bluuebunny/crossref_metadata_2025 huggingface dataset:
Apoptosis is induced by transforming growth factor-β1 within 5 hours in regressing liver without significant fragmentation of the DNA
Authors: Oberhammer Franziska , Bursch Wilfried , Tiefenbacher Roman , Fröschl Gertraud , Pavelka Margit , Purchio Tonio , Schulte-Hermann Rolf
Published: March 2007

In previous studies we showed that transforming growth factor-β1 induces apoptosis in hepatocyte cultures and regressing livers, the mature form being more potent than the transforming growth factor-β1 latency-associated protein. In this study we addressed the question of whether apoptosis can be induced within a short time after administration of transforming growth factor-β1. Five hours after a single intravenous injection of 25 μg mature transforming growth factor-β1/kg body weight, apoptosis is augmented ninefold in the regressing rat liver. A second preceding application induces no further augmentation. Transforming growth factor-β1 latency-associated protein shows no effect with either regimen. Morphological evaluation shows that 5 hr after injection of transforming growth factor-β1 nearly all apoptotic bodies are already engulfed by their neighbor cells. After homogenization of the transforming growth factor-β1-treated livers, the condensed apoptotic bodies are not destroyed and remain in the nuclear pellet. No DNA fragmentation into oligosomes could be detected after purification of the DNA from the nuclear pellet and application to conventional gel electrophoresis. Application of in situ nick translation, which allows detection of DNA single and double-strand breaks in individual apoptotic bodies, also revealed no substantial fragmentation of the DNA in apoptotic bodies. These studies show that transforming growth factor-β1 is able to induce apoptosis within a rather short time and also suggest that in vivo digestion of the DNA does not lead to chromatin condensation. (HEPATOLOGY 1993;18:1238-1246).
Alcohol consumption enhances fatty acid ω-oxidation, with a greater increase in male than in female rats
Authors: Ma Xiaoli , Baraona Enrique , Lieber Charles S.
Published: March 2007

Because ethanol inhibits mitochondrial fatty acid oxidation, with substantial accumulation of fatty acids in the livers of female (but not male) rats, and induces microsomal activities, we assessed possible changes in ω-oxidation. To study this, we pair-fed 24 male and 24 female littermate rats of the same age liquid diets containing 36% of energy either as ethanol or as additional carbohydrate for 4 wk. In controls, the microsomal ω-hydroxylation of lauric acid was 28% greater in female than in male rats (p < 0.05). Ethanol feeding significantly increased this activity in both genders (p < 0.01), but the rise in male rats (89%) was significantly higher than that in female rats (24%). This activity was unaffected by the presence of ethanol in the assay. The effects of ethanol were associated with increases in the content of cytochrome P-450 4A1 (as assessed in Western blots by the reactivity against a sheep antibody against P-450 4A1), and more so in male than in female rats. Despite possible competition by ethanol with the hydroxy fatty acid oxidation to dicarboxylic acids through alcohol dehydrogenase, suberic and sebacic acids accumulated significantly in the livers of alcohol-fed male rats. These effects of ethanol and gender on ω-oxidation paralleled those on the hepatic cytosolic fatty acid-binding protein and fatty acid esterification previously reported in similarly treated rats. Dicarboxylic acid products of ω-oxidation have been incriminated as mediators of similar effects by other drugs. Thus the increase in ω-oxidation may compensate, at least in part, for the deficit in fatty acid oxidation caused by the ethanol-induced injury of the mitochondria by promoting fatty acid binding, esterification and oxidation through alternate pathways. This compensatory effect is less efficient in female than in male rats, which may explain why alcohol administration results in a striking and potentially deleterious accumulation of fatty acids in the livers of female but not of male rats. (HEPATOLOGY 1993;18:1247-1253).
True transport: One or more sodium-dependent bile acid transporters?
Authors: Lester Roger , Zimniak Piotr
Published: March 2007

Sodium-dependent bile acid transport is a well-established function of the sinusoidal segment of the hepatocyte plasma membrane. Evidence has been provided previously by the authors for the existence of a putative sinusoidal plasma membrane sodium-dependent bile acid transporter with a mass of 49 kD. This protein has been partially characterized with a monoclonal antibody and by reconstitution in proteoliposomes. Further characterization is provided in the paper under discussion. The transporter was isolated from plasma membranes and from endoplasmic reticulum by immunoprecipitation with a monoclonal antibody. The protein was separated from antibody by means of HPLC and SDS-PAGE with electroelution. Amino acid analysis and NH2-terminal amino acid sequencing were performed. For both preparations, it was shown that the NH2-terminal amino acid sequence was identical, and the amino acid composition was similar to results obtained for the microsomal enzyme epoxide hydrolase. Purified mEH was immunoprecipitated with the monoclonal antibody which precipitated the plasma membrane protein. Isoelectric focusing established that the isoelectric points for epoxide hydrolase and the proteins isolated from the endoplasmic reticulum and plasma membranes were identical. The peptide maps prepared by subjecting the three proteins to enzymatic fragmentation with subtilisin were essentially identical. Because of the known contamination of plasma membrane fractions with endoplasmic reticulum and because of the high specific activity of epoxide hydrolase in endoplasmic reticulum, special attention was devoted to the question of the localization of the enzyme/transporter in plasma membrane. It was possible to show that virtually all of the immunoprecipitated protein obtained from intact cells derived from plasma membrane and that only a small fraction could be explained as having originated in endoplasmic reticulum. Finally, a hydrophobic plot of the amino acid sequence of epoxide hydrolase suggested that the protein contained four potential transmembrane domains. It could be concluded that a/the 49-kD sinusoidal plasma membrane sodium-dependent bile acid transporter protein was closely similar or identical to epoxide hydrolase. In view of the relatively small mass of the protein and its limited number of potential transmembrane domains, it appeared probable that the physiological membrane transporter consists of an assembly of 49-kD monomers or an assembly of heterologous proteins including the 49-kD protein.
Long-term follow-up of patients with chronic hepatitis C treated with different doses of interferon-α2b
Authors: Saracco Giorgio , Rosina Floriano , Abate Maria Lorena , Chiandussi Livio , Gallo Vittorio , Cerutti Elena , Napoli Angelo Di , Solinas Antonio , Deplano Angelo , Tocco Andreina , Cossu Pierangela , Chien David , Kuo George , Polito Alan , Weiner Amy J. , Houghton Michael , Verme Giorgio , Bonino Ferruccio , Rizzetto Mario
Published: March 2007

