bluuebunny/crossref_metadata_2025 huggingface dataset:
Hepatic fibrosis as a predictor of hepatic regenerative activity after partial hepatectomy in the rat
Authors: Macintosh Ethel L. , Gauthier Tony , Pettigrew Norman M. , Minuk Gerald Y.
Published: March 2007

Liver regeneration is an essential component of the recovery period after partial hepatectomy. Unfortunately, tests that accurately predict regenerative activity in the postoperative period have yet to be described. This study was designed to determine whether the extent of hepatic fibrosis correlates with liver regeneration activity after partial hepatectomy in rats with carbon tetrachloride-induced liver disease. Two groups of adult male Sprague-Dawley rats (12 to 30/group) were treated for 20 to 22 wk with weekly intragastric doses of carbon tetrachloride or vehicle. All rats then underwent 70% hepatectomy while under ether anesthesia. Liver regeneration activity was determined at 24 and 48 hr by [3H]thymidine incorporation into DNA. Hepatic fibrosis was calculated at the time of partial hepatectomy by automated image analysis on Van Gieson-stained liver tissue. Although a significant inverse correlation was found between the extent of hepatic fibrosis and DNA synthesis when all rats were considered (carbon tetrachloride-treated and vehicle-treated) at 24 and 48 hr after partial hepatectomy (r = -0.4943 and -0.7396, respectively; p < 0.05), no such correlation existed when carbon tetrachloride-treated rats were considered independently (r = -0.3231 and -0.0910 at 24 and 48 hr, respectively). In conclusion, we believe that in diseased livers, preoperative quantitation of hepatic fibrosis on automated image analysis does not serve as a useful predictor of liver regeneration activity. (Hepatology 1992;17:307-309.)
Aplastic anemia and hepatitis C: Molecular biology exonerates another suspect
Authors: Gumucio Jorge J. , Bachwich Dale , Dienstag Jules
Published: March 2007

Objective. - To test the hypothesis that the rare, often fatal, syndrome of hepatitis-associated aplasia is associated with hepatitis C virus infection.

Design. - Case series.

Setting.-Tertiary referral centers in the United States, Japan, Italy, and Germany.

Patients. - Twenty-eight patients with onset of aplastic anemia within 90 days after seeking medical attention for jaundice, or having serum transaminase levels 150% or more of normal (hepatitis-associated aplasia patients) and three patients who developed aplastic anemia following liver transplantation for non-A, non-B, hepatitis.

Outcome Measures. - Presence of hepatitis C in serum, bone marrow, and liver samples, detected by the polymerase chain reaction; antibody testing; and percentage of activated peripheral cytotoxic T lymphocytes determined by immunophenotyping.

Results. - Hepatitis ribonucleic acid was present in the serum samples of 10 (36%) patients with hepatitis-associated aplasia. However, hepatitic C virus viremia was associated with transfusions received after the onset of aplasia: seven (58%) of 12 patients with hepatitis-associated aplasia who had received 21 or more units of blood products at the time of serum sampling were viremic, compared with only three (19%) of 16 patients with hepatitis-associated aplasia who had received 20 or less units of blood products (P <.05). Hepatitis C virus was not found in blood and bone marrow samples of three National Institutes of Health case patients tested at the time of diagnosis. None of three livers from non-A, non-B hepatitis patients who developed aplastic anemia after liver transplantation contained hepatitis C virus ribonucleic acid. Activated CD8+ T lymphocytes were elevated three- to 20-fold early in the course of hepatitis-associated aplasia.

Conclusions. - Our results implicate a novel, non-A, non-B, and non-C agent in both hepatitis-associated aplasia and fulminant hepatitis.
The Self-sustained sequence replication amplification system-move over, PCR
Authors: Lown Kenneth S.
Published: March 2007

Mutations at amino acid positions 67, 70, 215, and 219 in the human immunodeficiency virus type 1 (HIV-1) pol gene correlate with the emergence of resistance to zidovudine (AZT). These four positions were monitored in viral RNA extracted from infected peripheral blood mononuclear cells (PBMC) and viral stocks obtained after coculture with uninfected lymphocytes. Genotype determinations were made using the selfsustained sequence replication (3SR) and differential bead-based sandwich hybridization (BBSH) assay. The hybridization results obtained by 3SR and BBSH analyses were verified by dideoxynucleotide sequencing of the 3SR products. Correlation of 3SR and BBSH with the polymerase chain reaction and Southern hybridization analyses of the PBMC and corresponding viral isolates indicated that PBMC and corresponding HIV-1 isolates may differ in their genotypes at the monitored amino acid positions, variations from the wild-type nucleotide sequence may occur proximal to the codons being monitored, and viral isolates possessing the same genotypes at the four monitored amino acid positions showed a threefold variation in their ID50 measurements.
Hemodynamic and humoral changes after liver transplantation in patients with cirrhosis
Authors: Navasa Miquel , Feu Faust , García-Pagan Joan Carles , Jiménez Wladimiro , Llach Josep , Rimola Antoni , Bosch Jaume , Rodés Joan
Published: March 2007

