bluuebunny/crossref_metadata_2025 huggingface dataset:
Hepatitis B virus, alpha‐fetoprotein synthesis, and hepatocellular carcinoma in Zaire
Authors: Kashala Lukumuena O. , Conne Béatrice , Kapanci Yusuf , Frei Philippe C. , Lambert Paul H. , Kalengayi Mbowa R. , Essex Max
Published: August 2010

ABSTRACT— We investigated the epidemiology of hepatocellular carcinoma (HCC) in Zaire, and evaluated the association between exposure to hepatitis B virus (HBV) and the development of HCC. Two hundred and twenty‐three consecutive cases of HCC diagnosed over 19 years (1966–1985) were reviewed. HCC represented 8.32% of all carcinomas and 5.56% of all cancers. Frequency was higher in males (75.7%) than in females (24.3%); a sex ratio of 3/1. The majority (82.1%o) of patients were aged 14 to 55 years with a peak occurrence in the fourth decade (28.6%). The mean age in males (41.27 ± 17.5 years) and females (37.40 ± 15.16 years) was significantly different (p<0.02). Sera from 40 patients and 68 age and sex‐matched controls were analyzed for markers of HBV infection: patients and controls had comparable rates of exposure (96%) vs 72.1%, respectively). However, patients had significantly higher HBsAg carrier rates (56.7% vs 7.35%; p < 0.001), and lower anti‐HBsAg seroconversion rates (25% vs 63.2%, p < 0.05). Using immunohistochemical analysis, the livers of patients were evaluated for HBsAg and HBcAg. These HBV antigens were more frequent in non‐tumorous hepatocytes (53.3% vs 23.3%, respectively) than in HCC cells (13.3% vs 3.3%). Serum alpha‐fetoprotein (AFP) was abnormal (>20 ng/ml) in 90%) of patients. The geometric mean (GM) AFP was 7273.8 ng/ml. AFP levels were significantly higher in HBsAg‐positive HCC cases (GM: 19 322.6 ng/ml; 95% confidence interval (CI): [3639.2, 102 565.2]) than in antigen negative cases (GM: 1939.5 ng/ml; 95%) CI: [182.8, 19952.6]), but did not correlate with HBV replication. Immunohistochemical detection of AFP revealed a similar correlation between AFP and HBsAg. Neither AFP level nor HBsAg production correlated with cellular atypia or tumor grade.
Hepatic endoplasmic reticulum storage diseases
Authors: Callea Francesco , Brisiptti Massimo , Fabbretti Giovanna , Bonino Ferruccio , Desmet Valeer J.
Published: August 2010

ABSTRACT— Endoplasmic Reticulum Storage Diseases (ERSD) represent a novel group of inborn errors of metabolism affecting secretory proteins and resulting in hepatocytic storage and plasma deficiency of the corresponding protein. The hepatocellular storage is due to a molecular abnormality hindering the translocation of the abnormal protein from the rough (RER) to the smooth endoplasmic reticulum (SER). The molecular abnormality is genetically determined; hence it is hereditary, congenital, familial and permanent. The storage is selective and exclusive for the mutant protein and predisposes to the development of chronic cryptogenic liver disease. ERSD include alpha‐1‐antitrypsin deficiency, fibrinogen storage and alpha‐1‐antichymotrypsin deficiency. Basically, the diagnosis of ERSD is a morphological one: immunohistochemistry and electron microscopy are essential tools for their identification.
Vitamin A abuse: development of cirrhosis despite cessation of vitamin A. A six‐year clinical and histopathologic follow‐up
Authors: Jorens Philippe G. , Michielsen Peter P. , Pelckmans Paul A. , Fevery Johan , Desmet Valeer J. , Geubel André P. , Rahier Jacques , Van Maercke Yvan M.
Published: August 2010

ABSTRACT— We report the case of a 35‐year‐old man who contracted vitamin A‐induced liver cirrhosis. Five years before, he had been investigated for vitamin A‐induced non‐cirrhotic portal hypertension. In this case, the clinical and histopathologic evolution from non‐cirrhotic portal hypertension to cirrhosis was documented. In spite of the cessation of pharmaceutical vitamin A intake, the disease progressed. Therapy with colchicine and phenobarbital apparently did not influence evolution to cirrhosis. This suggests that vitamin A can trigger largely unknown mechanisms of liver fibrosis which seem to be self‐perpetuating.
The use of radioactively labelled riboprobes for in situ hybridizaiotn; background and examples of applicaiton
Authors: Biffo Stefano , Tolosano Emanuela
Published: August 2010

Abstract: An overivew is provided concerning the preparation and use of radiolabelled single‐stranded RNA probes (ribooprobes) for in situ hybridization. All theoretical aspects are covered as weel as some practical examples from the authors' experience. The use of riboprobes for in situ hybridizaiton represents a valid and highly sensitive alternative to oligonucleotide probes. A sampel protocol is finally described in detail: this should enable any investigators to design the ISH assay that best fits their particular needs.
Bile duct changes in alcoholic liver disease
Authors: Ray Mukunda B. , Mendenhall Charles L. , French Samuel W. , Gartside Peter S. ,   Veterans Administration Cooperative Study Group
Published: August 2010