Eighty patients with chronic hepatitis C who completed a previously reported randomized controlled trial on the efficacy of interferon-α2b were followed up for at least 36 mo after therapy discontinuation. Seventeen patients (21.2%) maintained normal ALT values throughout the follow-up; 63 (78.8%) either did not normalize the levels of ALT or relapsed during the follow-up. A significantly greater proportion of patients treated with 3 million units of interferon three times a week subcutaneously for 48 wk were long-term responders compared with patients treated for 24 wk. Sex, age, hepatitis C virus antibody status, source of infection and pretreatment levels of ALT were not predictive of long-term response. Cirrhosis was found to be an unfavorable predictive factor. After 3 yr of follow-up, clearance of viremia was observed in 58.9% of the 17 long-term responders but in none of the nonresponders (p = 0.002). E2-NS1 antibody tested negative in 88.2% of long-term responders and in 14.3% of nonresponders (p = 0.001). Fifty-nine percent of long-term responders tested negative for C100-NS4 antibody compared with 14.3% of nonresponders (p = 0.031). No significant change was observed in other antibodies. Four long-term responders underwent liver biopsy 2 yr after discontinuation of therapy. All four patients had normal liver histology compared with baseline assessment of chronic active hepatitis in three and chronic persistent hepatitis in the other. Three of the four were negative for serum hepatitis C virus RNA. (HEPATOLOGY 1993; 18:1300-1305.)
Monitoring of antiviral therapy with quantitative evaluation of hbeag: A comparison with HBV DNA testing
Authors: Perrillo Robert , Mimms Larry , Schechtman Kenneth , Robbins David , Campbell Carolyn
Published: March 2007

The serological endpoint of response in the treatment of chronic hepatitis B is the loss of hepatitis B virus DNA and HBeAg. Because the quantitative measurement of hepatitis B virus DNA in serum has been shown to be useful for monitoring and predicting response to interferon-α therapy, we decided to evaluate whether changes in HBeAg concentration could also be used in this manner. Twenty-nine patients who were initially positive for HBeAg and HBV DNA were serially evaluated for HBeAg concentration with a microparticle-capture enzyme immunoassay. HBeAg levels in serum were calculated by means of comparison with a standard curve of fluorescence rate vs. HBeAg concentration. The results, expressed in milliunits per milliliter, were compared with hepatitis B virus DNA levels determined by means of solution hybridization. The baseline HBeAg concentration proved to be the best independent predictor of response on stepwise Cox regression analysis (p = 0.026). Similar disappearance curves were observed for the two markers, although hepatitis B virus DNA became undetectable at an earlier interval in 13 of 16 cases (81%). In the 16 responders, a decline in HBeAg concentration of more than 90% was observed by wk 12 of therapy (mean ± S.D., 95% ± 13%). Nonresponders did not demonstrate such steep declines in HBeAg values by wk 12 (mean ± S.D., 45% ± 27%), and levels tended to increase at subsequent time points. We conclude that serial monitoring of HBeAg concentration with a technique that should be readily adaptable to clinical laboratories may be useful in the initial evaluation and monitoring of patients undergoing antiviral therapy. (HEPATOLOGY 1993;18:1306-1312.)
Interferon-α2b therapy reduces liver fibrosis in chronic non-A, non-B hepatitis: A quantitative histological evaluation
Authors: Manabe Noboru , Chevallier Michèle , Chossegros Philippe , Causse Xavier , Guerret Sylviane , Trépo Christian , Grimaud Jean-Alexis
Published: March 2007

The aim of this study was to evaluate the effect of interferon-α on liver fibrosis with an established quantitative histochemical method for determining collagen as a marker. 59 patients (31 men, 28 women; 47 ± 14 yr) with chronic non-A, non-B hepatitis (92% with hepatitis C virus antibody) received subcutaneous injections of 3 or 1 MU recombinant interferon-α2b or placebo thrice weekly for 24 wk. Needle-biopsy sections taken before and after interferon treatment were examined for histological evaluation and collagen quantitation. Values were compared with results obtained by means of morphometrical analysis of liver collagen and Knodell scoring histological index. The index of periportal and/or bridging necrosis was the only component of Knodell’s histological score significantly decreased (p < 0.05) in patients treated with 3 MU interferon compared with placebo-treated controls. The fibrosis score was not significantly changed. In contrast, liver total collagen variations measured colorimetrically and morphometrically were significantly decreased in patients treated with 3 MU and 1 MU compared with the increase observed in the placebo-treated controls (p < 0.05). From these results, we conclude that a 6-mo course of 3 MU or 1 MU interferon-α2b causes slight but nonetheless significant regression of liver fibrosis as assessed on the basis of quantitative estimation of liver collagen, irrespective of other response criteria, whereas progression of liver fibrosis can be observed in the absence of treatment. (HEPATOLOGY 1993;18:1344-1349.)
Erythropoietin production in hepatocellular carcinoma cells associated with polycythemia: Immunohistochemical evidence
Authors: Sakisaka Shotaro , Watanabe Masahide , Tateishi Hideo , Harada Masaru , Shakado Satoshi , Mimura Yoshihiro , Gondo Kazuhisa , Yoshitake Masao , Noguchi Kazunori , Hino Teruko , Nohno Ryuichi , Majima Yasuo , Hirai Kenji , Sata Michio , Yoshida Hiroshi , Tanikawa Kyuichi
Published: March 2007