Splanchnic and systemic hemodynamics and plasma levels of aldosterone, glucagon and plasma renin were investigated in 12 patients with advanced cirrhosis before and 2 wk (14.6 ± 2.8 days) and 2 mo (60.8 ± 10.5 days) after orthotopic liver transplantation. Liver transplant was followed by significant (p < 0.01) changes in systemic hemodynamics at 2 wk, with a marked reduction in cardiac index (4.9 ± 0.8 vs. 3.7 ± 0.7 L/min ± m2) and increases in mean arterial pressure (79 ± 8 vs. 101 ± 11 mm Hg) and peripheral vascular resistance (721 ± 149 vs. 1,274 ± 253 dyn ± sec ± cm−5). Two months after liver transplant, we saw further significant increases in peripheral vascular resistance (1,700 ± 341 dyn ± sec ± cm−5; p<0.05) without changes in cardiac index. Hepatic venous pressure gradient, very high before transplantation, was normal 2 wk after liver transplant (18.7 ± 3.0 vs. 2.1 ± 0.8 mm Hg; p < 0.01). Hepatic blood flow rose markedly from 1.03 ± 0.46 to 2.25 ± 0.79 L/min (p < 0.01) and was still elevated at 2 mo (1.84 ± 0.74 L/min). Azygos blood flow had not changed after 2 wk with respect to pretransplant values (0.65 ± 0.26 vs. 0.69 ± 0.39 L/min) but had decreased significantly at 2 mo (0.39 ± 0.16 L/min; p < 0.05). The elevated aldosterone, plasma renin and glucagon levels found in our cirrhotic patients before transplantation decreased to near-normal values 2 wk after the procedure. These results suggest that most of the hemodynamic and humoral abnormalities characteristic of advanced cirrhosis are reversed after liver transplant. However, azygos blood flow remains high early after liver transplant but is decreased at 2 mo, indicating that the closure of porto-collateral vessels occurs slowly despite the rapid normalization of portal pressure. The increase in hepatic blood flow after liver transplant may be a consequence of hepatic denervation. (Hepatology 1993;17:355-360.)
Prevalence and characterization of neutrophil cytoplasmic antibodies in autoimmune liver diseases
Authors: Mulder Leontine A. H. , Horst Gerda , Haagsma Elizabeth B. , Limburg Pieter C. , Kleibeuker Jan H. , Kallenberg Cees G. M.
Published: March 2007

To evaluate the diagnostic significance of neutrophil cytoplasmic antibodies in chronic liver diseases, we assessed the prevalence of neutrophil cytoplasmic antibodies in autoimmune liver diseases, in particular in primary sclerosing cholangitis, autoimmune chronic active hepatitis and primary biliary cirrhosis, and we also determined the specificity of perinuclear-pattern neutrophil cytoplasmic antibodies for these autoimmune liver diseases by testing sera from patients with nonautoimmune chronic liver diseases. Neutrophil cytoplasmic antibodies were detected in 79% of sera from patients with primary sclerosing cholangitis (n = 24), in 88% of sera from patients with autoimmune chronic active hepatitis (n = 24) and in 28% of sera from patients with primary biliary cirrhosis (n = 25). The presence of neutrophil cytoplasmic antibodies in these diseases correlated significantly (p<0.008) with the presence of cirrhosis. Neutrophil cytoplasmic antibodies were not detected in nonautoimmune liver diseases. All neutrophil cytoplasmic antibody-positive sera produced a perinuclear fluorescence pattern on ethanol-fixed granulocytes. On neutrophils fixed with paraformaldehyde, a granular cytoplasmic immunofluorescence pattern was observed, demonstrating the cytoplasmic nature of the antigen or antigens involved. Further characterization studies showed that neutrophil cytoplasmic antibodies in autoimmune liver diseases are not directed against myeloperoxidase, proteinase 3 or elastase, the neutrophil cytoplasmic antibody specificities associated with necrotizing vasculitis, glomerulonephritis or both. On Western blots neutrophil cytoplasmic antibodies in autoimmune liver diseases showed reactivity with either lactoferrin, a 67/66-kD protein combination or a 40-kD polypeptide. Reactivity with either of these proteins was observed in sera from patients with primary sclerosing cholangitis (38%), autoimmune chronic active hepatitis (17%) and primary biliary cirrhosis (20%). Because the same specificities have been found in neutrophil cytoplasmic antibody-positive sera from patients with inflammatory bowel disease and rheumatoid arthritis, these data suggest that these neutrophil cytoplasmic antibodies are a corollary of chronic inflammation. (Hepatology 1993;17:411-417.)
Mitochondrial dysfunction in alcoholic patients as assessed by breath analysis
Authors: Lauterburg Bernhard H. , Liang Dana , Schwarzenbach Felix A. , Breen Kerry J.
Published: March 2007

Mitochondria of patients with alcoholic liver disease are morphologically abnormal, and mitochondria isolated from animals exposed to ethanol exhibit functional deficiencies in vitro. Because the functional consequences of the morphological alterations and the relevance of in vitro observations to mitochondrial function in alcoholic subjects are not clear, we assessed mitochondrial function noninvasively with a breath test. Mitochondrial function was assessed by measuring the exhalation of 14CO2 after administration of 1 μCi 2-keto[1-14C]isocaproic acid, the decarboxylation of which occurs in mitochondria. The results of the 2-keto[1-14C]isocaproic acid breath test in 17 alcoholic subjects were compared with the results in healthy controls and patients with nonalcoholic liver disease. The peak exhalation of 14CO2 and the fraction of the administered dose decarboxylated in 60 min were significantly lower in alcoholic patients than in healthy controls or patients with nonalcoholic liver disease. In alcoholic patients 2-keto[1-14C]isocaproic acid decarboxylation was impaired in the presence of normal conventional and quantitative liver function as assessed by aminopyrine breath test and galactose elimination capacity, indicating that 2-keto[1-14C]isocaproic acid decarboxylation does not simply reflect decreased functional liver mass. We conclude that mitochondrial function as reflected by 2-keto[1-14C]isocaproic acid decarboxylation is impaired in chronic alcoholic patients. The functional impairment is specific for excessive ethanol consumption and not a reflection of decreased global liver function or the presence of cirrhosis. 2-Keto[1-14C]isocaproic acid decarboxylation could thus be useful as a marker of excessive ethanol consumption. (Hepatology 1993;17:418-422.)
Accumulation of organic anion in intracellular vesicles of cultured rat hepatocytes is mediated by the canalicular multispecific organic anion transporter
Authors: Oude Elferink Ronald P. J. , Bakker Conny T. M. , Roelofsen Han , Middelkoop Esther , Ottenhoff Roelof , Heijn Marc , Jansen Peter L. M.
Published: March 2007