ABSTRACT— The histologic significance of various changes in the bile ductal structures as observed by cytokeratin immunoperoxidase assay was studied in 122 patients with alcoholic liver disease (ALD) as a part of a large Veterans Administration Cooperative Study on alcoholic hepatitis. Four types of morphologic changes in the biliary structures were observed: 1) proliferation of interlobular bile ducts in the portal tracts; 2) marginal bile ductular proliferation at the periphery of the portal tracts; 3) appearance of bile duct type cells (“oval cells”) in the liver parenchyma; and 4) metaplasia of bile duct epithelium to cells resembling hepatocytes. These bile ductal changes correlated strongly with liver fibrosis (p = 0.0003; 0.0003; 0.05; 0.0035, for 1, 2, 3, and 4, respectively), cirrhosis (p<0.0001 for all four parameters), portal inflammation (p<0.0001 for 1, 2, and 4; p = 0.0024 for “oval cells“), and with overall histologic severity scores (p<0.0001; p = 0.0001; p = 0.0017; p = 0.0005, respectively). However, these changes did not correlate significantly with fatty change, parenchymal degeneration and necrosis, cellular infiltrate or Kupffer cell hyperplasia, suggesting that they are probably not the direct consequences of liver cell necrosis. Periportal piecemeal necrosis correlated significantly with both portal bile duct (p = 0.0041) and marginal (p = 0.0078) bile ductular proliferation. Among all these changes, only marginal bile ductular proliferation correlated significantly with Mallory bodies present both in the hepatocytes (p = 0.05) and the bile ducts (p = 0.01). Both marginal bile ductular proliferation and “oval cells” correlated significantly with clinical severity of the liver disease as determined by high serum bilirubin and prolonged prothrombin time (Maddrey's discriminant function). This study suggests that: 1) a variety of important morphologic changes occur in the bile ductal structures in ALD, particularly when fibrosis and cirrhosis develop; 2) “oval cells” and bile duct metaplasia are frequent in ALD and appear to be significantly related to the development of fibrosis and cirrhosis; 3) bile ductules are an integral component of piecemeal necrosis in ALD; 4) some of the bile ductal changes may influence the clinical outcome of patients with ALD.
Hepatocyte‐conditioned medium potentiates insulin‐like growth factor (IGF) 1 and 2 stimulated DNA synthesis of cultured fat storing cells
Authors: Gressner Axel M. , Brenzel Arnfried , Vomeyer Tobias
Published: August 2010

ABSTRACT— IGF‐1 and IGF‐2 stimulate dose‐dependently DNA synthesis of nonconfluent cultures of rat fat storing cells, a nonparenchymal type of liver cells pathogenetically involved in the generation of liver fibrosis. Maximum stimulation of [3H] thymidine incorporation of about 2.6‐fold above control was reached with 100 ng/ml IGF‐1 and 500 ng/ml IGF‐2, respectively. The DNA synthesis promoting action of both IGF‐1 and IGF‐2 was most efficiently potentiated by hepatocyte‐conditioned medium raising the stimulatory effect up to 21‐fold above control cultures. Lysate of hepatocytes (up to 15 μg protein/ml) was not effective in potentiating the effect of IGF‐1. IGF‐1 is bound to free carrier protein(s) present in the medium of hepatocytes, but obviously absent in cell lysate. Three molecular weight fractions in the ranges of 67 kd, 35 kd, and 25 kd could be identified in the medium, which potentiate the growth‐promoting effect of IGF‐1. Applying Western ligand blot analysis, three molecular size classes of IGF‐1 binding proteins in the conditioned media of rat hepatocytes were determined. The major binding protein had a Mr of 28–34 kd, a minor portion was localized at Mr 24 kd, whereas trace binding affinities were found at Mr of about 95 kd. It is suggested that IGF‐1, IGF‐2 and the complex array of IGF‐binding proteins secreted by hepatocytes might be involved in the paracrine regulation of growth of fat storing cells.

Nonstandard abbreviations: BSA, bovine serum albumin; FCS, fetal calf serum; FSC, fat scoring cells; IGF, insulin‐like growth factor; IGFBP, IGF‐binding protein; Mr, relative molecular weight; PAGE, polyacrylamide gel electrophoresis; PBS, phosphate‐buffered saline; PCcM, hepatocyte‐conditioned medium; SDS, sodium dodecyl sulfate; SI, stimulation index.
Activation of nuclear protooncogenes and alpha‐fetoprotein gene in rat liver during the acute inflammatory reaction
Authors: Bernuau Dominique , Moreau Alain , Tournier Isabella , Legres Luc , Feldmann Gerard
Published: August 2010

ABSTRACT— Nuclear protooncogene and alpha‐fetoprotein gene expression is stimulated in hepatocytes during liver regeneration and by various growth factors in vitro. Metabolic adaptation of hepatocytes has been implicated in such gene reprogrammation. We examine here whether induction of an acute inflammation, a physiological situation of important metabolic adjustments, also triggers activation of nuclear oncogenes and of the AFP gene in rat liver. C‐fos, c‐jun and c‐myc mRNA accumulated on Northern blots between 4–12 h of inflammation and the steady‐state level of two small alpha‐fetoprotein transcripts characteristic of the adult liver increased at 4 h and 24 h of inflammation. In situ hybridization showed accumulation of the mRNA of the four genes studied in all hepatocytes, without any zonal lobular heterogeneity. 3H‐histoautoradiography and mitotic counts indicated an inhibition of DNA synthesis and mitosis, prolonged for at least 48 h after inflammation. Thus acute inflammation triggers the activation of nuclear protooncogenes and alpha‐fetoprotein gene in hepatocytes, but this activation is not followed by passage into the replicative cycle.
A case of drug‐induced ductopenia resulting in fatal biliary cirrhosis
Authors: Ishii Motoyasu , Miyazaki Yutaka , Yamamoto Takeshi , Miura Masahito , Ueno Yoshiyuki , Takahashi Tohru , Toyota Takayoshi
Published: August 2010

