bluuebunny/crossref_metadata_2025 huggingface dataset:
Predictive factors in ursodeoxycholic acid-treated patients with primary biliary Cirrhosis: Role of serum markers of connective tissue†
Authors: Poupon Renée E. , Balkau Beverley , Guéchot Jérôme , Heintzmann François
Published: March 2007

The aim of this study was to define factors predictive of the onset of the terminal phase, defined by hyperbilirubinemia or the occurrence of a severe clinical complication, in patients with primary biliary cirrhosis treated with ursodeoxycholic acid. The 97 primary biliary cirrhosis patients in this study participated in a 2-yr clinical trial. Four of the 49 patients treated with ursodeoxycholic acid (13 to 15 mg/kg/day) entered the terminal phase of the disease, compared with 9 of the 48 patients assigned to placebo. In addition to clinical, conventional biological and histological parameters, we analyzed three serum markers of connective tissue components-type III procollagen aminoterminal peptide, hyaluronic acid and laminin. In the ursodeoxycholic acid-treated group, hyaluronic acid, type III procollagen aminoterminal peptide, bilirubin and splenomegaly were the factors most closely associated with entry into the terminal phase of the disease. In multivariate analysis, after adjustment for age, the hyaluronic acid level was the only predictive factor. In the placebo-treated group, the bilirubin level, total bile acid level, Mayo score, type III procollagen aminoterminal peptide, hyaluronic acid, splenomegaly and pruritus were associated with aggravation of disease. In multivariate analysis, high bilirubin level, high type III procollagen aminoterminal peptide or hyaluronic acid levels and low prothrombin time independently implied poor prognosis. In conclusion, when patients with primary biliary cirrhosis are treated with ursodeoxycholic acid, bilirubinemia loses, in part, its predictive value. It is replaced by hyaluronic acid and type III procollagen aminoterminal peptide. This suggests that models used in deciding on the need for liver transplantation require adaptation for patients receiving ursodeoxycholic acid. (Hepatology 1994;19:635-640).
Endotoxin-induced nitric oxide synthesis in the perfused rat liver: Effects of l-arginine and ammonium chloride
Authors: Wettstein Matthias , Gerok Wolfgang , Häussinger Dieter
Published: March 2007

We used the single-pass-perfused rat liver model to study short-term regulation of endotoxin-inducible nitric oxide synthesis by following the release of nitrite and nitrate, the oxidation products of nitric oxide, into the effluent perfusate. In endotoxin-pretreated livers, the basal nitrite + nitrate release was 5.3 ± 1.2 nmol·gm liver−1·min−1. Nitrite and nitrate release was stimulated by L-arginine in a dose-dependent and saturable fashion. Maximal nitrite + nitrate release with 1 mmol/L L-arginine infused to the influent perfusate was 10.2 ± 1.1 nmol·gm liver−1·min−1, with a half-maximal effect at 53 μmol/L L-arginine. In the absence of molecular oxygen, nitric oxide synthesis was inhibited. Ammonium chloride infusion also stimulated nitrite and nitrate release to a maximal rate of 9.2 ± 0.8 nmol·gm liver−1 min−1 with half-maximal effects at 60 μmol/L ammonium chloride. Ammonium chloride-stimulated nitrite and nitrate release was abolished when urea synthesis was inhibited by bicarbonate-free liver perfusion. Citrulline and ornithine (200 μmol/L each) were without effect on nitrite and nitrate release. L-Nitroarginine methyl ester inhibited both, L-arglnine-and ammonium chloride-induced nitrite and nitrate release. Stimulation of nitric oxide synthesis by L-arginine and ammonium chloride addition (1 mmol/L each) was accompanied by a threefold-to-fourfold increase of cyclic GMP release into the effluent perfusate. In livers of endotoxin-pretreated rats the urea production from L-arginine was higher than that in untreated livers, suggesting induction of an L-arginine transport system in hepatocytes by endotoxin. The regulation of hepatic nitric oxide production by physiological concentrations of L-arginine and ammonia in the portal vein may be of importance in cirrhosis. (Hepatology 1994;19:641-647).
Biosynthesis and intracellular transport of a bile canalicular plasma membrane protein: Studies in vivo and in the perfused rat liver
Authors: Maurice Michele , Schell Michael J. , Lardeux Bernard , Hubbard Ann L.
Published: March 2007

B10 is an integral glycoprotein of the plasma membrane that is exclusively localized to the canalicular (apical) domain in normal rat hepatocytes but may be expressed on the basolateral (sinusoidal and lateral) membrane in pathophysiological situations. To understand how B10 may be localized to the basolateral surface, we studied the biosynthesis and transport of this apical protein. In vivo pulse-chase experiments, followed by subcellular fractionation of the liver and immunoprecipitation, showed that B10 is first synthesized as a high-mannose form of 123 kD and then matured to a complex glycosylated form of 130 kD, which peaks in the Golgi apparatus after 15 min of chase and reaches the plasma membrane with a half-time of 30 to 45 min. Analysis of the protein in plasma membrane domain fractions showed that most of the newly synthesized molecule was localized in basolateral fractions after 30 min of chase and subsequently appeared in apical fractions. After 90 min of chase, most of the radiolabeled protein had reached its steady-state apical distribution. The same experiments performed in the perfused rat liver, in which the chase can be improved, gave similar results, except that the apical distribution of the radioactive molecule was attained more quickly. Thus B10, like all apical plasma membrane proteins studied so far in hepatocytes, is first transported to the basolateral surface and then reaches the membrane of the bile canaliculi. Alterations of the transcytotic step from the basolateral to the apical surfaces may result in abnormal basolateral localization. (Hepatology 1994;19:648-655).
Structure and localization of the IGFBP-1 gene and its expression during liver regeneration
Authors: Lee Jehyuk , Greenbaum Linda , Haber Barbara A. , Nagle Deborah , Lee Victoria , Miles Vashti , Mohn Kenneth L. , Bucan Maja , Taub Rebecca
Published: March 2007