Transport of organic anions within hepatocytes and the possible involvement of intracellular vesicles were studied with fluorescence microscopy. For this purpose monochlorobimane, a nonfluorescent hydrophobic compound that readily permeates into cells and is conjugated with glutathione to form the fluorescent glutathione bimane, was used. In the isolated perfused livers of normal rats, glutathione bimane is rapidly secreted into bile. In contrast, in our study of livers from mutant TR− rats, a 100-fold reduction in glutathione bimane secretion into bile occurred. Mutant TR− rats have an inherited defect in the canalicular multispecific organic anion transporter, which mediates the ATP-dependent secretion of a wide range of organic anions over the canalicular membrane into bile. When cultured Wistar and TR− hepatocytes were loaded with glutathione bimane, both cell types displayed a strong cytosolic fluorescence. Wistar cells completely lost this cytosolic fluorescence at incubation on monochlorobimane-free medium because of secretion of glutathione bimane. A clear punctate fluorescence remained, however, which was scattered through the cell with some perinuclear concentration. In some cells vesicular fluorescence was also concentrated around a canaliculus. In contrast, TR− cells lost their cytosolic fluorescence more slowly and completely lacked the vesicular fluorescence. Making cells selectively permeable with digitonin directly after loading them with glutathione bimane to remove cytosolic fluorescence again revealed the presence of fluorescent vesicles in Wistar cells and their absence in TR− cells. In Wistar cells vesicular fluorescence could be increased by preincubation with monensin or methylamine, compounds that have been shown to interfere with plasma membrane recycling. In conclusion, these results suggest that apart from secretion over the plasma membrane, the canalicular multispecific organic anion transporter may be involved in accumulation of organic anion in intracellular vesicles. It is hypothesized that this intracellular localization of the canalicular multispecific organic anion transporter is caused by recycling of the transporter between the plasma membrane and intracellular membranes. (Hepatology 1993;17:434-444.)
A rapid method to study heterogeneous gene expression in liver by direct assay of messenger RNA from periportal and perivenous cell lysates
Authors: Saarinen Juhani , Saarelainen Ritva , Lindros Kai O.
Published: March 2007

To study zonation of liver gene expression, we obtained periportal or perivenous rat liver cell lysates virtually devoid of nuclear material by site-directed digitonin infusion in situ. Total RNA was isolated, messenger RNAs were reverse transcribed and complementary DNAs were assayed after polymerase chain reaction-mediated amplification. The zonal distribution of messenger RNAs of alcohol dehydrogenase (little zonation), glutamine synthetase (perivenous) and cytochrome P-450 2E1 (perivenous) messenger RNAs, as analyzed by this technique, were found to be similar to the distribution of corresponding apoproteins. Using appropriate primers or complementary DNAs, zonation of many different messenger RNAs can be determined from the same sample by this simple and rapid method. (Hepatology 1993;17:466-469.)
Acute Hepatitis A: An endangered species
Authors: Sjogren Maria H.
Published: March 2007

Background:

Although inactivated hepatitis A vaccine is known to be well tolerated and immunogenic in healthy children and adults, its efficacy has yet to be established.

Methods:

To evaluate the efficacy of the hepatitis A vaccine in protecting against clinically apparent disease, we conducted a double-blind, placebo-controlled trial in a Hasidic Jewish community in upstate New York that has bad recurrent outbreaks of hepatitis A. At the beginning of a summer outbreak, 1037 healthy seronegative children 2 to 16 years of age were randomly assigned to receive one intramuscular injection of a highly purified, formalin-inactivated hepatitis A vaccine or placebo. A case was defined by the presence of typical signs and symptoms, a diagnostic increase in IgM antibody to hepatitis A, and a serum concentration of alanine aminotransferase at cast twice the upper limit of normal. Cases occurring ≥ 50 days after the injection were included in the evaluation of efficacy. The children were followed for a mean of 103 days.

Results:

A total of 519 children received vaccine, and 518 received placebo. The vaccine was well tolerated, with no serious adverse reactions. From day 50 after the injection, 25 cases of clinically apparent hepatitis A occurred in the placebo group and none in the vaccine group (P < 0.001), confirming that the vaccine had 100 percent protective efficacy. Before day 21, seven cases occurred in the vaccine group and three cases in the placebo group. After that time, there were no cases among vaccine recipients and 34 cases among placebo recipients.

Conclusions:

The inactivated purified hepatitis A vaccine that we tested is well tolerated, and a single dose is highly protective against clinically apparent hepatitis A.
Histological evaluation of the intrahepatic biliary tree in intrahepatic cholesterol stones, including immunohistochemical staining against apolipoprotein A-1
Authors: Ohta Tetsuo , Nagakawa Takukazu , Takeda Toshiya , Fonseca Luis , Kanno Masahiro , Mori Kazuhiro , Kayahara Masato , Ueno Keiichi , Miyazaki Itsuo , Terada Tadashi
Published: March 2007