ABSTRACT— A 50‐year‐old woman suffered from a diffuse skin rash, high fever and jaundice immediately after a second injection of glutathion and Stronger Neo‐minophagen C which contains glycyrrhizin. Liver biopsy performed 11 months after the onset showed mild spotty hepatocyte necrosis, marked cholestasis in parenchyma, and some lymphocyte infiltration in the portal area. Interlobular bile ducts had undergone vacuolar degeneration or were absent in some portal tracts. In her hospital course, unremitting jaundice persisted and biliary cirrhosis developed with signs of portal hypertension; she died from liver failure 26 months after the onset. A liver specimen at her death revealed that most of the interlobular bile ducts had vanished. Based on the clinical course and pathology, drug‐induced ductopenia, possibly due to an adverse reaction to glycyrrhizin, is the most likely diagnosis. While drug‐related biliary cirrhosis is rarely fatal, this case presented an unusually rapid course of fatal biliary cirrhosis.
Epidemiology of hepatitis D virus (delta) infection in Yugoslavia
Authors: Delić Dragan , Gotić Mira , Oštrić Vladimir , Fridman Vesna , Nikolić Predrag , Jemuović Ljiljana , Nikolov Vladimir , Žerjav Sonja , Groza Šandor
Published: August 2010

ABSTRACT— In 614 HBsAg‐positive Yugoslavian patients, radioimmunoassay testing for anti‐delta showed the presence of this antibody in serum in 11.2%. Of the patients, 213 belonged to a risk group (i.v. drug users, hemophiliacs, hemodialysed patients and patients with posttransfusion hepatitis); a significant number of these patients (63; 29.6%) were found to have anti‐delta. A second group was composed of 401 HBsAg‐positive patients from the general population (patients with acute hepatitis B, with fulminant hepatitis B and patients with chronic HBV infection); delta infection was found only in six (1.5%). Immunohistochemical methods failed to demonstrate the delta antigen in the livers of 73 patients with chronic HBV infection. Testing the liver of 36 patients with fulminant hepatitis B for delta antigen demonstrated this reactivity in only one (2.8%) liver sample. Delta antigen was also found in the liver of a female patient who underwent biopsy in 1972. The results of this study suggest the HDV is not endemic in Yugoslavia; however, it is frequently found in patients at risk of blood exposure, primarily i.v. drug users.
Deposition of C3, the terminal complement complex and vitronectin in primary biliary cirrhosis and primary sclerosing cholangitis
Authors: Garred Peter , Lyon Hans , Christoffersen Per , Mollnes Tom Eirik , Tranum‐Jensen Jergen
Published: August 2010

ABSTRACT— Characteristics of primary biliary cirrhosis (PBC) and primary sclerosing cholangitis (PSC) are bile duct destruction and portal inflammation. Increased levels of circulating complement activation products are also present. This raises the possibility of involvement of complement‐dependent cytotoxic mechanisms in the pathogenesis. Therefore, we investigated liver biopsy specimens from 21 patients with PBC, six patients with PSC and six controls for complement deposits by immunohistochemistry using polyclonal and monoclonal antibodies against C3d, the terminal complement complex (TCC) and vitronectin (S‐protein). We found C3d, TCC and vitronectin deposits only in the portal tracts. C3d and TCC were present in the walls of the hepatic arteries and in the connective tissue stroma but never around the bile ducts. We found vitronectin deposits throughout the connective tissue, often independent of the TCC deposits. When vitronectin and TCC were co‐localized, the staining patterns were inverse; that is, intense staining for TCC accompanied weak staining for vitronectin and vice versa. Occasionally complete dissociation between TCC and vitronectin staining was observed. Deposits of TCC and vitronectin showed a focal distribution leaving many portal tracts free of TCC. Our results question whether complement‐dependent cytotoxic mechanisms take part in the bile duct destruction in PBC and PSC.
Hepatitis C virus infection in anti‐HIV positive and negative French homosexual men with chronic hepatitis: comparison of second‐ and third‐generation anti‐HCV testing
Authors: Marcellin Patrick , Colin Jean‐François , Martinot‐Peignoux Michèle , Pham Bach‐Nga , Lefort Valérie , Picault Anne‐Blandine , Degott Claude , Erlinger Serge , Benhamou Jean‐Pierre
Published: August 2010

ABSTRACT— To determine the prevalence of hepatitis C virus (HCV) infection in homosexuals with chronic hepatitis, we tested for anti‐HCV antibodies 113 (47 anti‐HIV positive) French non‐drug‐addicted homosexual men admitted for chronic viral hepatitis. Anti‐HCV were detected with second‐ and third‐generation ELISAs (ELISA2 and ELISA3) and RIBAs (RIBA2 and RIBA3). Chronic hepatitis was related to non‐A, non‐B infection in four, to hepatitis D virus (HDV) infection in five and to hepatitis B virus (HBV) infection in 104 patients. Anti‐HCV positivity was found in 50.4% and 12.4% of the 113 patients, with ELISA2 and ELISA3, respectively. Positivity with RIBA2 and RIBA3 was found in only six of the 57 ELISA2 positive patients (all six were ELISA3 positive). The high prevalence of positivity with ELISA2 not confirmed by RIBA2 or RIBA3 suggests false‐positive results. ELISA2 positive results were more frequent with frozen serum samples than with fresh serum samples (62% vs 23.5%, p = 0.0003). However, even with fresh serum, ELISA2‐positive RIBA‐negative results remained frequent in anti‐HIV positive patients. ELISA3 seems to give more specific results. We conclude that the prevalence of HCV infection, as assessed with RIBA, was 5.3% among French homosexual men with chronic hepatitis (3.8% after exclusion of transfused patients). This low prevalence suggests that homosexual transmission of HCV is relatively uncommon.
Immunohistochemical and ultrastructural study of hepatic sinusoidal linings during dichloropropanol‐induced acute hepatic necrosis
Authors: Haratake Joji , Furuta Akiko , Hashimoto Hiroshi
Published: August 2010

