scanning electron microscopy with windowless energy-dispersive x-ray microanalysis to determine the prevalence of calcium salts in a series of cholesterol (n=105), black pigment (n=35) and brown pigment (n=6) gallstones obtained from 146 consecutive patients undergoing cholecystectomy. These techniques provide specific identification of cholesterol and individual calcium salts as they occur within the core, periphery and shell of gallstones without destroying stone structure. Calcium precipitates more than 0.5 μm in diameter can be detected in a cholesterol background at a detection limit of 0.01% by weight. Calcium salts were detected in the centers of 88% of cholesterol and 100% of black (p<0.05 vs. cholesterol) and brown pigment stones. Calcium bilirubinate was identified in the cores of 54% of cholesterol and in all pigment stones (p<0.001 black pigment vs. cholesterol). Calcium palmitate was detected in all brown pigment stones, in 39% of cholesterol stones (p<0.001 vs. brown pigment) and in 31% of black stones (p<0.01 vs. brown pigment). Peripheral calcium salts were detected less in cholesterol (19%) than in black or brown stones (100%, p<0.05). Fourteen percent of cholesterol and black pigment stones were surrounded by shells containing mostly calcium carbonate. The range of stone cholesterol contents (0% to 100%) and frequency of calcium salt precipitates suggest that pigment and cholesterol gallstones represent a spectrum of disease differing more in their peripheral compositions than in their central regions. Cholesterol and pigment gallstones may differ more in growth than in the initial events of stone nidation. (Hepatology 1994;19:1124-1132.)