Apolipoprotein A-1 is known to be one of inhibiting factors of cholesterol nucleation in bile, and decreased activity of apolipoprotein A-1 is considered to predispose cholesterol-supersaturated bile to formation of cholesterol crystals. To study the pathogenesis of the intrahepatic formation of cholesterol stones, we examined surgically resected liver specimens from six patients with intrahepatic cholesterol stones and compared the characteristic histopathological features with those of intrahepatic calcium bilirubinate stones, using morphological examination and immunohistochemical staining against apolipoprotein A-1. Morphologically, in all six patients with cholesterol stones the severity of chronic proliferative cholangitis with proliferation of the mucus-producing glandular elements in the walls of the large bile duct or periductal tissues was less extensive than that seen with calcium bilirubinate stones, and cholesterol crystals had formed in the septal and interlobular bile ducts. Immunohistochemically, unlike the normal liver and calcium bilirubinate stone-containing lobes, the hepatocytes and the epithelial lining of the bile ducts and peribiliary glands of the cholesterol stone-containing lobes did not react completely (some of the epithelial cells reacted only faintly) with apolipoprotein A-1 antibody. These findings suggest that an abundance of mucous substance and bacterial infection of the biliary tree may not be necessary for the formation of cholesterol stones, compared with findings in cases of calcium bilirubinate stones. We suggest that cholesterol crystals may be produced in the septal and interlobular bile ducts in the microenvironment of cholesterol-supersaturated bile and decreased activity of apolipoprotein A-1. (Hepatology 1993;17:531-537.)
Hepatitis B virus DNA in serum and liver is commonly found in chinese patients with chronic liver disease despite the presence of antibodies to HBsAg
Authors: Zhang Yong-Yuan , Hansson Bengt Göran , Kuo Lin Sheng , Widell Anders , Nordenfelt Erik
Published: March 2007

Sera from 410 patients from the Wuhan area in the central part of China with the diagnosis of chronic liver disease were analyzed for markers of hepatitis B, C and D virus infections. All sera, plus liver biopsy specimens from 188 of the patients, were also tested for hepatitis B virus DNA by polymerase chain reaction. Sixty-eight percent were HBsAg positive in serum, whereas 29% showed markers of past hepatitis B virus infection. Hepatitis B virus DNA was detected in all HBeAg-positive sera but also in 58% of patients with HBe antibody. In the liver specimens of the corresponding patient groups, 97% and 78%, respectively, were hepatitis B virus DNA positive. However, more noteworthy was that of the HBsAg-negative/HBsantibody positive patients 30% had detectable hepatitis B virus DNA in serum and 32% had hepatitis B virus DNA in liver tissue, whereas in a control group of healthy blood donors, of which 90% had HBs antibody, none was hepatitis B virus DNA positive. Our results demonstrate that among patients with chronic liver disease, infections with hepatitis B virus or hepatitis B virus-related virus(es) may frequently occur without being revealed by conventional serological methods. Hepatitis C and D viruses seem to be of only minor importance in the pathogenesis of chronic liver disease in this part of China. (Hepatology 1993;17:538-544.)
Quantitation of hepatitis C virus RNA in serum of asymptomatic blood donors and patients with type C chronic liver disease
Authors: Hagiwara Hideki , Hayashi Norio , Mita Eiji , Naito Masafumi , Kasahara Akinori , Fusamoto Hideyuki , Kamada Takenobu
Published: March 2007

We describe a method for quantifying hepatitis C virus RNA in serum. This competitive assay combines reverse transcription and polymerase chain reaction and is based on coamplification of the target RNA with known amounts of synthetic mutated RNA. We tested serum samples from 104 hepatitis C virus carriers (9 asymptomatic blood donors and 95 patients with type C chronic liver disease) to determine the relationship between the replicative level of hepatitis C virus and various stages of the carrier states. The amount of circulating hepatitis C virus RNA ranged from 104 to 109.5 genomes/ml serum. The titer of hepatitis C virus RNA (logarithmic transformed copy numbers of RNA per milliliter of serum) was lower in asymptomatic blood donors (5.4 ± 2.0) and in patients with chronic persistent hepatitis (7.3 ± 1.1) than in patients with chronic active hepatitis (7.9 ± 0.8), cirrhosis (7.8 ± 0.7) or hepatocellular carcinoma (7.9 ± 0.7). The titer of hepatitis C virus RNA was significantly lower in carriers younger than 40 yr old and correlated positively with the logarithmic transformed serum ALT level. Logistic regression showed that age and titer of hepatitis C virus RNA correlated independently with the stages of liver disease. These results showed that the replicative level of hepatitis C virus is higher in advanced liver disease and that elevation of viral replication may play an important role in liver injury and progression of liver disease. This competitive assay is useful in evaluating the state of viral replication in hepatitis C virus infection. (Hepatology 1993;17:545-550.)
Hepatitis C virus antibodies and liver disease in patients with porphyria cutanea tarda
Authors: Decastro Mar , Sánchez Javier , Herrera Jesús F. , Cháves Asunción , Durán Rafael , García-Buey Luisa , García-Monzón Carmelo , Sequí Julia , Moreno-Otero Ricardo
Published: March 2007