An acute phase of severe hepatic necrosis induced by dichloropropanol was examined immunohistochemically and ultrastructurally, in order to study chronological changes of sinusoidal morphology during acute hepatic injury. Male Wistar rats were injected with 1,3‐dichloro‐2‐propanol (DC2P) and sacrificed at various intervals after the injections. DC2P‐injected rats showed zonal necrosis of the centrilobular space with a peak from 24 to 48 h after the injection. Destruction of sinusoidal linings appeared at 4 h, and was gradually aggravated along the advancing hepatocytic necrosis. Monocytic influx into the necrotic areas was initiated at 6 h. At 48 h, collapsed centrilobular spaces showed a loss of most sinusoidal structures with active phagocytosis of macrophages, proliferation of perisinusoidal cells, and accumulation of collagen fibrils. At 72 h, there were many regenerating sinusoidal structures, which were composed of rather thick and less fenestrated endothelium and underlying multilayered processes of mesenchymal cells, along the regenerating hepatocytes. In these areas, occasional junctions between regenerating hepatocytes and mesenchymal cells were seen. Reconstruction of sinusoidal linings was closely related to the hepatocytic regeneration, and a hepatocytic‐mesenchymal interaction might participate in this morphodynamic course of the sinusoidal reconstruction.
Hepatitis B and C virus infection as risk factors for liver cirrhosis and cirrhotic hepatocellular carcinoma: a case‐control study
Authors: Tsai Jung‐Fa , Chang Wen‐Yu , Jeng Jen‐Eing , Ho Mei‐Shang , Lin Zu‐Yau , Tsai Jeui‐Hsiung
Published: August 2010

To investigate whether hepatitis B virus (HBV) and hepatitis C virus (HCV) infection are risk factors for liver cirrhosis and hepatocellular carcinoma (HCC), a case‐control study of 102 cirrhotic HCC patients, 102 sex‐matched and age‐matched patients with liver cirrhosis, and 102 matched patients with non‐hepatic disease controls was performed. The prevalences of hepatitis B surface antigen (HBsAg) and antibody to HCV (anti‐HCV) in HCC (70.5%, 39.2%) and liver cirrhosis (74.5%, 27.4%) were higher than controls (16.6%, 10.5%) (P = 0.0001). In HBsAg‐negative patients, the prevalence of anti‐HCV in cirrhotic HCC (66.6%) and liver cirrhosis (46.1%) was higher than in controls (10.5%; P = 0.0001). There was no such difference in HBsAg‐positive patients. Multivariate analysis revealed that both HBsAg and anti‐HCV were important risk factors for HCC (odds ratio, 6.52 and 4.59, respectively) and liver cirrhosis (odds ratio, 4.22 and 2.29, respectively). There was no difference in odds ratio when HCC and liver cirrhosis were compared. Our result implies that both HBV and HCV are independent risk factors for cirrhotic HCC and liver cirrhosis in Taiwan.
Endothelial cell heterogeneity in the normal human liver acinus: in situ immunohistochemical demonstration
Authors: Scoazec Jean‐Yves , Racine Lorraine , Couvelard Anne , Flejou Jean‐Francois , Feldmann Gérard
Published: August 2010

While a certain degree of structural and functional intra‐lobular heterogeneity of sinusoidal endothelial cells has been observed in rodents, little information is available about the zonal characteristics of sinusoidal endothelial cells in the human liver acinus. We have therefore examined the intra‐acinar distribution of a panel of endothelial markers in the normal human liver, including: (a) structural markers of continuous and sinusoidal endothelia (PECAM‐1, CD‐34 protein, VE‐cadherin, 1F10 antigen), (b) functional markers specific for sinusoidal endothelial cells, as previously determined in the laboratory (CD4 protein, the lipo‐polysaccharide‐binding protein receptor (CD 14), aminopeptidase N, ICAM‐1, receptors II and III for the Fc fragment of immunoglobulins G), (c) endothelial cell‐matrix adhesion proteins and leukocyte‐endothelial cell adhesion molecules. We observed a heterogeneous distribution for: (a) the 1F10 antigen, whose distribution in the human liver acinus was restricted to vessels situated along the axis of acinar zone 1, (b) the lipopolysaccharide‐binding protein receptor and the receptor III for the Fc fragment of IgG, not expressed or only barely expressed in acinar zone 1. The distribution of the other markers tested did not display significant intra‐lobular variation. Our in situ results suggest the existence of a degree of zonal heterogeneity in the structural and functional characteristics of sinusoidal endothelial cells in the human liver acinus. This might contribute to the constitution of distinct microenvironments within the human liver parenchyma.
Testing for hepatitis C virus sequences in peripheral blood mononuclear cells of patients with chronic hepatitis C in the absence of serum hepatitis C virus RNA
Authors: Muratori Luigi , Giostra Fabrizio , Cataleta Michela , Francesconi Raffaella , Ballardini Giorgio , Cassani Fabio , Lenzi Marco , Bianchi Francesco B.
Published: August 2010