Insulin-like growth factor-binding protein-1s are important modulators of the insulin-like growth factors that may have both positive and negative effects on the ability of insulin-like growth factors to stimulate cell growth. The IGFBP-1 gene is one of the most highly induced immediate-early genes after partial hepatectomy. The IGFBP-1 gene is also expressed at a high level during fetal liver development and in response to nutritional changes and diabetes. Therefore it may have important roles in liver growth and metabolism. To begin to examine the regulation of this gene, we cloned and sequenced the entire mouse IGFBP-1 gene. Its structure is highly similar to that of the human gene, and, in addition to the exonic regions, the two genes are highly conserved in specific regions in the promoter and first intron. Analysis of this conservation allows us to predict important regulatory sites that define the tissue specific and insulin-mediated regulation of the gene and identify potential sites that might be important for the transcriptional induction during liver regeneration. The mouse gene is located on mouse chromosome 11; it is found at the boundary between regions in the mouse genome homologous to human chromosomes 22 and 7. We found IGFBP-1 mRNA in both parenchymal and nonparenchymal RNA after partial hepatectomy. Using in situ hybridization of IGFBP-1 mRNA in regenerating rat liver tissue, we demonstrated IGFBP-1 transcripts in several cell types. We found that IGFBP-1 gene induction after partial hepatectomy is paralleled by protein expression. However, on immunohisto-chemical study, insulin-like growth factor-binding protein-1 protein is found only in hepatocytes after hepatectomy. Unlike IGFBP-1 mRNA, serum levels of insulinlike growth factor-binding protein-1 are increased for a relatively short time with a peak at 2 to 3 hr after hepatectomy. Increased levels of insulinlike growth factor-binding protein-1 could be important in modulating insulinlike growth factor-1 effects on metabolism and growth during liver regeneration. (Hepatology 1994;19:656-665).
Proliferation of rat liver macrophages in vitro : Influence of hemopoietic growth factors
Authors: Hoedemakers Rein M. J. , Scherphof Gerrit L. , Daemen Toos
Published: March 2007

We examined the effects of several hemopoietic growth factors on proliferation of rat liver macrophages in vitro. The proliferative response of liver macrophages to hemopoietic growth factors was assayed on the basis of [methyl-3H]thymidine uptake. Macrophage colony-stimulating factor and recombinant murine granulocyte-macrophage colony-stimulating factor stimulated [methyl-3H]thymidine incorporation in a concentration-dependent manner. With granulocyte-macrophage colony-stimulating factor, maximum incorporation was observed at 50 U/ml, whereas with macrophage colony-stimulating factor no incorporation plateau was observed up to 50 L929-conditioned medium. Incubation of liver macrophages with various concentrations of recombinant human interleukin-2, recombinant murine interleukin-3 and recombinant human interleukin-6 or culture medium alone did not result in significant incorporation of [methyl-3H]thymidine. When liver macrophages were fractionated according to cell size, highest incorporation was observed in the large macrophages. Proliferating cells in cultures of all subfractions were microscopically identified as typical macrophages by the use of macrophage-specific monoclonal antibodies. After 6 days in culture, these macrophages had functional properties similar to those of resident liver macrophages with respect to phagocytosis and in vitro activation with immunomodulators to tumorcytotoxicity and secretion of nitric oxide and tumor necrosis factor-α. These results suggest that macrophage colony-stimulating factor and granulocyte-macrophage colony-stimulating factor play important roles among the regulatory factors that support local proliferation of rat liver macrophages. (Hepatology 1994;19:666-674).
Endothelin-1 is involved in the pathogenesis of ischemia/reperfusion liver injury by hepatic microcirculatory disturbances
Authors: Goto Moritaka , Takei Yoshiyuki , Kawano Sunao , Nagano Kouichi , Tsuji Shingo , Masuda Eiji , Nishimura Yoshiya , Okumura Shigetoshi , Kashiwagi Toru , Fusamoto Hideyuki , Kamada Takenobu
Published: March 2007