The recent identification of the hepatitis C virus and development of assays to detect antibodies to hepatitis C virus has allowed assessment of the prevalence of hepatitis C virus infection in patients with a variety of liver and other diseases. The aim of this study was to investigate the prevalence of hepatitis C virus antibodies and severity of liver injury in patients with porphyria cutanea tarda. Sixty-two patients were studied. Serum samples were analyzed for liver function parameters and markers of hepatitis B virus infection. Frozen serum samples from 34 patients with porphyria cutanea tarda, obtained when patients were seen at the hospital for the first time, were analyzed for hepatitis C virus antibodies with enzyme-linked immunosorbent assays (first- and second-generation) and a recombinant immunoblot assay. As controls, serum samples from 19,788 blood donors, 40 patients with alcoholic liver disease and 138 hospitalized patients without liver disease were also tested for hepatitis C virus antibodies. Liver biopsy was performed in 42 porphyria cutanea tarda patients. Specimens were evaluated for steatosis, siderosis, fibrosis, severity of inflammation and the presence of cirrhosis. In addition, the degree of necroinflammatory change and fibrosis were quantitated with the histologic activity index described by Knodell et al. The prevalence of hepatitis C virus antibodies in patients with porphyria cutanea tarda (62%) was higher than that in blood donors (0.79%), patients with alcoholic liver disease (17.5%) or hospitalized patients without liver disease (5.8%). Among porphyria cutanea tarda patients, those who were positive for hepatitis C virus antibodies by recombinant immunoblot assay had mean serum ALT levels (114 IU/L) significantly higher (p < 0.05) than those in patients negative by recombinant immunoblot assay (mean = 54 IU/L). Of 42 patients who underwent liver biopsies, 20 had been tested for hepatitis C virus antibodies. Of these patients, 15 were recombinant immunoblot assay positive (11 had chronic hepatitis or cirrhosis and 4 had steatosiderosis) and 5 were recombinant immunoblot assay negative (all had steatosiderosis). In conclusion, these results strongly suggest that hepatitis C virus infection may contribute to liver damage in patients with porphyria cutanea tarda. (Hepatology 1993;17:551-557.)
Cholesterol-lowering effect of ursodeoxycholic acid in patients with primary biliary cirrhosis
Authors: Poupon Renée E. , Ouguerram Khadija , Chrétien Yves , Verneau Claudine , Eschwège Eveline , Magot Thierry , Poupon Raoul
Published: March 2007

We have previously shown in a 2-yr controlled trial that hypercholesterolemia, frequent in primary biliary cirrhosis, is lowered by ursodeoxycholic acid (13 to 15 mg daily). To further investigate this effect, we analyzed the influence of long-term ursodeoxycholic acid administration on serum lipids, lipoproteins and bile acids. The study involved a subgroup of 33 noncirrhotic patients (17 received ursodeoxycholic acid and 16 received a placebo) analyzed at inclusion and after 2 yr. The total serum cholesterol concentration was markedly reduced in the ursodeoxycholic acid-treated patients in comparison with the controls (mean ± S.E.M. = 7.49 ± 0.42 mmol/L and 7.07 ± 0.23 mmol/L at entry and 4.44 ± 0.40 mmol/L and 6.89 ± 0.27 mmol/L at 2 yr in the ursodeoxycholic acid and placebo groups, respectively; p < 0.02). Quantitatively, this decrease was mainly caused by a fall in low-density-lipoprotein cholesterol, but very low density-lipoprotein cholesterol levels also fell significantly. High-density-lipoprotein cholesterol levels remained stable in both groups, but the high-density-lipoprotein2/high-density-lipoprotein3 cholesterol ratio fell significantly during ursodeoxycholic acid treatment. No significant change occurred in total triglyceride or total phospholipid levels. In the treated group, the proportion of ursodeoxycholic acid increased (up to 60% of total circulating bile acids), whereas that of cholic and chenodeoxycholic acids fell significantly. In conclusion, the cholesterol-lowering effect of ursodeoxycholic acid could be related to an improvement of cholestasis, modifications in cholesterol metabolism or both. Changes in endogenous bile acid composition induced by ursodeoxycholic acid might be the common denominator of these two mechanisms. (Hepatology 1993;17:577-582.)
Compression of the common bile duct due to portal-vein thrombosis in polycythemia vera
Authors: Löhr Matthias J. , Kuchenreuter Stefan , Grebmeier Hans , Hahn Eckhardt G. , Fleig Wolfgang E.
Published: March 2007

The case of a 56-yr-old patient with polycythemia vera who was initially seen with jaundice is reported. Compression of the common bile duct by choledochal varices was found to be the cause of the obstructive jaundice. This compression was successfully treated with placement of a plastic endoprosthesis across the stenosis during endoscopic retrograde cholangiopancreatography. The significance of choledochal varices in the evaluation of obstructive jaundice is emphasized. (Hepatology 1993;17:586-592.)
Cirrhosis of undefined pathogenesis: Absence of evidence for unknown viruses or autoimmune processes
Authors: Greeve Martina , Ferrell Linda , Kim Michael , Combs Connie , Roberts John , Ascher Nancy , Wright Teresa L.
Published: March 2007

To examine whether unknown viruses or autoimmune processes contribute to the development of cryptogenic liver disease, we studied 48 patients undergoing liver transplantation who had non-A, non-B cirrhosis; non-blood-borne cirrhosis of unknown etiology; or autoimmune cirrhosis. After the diagnosis of hepatitis C virus infection was established by the presence of viral antibodies or viral RNA, patients were reclassified into three groups: hepatitis C virus infection, autoimmune cirrhosis and cryptogenic cirrhosis. Explant histological appearance, incidence of posttransplant hepatitis and immunological features were compared in the three groups. Thirty-one percent of patients had neither hepatitis C virus infection nor classical autoimmune cirrhosis and were classified as having cryptogenic cirrhosis. Unlike histological appearance in hepatitis C virus infection but similar to that in autoimmune cirrhosis, explant histological appearance of cryptogenic cirrhosis showed inactive cirrhosis with little inflammation. After transplantation, histological hepatitis of the allograft was demonstrated in 44% of patients with hepatitis C infection but in no patient with autoimmune or cryptogenic cirrhosis. The autoimmune score, developed from clinical criteria associated with autoimmune liver disease, was significantly lower in cryptogenic cirrhosis and hepatitis C virus infection than in autoimmune cirrhosis. Autoantibodies-including antinuclear antibodies, smooth muscle antibodies and liver-kidney microsomal antibodies-were not commonly present in serum of patients with cryptogenic cirrhosis, whereas antibodies to soluble liver antigens were found with increased frequency in this entity. We conclude that in many patients with liver disease, no pathogenesis can be identified. The mild inflammation of the explant on histological study and the low incidence of posttransplant hepatitis in cryptogenic cirrhosis suggest that chronic active viral infection is unlikely to be implicated in the pathogenesis. Although the presence of antibodies to soluble liver antigens implies an immune component to the pathogenesis of this disease, low autoimmune score and infrequent presence of other autoantibodies suggest that cryptogenic cirrhosis is distinct from classical autoimmune liver disease. (Hepatology 1993;17:593-598.)
Diagnostic features and clinical outcome of ischemic-type biliary complications after liver transplantation
Authors: Sanchez-Urdazpal Luis , Gores Gregory J. , Ward Ellen M. , Maus Timothy P. , Buckel Erwin G. , Steers Jeffrey L. , Wiesner Russell H. , Krom Ruud A. F.
Published: March 2007

