bluuebunny/crossref_metadata_2025 huggingface dataset:
Histological changes of the liver in experimental graft‐versus‐host disease across minor histocompatibility barriers. VIII. Role of eosinophil infiltration
Authors: Nonomura Akitaka , Kono Naoko , Mizukami Yuji , Nakanuma Yasuni
Published: August 2010

Abstract: Although eosinophil infiltrate has been recognized in hepatic graft‐versus‐host disease, its significance in relation to hepatic graft‐versus‐host disease lesions is unknown. In the present study, we analyzed hepatic eosinophil infiltration in relation to bile duct damage in experimental mouse graft‐versus‐host disease across minor histocompatibility barriers up to 14 months after transplantation. Portal eosinophil infiltration was found from 1 week after transplantation throughout the entire 14‐month observation period. It was most striking during the early chronic stage of hepatic graft‐versus‐host disease between 2 to 7 months, with a peak at 5 months after transplantation. Microscopic and electron microscopic study revealed eosinophils infiltrated around the bile duct as well as in the bile duct epithelial layer. They were commonly found together with lymphocytes but were also occasionally found singly around the bile duct and in the bile duct epithelial layer. Bile duct epithelial cells in contact with and in the vicinity of eosinophils showed a variety of degenerative changes, occasionally associated with the presence of extracellular eosinophil granules. Bile duct epithelial cells with eosinophil infiltration just beneath the basement membrane frequently showed further characteristic severe degenerative changes with shedding or dropping‐off into the lumen, which features were quite similar to those seen in the bronchial epithelium in asthma patients. These results indicate that not only lymphocytes but also eosinophils may be involved in the production of the bile duct injury in hepatic graft‐versus‐host disease, especially in its early chronic stage.
Hyaluronate levels in donor organ washout effluents: a simple and predictive parameter of graft viability
Authors: Rao Prakash N. , Bronsther Oscar L. , Pinna Antonio D. , Snyder James T. , Cowan Scott , Sankey Scott , Kramer David , Takaya Shunichi , Starzl Thomas
Published: August 2010

Abstract: The principal cause of primary non‐function in orthotopic liver transplantation is thought to be preservation injury to the microvasculature. We, therefore, evaluated if effluent levels of hyaluronate, whose uptake is an endothelial cell marker, could predict early graft function and ultimate graft outcome in orthotopic liver transplantation. A total of 102 cases were studied in two phases. In the first phase, we attempted to determine if a correlation existed between effluent hyaluronate levels, early graft function and ultimate graft outcome. This phase of the study was also used to determine hypothetical cut‐off values for hyaluronate which could discriminate between good and bad livers. Thirty‐two livers orthotopically transplanted to randomly selected primary recipients were studied. After varying periods of static cold storage (4°C) in University of Wisconsin solution, the livers were reinfused with cold (4°C) lactated Ringer's solution. The first 50 ml of the reperfusion effluent was collected from the infrahepatic vena cava. Effluent samples were analyzed for hyaluronate. Linear regression analysis demonstrated a significant correlation between effluent hyaluronate levels and post‐operative aspartate and alanine aminotransferase levels (p>0.001 for both). Logistic regression demonstrated a highly significant correlation (p=0.0056) between effluent hyaluronate levels and ultimate graft outcome. Generation of Receiver Characteristics Curves indicated that a level between 400 and 430 μg l‐1 could possibly discriminate between good livers and those at risk of early graft failure. The authenticity of this hyaluronate cut‐off level was further confirmed in the second phase of the study where 70 consecutive primary crossmatch‐negative transplants were performed. A highly significant difference was observed in peak aspartate and alanine aminotransferase levels in the first week (p>0.0006 and p>0.0005, respectively) between livers with effluent hyaluronate levels 400 μg · l‐1 and livers with hyaluronate levels higher than 400 μg l‐1. Logistic regression revealed a highly significant correlation between effluent hyaluronate levels and graft success (p=0.0001). Since hyaluronate uptake by the microvascular endothelial cell is significantly greater than production, high hyaluronate effluent levels in failed livers would be due to decreased hyaluronate uptake by the injured microvascular endothelial cell. We therefore conclude that effluent hyaluronate levels may prove to be a reliable preoperative test to assess early graft function and outcome in clinical orthotopic liver transplantation.
Portal vein thrombosis complicating hepatocellular carcinoma. Value of ultrasound‐guided fine‐needle biopsy of the thrombus in the therapeutic management
Authors: Cedrone Augusto , Rapaccini Gian Ludovico , Pompili Maurizio , Aliotta Antonio , Trombino Concetta , De Luca Francescantonio , Caturelli Eugenio , Caputo Salvatore , Gasbarrini Giovanni
Published: August 2010