Hepatic microcirculatory perturbation is observed after ischemia/reperfusion. Endothelin-1, a potent vasoconstrictive peptide, is known to modulate local circulation. This study was designed to examine whether endothelin-1 participates in the mechanism of microcirculatory disturbance and damage of the liver after ischemia/reperfusion. Ischemia in the median and left lateral lobes of the liver was induced for 60 min; it was followed by reperfusion for 24 hr. In some rats, endothelin-1 antiserum or control serum without endothelin-1-blocking activity was administered intravenously just before reperfusion. Rats were divided into three groups: an ischemia/reperfusion group that was injected with control serum, an endothelin-1 antiserum-treated group and a sham-operated group. Endothelin-1 concentrations in blood collected from the suprahepatic vena cava were measured before and after ischemia/reperfusion by use of a sandwich enzyme immunoassay. Index of blood volume in regional hepatic tissue and index of blood oxygenation in regional hepatic tissue were assessed with an organ reflectance spectrophotometry system before and at 5 min and 1, 2, and 24 hr after reperfusion. The endothelin-1 concentration in the ischemia/reperfusion group started to rise immediately at onset of reperfusion from basal values around 1 pg/ml and reached a value of 5 to 6 pg/ml 5 min after reperfusion; it was maintained at significantly high levels during the reperfusion period compared with the sham-operated group. Hepatic microcirculatory disturbance indicated by lowered index of blood volume in regional hepatic tissue and index of blood oxygenation in regional hepatic tissue levels was observed in the early phase of reperfusion in the ischemia/reperfusion group. Endothelin-1 antiserum injection diminished significantly the microcirculatory disturbance in the liver after ischemia/reperfusion. The serum ALT levels in the endothelin-1 antiserum-treated group 2 hr after reperfusion were decreased by 40 to 50 compared with the nontreated group. Moreover, liver damage assessed histologically was reduced dramatically in the endothelin-1 antiserum-treated group. In conclusion, endothelin-1 appears to be involved in the microcirculatory disturbance and histological damage in the liver after ischemia/reperfusion. Blockade of endothelin-1 action may prevent liver injury after ischemia/reperfusion. (Hepatology 1994;19:675-681).
Decreased expression of liver glutathione peroxidase in Long-Evans cinnamon mutant rats predisposed to hepatitis and hepatoma
Authors: Suemizu Hiroshi , Yoshimura Shinichi , Takeichi Noritoshi , Moriuchi Tetsuya
Published: March 2007

The Long-Evans Cinnamon rat is a mutant strain that contracts hereditary hepatitis and, eventually, spontaneous hepatoma. Recently, abnormal copper accumulations in Long-Evans Cinnamon rat livers were shown to be genetically linked to the development of hepatitis. Because reduced glutathione and glutathione-related enzymes are known to play important roles in cellular resistance to transition metal toxicity, we determined the levels of reduced glutathione and glutathione-related enzymes in seven different tissues of Long-Evans Cinnamon and control Long-Evans Agouti rats. Of the enzymes examined, only hepatic glutathione peroxidase was markedly decreased in Long-Evans Cinnamon rats. Glutathione peroxidase content in the liver of Long-Evans Cinnamon rats was 39%, 53% and 58% of the control values at 9 (normal stage), 19 (acute hepatitis stage) and 27 (chronic hepatitis stage) wk of age, respectively. Northern-blot analysis revealed that messenger RNA levels of glutathione peroxidase in the livers of Long-Evans Cinnamon rats were about 40% of the control levels. The activity of glutathione S-transferase was slightly decreased in the livers of Long-Evans Cinnamon rats. These data suggest that the liver of the Long-Evans Cinnamon rat is poorly protected against active oxygen species, the production of which is enhanced in the presence of excess copper. Glutathione-reductase activity in the livers of Long-Evans Cinnamon rats increased to 166% and 148% of the control levels at 19 and 27 wk of age, respectively. No significant changes were observed in the activity of γ-glutamylcysteine synthetase or in the content of total reduced glutathione in the liver of the Long-Evans Cinnamon rat. The changes in activity of glutathione peroxidase and glutathione reductase in the Long-Evans Cinnamon rat livers resembled those observed during chemical hepatocarcinogenesis. (Hepatology 1994;19:694-700).
Expression of platelet-derived growth factor in a model of acute liver injury†
Authors: Pinzani Massimo , Milani Stefano , Grappone Cecilia , Weber Frederick L. , Gentilini Paolo , Abboud Hanna E.
Published: March 2007

Platelet-derived growth factor has been shown to play an important role in the repair process after acute tissue injury and in the pathogenesis of several fibrogenic disorders. The aim of this study was to evaluate whether increased expression of platelet-derived growth factor and its β-receptor subunit occurs in a model of acute liver injury. Male Sprague-Dawley rats were given a single intragastric dose of carbon tetrachloride and killed at intervals of 24, 48 and 72 hr and 1 wk. Control animals were included in each group. Platelet-derived growth factor-B chain mRNA expression, analyzed by RNase protection assay, was not detectable in control samples or in samples obtained 24 hr or 1 wk after carbon tetrachloride. However, the presence of protected fragments of 130 kb was clearly detected after 48 hr and was still present, although less abundant, after 72 hr. The distribution of platelet-derived growth factor protein in liver tissue sections, evaluated by immunohistochemistry, was restricted to centrilobular veins and portal tracts in normal liver. In carbon tetrachloride-treated rats, prominent staining was observed in areas corresponding to hepatocellular necrosis and inflammatory infiltration. This feature, already present at 24 hr after carbon tetrachloride, became more marked at 48 hr with a gradual resolution after 72 hr. The expression of platelet-derived growth factor-receptor β-subunit mRNA, evaluated by in situ hybridization, was markedly increased after carbon tetrachloride with a peak at 24 hr and was mainly localized over mesenchymal cells in the hepatic sinusoids. The overall analysis of these results suggests that platelet-derived growth factor may play a positive role in the process of tissue repair after acute liver injury by promoting effective necrotic tissue removal and the reconstruction of an adequate extracellular matrix network. (Hepatology 1994;19:701-707).
Enhanced hepatic collagen type I mRNA expression into fat-storing cells in a rodent model of hemochromatosis†
Authors: Pietrangelo Antonello , Gualdi Rossana , Casalgrandi Giovanna , Geerts Albert , de Bleser Pieter , Montosi Giuliana , Ventura Ezio
Published: March 2007