The clinical significance and outcome of nonanastomotic strictures and dilatations involving only the biliary tree of the graft with a radiological appearance of biliary ischemia is unknown. Therefore we analyzed the grafts of 128 patients to evaluate the biochemical, radiological and histological features that prompted the diagnosis of ischemic-type biliary stricture and the clinical outcome of this complication. Ischemic-type biliary strictures were diagnosed in 25 patients (19%). Initial graft function was similar in all patients, whether or not this complication developed. Most ischemic-type biliary strictures occurred between 1 and 4 mo after orthotopic liver transplantation. However, the appearance of ischemic-type biliary stricture in the month after transplantation was predictive for a poor outcome in all six grafts with early onset of ischemic-type biliary strictures. Eighteen patients (72%) were treated with biliary stents and repeated dilatations. Long-term patency was achieved in 88% of these patients. Repeat transplantation was performed in six patients (24%); five survived. Finally, patients with ischemic-type biliary strictures spent more time in the hospital during the first year after orthotopic liver transplantation than did patients without the complication (62 ± 27 days vs. 37 ± 20 days; p ± 0.001). This was due to repeated hospitalizations and a higher incidence of retransplantation. One-year graft survival was lower in patients with ischemic-type biliary strictures than in patients without ischemic-type biliary strictures (69% vs. 88%; p = 0.006). However, 1-yr patient survival was similar in the two groups (91% vs. 89%). In conclusion, early appearance of ischemic-type biliary stricture features is associated with poor graft prognosis. The occurrence of ischemic-type biliary stricture after transplantation is associated with increased morbidity, extended hospitalization and a higher incidence of repeat transplantation. (Hepatology 1993;17:605-609.)
Regulation of glycogen phosphorylase activity in isolated human hepatocytes
Authors: Keppens Stefaan , Vandekerckhove Ann , Moshage Han , Yap Sing Hiem , Aerts Raymond , de Wulf Henri
Published: March 2007

Hepatocytes were isolated from human liver tissue by a two-step perfusion technique. They were treated with vasopressin, angiotensin, ATP and phenylephrine, which are known to be Ca2+-mediated glycogenolytic agents in rat liver tissue, and as a control, they were treated with the cyclic AMP-mediated hormones glucagon and isoproterenol. All agonists induce a timedependent activation of glycogen phosphorytase. Glucagon and isoproterenol induce a somewhat higher degree of phosphorylase activation compared with vasopressin, angiotensin, ATP and phenylephrine, which all increase inositol tris-phosphate levels and have no effect on the cyclic AMP levels. The total activity of glycogen phosphorylase (a + b), amounting to 30 to 35 mU/mg protein, is found to be much lower than that found in rat liver tissue. Because only minor differences could be found, we conclude that the regulation of glycogen phosphorylase in human liver tissue is basically the same as that found in rat liver tissue. (Hepatology 1993;17:610-614.)
Augmented glucose use and pentose cycle activity in hepatic endothelial cells after in vivo endotoxemia
Authors: Spolarics Zoltán , Spitzer John J.
Published: March 2007

Glucose use and pentose cycle activity were determined in freshly isolated rat hepatic endothelial cells 3 hr after an intravenous injection of Escherichia coli lipopolysaccharide (0.1 mg/kg body weight), by use of (1-14C)glucose, [6-14C]glucose and [2-H]glucose. Lipopolysaccharide treatment in vivo increased glucose use fivefold, whereas glucose oxidation in the pentose cycle was elevated from 0.2 to 1.5 nmol/hr/107 cells. In vitro incubation of endothelial cells from saline- and lipopolysaccharide-treated animals in the presence of phorbol 12-myristate 13-acetate (10−6 mol/L) increased pentose cycle activity twofold and eightfold, respectively. Phorbol 12-myristate 13-acetate caused only a 40% to 60% increase in glycolysis in both groups. Addition of t-butyl hydroperoxide (0.5 mmol/L), a substrate for gluathione peroxidase, caused a 24-fold and 16-fold increase in the glucose flux through the pentose cycle in cells from saline- and lipopolysaccharide-treated rats, respectively. Oxidation of glucose through the Krebs cycle was also increased several-fold after t-butyl hydroperoxide administration. Depletion of cellular glutathione by N-ethylmaleimide (0.1 mmol/L) inhibited the phorbol 12-myristate 13-acetate-induced or t-butyl hydroperoxide-induced increase in the pentose cycle activity with no marked effects on glycolysis. Diphenyleneiodonium (0.1 mmol/L), an inhibitor of superoxide and nitric oxide synthesis inhibited the phorbol 12-myristate 13-acetate-induced increased pentose cycle activity with no effects on the t-butyl hydroperoxide-induced response. Endothelial cells from control animals treated with either 12-myristate 13-acetate or t-butyl hydroperoxide in the presence of exogenous glucose 20 mmol/L showed a similar increase in glycolysis but less increase in the pentose cycle activity as found after lipopolysaccharide treatment in the presence of glucose 5 mmol/L. This finding suggests that glucose-6-phosphate dehydrogenase or pathways dependent on pentose cycle intermediates became up-regulated after lipopolysaccharide administration. The lipopolysaccharide-induced elevated glucose use, accompanied by an increased activity of the pentose cycle, may also represent a potentiated mechanism for eliminating hydrogen peroxide derived from intracellular sources or from activated Kupffer cells and sequestered neutrophils in the hepatic sinusoid. (Hepatology 1993;17:615-620.)
DNA image cytometric analysis of macroregenerative nodules (adenomatous hyperplasia) of the liver: Evidence in support of their preneoplastic nature
Authors: Orsatti Giulia , Theise Neil D. , Thung Swan N. , Paronetto Fiorenzo
Published: March 2007