Hepatitis C virus (HCV) is able to replicate in peripheral blood mononuclear cells (PBMC) of HCV‐infected patients. Few data are available on PBMC testing for HCV RNA in serum HCV RNA negative patients, positive for anti‐HCV and with histological evidence of chronic hepatitis. Twenty such patients were studied; of these, 11 were tested during interferon α (IFN) treatment, at the time of serum HCV RNA clearance and ALT normalisation: only one was found to be positive for HCV sequences in PBMC. Within 3 months of IFN withdrawal all 11 patients relapsed with high ALT and recurrence of serum HCV RNA. Of nine serum HCV RNA negative patients with chronic hepatitis C who were not receiving IFN when tested (four untreated patients and five patients who had already completed IFN schedule), PBMC HCV RNA was detected in four. Evidence of active HCV replication (presence of the minus strand genome) in PBMC was also observed in two cases. Thus, five of the 20 patients without detectable serum HCV RNA turned out to be carriers of HCV sequences in PBMC. These data indicate that: 1. PBMC are an extrahepatic replication site of HCV; this is true also in the absence of serum HCV RNA; 2. the role of PBMC as a “viral reservoir” after IFN‐induced serum HCV RNA clearance is questioned; 3. the absence of both serum and PBMC HCV RNA in patients under IFN is not predictive of sustained viral loss; 4. testing for PBMC viral sequences might enhance the chances of detecting HCV infection.
Iron concentration and distribution in the newborn liver
Authors: Faa G. , Sciot R. , Farci A. M. G. , Callea F. , Ambu R. , Congiu T. , van Eyken P. , Cappai G. , Marras A. , Costa V. , Desmet V. J.
Published: August 2010

Recent observations on a correlation between fetal serum ferritin and gestational age, consistent with an increase in fetal iron stores during pregnancy, led us to study liver iron content in 22 human stillborns, newborns and infants of different gestational and postnatal age. At autopsy, a longitudinal liver slice was subdivided into ten blocks. Each sample was analyzed for iron content by atomic absorption spectroscopy. The mean iron concentration in the studied livers was 21.6 μM/g dry tissue (d.t.). A striking interindividual variability in iron content was observed: the hepatic concentration of the metal ranged from 3.3 to 64.4μM/g d.t. No correlation was found between the hepatic iron concentration and gestational age or other clinical parameters of the patients studied. Moreover, the total storage iron of the liver did not appear to be correlated with the gestational age. The analysis of iron concentration in ten blocks in each liver revealed an irregular distribution of the metal. Lobar differences were observed, with a tendency of the left lobe to accumulate more iron than the right one. Furthermore, striking differences in iron content were found between adjacent liver samples, ranging in one instance from 4.5 up to 109.0 μM/g of dry tissue. Perls' stain for iron was positive in 7 out of the 22 livers examined, showing an irregular acinar distribution, with preferential periportal localization. Our data show that the newborn liver can be considered an interesting model for the study of iron storage. The observed uneven distribution of the metal, even in the absence of fibrosis normally associated with iron storage in adults, casts doubt on the interpretation of iron concentration of a small liver sample as representative of the mean hepatic iron concentration.
Ca2+influx initiates death of hepatocytes injured by activation of complement
Authors: Nishikawa Yoshiyuki , Ukida Minoru , Matsuo Ryuichi , Omori Nobuhiko , Tsuji Takao
Published: August 2010

To clarify the role of cytosolic Ca2+in hepatocellular death, we exposed cultured hepatocytes to human serum and a monoclonal antibody directed against rat liver plasma membranes to produce complement‐mediated cell injury. The change in cytosolic Ca2+concentration was measured by fura2 and fluo3 fluorescence. With the addition of monoclonal antibody, an increase in cytosolic Ca2+was observed, followed by cell death. Both the increase in intracellular Ca2+and cell death were prevented by intracellular Ca2+chelation or removal of extracellular Ca2+. We conclude that an increase in cytosolic Ca2+plays a major role in hepatocellular injury induced by exposure of the cell membrane to monoclonal antibody.
Serum markers of hepatic fibrogenesis in chronic hepatitis type C treated with alfa‐2A interferon
Authors: Gallorini A. , Plebani M. , Pontisso P. , Chemello L. , Masiero M. , Mantovani G. , Alberti A.
Published: August 2010

Hepatic fibrogenesis is a dynamic process which characterizes the course of chronic hepatitis. It has stimulated interest in the possible effect of interferon therapy on liver fibrosis. We have evaluated a panel of serum markers of fibrogenesis, namely N‐terminal procollagen III peptide (PII‐INP), C‐terminal procollagen I peptide (PICP), laminin and hyaluronate in 35 patients with chronic hepatitis type C, before, during and after interferon treatment. Before treatment, PIIINP was elevated in 8.5%, 44% and 71% of patients with chronic persistent hepatitis, chronic active hepatitis and cirrhosis, respectively, while the corresponding figures for PICP were 0%, 50% and 46%, and for laminin 16.5%, 70% and 71%; hyaluronate was elevated in only five out of seven patients with cirrhosis. Patients with high PIIINP levels at presentation and a persistent response to treatment showed persistent normalization of this parameter, which was not observed in non‐responders. In contrast, the other markers showed no significant correlation with interferon response. These results indicate that PIIINP correlates with interferon response in chronic hepatitis type C.
An immunohistochemical analysis of 13 cases with combined hepatocellular and cholangiocellular carcinoma
Authors: Haratake Joji , Hashimoto Hiroshi
Published: August 2010