In recent years, identifying the hepatic cell type responsible for collagen synthesis in experimental models of postnecrotic or inflammatory fibrosis has been the subject of active investigation. In primary iron overload states, however, hepatic fibrosis and cirrhosis occur without accompanying necroinflammatory phenomena. In this study, we combined morphological, immunological, cell isolation and purification and molecular biological techniques to identify the hepatic cell responsible for enhanced collagen type I gene expression during chronic enteral iron overload in the rat. Ultrastructural analysis of liver tissue sections from iron-loaded rats specifically revealed an altered appearance of fat-storing cells, which showed few if any fat droplets left and increased rough endoplasmic reticulum. In situ hybridization analysis with specific complementary RNA probes identified enhanced signal for collagen type I into nonparenchymal cells in zones 1 and 2, without signal over the background onto iron-laden hepatocytes. Immunocytochemistry with desmin antibodies combined with in situ hybridization on the same tissue sections identified the cells expressing high level of collagen type I transcripts as fat-storing cells. Northern-blot analysis on RNA extracted from various purified cell isolates, confirmed the presence of collagen type I mRNA signal only into the fat-storing cells isolate. Our study shows that in an experimental model of metabolic fibrosis in which the hepatotoxin selectively accumulates into parenchymal cells, fat-storing cells are the main source of enhanced collagen type I gene expression. (Hepatology 1994;19:714-721).
Expression of spermidine/spermine N1-acetyltransferase in growing Yoshida AH-130 hepatoma cells
Authors: Desiderio Maria Alfonsina , Bardella Lidia
Published: March 2007

Activity and messenger RNA levels of spermidine/spermine N1-acetyltransferase, the rate-limiting enzyme of the polyamine interconversion pathway, were investigated in host liver and in Yoshida AH-130 ascites hepatoma cells as a function of tumor growth phases. Enzyme activity reached maximal values at day 10 in host liver (2.0-fold increase) and at days 10 and 14 in hepatoma cells (4.2- and 5.4-fold increases)- that is, when the cellular growth was nearly arrested. At day 10 the messenger RNA levels of spermidine/spermine N1-acetyltransferase were augmented concomitantly; they were about two and four times higher, respectively, in host liver and tumor cells than in control liver. The in vitro transcription rate seemed to be constant during hepatoma cell growth. Treatment of the animals with N1, N2-bis-(2,3-butadienyl)-1,4-butanediamine (MDL 72527), a specific inhibitor of polyamine oxidase, caused large accumulation of N1-acetylspermidine in hepatoma cells and in the ascitic fluid; the maximal values were reached at day 14. The levels of putrescine in inhibitor-treated rats decreased in hepatoma cells (day 5) and in ascitic fluid (days 5 and 14), whereas values of spermidine and spermine remained unchanged. The proposed role for spermidine/spermine N1-acetyltransferase-enhanced expression is to regulate the cellular polyamine pool by causing their excretion as acetylderivatives from tumor cells into the ascitic fluid, even if putrescine seems also to be excreted. Eventual repeat uptake of putrescine by hepatoma cells could contribute to the control of cellular polyamine levels. (Hepatology 1994;19:728-734).
Effect of chemotherapy on the larval mass and the long-term course of alveolar echinococcosis
Authors: Ammann Rudolf W. , Ilitsch Nicolas , Marincek Borut , Freiburghaus Andreas U.
Published: March 2007

The efficacy of long-term chemotherapy in nonresectable alveolar echinococcosis is debated, particularly because of the difficulty in defining therapeutic success. In this study the effect of chemotherapy on the parasitic mass was evaluated in a series of 37 patients. The patients had larval lesions documented by serial computed tomography studies at least 1.5 yr after chemotherapy (mean = 6.4 yr, range = 1.5 to 10.7 yr). The therapeutic regimen consisted of mebendazole (n = 34) or albendazole (n = 3) as previously described. The maximal areas of the parasitic lesions were assessed morphometrically by means of digital image analysis, utilizing the point-integration method, before and after chemotherapy. Marked regression of larval tissue occurred in 18 patients (group A; 48.6%), stationary lesions were noted in 13 patients (group C; 35.1%) and progression was found in 6 patients (group B; 16.2%). The three groups did not differ significantly with regard to age, plasma drug levels, duration of chemotherapy or initial size and composition of lesions (e.g., cystic cavities, degree of calcification). Despite morphologically successful chemotherapy in most patients (e.g., 84%; groups A and C), late cholestatic complications after 1.5 to 11 yr of chemotherapy occurred in 10 patients (group A, n = 7; group C, n = 3; 4 of them died) and esophageal variceal bleeding occurred in 3 patients (relieved by sclerotherapy). These late complications were probably mainly due to posttherapy fibrosis of hilar structures. In conclusion, our data support the efficacy of chemotherapy. However, chemotherapy is not curative, and severe late complications were observed in patients with hilar (fibrotic) involvement. (Hepatology 1994;19:735-742).
Biliary phospholipases in the prairie dog model for cholesterol cholelithiasis
Authors: Booker Michael L. , Lamorte Wayne W.
Published: March 2007