Twenty-eight macroregenerative nodules from 14 cirrhotic patients who underwent orthotopic liver transplantation were evaluated for DNA ploidy by means of image analysis of Feulgen-stained tissue sections. The lesions were classified as type 1 (16 cases) or type 2 (12 cases) on the basis of the absence or presence of cellular or architectural atypia in the nodules. The surrounding cirrhotic nodules were evaluated for liver cell dysplasia. Aneuploid peaks were significantly more frequent in type 2 macroregenerative nodules (58.3%) than in the cirrhotic regenerative nodules (7.1%) (p < 0.007). In addition, aneuploid peaks occurred with increased frequency in type 2 nodules (58.3%) than in type 1 macroregenerative nodules (6.2%) (p < 0.02). Only two aneuploid peaks (14.2%) were found in dysplastic cirrhotic livers. The nuclear area of aneuploid hepatocytes (71.6 μm ± 10.1%, mean ± S.D.) differed significantly from that of diploid liver cells (45.4 μm ± 6.5%) (p < 0.0001). Tetraploid peaks occurred in three type 2 lesions (25%); they were also found in one type 1 macroregenerative nodule (6.2%), one cirrhotic liver without dysplasia (7.1%) and three cirrhotic livers with dysplasia (21.4%). These findings support the notion that macroregenerative type 2 nodules are directly implicated in hepatocarcinogenesis and that their presence should be sought as an indicator of malignant potential in cirrhotic livers. (Hepatology 1993;17:621-627.)
Hepatic stimulator substance protects against acute liver failure induced by carbon tetrachloride poisoning in mice
Authors: Mao-Hua Mei , Wei An , Bao-Hong Zhang , Qing Shao , De-Zheng Gong
Published: March 2007

Hepatic stimulator substance was extracted from the liver of weanling Sprague-Dawley rats according to the method of LaBrecque. Quang-Ming mice were injected with carbon tetrachloride to induce acute liver failure. Hepatic stimulator substance suppressed the elevation of ALT and AST induced by carbon tetrachloride in a dose-dependent manner. Hepatic histological changes indicated that hepatic stimulator substance reduced the severity of hepatic lesion induced by carbon tetrachloride and reversed carbon tetrachloride-induced reduction of hepatic mitochondrial succinic dehydrogenase activity. In attempting to elucidate the mechanism or mechanisms of this protective effect, we found that hepatic stimulator substance significantly restored the carbon tetrachloride-induced decrease of hepatocyte plasmalemma and mitochondrial and microsomal membrane fluidity. Hepatic stimulator substance also decreased the malondialdehyde content of carbon tetrachloride-intoxicated mice; restored the liver-reduced glutathione content, which was lowered by carbon tetrachloride intoxication; stimulated liver regeneration, as shown by enhanced DNA synthesis; and increased the H-thymidine incorporation into DNA of hepatocytes. We propose that hepatic stimulator substance protects the liver against acute liver failure induced by carbon tetrachloride poisoning, probably by an antioxidative effect on hepatocyte membrane lipid peroxidation, which was increased by free radicals produced from carbon tetrachloride. In addition, hepatic stimulator substance stimulates hepatocyte proliferation. These protective mechanisms may act in concert to protect against carbon tetrachloride injury. (Hepatology 1993;17:638-644.)
Cellular heterogeneity in binding and uptake of low-density lipoprotein in primary rat hepatocytes
Authors: Corsetti James P. , Sparks Janet D. , Sikora Barbara , Sparks Charles E.
Published: March 2007

Hepatocellular heterogeneity of biochemical function is well established for many aspects of liver metabolism. This study addresses the question of cellular heterogeneity in the catabolism of low-density lipoprotein by rat hepatocytes. Low-density lipo-protein binding (4° C) and uptake (37° C) by rat hepatocytes were studied by use of human low-density lipoprotein labeled with a highly fluorescent lipophilic probe, N, N-dipentadecylaminostyrylpyridinium iodide, recently developed by us. Single-cell suspensions derived from rat hepatocytes in primary culture and from liver perfusion were studied with flow cytometry with and an approximation algorithm for data analysis. These studies show subpopulations of cells negative and positive for the specific binding and uptake of low-density lipoprotein. Dissociation constants for low-density lipoprotein binding and uptake were determined for the total population (18 μg/ml, binding; 12 μg/ml, uptake) and found to be in good agreement with previously reported values. Additionally, the dissociation constant for binding for the positive subpopulation was determined and found to be 3 μg/ml. This lower value is more typical of the values seen in other cell types. These findings are strongly suggestive of functional heterogeneity in the hepatic catabolism of low-density lipoprotein. (Hepatology 1993;17:645-650.)
Decrease in the hepatic clearance of hepatocyte growth factor in carbon tetrachloride-intoxicated rats
Authors: Liu Ke-Xin , Kato Yukio , Yamazaki Masayo , Higuchi Osamu , Nakamura Toshikazu , Sugiyama Yuichi
Published: March 2007