Thirteen cases of combined hepatocellular (HCC) and cholangiocellular carcinoma (CCC) were examined. In addition to routine pathology, immunoreactivities for carcinoembryonic antigen, α‐fetoprotein (AFP), cytokeratin (Cam 5.2 and AE1), epithelial membrane antigen (EMA) and tumor‐associated glycoprotein 72 (B72.3) were also examined. The average age of the 13 cases was 64.8 years, which lay between the average ages of pure HCC and CCC cases. They were categorized as separate type (2), collision type (6), and intermingled type (5). AE1 and EMA were the best markers to differentiate the CCC from the HCC area. B72.3 immunoreactivity was detected only in CCC (46%). There were no transitional features between HCC and CCC in two cases of the separate type and two cases of the collision type. However, focal transitional features from HCC to CCC were observed in all cases of the intermingled type and in four of six cases of the collision type. In one case of the intermingled type, many cancer cells contained both bile and mucus simultaneously, and revealed dual immunoreactivities. The conclusions are: 1) the combined type is generated from two sources; one is the intrahepatic double cancer (thoroughly separate type and a part of the collision type) and another is the stem cell origin with diverse phenotypes (intermingled type and a part of the collision tumor); and 2) AE1 was the most helpful marker to differentiate the CCC area from HCC, and other markers, e.g. AFP for HCC and EMA, CEA, and B72.3 for CCC, were also supportive but somewhat limited in the differential diagnosis.
Severe hepatotoxicity related to benzarone: a report of three cases with two fatalities
Authors: Hautekeete Marc Leopold , Henrion Jean , Naegels Serge , DeNeve André , Adler Michael , Deprez Carlne , Devis Ghislain , Klöppel Günter
Published: August 2010

We report three cases of severe hepatotoxicity related to benzarone, a benzofuran derivative. Our cases include a 35‐year‐old woman with (sub)fulminant hepatitis, a 67‐year‐old woman with macronodular cirrhosis, and a 68‐year‐old man with severe chronic active hepatitis and cirrhosis, with positivity of anti‐smooth muscle antibodies. Two patients died. We stress the potential of benzarone to cause hepatotoxicity, which usually resembles severe chronic active hepatitis. Our cases constitute the most severe cases of benzarone hepatotoxicity reported so far, and comprise the first cases of (sub)fulminant hepatitis and cirrhosis related to benzarone.
Dose‐related opposite effects of hydrocortisone on hepatocyte proliferation in the rat
Authors: Nadal C.
Published: August 2010

To evaluate the role of glucocorticoids in the regulation of hepatocyte proliferation, high doses (6.25 mg/kg or more) and low doses (1.25 mg/kg or less) of hydrocortisone were injected into various categories of rats which had very different levels of proliferation. We used: (a) suckling rats whose stable unsynchronized proliferation can be stimulated and synchronized by acute inflammation or cyproterone feeding; (b) adult rats with a low level of proliferation, and (c) 2/3 hepatectomized rats during the first period of synchronized proliferation or during the second period of lower and unsynchronized proliferation. The kinetics of the labelling index and mitotic index were observed after hydrocortisone injection. High doses inhibited proliferation, as already found by many investigators. This effect was observed in suckling and hepatectomized rats and during acute inflammation in suckling or adult rats. Low doses had no inhibitory activities and, on the contrary, stimulated proliferation in some situations. The highest stimulations were observed during the early period after 2/3 hepatectomy or in combination with acute inflammation either in suckling rats or in 2/3 hepatectomized rats during the late period of regeneration. In all these situations hydrocortisone was a cofactor active during the early G1‐phase. In contrast, in normal suckling rats a slight stimulation was observed due to an action at the G0‐G1 transition. No such effect was found in adult rats. Our data show that doses of hydrocortisone in the therapeutic range or close to physiological values do not inhibit and, on the contrary, can stimulate hepatocyte proliferation.
Liver disease and serum hexosaminidase levels. Studies in a human hepatoma cell‐line (Hep G2 cells)
Authors: Hultberg B. , Floren C. H. , Isaksson A. , Jensen E.
Published: August 2010

In various forms of liver disease, increased levels of the lysosomal enzyme β‐hexosaminidase (Hex) occur in serum. This may be caused by disturbances of the hepatocytic function, and we therefore studied the intracellular and extracellular isoenzyme pattern of Hex in a human hepatoma cell‐line (Hep G2), using an immunoassay method, which separates Hex A and Hex B isoforms. This cell‐line synthesizes and secretes Hex. The cumulative release of extracellular activity was about 3–10% of the intracellular activity. B‐isoforms comprised one‐third of intracellular activity but only 20% of extracellular activity. The proportion of extracellular B‐isoforms increased with time, presumably due to instability of A‐isoforms at 37°C. Cycloheximide inhibited the release of Hex activity, whereas NH4Cl increased the extracellular fraction of Hex, even at a concentration of 1 mmol/l. We speculate that the increased concentration of NH4+ in patients with liver disease interferes with the distribution pathway of the lysosomal enzymes. This might be one reason for the increased serum Hex activity found in patients with liver disease.
HLA‐DR expression in liver in human HIV infection
Authors: Guido Maria , Rugge Massimo , Rocchetto Piero , Fattovich Giovanna , Murer Bruno
Published: August 2010

Abstract: In order to clarify the relationship between HLA‐DR antigen expression in the liver and human HIV infection, we studied 66 non‐autopsy specimens obtained from HIV‐infected patients with and without AIDS; 33 biopsies from HIV‐negative patients were considered for control purposes. All biopsies were immunohistochemically tested with monoclonal antibody TAL‐1B5, reacting with the α‐chain subunit of HLA‐DR in the avidin biotin complex method. We found occasional and usually weak HLA‐DR expression on hepatocytes and biliocytes. By comparison with controls, HIV‐positive cases showed a significantly higher percentage of HLA‐DR immunoreactivity on hepatic artery and portal vein endothelial cells. No relationship was demonstrated between HLA‐DR endothelial expression and the CD4+ lymphocyte count or grade of portal/periportal inflammation in the liver. The biological significance of HLA‐DR expression on intrahepatic vascular endothelial cells in HIV infection still remains to be clarified.
Long interval between HCV infection and development of hepatocellular carcinoma
Authors: Castells Luis , Vargas Victor , Gonzalez Antonio , Esteban Juan , Esteban Rafael , Guardia Jaume
Published: August 2010