Abstract: During a 4‐year period portal vein thrombosis was diagnosed in 20 Child class A patients with cirrhosis by means of ultrasound and ultrasound‐Doppler study. Seventeen of them showed single or multiple focal liver lesions diagnosed as hepatocellular carcinoma by ultrasound‐guided fine‐needle biopsy and the remaining three a coarse liver echo‐pattern without focal lesions. One patient was found to have developed portal vein thrombosis after the fifth ethanol injection of a single hepatocellular carcinoma lesion 17 mm in diameter. Ultrasound‐guided fine‐needle biopsy of the thrombus was performed on all the patients: portal vein thrombosis was neoplastic in 13 cases and non‐neoplastic in seven cases (five patients with a single lesion; one with two lesions; one with coarse liver echo‐pattern). Among the five patients with a single lesion, one had already been treated by percutaneous ethanol injection therapy. There were no complications related to the biopsy procedures. The diagnosis of non‐neoplastic thrombosis allowed five new patients to be recruited for percutaneous ethanol injection treatment and allowed it to continue in the patient with portal vein thrombosis occurring after the fifth ethanol injection. The routine use of ultrasound‐guided fine‐needle biopsy of portal vein thrombosis yields an accurate diagnosis of the nature of the thrombus and can improve the selection for percutaneous ethanol injection treatment of patients with cirrhosis with hepatocellular carcinoma lesions.
Apoptosis and proliferation of human hepatocellular carcinoma
Authors: Hino Naoki , Higashi Toshihiro , Nouso Kazuhiro , Nakatsukasa Harushige , Tsuji Takao
Published: August 2010

Abstract: To investigate the contribution of apoptosis, a major mechanism of cell death, in the growth of hepatocellular carcinoma, we analyzed both apoptosis and cell proliferation in human hepatocellular carcinoma. We used the terminal deoxynucleotidyl transferase‐mediated dUTP‐biotin nick end labeling method and proliferative cell nuclear antigen staining, respectively. Among 21 hepatocellular carcinoma specimens examined, four were well, ten were moderately, and seven were poorly differentiated hepatocellular carcinoma. Terminal deoxynucleotidyl transferase‐mediated dUTP‐biotin nick end labeling‐positive cells in hepatocellular carcinoma were scattered individually or were sometimes clustered in the tumors. The terminal deoxynucleotidyl transferase‐mediated dUTP‐biotin nick end labeling indices were 0.35±0.09, 0.81±0.29, and 1.9±0.94 in well, moderately, and poorly differentiated hepatocellular carcinoma, respectively. The proliferative cell nuclear antigen labeling indices were 6.6±0.9, 13.1±3.5, and 26.7±6.3 in hepatocellular carcinoma in the same respective order of differentiation. The differences in both terminal deoxynucleotidyl transferase‐mediated dUTP‐biotin nick end labelling indices and proliferative cell nuclear antigen labeling indices (p>0.05) were significant between well, moderately and poorly differentiated hepatocellular carcinoma. There was a positive correlation between the terminal deoxynucleotidyl transferase‐mediated dUTP‐biotin nick end labeling and proliferative cell nuclear antigen labeling indices in hepatocellular carcinoma (r=0.84, p>0.001). This study showed that the proliferation rate and the incidence of apoptosis increased as the differentiation grade of hepatocellular carcinoma was lowered, suggesting a rapid turnover of cancer cells in the lower differentiation grades. Apoptosis may thus play an important role in the growth of hepatocellular carcinoma.
Serum clusterin and vitronectin in alcoholic cirrhosis
Authors: Høgåsen Kolbjørn , Homann Christian , Mollnes Tom Eirik , Graudal Niels , Høgåsen Anne Kirsti Myrvang , Hasselqvist Philip , Thomsen Åge Chr. , Garred Peter
Published: August 2010

Abstract: Clusterin and vitronectin are multifunctional regulatory proteins which both serve as complement lysis inhibitors. Previous data have strongly suggested that serum vitronectin is mainly produced in the liver, whereas the biosynthetic origin for serum clusterin has not been determined. In the present study we aimed to determine the role of the liver in producing these proteins and to evaluate the proteins as possible markers of liver failure. We therefore quantified clusterin and vitronectin in serum from patients suffering from alcoholic liver cirrhosis (n=83), and in serum‐free culture supernatants from the hepatoma cell line HepG2. The median clusterin concentration was 0.20 g/l in cirrhosis and 0.37 g/l in the controls, whereas corresponding vitronectin values were 0.19 and 0.26 g/l, respectively. The concentration of both proteins showed significant correlation (p>0.0001) with disease severity and with established plasma markers of hepatic synthetic function, such as albumin and prothrombin complex. The clusterin level, but not the vitronectin level, correlated with survival (p=0.005). The rates of synthesis of clusterin, vitronectin and C3 from HepG2 cells were 0.02, 0.21 and 1.9 μg/106 cells/24 h, respectively. From the present data we conclude that clusterin (as vitronectin and C3) is mainly produced in the liver and may be a useful marker in the evaluation of severity of liver disease and prognosis of patients with alcoholic cirrhosis.
Liver‐infiltrating and circulating CD4+ T cells in chronic hepatitis C: immunodominant epitopes, HLA‐restriction and functional significance
Authors: Löhr Hanns F. , Schlaak Jörg F. , Kollmannsperger Stefan , Dienes Hans‐Peter , zum Büschenfelde Karl‐Herman Meyer , Gerken Guido
Published: August 2010