To examine whether a decrease in hepatic uptake, clearance or both of hepatocyte growth factor contributes to increased plasma hepatocyte growth factor levels, we kinetically analyzed hepatic hepatocyte growth factor handling using rats with carbon tetrachloride-induced liver injury in both in vivo and perfused liver systems. After the intravenous administration of tracer 125I-hepatocyte growth factor, the time profile of trichloroacetic acid-precipitable 125I-hepatocyte growth factor was analyzed, and tissue clearance and total body plasma clearance were determined. For the tissues examined (liver, kidney, lung, spleen and adrenal), liver and adrenal clearance of 125I-hepatocyte growth factor decreased significantly. It was found that the hepatic clearance explains the bulk of the total body plasma clearance. The hepatic clearance and the total body clearance decreased to minimums (approximately 40% of control) 24 hr after carbon tetrachloride administration and recovered to near-control values over a 6-day period. At 24 hr after carbon tetrachloride administration, a single-pass liver perfusion of 125I-hepatocyte growth factor was performed, and its results were compared with the control results. After a 15-min perfusion of 125I-hepatocyte growth factor, we washed the liver sequentially with heparin and then with acid buffer to separately determine the cell-surface-bound and internalized 125I-hepatocyte growth factor. In carbon tetrachloride-intoxicated rats, both the acid-washable binding and the internalized 125I-hepatocyte growth factor dropped to almost half of the control values, but the decrease in heparin-washable binding was minimal. In contrast, when 125I-hepatocyte growth factor was perfused with excess unlabeled hepatocyte growth factor (135 pmol/L), mostly saturating the cell-surface receptors, the change in cell-surface-bound 125I-hepatocyte growth factor and internalized 125I-hepatocyte growth factor in carbon tetrachloride-intoxicated rats was minimal. This finding, along with our previous finding that the cell-surface hepatocyte growth factor receptors are greatly down-regulated in carbon tetrachloride-intoxicated rats, suggests that the hepatic clearance of hepatocyte growth factor through receptor-mediated endocytosis decreases in carbon tetrachloride-intoxicated rats. The decrease in the hepatic clearance of hepatocyte growth factor could be one of the causes of the elevated hepatocyte growth factor level in the circulating blood in liver diseases. (Hepatology 1993;17:651-660.)
Time course of ethanol-induced impairment in fluid-phase endocytosis in isolated rat hepatocytes
Authors: Camacho Kenneth B. , Casey Carol A. , Wiegert Robert L. , Sorrell Michael F. , Tuma Dean J.
Published: March 2007

The time-course effects of long-term ethanol administration on fluid-phase endocytosis were studied in isolated rat hepatocytes. Rats were pair-fed an ethanol-supplemented liquid diet or an isocaloric control diet for 3 days, 1 wk, 2 wk or 5 wk. Hepatocytes were isolated and incubated at 37° C with various concentrations of the fluid-phase marker Lucifer yellow.-Net internalization of the marker dye was determined. After as little as 1 wk, ethanol-fed rats demonstrated marked decreases in the net internalization of dye compared with pair-fed controls; these changes persisted throughout 5 wk of feeding. Because net internalization is the balance between uptake into the cells vs. efflux from the cells, these components were examined individually. Early uptake was not significantly decreased by ethanol feeding; however, efflux of preloaded Lucifer yellow from cells from the ethanol-fed animals was markedly faster than efflux from pair-fed controls. This increased efflux was more prominent in the longer preload time (90 min) compared with a shorter preload time (15 min), indicating an alteration in dye distribution among various intracellular pools. These ethanol-induced changes in fluid-phase endocytosis were apparent for 1 wk through 5 wk of feeding and were similar for all Lucifer yellow concentrations examined. These results indicate that the decreased net internalization of Lucifer yellow through fluid-phase endocytosis is mainly a result of an ethanol-induced increase in efflux possibly caused by altered intracellular trafficking rather than by reduction in uptake. (Hepatology 1993;17:661-667.)
Effect of hypoxia on the hepatic metabolism of lidocaine in the isolated perfused pig liver
Authors: Mets Berend , Hickman Rosemary , Allin Rosemary , van Dyk Jean , Lotz Zoe
Published: March 2007

The metabolism of lidocaine to monoethylglycinexylidide has been found useful as an indicator of liver function in association with liver transplantation. It has been postulated that this is due to the common effect of hypoxic damage on liver function and lidocaine metabolism. The effects of hypoxia on the elimination of lidocaine and the formation of mono-ethylglycinexylidide and on indexes of liver function were investigated with the isolated perfused pig liver preparation. This study was performed at similar hepatic effluent lidocaine concentrations of approximately 5 μg ml−1 in normoxic (n = 7) and hypoxic (n = 8) livers of similar mass harvested from male Landrace x Large White pigs and perfused at standard unit hepatic flow rates. Whole blood lidocaine extraction ratio was 0.63 ± 0.02 in normoxic livers 30% O2 at oxygenator inflow. It was significantly less (0.23 ± 0.03) in livers subjected to hypoxia (2% O2 at oxygenator inflow), as were hepatic clearance (57.1 ± 2.1 vs. 20.3 ± 3.1 ml ± min−1 ± 100 gm−1), intrinsic clearance (1,706 ± 182 vs. 284 ± 53 ml ± min−1 ± 100 gm−1) and monoethylglycinexylidide formation as indicated by monoethylglycinexylidide/lidocaine ratios in the hepatic venous effluent (0.379 ± 0.061 vs. 0.073 ± 0.014) (p < 0.01). Hepatic oxygen consumption, adenine nucleotide status and bile flow were significantly impaired by hypoxia. Whereas perfusate potassium concentration increased early, AST levels showed delayed increases and ALT levels showed no changes. These changes correlated strongly with hepatic lidocaine elimination (p < 0.01). We conclude that lidocaine metabolism may be an early indicator of severe hepatic hypoxia. (Hepatology 1993;17:668-676.)