Abstract: A high prevalence of HCV infection has been reported in patients with hepatocellular carcinoma. The progression from acute transfusion‐associated hepatitis to hepatic cirrhosis and hepatocellular carcinoma has been suggested in several studies to be very long. We have investigated the prevalence of anti‐HCV and the interval between HCV infection and hepatocellular carcinoma among 191 consecutive patients with cirrhosis and liver‐cell carcinoma. Serum samples from 191 patients with cirrhosis and hepatocellular carcinoma, consecutively diagnosed in our hospital between 1988 and 1993, were tested for serological markers of HBV and HCV infection. One hundred and forty‐eight patients (77.5%; 95% confidence interval (c.i): 76% to 80%) were anti‐HCV positive by 2nd generation enzyme immunoassay (confirmed by 2nd generation recombinant immunoblot assay) and 152 patients (79.5%; 95% c.i: 76% to 80%) were anti‐HCV positive by 3rd generation enzyme immunoassay, while only 14 (7.4%; 95% c.i: 5% to 10%) were HBsAg positive. Of the 29 anti‐HCV positive patients with previous transfusion, the interval between the date of blood transfusion and the diagnosis of hepatic cirrhosis was 24±12.5 years and that of hepatocellular carcinoma was 26.8±12.4 years. These results confirm the high prevalence of HCV infection in patients with hepatocellular carcinoma and the slow sequential progression from HCV infection through cirrhosis and hepatocellular carcinoma.
Hepatocyte and Kupffer cell function after liver transplantation in the rat –in vivo evaluation with dynamic scintigraphy
Authors: Svensson Gunnar , Friman Styrbjörn , Jacobsson Lars , Holmberg Stig B.
Published: August 2010

Abstract: In vivo physiological measurements of hepatocyte and Kupffer cell function after liver transplantation are desirable. Orthotopic liver transplantation was performed in 54 rats. Hepatocyte and Kupffer cell function were measured with dynamic liver scintigraphy. Hepatic clearance of 99mTc‐Nanocoll (%/min), an albumin colloid phagocytosed by the Kupffer cells, was used to evaluate Kupffer cell function. Hepatic clearance of 99mTc‐IODIDA (%/min), an imino‐diacetic‐acid taken up and secreted by the hepatocytes, was used to evaluate the hepatocyte function. Hepatic clearance in control rats was 27±2 %/min for Nanocoll and 30±3 %/min for IODIDA. After syngenic liver transplantation, without rejection, there was a rise in Nanocoll clearance (34±2 %/min p<0.01) after 3 weeks, but no change in IODIDA clearance (32±3 %/min N.S.). After syngenic liver transplantation with preservation time prolonged to 16 h, there were no changes in IODIDA or Nanocoll clearance 1 day after transplantation. Both IODIDA (11±2 %/min) and Nanocoll clearance (22±2 %/min) were decreased (p<0.001) during rejection after allogenic transplantation. An in vivo method of measuring the hepatocyte and Kupffer cell function in the transplanted liver is described. Kupffer cell function was increased after syngenic liver transplantation. Kupffer cell and hepatocyte function were decreased during rejection. Dynamic liver scintigraphy seems a suitable procedure for examining liver injury after liver transplantation in the experimental setting.
Manifestation late in life of idiopathic adulthood ductopenia
Authors: Müller Christian , Ulrich Walter , Penner Edward
Published: August 2010

Abstract: Idiopathic adulthood ductopenia is a chronic cholestasic liver disease of unknown etiology characterized by the loss of interlobular bile ducts. We describe three patients who fulfilled the diagnostic criteria of idiopathic adulthood ductopenia, but differed from the cases reported so far in late manifestation of the disease and a benign clinical course despite histologic evidence of ongoing cholangitis. Treatment with ursodeoxycholic acid in one patient resulted in improvement of biochemical markers of cholestasis, suggesting that chronic cholestasis in idiopathic adulthood ductopenia can be influenced beneficially.
Autoimmune hepatitis versus viral hepatitis C
Authors: Strassburg C. P. , Manns M. P.
Published: August 2010

Abstract: Numerous viruses are capable of inducing the syndrome of chronic hepatitis. Among them are the hepatitis B, C and D viruses. Out of the most common agents of chronic hepatitis, the hepatitis C virus has been found to be strikingly associated with autoimmune diseases and serological markers of autoimmunity. Conversely, the syndrome of genuine autoimmune hepatitis lacks evidence of previous or ongoing virus infection and is diagnosed by additionally excluding metabolic, toxic, and genetic causes of chronic hepatitis, and by the response to immunosuppressive treatment. This review article summarizes the current knowledge of hepatotropic virus‐induced autoimmunity. It focuses on the present molecular and immunological definitions, the clinical and molecular distinction between autoimmune hepatitis and chronic viral hepatitis and the implications for the safe and efficacious therapy of these disease entities.
Biochemical and clinical study of muscle atrophy at thenar and hypothenar eminences in patients with cirrhosis
Authors: Nambu Masaji , Kaneok Kazuhiro , Lijima Toshihiko
Published: August 2010

Abstract: The results of various biochemical examinations in 14 patients with cirrhosis (6 males and 8 females) with muscle atrophy at the thenar and hypothenar eminence (muscle atrophy group; mAG) were compared with those in 13 patients (8 males and 5 females) with cirrhosis without muscle atrophy at these sites (non‐muscle atrophy group; NmAG). All patients were elderly men and women (mAG and NmAG, mean age, 69±3 years and 60±7, respectively). In most mAG patients, muscle atrophy was accompanied by palmar erythema. Muscle atrophy was histologically demonstrated by biopsy. Furthermore, electromyography and magnetic resonance study of the cervical spinal cord revealed that the atrophy was of myogenic rather than neurogenic origin. The Child‐Pugh score, body mass index and sex hormone level in urine (total 24 h) in the two groups were compared along with the biochemical results. There were no significant differences between the two groups in urine estrogen and testosterone levels. The urinary creatinine excretion was significantly reduced in mAG. The creatine phosphokinese, lactate dehydrogenase isoenzyme and aldolase levels in serum did not differ significantly in the two groups, whereas the serum albumin level was significantly increased in NmAG. Significant differences were observed only for the serum albumin level, age and body mass index. Thus, we consider that palmar muscle atrophy in patients with cirrhosis is not due to hormonal excess in serum, but may be attributable to advanced age and diminished physical strength.
Liver abscesses due to Listeria monocytogenes
Authors: Marino Pietro , Maggioni Marco , Preatoni Armando , Cantoni Alessandra , Invernizzi Fulvio
Published: August 2010