Lysolecithin has been implicated as a contributing factor in the pathogenesis of cholecystitis and cholesterol cholelithiasis. The phospholipases are key enzymes in the generation of a number of metabolites including lysolecithin, but conflicting reports exist concerning the presence of these enzymes in the biliary tract. In this study, measurement of phosphatidylcholine-specific phospholipase activity by means of the hydrolysis of radiolabeled phosphatidylcholine (100 nmol) by 90 μg of homogenate protein during a 60-min incubation demonstrated substantial enzyme activities in gastric fundus and distal ileum (90% and 70% hydrolysis, respectively), whereas activity was virtually undetectable in gallbladder mucosa (0.7% hydrolysis). Additional studies were conducted in prairie dogs fed diets high in cholesterol or with trace amounts of cholesterol using homogenates of gallbladder mucosa, seromuscularis and full-thickness tissue, as well as samples of hepatic and gallbladder bile. The only hydrolytic activity in excess of blank values that was detected was a highly variable phospholipase A2 activity in several gallbladder biles from animals given diets with both low levels and high levels of cholesterol, with the enzyme activities of the two dietary groups being similar. These results demonstrate that prairie dog gallbladder contains extremely low levels of phospholipase activity, in marked contrast to other gastrointestinal tissues. However, there was evidence of a phospholipase A2 activity in gallbladder bile. In light of the low activity in gallbladder tissue, the source of this enzyme appears to be the liver and not the gallbladder. The fact that hydrolytic activity after cholesterol feeding was similar to that in control animals suggests that the increases in lysolecithin concentration in gallbladder bile that occur during early cholelithiasis may result from stasis rather than from an increased hepatic secretion of phospholipase. (Hepatology 1994;19:743-749).
Partial cloning of the M subunit of laminin from adult rat lipocytes: Expression of the M subunit by cells isolated from normal and injured liver†
Authors: Maher Jacquelyn J. , Tzagarakis Christina
Published: March 2007

Laminin is a heterotrimeric glycoprotein found in the perisinusoidal space of adult rat liver. The principal cellular source of laminin in liver is the lipocyte, with its three subunits measuring 324, 200 and 200 kD. The large subunit of lipocyte-derived laminin is distinct from the A subunit of murine laminin (440 kD); its size suggests that it represents a peptide, called M, recently cloned from human placenta. Using oligonucleotide primers derived from the human M-subunit cDNA, we amplified a 445-bp sequence encoding a fragment of M-laminin from adult rat lipocytes. The rat cDNA is 90% homologous to the human M-subunit cDNA and recognizes an mRNA in lipocytes measuring about 10 kb. M-subunit transcripts were identified only in lipocytes from normal adult liver; they could not be identified in hepatocytes, endothelial cells or Kupffer cells. Lipocytes were screened for M-subunit protein with a polyclonal M antiserum. Cells stained specifically for the M-subunit after 36 hr in primary culture; the protein was also identified in freshly isolated cells by means of immunoblotting. To determine whether lipocytes alter their expression of the laminin M subunit during liver injury, we monitored M-subunit mRNA in these cells at various intervals after carbon tetrachloride administration. M-subunit transcripts increased twofold within 12 hr of toxin exposure, returning to below baseline by 48 hr. The results indicate that lipocytes produce the M subunit of laminin in place of A. Production of this subunit by lipocytes may facilitate cell growth and reorganization during liver regeneration. (Hepatology 1994;19:764-770).
Immunohistochemical study of hepatic fibrosis induced in rats by multiple galactosamine injections
Authors: Jonker Mieke A. , Dijkhuis Freke W. J. , Hardonk Machiel J. , Moerkerk Peter , Kate Joop Ten , Grond Joris
Published: March 2007

Multiple injections of D-galactosamine induce liver fibrosis and cirrhosis in rats. The purpose of this immunopathological study was to correlate inflammation and hepatic extracellular matrix remodeling after repeated administration of galactosamine. Rats were given 10, 20, 30, 40, 60, 80, 100 and 140 intraperitoneal injections of D-galactosamine (500 mg/kg body wt, three times weekly). Controls received injections of saline solution. Cryostat sections of liver tissue obtained on biopsy or autopsy were immunostained with a panel of monoclonal and polyclonal monospecific antibodies reactive with macrophages, T and B lymphocytes, desmin, the extracellular matrix components fibronectin; laminin; collagen types I, III and IV; and the fibronectin receptor α5β1. Total RNA was extracted and Northern-blot analysis was performed with a specific cDNA probe for rat collagen type III. Spotty liver cell necrosis and mild portal and parenchymal inflammation was seen after 10 injections of galactosamine. After 20 to 40 injections, expansion of portal tracts, prominent bile ductular proliferation and gradual formation of fibrous septa were encountered with the development of cirrhosis at later intervals. These progressive histological changes were paralleled by a gradual increase of desmin-positive cells in developing septa with deposition of fibronectin; collagen types I, III and IV; and laminin. Northern-blot analysis showed that this accumulation of extracellular matrix was not accompanied by increase of mRNA for collagen type III. In conclusion, repetitive administration of galactosamine causes progressive liver disease with prominent bile ductule proliferation and development of fibrous septa. These pathological alterations bear some resemblance to the morphological changes in chronic biliary disease in human beings. (Hepatology 1994;19:775-781).
Activin A and hepatocyte growth regulation
Authors: Dubois Raymond N.
Published: March 2007