Patients with hepatocellular carcinoma sometimes have erythrocytosis and high plasma erythropoietin levels. However, previous studies have not revealed direct evidence that the carcinoma cells produce the erythropoietin. To address this question, we carried out light and electron microscopic immunohistochemical studies, using a human erythropoietin antibody to the liver in three male patients with hepatocellular carcinoma and erythrocytosis. α-Fetoprotein localization was also examined in serial liver sections by light microscopic immunohistochemistry with an antibody to α-fetoprotein. All three patients demonstrated high hemoglobin levels (16.7, 17.6 and 18.1 gm/dl) and high plasma erythropoietin levels (227, 266 and 280 mU/ml). In one patient the plasma erythropoietin level in the hepatic vein was significantly higher than that in the hepatic artery. The levels of plasma erythropoietin, as well as such tumor markers for hepatocellular carcinoma as serum α-fetoprotein and plasma des-γ-carboxyprothrombin, were significantly reduced after treatment with an anticancer drug, cisplatin. Light microscopic immunohistochemistry showed that erythropoietin was definitely present in the cytoplasm of the hepatocellular carcinoma cells, but not in normal hepatocytes around the carcinoma lesion or in other nonparenchymal cells such as vascular endothelial cells and Kupffer cells. In electron microscopic immunohistochemistry, reaction products for erythropoietin were revealed in the cisternae of the endoplasmic reticulum in the carcinoma cells, suggesting the production of erythropoietin by these cells. Light microscopic immunohistochemistry showed that α-fetoprotein was localized in the hepatocellular carcinoma cells that were erythropoietin positive in the serial sections. These findings indicated that hepatocellular carcinoma cells produced erythropoietin as well as α-fetoprotein in these cases, leading to the complication of erythrocytosis. (HEPATOLOGY 1993;18:1357-1362.)
High plasma concentration of myeloperoxidase in cirrhosis: A possible marker of hypersplenism
Authors: Nakamuta Makoto , Ohashi Masao , Tanabe Yuichi , Hiroshige Kaichiro , Nawata Hajime
Published: March 2007

Plasma myeloperoxidase levels in patients with cirrhosis were compared with those in patients with chronic hepatitis and healthy controls by means of a specific radioimmunoassay for myeloperoxidase. The mean concentration of plasma myeloperoxidase in cirrhotic patients (309.1 ± 17.2 ng/ml, n = 41) was markedly higher than that in chronic hepatitis patients (222.6 ± 17.2 ng/ml, n = 21) (p < 0.01) and normal controls (219.5 ± 5.7 ng/ml, n = 50) (p < 0.01). Plasma myeloperoxidase showed good negative correlations with neutrocyte count (r = −0.32, p < 0.01), thrombocyte count (r = −0.40, p < 0.01), red blood cell count (r = −0.32, p < 0.01), serum albumin (r = −0.35, p < 0.01), and cholinesterase (r = −0.32, p < 0.02) and positive correlations with serum alkaline phosphatase (r = 0.49; p < 0.01) and lactate dehydrogenase (r = 0.31, p < 0.01) in patients with cirrhosis or chronic hepatitis. Among lactate dehydrogenase isozymes, a good positive correlation was seen between plasma myeloperoxidase and lactate dehydrogenase-2 (r = 0.40, p < 0.01) and lactate dehydrogenase-1 (r = 0.03, p < 0.02). Plasma myeloperoxidase was significantly higher in the cirrhotic and chronic hepatitis patients with splenomegaly (341.1 ± 19.4 ng/ml, n = 31) than in those without splenomegaly (217.4 ± 12.2 ng/ml, n = 29) (p < 0.01). We also examined the difference between plasma levels of myeloperoxidase in the portal and peripheral blood. In the patients with nonhepatic disease, we found a significant portalsystemic difference (portal myeloperoxidase, 221.5 ± 19.7 ng/ml, vs. systemic myeloperoxidase, 179.1 ± 10.8 ng/ml; n = 15) (p < 0.01), whereas there was no such difference in the cirrhotic patients (portal myeloperoxidase, 281.5 ± 15.2 ng/ml vs. systemic myeloperoxidase, 268.0 ± 28.2 ng/ml; n = 7). A significantly high concentration of plasma myeloperoxidase was evident in the case of esophageal varices, which is a sign of portalsystemic shunting in human beings. The results of our study imply that the release of myeloperoxidase from destroyed neutrocytes, in addition to its decreased hepatic clearance, are responsible for the high concentrations of plasma myeloperoxidase in cirrhotic patients. Therefore measurement of plasma myeloperoxidase may be useful in assessing hypersplenic function in liver disease. (HEPATOLOGY 1993;18:1377-1383.)
Histological evidence for recurrence of primary biliary cirrhosis after liver transplantation
Authors: Balan Vijayan , Batts Kennetii P. , Porayko Michael K. , Krom Ruud A. F. , Ludwig Jurgen , Wiesner Russell H.
Published: March 2007