Abstract: Involvement of the liver in cases of Listeria monocytogenes is uncommon. We report a case of hepatitis due to L. monocytogenes, with a pathological finding of multiple abscesses. Blood cultures yielded L. monocytogenes. The patient died a few days after admission.
Antimitochondrial antibodies in patients with chronic hepatitis C
Authors: Grimbert Sylvie , Johanet Catherine , Bendjaballah Fadila , Homberg Jean‐Claude , Poupon Raoul , Beaugrand Michel
Published: August 2010

Abstract: Although autoantibodies have been found in the serum of patients with chronic hepatitis C virus (HCV) there has been no convincing evidence of the presence of antimitochondrial antibodies, until now. Sera from 460 untreated patients with chronic hepatitis C were tested for antimitochondrial antibodies, using an indirect immunofluorescence technique; and if they tested positive for the antibodies (titer more than 1:50), they also were treated by Western blot analysis. Seven (1.5%) sera were positive. None of the patients had biological or histological evidence of primary biliary cirrhosis. Antimitochondrial antibodies recognized one of the oxo‐dehydrogenase multienzyme complexe's epitopes by Western blot assay in three patients only. All seven patients were then treated by interferon alpha for six months. None showed exacerbation of liver disease during treatment. HCV‐RNA disappeared from the serum in one patient who became negative for anti‐M2 antibodies. The four patients who did not respond to interferon‐alpha therapy, and the two who relapsed after treatment withdrawal, had sustained positive antimitochondrial antibodies. These data suggest that: 1) antimitochondrial antibodies present in patients with chronic hepatitis C do not always recognize the same epitopes as in primary biliary cirrhosis; 2) these antibodies may disappear after eradication of HCV, suggesting that the production of antimitochondrial antibodies is linked to the presence of the virus and 3) the clinical and biological course of chronic hepatitis C, and the response to interferon‐alpha therapy, does not seem to be different in patients who are positive for antimitochondrial antibodies.
Strain‐ and sex‐specific variations in hepatic glutamine synthetase activity and distribution in rats and mice
Authors: Sirma Hüseyin , Williams Gary M. , Gebhardt Rolf
Published: August 2010

Abstract: The distribution of glutamine synthetase (GS) in a mammalian liver is restricted to a small zone of hepatocytes surrounding the central veins. The determination of the size of the GS+ zone in rats by immunohistochemistry revealed that it differed between rat strains and was larger in males than in females of each strain. Accordingly, the means of the relative mean width (RMW) values that characterize the size of the GS+ zone were 19%, 26%, and 39% lower in females than in males of Sprague‐Dawley, Wistar, and Fischer rats, respectively. Upon orchidectomy of male rats, the size of the GS+ zone diminished towards the value found in females, while ovariectomy was without effect. This orchidectomy‐induced reduction was reflected in corresponding changes of the RMW values as well as in the number of GS+ cells per pericentral field and was not due to the slightly smaller size of the GS+ hepatocytes in the orchidectomized males. No such sex difference was found in M775 mice. Biochemical GS activity was higher in the male rats than in the female rats and changed correspondingly to the distribution after gonadectomy. In the mice, only the specific activity of GS dropped after orchidectomy. In primary cultures of rat hepatocytes, no influence of testosterone or estrogen on GS activity and cellular distribution was observed, even after stimulation of GS activity with dexamethasone and growth hormone. Both sex hormones, however, were able to affect the activity of glucose‐6‐phosphate dehydrogenase (G6PD). The observed sex differences in the activity and distribution of GS in rat livers suggest that sex hormones not only modulate the level of this enzyme but are at least partially involved in the determination of the size of the compartment of GS expression. According to the results in the cell cultures, the effects of the sex hormones appear indirect rather than direct.
Pathogenesis of focal and random hepatocellular necrosis in endotoxemia: microscopic observation in vivo
Authors: Asaka Shosaku , Shibayama Yuro , Nakata Katsuji
Published: August 2010

Abstract: The present study was undertaken in rats to clarify the role of sinusoidal circulatory disturbances due to fibrin thrombi in the development of focal and random hepatocellular necrosis in endotoxemia. Sinusoidal circulation was examined microscopically in vivo in rats injected with endotoxin or heparin, or both. The sinusoids in places were occluded by adherent fibrin and neutrophils soon after endotoxin injection, and subsequently the sinusoidal blood flow stagnated, reversed, or detoured. Most of these sinusoidal circulatory disturbances recovered in a few hours. However, when the sinusoidal occlusion developed simultaneously in clusters of adjacent sinusoids, the sinusoidal circulatory disturbance persisted and induced ischemic foci and then hepatocellular coagulative necrosis. Pretreatment with heparin definitely prevented the adherence of fibrin and neutrophils to the sinusoidal walls, and focal hepatocellular necrosis did not appear. These results suggest that focal and random hepatocellular necrosis in endotoxemia is caused by circulatory disturbances due to fibrin thrombi in clusters of adjacent sinusoids.