The present study was conducted to examine the effect of activin A on growth of rat hepatocytes. EGF induced a 10-fold increase in DNA synthesis as assessed by [3H]thymidine incorporation in cultured hepatocytes. When activin A was added together with EGF, DNA synthesis induced by EGF was markedly inhibited. Inhibition was detected at a concentration of 10−10 M, and 5×10−9 M activin A almost completely blocked EGF-mediated DNA synthesis. Similarly, activin A completely blocked DNA synthesis induced by hepatocyte growth factor/scatter factor. Activin A was capable of inhibiting EGF-mediated DNA synthesis, even when added 36 h after the addition of EGF. With the same time interval, TGF-β also blocked EGF-induced DNA synthesis. Although both activin A and TGF-β inhibited growth of hepatocytes in a similar manner, either activin A or TGF-β did not compete with each other in their binding when assessed by competitive binding using an iodinated ligand. When hepatocytes were incubated with EGF, release of bioactivity of activin A into culture medium was detected after 48 h or later. Activity of activin A was released from parenchymal cells but not from non-parenchymal cells. mRNA for βA subunit of activin was detected only slightly in unstimulated hepatocytes, but markedly increased at 48 h after the addition of EGF. To determine whether endogenously produced activin A affects DNA synthesis, we examined the effect of follistatin, an activin-binding protein that blocks the action of activin A. An addition of follistatin significantly enhanced EGF-induced DNA synthesis. Finally, in partial hepatectomized rat, expression of mRNA for βA subunit in liver was markedly increased 24 h after the partial hepatectomy. These results indicate that activin A inhibits initiation of DNA synthesis in hepatocytes by acting on its own receptor and that activin A acts as an autocrine inhibitor of DNA synthesis in rat hepatocytes.
Genes that control the formation of the liver
Authors: Zaret Ken
Published: March 2007

The proto-oncogene c-jun is the cellular homologue of v-jun, the transforming oncogene of the avian sarcoma virus 17. c-jun encodes one major component of the AP-1 transcription factor complex and is expressed in many organs during mouse development and in the adult. Because of its rapid induction in cells following growth stimulation and the presence of AP-1 binding sites in the promoter regions of many genes, the c-Jun protein is thought to have important functions in cell proliferation and differentiation. But embryonic stem (ES) cells lacking c-Jun are viable and have a normal in vitro differentiation capacity, although c-Jun appears to be important for growth of teratocarcinomas in vivo. To define the function of c-jun better, targeted ES cells were used to generate mice lacking c-Jun. Here we report that heterozygous mutant mice appear normal, but embryos lacking c-Jun die at mid- to late-gestation and exhibit impaired hepatogenesis, altered fetal liver erythropoiesis and generalized oedema. Interestingly, c-jun
−/- ES cells can participate efficiently in the development of all somatic cells in chimaeric mice except liver cells, further suggesting an essential function of c-Jun in hepatogenesis.
High-level expression of hepatitis B virus HBx gene and hepatocarcinogenesis in transgenic mice
Authors: Koike Kazuhiko , Moriya Kyoji , Iino Shiro , Yotsuyanagi Hiroshi , Endo Yasuo , Miyamura Tatsuo , Kurokawa Kiyoshi
Published: March 2007

We studied the development of liver tumors in male HBx gene transgenic mice. Of two lineages studied, in the lineage with the lowest HBx gene expression liver tumors developed only in an incidence comparable with that in normal CD-1 strain, whereas 84% of male mice with a high level of the HBx gene product succumbed to liver neoplasia, indicating that continued HBx gene expression higher than a certain threshold level may be necessary for the development of hepatic neoplasia. Sixty-five mice from a lineage with a high level of HBx expression were then followed throughout their 24-mo lifespan. The livers of transgenic mice showed foci of cellular alteration with cytoplasmic vacuolations around the central veins from the age of 2 mo, but these foci did not expand progressively by the age of 12 mo. Immunostaining demonstrated such hepatocytes had higher expression of HBx protein than surrounding cells. Neoplastic lesions including liver cell adenomas and hepatocellular carcinomas developed from the age of 13 mo. By bromodeoxyuridine labeling analysis, hepatocytes in altered foci were found to have increased DNA synthesis, whereas no labeling was observed in age- and sex-matched nontransgenic littermate controls. Furthermore, DNA content analysis revealed the existence of several small aneuploid peaks in the transgenic liver before the age of tumor development. These results suggest that the continued expression of HBx gene may initiate a complex process to hepatocellular carcinoma by inducing DNA synthesis and placing large numbers of hepatocytes subjective to secondary events for transformation. (Hepatology 1994;19:810-819.)
Interferon-α for chronic hepatitis C: An analysis of pretreatment clinical predictors of response
Authors: Pagliaro Luigi , Craxí Antonio , Cammaá Calogero , Tiné Fabio , Di Marco Vito , Iacono Oreste Lo , Almasio Piero
Published: March 2007