Whether primary biliary cirrhosis recurs after orthotopic liver transplantation remains a controversial issue. Sixty consecutive patients with primary biliary cirrhosis with at least 1 yr of follow-up after liver transplantation were studied. All patients were treated with triple-drug immunosuppression (cyclospoine, prednisone, azathioprine). Hepatic biochemical parameters and protocol liver biopsy specimens were evaluated 1 wk, 3 wk, 4 mo and yearly after orthotopic liver transplantation and at times of liver dysfunction. Antimitochondrial antibody titers and IgM levels were determined at 4 mo and yearly. At the time of last follow-up, all patients had marked symptomatic improvement compared with their pretransplant condition, and 91% of the patients had normal hepatic biochemical parameters, including serum levels of alkaline phosphatase, bilirubin, ALT and IgM. In addition, all patients had significant decreases in antimitochondrial antibody titer (p = 0.0001) and significant decreases in serum levels of IgM (p = 0.0001). Forty-one of the 60 patients had near-normal liver histological appearance. Of those with abnormal histological appearance, five patients, 2 to 6 yr after orthotopic liver transplantation, had histological features typical of a florid duct lesion, suggesting recurrent primary biliary cirrhosis. All five patients with portal granulomas had normal hepatic biochemical values and were clinically asymptomatic. Two of the five patients had persistent antimitochondrial antibody titers. We consider the documented histological changes highly suggestive of recurrence of primary biliary cirrhosis after liver transplantation but, so far, have no evidence that the condition in these patients is progressive. (HEPATOLOGY 1993;18:1392-1398.)
Role of ethanol-inducible cytochrome p-450 2E1 in the development of hepatocellular carcinoma by the chemical carcinogen, N-nitrosodimethylamine
Authors: Tsutsumi Mikihiro , Matsuda Yoshiro , Takada Akira
Published: March 2007

Cytochrome P-450 2E1 is a specific isozyme of cytochrome P-450 induced by ethanol. P-450 2E1 may also be the only enzyme that metabolizes N-nitrosodimethylamine at a very low concentration. Because N-nitrosodimethylamine is a procarcinogen, the possibility that induction of P-450 2E1 by alcohol abuse may accelerate the carcinogenic action of a very small dose of N-nitrosodimethylamine should be considered. To investigate this possibility, effects of ethanol on the hepatic carcinogenic action of a very small dose of N-nitrosodimethylamine were analyzed in rats. Sixty Wistar male rats were divided into two groups (ethanol and control) according to the liquid diets they were fed. After preliminary feeding, N-nitrosodimethylamine in two concentrations (1.5 ng/ml and 3.0 μg/ml) in drinking water was given with the diets for 40 or 60 wk. To avoid metabolic competition between ethanol and n-nitrosodimethylamine, we gave N-nitrosodimethylamine and the diets alternately. The average amount of N-nitrosodimethylamine consumed per day was 11 to 37 ng/kg body weight in the 1.5-ng group and was 27 to 37 μg/kg body weight in the 3.0-μg group. Hepatic P-450 2E1 content was six times higher in the ethanol-treated groups than in the control groups throughout the experiment. Visible nodules and enzyme-altered foci were not found in rats in any group at the end of the fortieth week. These preneoplastic changes were found at the end of the sixtieth week only in rats in the groups treated with both ethanol and N-nitrosodimethylamine. However, these changes were not found in rats treated with N-nitrosodimethylamine or ethanol alone for the same period. These results suggest that long-term administration of ethanol may accelerate the development of liver cancer induced by N-nitrosodimethylamine, and this enhancing effect may be related to the induction of P-450 2E1 by ethanol. Enhancement was observed in rats given a small dose of N-nitrosodimethylamine, suggesting that in alcoholic individuals cancer may develop from exposure to small doses of carcinogen in normal life. (HEPATOLOGY 1993;18:1483-1489.)
Cholesterol carriers in human bile: Are “lamellae” involved?
Authors: Cohen David E. , Kaler Eric W. , Carey Martin C.
Published: March 2007

Cholesterol, a highly insoluble molecule, is transported in bile by specialized lipid aggregates. On the basis of extensive correlations between laboratory-prepared model biles and surgically harvested native biles, it has become generally accepted that biliary cholesterol is solubilized by simple and mixed micelles, single bilayered (unilamellar) vesicles and, under certain conditions, multilamellar vesicles (liposomes or liquid crystals) all composed of bile salts, lecithin and cholesterol in different proportions. Current concepts suggest that in lithogenic biles multilamellar vesicles result from aggregation and fusion of unilamellar vesicles and are a principal source from which cholesterol precipitates to form gallstones. Recent reports now challenge the prevailing paradigm by proposing that the principal cholesterol-carrying particles in human biles are not micelles but are “lamellae” composed of stacked membrane-like bilayers of lipids. In this article, we provide a critical overview of the experiments that led to the established views of biliary cholesterol transport and to the newer lamellae hypothesis. The principal evidence for lamellae stems from negative-stain electron microscopy, an artifactprone technique when used to study lipid-rich fluids such as bile. We show that lamellar structures represent both the electron microscopic analog of multilamellar vesicles in supersaturated biles that presage the nucleation of cholesterol crystals and an electron microscopic artifact of fossilized mixed micelles that are in fact very tiny (2 to 4 nm in radius) by state-of-the-art noninvasive techniques. We argue further that the lamellae nomenclature improperly equates two fundamentally distinct physical-chemical mechanisms for cholesterol solubilization and dispersion in bile on the basis of identically appearing electron microscopic images. (HEPATOLOGY 1993;18:1522-1532.)
Hepatocyte apoptosis: Is transforming growth factor-B1 the kiss of death?
Authors: Fausto Nelson
Published: March 2007