To identify predictors of short-term and sustained ALT normalization after interferon treatment in adult patients with chronic hepatitis C, we performed a metanalysis of individual patients’ data, with construction and cross-validation of a prediction rule, in 361 patients from two randomized trials. In one trial, 116 subjects with transfusion-related chronic hepatitis C were treated with lymphoblastoid interferon (5 MU/m2 three times a week for 2 mo, then 3 MU/m2 three times a week for 4 or 10 mo). In the other study, 245 patients with community-acquired chronic hepatitis C received recombinant interferon-α2b (10 MU three times a week for 2 mo, then 5 MU three times a week for 4 mo; then random allocation of subjects with normal aminotransferase levels to stop or continue interferon for a further 6 mo). Overall, 164 subjects (45%; 95% confidence interval, 40% to 50%) had short-term responses; 61 (18%; 95% confidence interval, 14% to 22%) maintained sustained responses. Sixty patients (17%; 95% confidence interval, 13% to 21%) withdrew from treatment because of side effects or subjective intolerance. Logistic regression analysis showed that short-term and sustained response were independently predicted by lobular structure on pretreatment liver biopsy (p < 0.0001) and by short disease duration, defined as the time elapsed since transfusion in posttransfusion cases or since the first observation of abnormal aminotransferase levels in cryptogenic disease (p < 0.01). Rules to predict short-term and sustained response to interferon were derived from these items, showing a discriminatory ability of 0.73 and 0.70. We conclude that, in patients with C hepatitis, presence of cirrhosis and long disease duration predict low likelihood of response to interferon. Although the predictive rule based on these items is not sufficiently accurate for decision-making in individual patients, it can definitely assist when applied to groups of patients for planning or interpreting therapeutic studies. The addition of virological features such as quantitation of viremia and hepatitis C virus genotypes may improve the ability to predict treatment outcomes. (Hepatology 1994;19:820-828.)
Contribution of hepatitis C virus to non-A, non-B fulminant hepatitis in Japan
Authors: Yoshiba Makoto , Dehara Kazuhiko , Inoue Kazuaki , Okamoto Hiroaki , Mayumi Makoto
Published: March 2007

To assess the contribution of hepatitis C virus to non-A, non-B fulminant hepatitis in Japan, we compared 10 major clinical features among 7 patients with type B fulminant hepatitis (type B group), 13 patients with non-A, non-B fulminant hepatitis with evidence of hepatitis C virus infection (type C group) and 10 patients without evidence of hepatitis C virus infection (NANB group). Duration from first symptom to coma and that from onset of jaundice to coma was significantly longer in the type C group (median = 39 and 25 days, respectively) and in the non-A, non-B group (median = 29 and 12 days, respectively) than in the type B group (median = 9 and 2 days, respectively) (p < 0.01). The maximum median AST level was significantly lower in the type C (1,689 U/L) and non-A, non-B groups (1,353 U/L) than in the type B group (5,780 U/L) (p < 0.05). Serum transaminase levels showed a single peak in six of seven of the type B patients, whereas they formed two or more peaks in all of the type C patients and in most of the non-A, non-B group (p < 0.05). Six of seven in the type B group, 6 of 13 in the type C group and 4 of 10 in the non-A, non-B group survived (p < 0.05). We found no significant difference in any of the 10 clinical features between the type C and non-A, non-B groups. Compared with type B fulminant hepatitis, type C and most cases of non-A, non-B fulminant hepatitis in Japan are, thus, characterized by slower and less severe but more persistent hepatocyte destruction. (Hepatology 1994;19:829-835.)
Multiple viral infection as the most common cause of fulminant and subfulminant viral hepatitis in an area endemic for hepatitis B: Application and limitations of the polymerase chain reaction
Authors: Wu Jaw-Ching , Chen Chih-Li , Hou Ming-Chih , Chen Trong-Zong , Lee Shou-Dong , Lo Kwang-Juei
Published: March 2007

We tested serum samples from 25 fulminant hepatitis and 7 subfulminant hepatitis patients for hepatitis A, B, C, D and E viral markers and nucleic acids by means of polymerase chain reaction to determine the role of each virus on such catastrophic events in an area endemic for hepatitis B. Of these 32 patients, 14 (44%) were hepatitis B virus carriers with hepatitis D virus superinfection (1 with hepatitis C virus infection), 3 others had coexisting hepatitis B virus and hepatitis C virus infections, 6 had reactivation of underlying chronic hepatitis B, 4 had acute hepatitis B, 2 had acute hepatitis C and 1 had acute hepatitis E. Pathogenesis in the remaining two cases was unclear. Serum hepatitis B virus DNA was detectable in most carriers without superinfection and in one third of those with superinfection detected on polymerase chain reaction (6 of 7 vs. 6 of 16, p < 0.05). Of the polymerase chain reaction-positive samples, only 17% yielded positive results on spot hybridization. Hepatitis B virus DNA was the only marker to indicate coexisting hepatitis B virus infection in one patient positive for hepatitis C virus antibody. Only three of the six hepatitis C virus-infected cases were positive for hepatitis C virus antibody; diagnoses in the remaining three were established by means of detection of hepatitis C virus RNA. Of the hepatitis D virus-infected patients, infection in only half was diagnosed by means of total hepatitis D virus antibody assay. Twelve (86%) were positive for anti-hepatitis D virus IgM and nine (64%) had detectable hepatitis D virus RNA on reverse transcription-polymerase chain reaction. Only one third of reverse transcription-polymerase chain reaction positive cases had positive results on Northern-blot hybridization. Hepatitis D virus RNA was the only marker indicating hepatitis D virus infection in two (14%) of the hepatitis D virus-infected patients. In conclusion, most fulminant and subfulminant viral hepatitis in Taiwan is caused by multiple viral infection (hepatitis B virus with hepatitis D virus or hepatitis C virus), which might be underestimated if only one kind of marker were used. (Hepatology 1994;19:836-840.)
Decrease in serum levels of markers of hepatic connective tissue turnover during and after treatment of chronic hepatitis B with interferon-α
Authors: Teran Carlos J. , Mullen Kevin D. , Hoofnagle Jay H. , McCullough Arthur J.
Published: March 2007