In previous studies hepatocytes undergoing cell death by apoptosis but not normal hepatocytes in rat liver showed immunostaining for transforming growth factor β1 (TGF-β1). Staining was much stronger with antibodies recognizing the pro-region of TGF-β1 than the mature peptide itself. Therefore we investigated the ability of both forms of TGF-β1 to induce apoptosis in primary cultures of rat hepatocytes. Mature TGF-β1 induced rounding up of the cells and fragmentation into multiple vesicles. As revealed by the DNA-specific stain H33258, the chromatin of these cells condensed and segregated into masses at the nuclear membrane; this was obviously followed by fragmentation of the nucleus. Ultrastructurally the cytoplasm was well preserved, as demonstrated by the presence of intact cell organelles. These features strongly suggest the occurrence of apoptosis. Quantification of nuclei with condensed chromatin revealed that mature TGF-β1 was 30-fold more effective than the TGF-β1 latency-associated protein complex. Finally, we administered TGF-β1 in vivousing an experimental model in which regression of rat liver was initiated by a short preceding treatment with the hepatomitogen cyproterone acetate. Two doses of TGF-β1, each 1 nM/kg, augmented the incidence of apoptotic hepatocytes 5-fold. Equimolar doses of TGF-β1 latency-associated protein complex were ineffective. These studies suggest that TGF-β1 is involved in the initiation of apoptosis in the liver and that the mature form of TGF-β1 is the active principle.
Pretransplantation interferon treatment and recurrence of hepatitis B virus infection after liver transplantation for hepatitis B-related end-stage liver disease
Authors: Marcellin Patrick , Samuel Didier , Areias Jorge , Loriot Marie-Anne , Arulnaden Jean-Louis , Gigou Michèle , David Marie-FrançOise , Bismuth Alain , Reynes Michel , Bréchot Christian , Benhamou Jean-Pierre , Bismuth Henri
Published: March 2007

Orthotopic liver transplantation in patients with hepatitis B-related cirrhosis is commonly complicated by reinfection with the hepatitis B virus, with rapidly progressive liver disease and poor survival rate. We assessed the efficacy of prior therapy with recombinant interferon-α on the prevention of posttransplantation hepatitis B virus reinfection. Twenty-two patients with hepatitis B-related cirrhosis waiting for liver transplantation received 3 MU (decreased to 1.5 MU in cases of intolerance) of recombinant interferon-α until transplantation. The rates of posttransplantation hepatitis B virus reinfection and survival in this group were compared with those in a group of 26 patients previously given transplants for the same disease but not given interferon therapy. The same protocol of HBs antibody passive immunoprophylaxis was applied after transplantation in both groups. Recombinant interferon-α was administered for 14 ± 7 wk. The treatment had an antiviral effect, with disappearance of serum hepatitis B virus DNA in seven of the eight patients initially positive for hepatitis B virus DNA and disappearance of HBeAg in two of the three patients initially positive for HBeAg. Serum hepatitis B virus DNA remained detectable with polymerase chain reaction at transplantation in 56 of the interferontreated patients. After transplantation, hepatitis B virus reinfection was more frequent in polymerase chain reaction-positive than in polymerase chain reaction-negative patients (78 vs. 17, p < 0.05). One patient’s condition deteriorated during interferon treatment; this patient was not given a transplant. Two patients (in whom hepatitis B virus DNA disappeared from serum) improved so markedly during treatment that they were not given transplants. The rates of posttransplant hepatitis B virus reinfection (44 and 58, respectively), the delay of hepatitis B virus reinfection (8 ± 5 mo and 6 ± 3 mo, respectively) and the survival rates (83 and 69, respectively) were not statistically different in the interferon-treated and untreated groups. We conclude that, in patients with hepatitis B-related cirrhosis, pretransplantation recombinant interferon-α, at the dosage and duration we used, has an antiviral effect but does not prevent posttransplantation hepatitis B virus reinfection. Detection of serum hepatitis B virus DNA with polymerase chain reaction before transplantation might be useful in predicting reinfection after transplantation. (Hepatology 1994;19:6-12).
A multicenter randomized controlled trial of recombinant interferon-α2b in patients with acute transfusion-associated hepatitis C
Authors: Lampertico Pietro , Rumi Mariagrazia , Romeo Raffaella , Craxi Antonio , Soffredini Roberta , Biassoni Daniela , Colombo Massimo
Published: March 2007

To assess whether interferon-α might prevent non-A, non-B hepatitis from becoming chronic, 45 consecutive patients with transfusion-associated hepatitis were enrolled in a randomized clinical trial. Thirty-eight patients had hepatitis C virus infection, and 7 had non-A, non-B, non-C hepatitis. Twenty-six patients (22 with HCV) were given 3 MU of recombinant interferon-α2b three times a week for 12 wk, whereas 19 (16 with HCV) were not. Biochemical and virological parameters were monitored at regular intervals during an 18-mo follow-up. At the end of the 3-mo therapy, 16 (73) patients with hepatitis C had normal serum ALT activity, compared with 7 (44) who were not treated (NS). Fifty-three percent of the treated patients and none of the untreated patients had normal ALT levels and no HCV RNA (p = 0.0087). At the end of the 18-mo follow-up, 13 (59) treated patients had normal ALT levels, compared with 6 (37) untreated controls (NS). Thirty-nine percent had normal ALT and no HCV RNA, compared with none of the controls (p = 0.035). Four patients (22) had had sustained complete responses to interferon, defined as normal ALT levels and no HCV RNA at the end of the 3-mo treatment period and the 18-mo follow-up period. All seven patients with non-A, non-B, non-C hepatitis, treated and untreated, recovered uneventfully from hepatitis. One major finding was that short-term treatment with interferon-α was effective in most patients with acute hepatitis C and led to complete recovery from hepatitis in 39 of the cases. (Hepatology 1994;19:19-22).
In vivo evidence of enhanced guanylyl cyclase activation during the hyperdynamic circulation of acute liver failure
Authors: Schneider Francis , Lutun Philippe , Boudjema Karim , Wolf Philippe , Tempé Jean-Daniel
Published: March 2007

Nitric oxide and atrial natriuretic peptides are the main activators of guanylyl cyclases, which transform ITP into cyclic GMP and thereby contribute to the decrease of vascular tone. To investigate the increase, if any, of plasma cyclic GMP concentrations in human patients with hyperdynamic circulation resulting from acute liver failure and to ascertain whether guanylyl cyclase activation is involved in the decline of systemic vascular resistance that occurs in this pathophysiological condition, we simultaneously recorded hemodynamic data and cyclic GMP levels in patients with fulminant liver failure before and after liver transplantation and in normokinetic patients undergoing abdominal nonseptic surgery. We also compared these data with those recorded in patients with hyperkinetic shock resulting from gram-negative sepsis or nitric oxide-independent vasomotor agent (carbamate) overdose. In all these patients we simultaneously studied kidney function, platelet counts and atrial natriuretic peptides. Patients with fulminant liver failure had higher cyclic GMP concentrations than did control patients undergoing abdominal surgery (11.02 ± 1.55 pmol·ml−1 vs. 1.77 ± 0.18 pmol · ml−1, P < 0.001). At similar heart-loading conditions these concentrations were lower than those in gram-negative septic shock (18.2 ± 1.35 pmol·ml−1, p < 0.05) but higher than those in carbamate-induced shock (3.6 ± 0.7 pmol·ml−1, p < 0.01). In addition, cyclic GMP concentrations significantly decreased from the fulminant liver failure period to the posttransplantation period, although atrial natriuretic peptide levels did not change significantly and kidney function worsened. During the period of fulminant liver failure, cyclic GMP levels correlated with the decline in peripheral resistance but not with other hemodynamic or biological data. With the improvement of peripheral resistance after the transplantation procedure, they did not further correlate with systemic vascular resistance, only with creatininemia. Together these results suggest that guanylyl cyclase activation is involved in the low vascular resistance observed during acute hepatocellular syndrome; furthermore, they indicate that particulate guanylyl cyclase activation only partially explains the increase in cyclic GMP observed during fulminant liver failure and put forward a role for soluble guanylyl cyclase activators such as nitric oxide. (Hepatology 1994;19;38-44).
Reduction of gastric hyperemia by glypressin and vasopressin administration in cirrhotic patients with portal hypertensive gastropathy
Authors: Panés Julián , Piqué Josep M. , Bordas Josep M. , Llach Josep , Bosch Jaime , Terés Josep , Rodés Juan
Published: March 2007

Gastric mucosal perfusion is increased in portalhypertensive gastropathy, and this may contribute to gastric bleeding from these lesions. Therefore drugs reducing gastric mucosal perfusion may be beneficial in the treatment of overt bleeding from portal-hypertensive gastropathy. In this study gastric mucosal perfusion was assessed in 28 cirrhotic patients with portal-hypertensive gastropathy under basal conditions and after double-blind intravenous administration of vasopressin (0.4 U/min), glypressin (2-mg injection) or placebo, with laser-Doppler flowmetry and reflectance spectrophotometry. Vasopressin and glypressin induced a significant increase in blood pressure and a decrease in heart rate. These effects were more pronounced in the vasopressin group. Both vasopressin and glypressin induced a sustained and similar reduction in gastric mucosal perfusion as assessed by laser-Doppler flowmetry (−36 ± 8 and −34 ± 6, respectively; p < 0.05 with respect to basal values and with respect to the control group), whereas placebo had no effect. Both drugs significantly reduced the oxygen content of the gastric mucosa; however, the impairment in mucosal oxygenation was greater (p < 0.05) in the vasopressin group (−17 ± 3) than in the glypressin group (−6 ± 0.1). We conclude that the increased gastric perfusion in cirrhotic patients with portal-hypertensive gastropathy may be reduced by either vasopressin or glypressin. These findings support the use of these drugs in clinical trials treating bleeding portal-hypertensive gastropathy. The lower reduction in gastric mucosal oxygen content observed with glypressin could decrease the incidence of ischemic adverse events associated with the use of vasopressin. (Hepatology 1994;19:55-60).
Prospective study of α-fetoprotein in cirrhotic patients monitored for development of hepatocellular carcinoma
Authors: Oka Hiroko , Tamori Akihiro , Kuroki Tetsuo , Kobayashi Kenzo , Yamamoto Sukeo
Published: March 2007

The usefulness of measurements of serum α-fetoprotein elevation for diagnosis of the development of hepatocellular carcinoma was evaluated by a prospective study of 260 patients with cirrhosis. Hepatocellular carcinoma was found in 55 patients during the 5-yr follow-up, excluding 7 found to have hepatocellular carcinoma in the first 6 mo. The cumulative incidence of hepatocellular carcinoma was 26 in the 185 patients who had α-fetoprotein levels below 20 ng/ml at the time of entry and 46 in the 68 patients who had α-fetoprotein levels of 20 ng/ml or more but below 200 ng/ml. In 169 of the patients, serum levels of α-fetoprotein were assayed regularly for at least 2 yr. The incidence of hepatocellular carcinoma development in the 36 patients who had repeated transient increases in α-fetoprotein to above 100 ng/ml was 36. This was significantly higher than the incidence in the 99 patients who had α-fetoprotein levels consistently below 20 ng/ml. Thus patients who had α-fetoprotein levels of 20 ng/ml or more, who had transient increases in α-fetoprotein or who had both should be treated as being in a super-high-risk group for hepatocellular carcinoma. Frequent and careful examination by ultrasonography of such patients is recommended. (Hepatology 1994;19:61-66).
Cerebral hemodynamic and metabolic changes in fulminant hepatic failure: A retrospective study
Authors: Aggarwal Shushma , Kramer David , Yonas Howard , Obrist Walter , Kang Yoogoo , Martin Maureen , Policare Raymond
Published: March 2007