Interferon-α induces remission in 30% to 40% of patients with chronic hepatitis B, but its effect on hepatic connective tissue turnover has not been well documented. We studied the changes in serum procollagen III propeptide and laminin-P1 peptide (Lam-P1) in 33 patients with chronic hepatitis B (11 nontreated controls and 22 treated patients) during a 4-mo randomized trial of interferon-α. Liver biopsy specimens were obtained at the start of treatment and 12 mo later. Liver biochemical tests, procollagen III propeptide, laminin-P1 peptide and hepatitis B virus DNA polymerase were determined before treatment with interferon was begun (mo -3), at the initiation (0 time) and completion of treatment (mo 4) and also at 8, 12 and 18 mo. Treated patients were classified as “responders” and “nonresponders” on the basis of clearance of HBV e antigen from serum. There were no significant changes in the control group, whereas the responders had persistent decreases in ALT, AST, hepatitis B virus dna polymerase, procollagen III propeptide and laminin-P1 peptide. The nonresponders had transient ALT, AST and hepatitis B virus dna polymerase reductions that returned toward baseline levels during follow-up, but procollagen III propeptide and laminin-P1 peptide persisted below the baseline at mo 18. Significant correlations between procollagen III propeptide and laminin-P1 peptide with ALT, AST and liver histologic specimens were noted at baseline but not after 12 mo. Changes in procollagen III propeptide levels also correlated with changes in AST, ALT and liver histologic specimens. On the basis of logistic regression, neither markers of connective tissue turnover nor histology improved the accuracy of AST, ALT and hepatitis B virus dna polymerase for predicting response to interferon. We conclude that interferon treatment induces a persistent suppression in serum markers of hepatic connective tissue turnover in chronic hepatitis B patients independent of its effect on viral replication and hepatic necroinflammation. (Hepatology 1994;19:849-856.)
Seroprevalence of antibody to hepatitis E virus among Chinese subjects in Taiwan
Authors: Lee Shou-Dong , Wang Yuan-Jen , Lu Ruey-Hwa , Chan Cho-Yu , Lo Kwang-Juei , Moeckli Randolph
Published: March 2007

Recently, with an available serological hepatitis E virus diagnostic kit, the prevalence of IgG antibody to hepatitis E virus among Chinese subjects in Taiwan was evaluated by means of a solid-phase enzyme-linked immunoassay based on two recombinant hepatitis E virus antigens. The overall prevalence of hepatitis E virus antibody was 10.7% among 384 healthy subjects older than 20 yr but only 0.3% among 600 school-children and young adolescents younger than 20 yr (p < 0.0001). Serial serum samples from 32 hepatitis E virus antibody-positive healthy subjects showed 84% of them to have antibodies persisting more than 3 to 8 yr. Among patients with viral hepatitis, IgG hepatitis E virus antibody was detected in 10% of 10 patients with acute hepatitis A, in 9.5% of 63 patients with acute hepatitis B and in 13.9% of 36 patients with acute posttransfusion hepatitis C. Of 77 patients with sporadic non-A, non-B hepatitis, IgG hepatitis E virus antibody was detected in 18.9% of 53 patients positive for antibody to hepatitis C virus and in 45.8% of 24 patients negative for hepatitis C virus antibody (p < 0.03). Most of our hepatitis E virus antibody-positive normal subjects and patients had never been abroad. These findings demonstrate that sporadic or subclinical hepatitis E virus infections also exist among the Chinese subjects in Taiwan. Hepatitis E virus infection may play an important role in patients with hepatitis C virus antibody-negative sporadic non-A, non-B hepatitis. IgG hepatitis E virus antibody in the sera of normal subjects may last for more than 8 yr. (Hepatology 1994;19:866-870.)
Serum hepatitis C virus RNA quantity and histological features of hepatitis C virus carriers with persistently normal ALT levels
Authors: Naito Masafumi , Hayashi Norio , Hagiwara Hideki , Hiramatsu Naoki , Kasahara Akinori , Fusamoto Hideyuki , Kamada Takenobu
Published: March 2007

We studied hepatitis C virus carriers with normal liver function to evaluate the histological features of their livers and the replicative levels of hepatitis C virus. Liver biopsies were performed in 22 hepatitis C virus carriers with persistently normal ALT levels. Hepatitis C virus RNA in serum was quantified with a competitive assay that combined reverse transcription and the polymerase chain reaction, which is based on coamplification of the target RNA with known amounts of synthetic mutated RNA. Three patients had normal livers on histological study, whereas the other 19 had chronic persistent hepatitis, with lymphoid infiltrates or aggregates in portal tracts commonly observed but intralobular inflammatory changes absent or minimal. The titer of hepatitis C virus RNA (logarithmic transformed copy number per milliliter of serum) varied from 4.0 to 8.0 (mean ± S.D.: 6.3 ± 1.1); it was significantly lower in the three patients with normal livers (4.3 ± 0.2) than in those with chronic persistent hepatitis with mild (6.4 ± 0.8, n = 11) or moderate (7.1 ± 0.5, n = 8) portal inflammation. The titer of hepatitis C virus RNA was correlated with the total score (r = 0.68) and the score for portal inflammation (r = 0.68) in the histological activity index. These results indicated that there seem to be “healthy carriers” of hepatitis C virus with extremely low levels of viral replication. However, in most hepatitis C virus carriers with persistently normal ALT levels, there are inflammatory changes in the portal tracts, with severity depending on the replicative levels of hepatitis C virus. (Hepatology 1994;19:871-875.)