The purpose of this retrospective study was to determine cerebral hemodynamic and metabolic changes in comatose patients with fulminant hepatic failure. Computerized tomography of the brain and cerebral blood flow measurements by the xenoncomputerized tomography scan or intravenous xenon-133 methods were obtained in 33 patients with fulminant hepatic failure. In a subgroup of 22 patients, arteriojugular venous oxygen content difference and cerebral metabolic rate for oxygen were determined. Carbon dioxide reactivity was tested in 17 patients, and intracranial pressure was recorded by an epidural monitor in 8 patients. Cerebral blood flow and arteriojugular venous oxygen content difference were adjusted to the average arterial carbon dioxide pressure of the sample (32 mm Hg). Adjusted cerebral blood flow varied from 16.5 to 94.7 ml/100 gm/min; 52 of the patients had reduced adjusted cerebral blood flows (less than 33 ml/100 gm/min), whereas 24 had hyperemic values (greater than 50 ml/100 gm/min). Patients with higher adjusted cerebral blood flows showed cerebral swelling on computerized tomography scan (p < 0.002), were in deeper coma (p < 0.05) and had greater mortality (p < 0.002). The adjusted arteriojugular venous oxygen content difference was negatively correlated with adjusted cerebral blood flow (r = −0.61, p < 0.002). The majority of patients with reduced adjusted cerebral blood flows had low adjusted arteriojugular venous oxygen content differences (less than 5 vol), indicating hyperemia rather than ischemia. The average cerebral metabolic rate for oxygen was 50 of normal (1.6 ± 0.4 ml/100 gm/min); even patients with low cerebral metabolic rates for oxygen recovered neurologically. Carbon dioxide reactivity was preserved in almost all patients, which argues for the effectiveness of hyperventilation in reducing blood flow. All four patients with elevated intracranial pressures (greater than 24 mm Hg) had increased adjusted cerebral blood flows; three of four patients with intracranial pressures less than 20 had reduced adjusted cerebral blood flows. These findings suggest that cerebral swelling, intracranial hypertension and cerebral hyperemia are poor prognostic signs in patients with fulminant hepatic failure. (Hepatology 1994; 19:80-87).
c-met mRNA overexpression in human hepatocellular carcinoma
Authors: Boix Loreto , Rosa Jose Luis , Ventura Francesc , Castells Antoni , Bruix Jordi , Rodés Joan , Bartrons Ramon
Published: March 2007

This study was aimed at assessing the presence of c-met overexpression in human hepatocellular carcinoma and at determining whether this feature is associated with a definite clinical or pathological characteristic. Expression of c-met was determined by Northern-blot hybridization of a specific probe (human met proto-oncogene) in 18 tumoral and nontumoral liver samples obtained in 18 cirrhotic patients with hepatocellular carcinoma submitted to surgical treatment. Eight of the 18 hepatocellular carcinomas exhibited c-met overexpression, with an increase ranging between 2-fold and 10-fold when compared by densi-tometry with the surrounding liver. By contrast, in the remaining 10 cases c-met expression was almost identical to that of the surrounding nontumoral liver tissue. Overexpression of c-met was not related to either the age, sex, etiology or functional status of the underlying liver disease, or to the size of the tumor, to its differentiation degree or to the presence of pseudocapsule invasion and existence of additional neoplastic nodules. These data indicate that almost half of the human hepatocellular carcinomas exhibit c-met overexpression. Nevertheless, the biological relevance of this characteristic is not known. (Hepatology 1994;19:88-91).
Androgen receptor in human liver: Characterization and quantitation in normal and diseased liver
Authors: Eagon Patricia K. , Elm Mary S. , Stafford Elizabeth A. , Porter Lynne E.
Published: March 2007

Liver is responsive to sex hormones. The role of androgens in normal human liver function is not well understood, although androgens have been implicated in several liver diseases. Because the human hepatic androgen receptor has not been adequately characterized, we analyzed cytosolic and nuclear fractions from normal human liver of both sexes for androgenbinding activity using multipoint saturation analysis with the androgenic radioligand methyltrienelone (R1881). Both cytosolic and nuclear fractions of both sexes displayed high affinity R1881 binding (dissociation constants = nanomolar range). The R1881 binding in both fractions is highly specific in that potent androgens compete well, and the antiandrogens hydroxyflutamide and cyproterone acetate show partial competition; other nonandrogenic steroid hormones do not compete. The cytosolic R1881 receptor displays physicochemical characteristics of androgen receptors in other tissues in that it is retained by heparin-Sepharose and by DNA cellulose after activation, and it displays a molybdate-stabilized 8S form on sucrose gradients and a 7.3-nm species on gel filtration chromatography. Receptor activity was also quantitated in specimens of hepatic adenoma, focal nodular hyperplasia and metastatic carcinoma to the liver and in samples of adjacent histologically normal specimens when available. In general, both the diseased and normal portions of the livers from the patients with hepatic adenoma and metastatic carcinoma to the liver, but not focal nodular hyperplasia, demonstrated reduced total androgen-receptor activity as compared with liver from normal individuals. However, in the tumor-containing specimens from all three liver diseases, the nuclear receptor (i.e., the biologically active form) represented a higher proportion of the total receptor activity than was evidenced in either the normal adjacent specimens or liver from normal individuals. This finding suggests that these tumors may differ from normal liver in their responses to androgens. (Hepatology 1994;19:92-100).
Relationship between plasma benzodiazepine receptor ligand concentrations and severity of hepatic encephalopathy
Authors: Basile Anthony S. , Harrison Phil M. , Hughes Robin D. , Gu Zi-Quang , Pannell Lewis , McKinney Angelique , Jones Anthony E. , Williams Roger
Published: March 2007

Levels of benzodiazepine receptor ligands were measured in plasma samples from 25 patients in various stages of hepatic encephalopathy due to fulminant liver failure who were not exposed to pharmaceutical benzodiazepines immediately before or during hospitalization. Chromatographic analysis of extracted plasma samples revealed one to nine fractions containing material that competitively inhibited [3H]flumazenil binding to benzodiazepine receptors with the pharmacological properties of benzodiazepine receptor agonists. Two of these peaks were positively identified as the 1,4-benzodiazepines diazepam and N-desmethyldiazepam on the basis of chromatographic, ultraviolet and mass spectral evidence. The plasma levels of diazepam and N-desmethyldiazepam were significantly increased above control values in stage 4 hepatic encephalopathy, whereas total benzodiazepine receptor ligand concentrations were increased above control in stages 1 through 4. A significant but weak linear correlation was found between the relative increase in the levels of diazepam, N-desmethyldiazepam and total benzodiazepine receptor ligands and the severity of hepatic encephalopathy. Thus increased concentrations of benzodiazepine receptor ligands appear to contribute to the enhancement of GABAergic neurotransmission in hepatic encephalopathy, particularly in stage 4. These results constitute further support for a role for benzodiazepine receptor ligands in the pathogenesis of hepatic encephalopathy associated with acute liver failure. (Hepatology 1994;19:112-121).
Disulfide bond formation between dimeric immunoglobulin A and the polymeric immunoglobulin receptor during hepatic transcytosis†
Authors: Chintalacharuvu Koteswara R. , Tavill Anthony S. , Louis Loizos N. , Vaerman Jean-Pierre , Lamm Michael E. , Kaetzel Charlotte S.
Published: March 2007

The polymeric immunoglobulin receptor on rat hepatocytes binds dimeric IgA on the sinusoidal surface and mediates its transport to the canaliculus, where the complex of dimeric IgA and secretory component, the cleaved extracellular domain of polymeric immunoglobulin receptor, is secreted into bile. This process is unique in that disulfide bonds are formed between dimeric IgA and polymeric immunoglobulin receptor during transcytosis, permanently preventing their dissociation. Here we present three lines of evidence that disulfide bonding between dimeric IgA and polymeric immunoglobulin receptor occurs predominantly in a late transcytotic compartment and that hepatic transcytosis can proceed in the absence of disulfide bond formation. First, throughout the course of transcytosis the percentage of intracellular dimeric IgA disulfide bonded to polymeric immunoglobulin receptor is less than half that in bile, suggesting that disulfide bond formation is a late event in transcytosis. Second, dimeric IgA that recycles from early endocytotic compartments into the circulation is mostly noncovalently bound to secretory component. Finally, the rate of transcytosis of dimeric IgA and its appearance in bile are not affected when disulfide bond formation with polymeric immunoglobulin receptor is inhibited by blocking of free thiol groups on dimeric IgA with iodoacetamide. These results are consistent with other findings in the literature and indicate that the main physiological role of disulfide bond formation between dimeric IgA and polymeric immuno-globulin receptor is not to facilitate transcytosis but, rather, to stabilize the dimeric IgA-secretory component complex after its release into external secretions such as bile and intestinal secretions. (Hepatology 1994;19:162-173).
An in vitro model of ethanol-dependent liver cell injury
Authors: Lamb Robert G. , Koch John C. , Snyder Jack W. , Huband Shiva M. , Bush Suzanne R.
Published: March 2007

Primary cultures of adult rat hepatocytes were incubated (6 to 96 hr) with 50 to 150 mmol/L ethanol, 0.5 mmol/L linoleate, 0.5 mmol/L palmitate, 0.5 mmol/L 4-methylpyrazole, 0 to 25 μmol/L vitamin E phosphate or selected combinations of these agents. Agent-dependent changes in liver cell viability (AST release and reduction of 3-(4,5-dimethylthiazol-2-yl)−2,5-diphenyltetrazolium bromide) and function (phospholipid peroxidation, hydrolysis, biosynthesis and triacylglycerol biosynthesis) were determined. The influence of ethanol on liver cell function and viability was dose and incubation time dependent. Short periods (24 hr or less) of exposure to 100 mmol/L ethanol increased liver cell triacylglycerol biosynthesis and phospholipid hydrolysis, peroxidation and biosynthesis without altering cell viability. However, longer periods (72 hr or more) of exposure to 100 or 150 mmol/L ethanol resulted in significant reductions (30 to 50) in cell viability, function and phosphatidylcholine biosynthesis and content. The ethanol-dependent decreases in cell function and viability were potentiated by linoleate and reduced by vitamin E phosphate, palmitate and 4-methylpyrazole. These results suggest that ethanol-induced liver cell injury in vitro is not a result of ethanol per se, but factors such as acetaldehyde or oxyradicals produced as a consequence of ethanol metabolism. Therefore the incubation of cultured hepatocytes with ethanol may be an appropriate model in vitro for determining the mechanisms by which ethanol intake disrupts liver cell function in vivo. (Hepatology 1994;19:174-182).