Abstract: The aim was to assess the specificity and functional significance of liver‐infiltrating and peripheral blood T cells in chronic hepatitis C. Peripheral blood mononuclear cells hepatitis C virus from 50 of 58 (86.2%) patients with chronic hepatitis C virus infection and 6 of 28 (21.4%) controls showed a proliferative T cell response to at least one of 16 synthetic peptides covering highly conserved regions of the core, envelope (El) and non‐structural regions (NS4) of hepatitis C virus. However, six immunodominant peptides were exclusively recognized by the proliferating blood mononuclear cells from 46 patients with chronic hepatitis C virus infection (79.3%). Fine specificity and HLA‐restriction were studied with 15 peptide‐specific CD4+ T cell lines and 23 T cell clones isolated from liver tissue and peripheral blood of 12 patients with chronic hepatitis C. It was demonstrated that the peptide‐specific response of CD4+ T cells was restricted to the presence of autologous accessory cells and HLA‐DR and ‐DP molecules. Eight peptide‐specific T cell lines and five T cell clones derived from liver tissue and peripheral blood, released interferon‐γ (200–6600 pg/ml) and tumor necrosis factor‐α (100–400 pg/ml) and no or little interleukin‐4 (<140 pg/ml) after peptide‐specific or mitogeneic stimulation, thus resembling a Th1‐like cytokine profile. Patients with active liver disease showed significantly higher proliferative responses to hepatitis C virus core peptides than asymptomatic hepatitis C virus carriers or complete responders to interferon therapy. In conclusion, class II‐restricted CD4+ T cell responses to some immunodominant epitopes within the hepatitis core region correlated with disease activity in chronic hepatitis C virus infection. Functionally, liver‐infiltrating and peripheral blood T cells released Th1‐like cytokines in response to the specific stimulus. Thus, it can be suggested that CD4+ T cells can mediate the pathogenesis of chronic hepatitis C virus induced liver disease.
Serum levels of endothelial injury markers creatine kinase‐BB and soluble thrombomodulin during human liver transplantation
Authors: Chazouilleres Olivier , Vaubourdolle Michel , Robert Annie , Fourel Valerie , Balladur Pierre , Laribi Abderrhamane , Giboudeau Jacqueline , Parc Rolland , Poupon Raoul , Hannoun Laurent
Published: August 2010

Abstract: Endothelial damage within the sinusoids of the liver probably plays a key role in primary liver dysfunction following transplantation. The aim of this work was to study the serum levels of two potential markers of endothelial damage, creatine kinase‐BB and soluble thrombomodulin, during human graft revascularization. Thirteen human liver grafts were preserved in UW solution (mean time: 13.8 h). Creatine kinase‐BB and transaminase activities and soluble thrombomodulin levels were measured: 1) in effluent and 2) in serum samples sequentially collected before revascularization, then during the first 120 min of revascularization and first post‐operative week. No correlation was observed between serum values (peak) and effluent values. In serum, pre‐operative creatine kinase‐BB activities were correlated with soluble thrombomodulin levels (p=0.01). Both increased significantly during the first minutes of the revascularization, then decreased markedly. In contrast, AST activity was maximal at day 1. This detectable and early release of creatine kinase‐BB and soluble thrombomodulin in blood is in keeping with the early occurence of endothelial damage. Together with previous data, these findings suggest that serum determination of these two markers may be a useful tool in the assessment of endothelial injury in liver transplantation.
A prospective study of hepatitis C virus infection after needlestick accidents
Authors: Arai Yumi , Noda Katsuhisa , Enomoto Norihiro , Arai Keiichi , Yamada Yukinori , Suzuki Kunio , Yoshihara Harumasa
Published: August 2010

Abstract: There have been few prospective studies of hepatitis C virus (HCV) infection after needlestick accidents in hospital employees. In the present study, the prevalence and features of HCV infection after needlestick accidents were evaluated prospectively measuring serum HCV‐RNA. Subjects were 56 employees who had HCV needlestick accidents. To monitor the development of hepatitis, the serum ALT levels and HCV‐related seromarkers, such as first generation anti‐HCV (RIA), second generation anti‐HCV (PHA) and HCV‐RNA (RT‐PCR) were measured every month for at least 12 months after the accidents. Three of 56 (5.4%) recipients developed HCV infection. HCV‐RNA was detected in all three recipients within 4 months after the exposure, and second‐generation HCV antibody was detected in two of three recipients. The detection of HCV‐RNA was earlier than that of HCV antibody. Two of three HCV‐infected recipients developed type C acute hepatitis and one of two received interferon therapy; however, the other case received no medication. The detection of HCV‐related seromarkers and the elevation of ALT levels were transient in these three recipients; thus, none developed chronic hepatitis. In conclusion, HCV infection developed in 5.4% of recipients within 4 months after HCV accidents. All of these HCV‐infected recipients showed fair prognosis. HCV‐RNA was a beneficial parameter for early detection of HCV infection.
Recurrent liver involvement in heatstroke
Authors: Sort Pau , Mas Antoni , Salmeron Joan Manel , Bruguera Miquel , Rodés Joan
Published: August 2010

Abstract: We report the case of a young patient who presented two self‐limited episodes of acute jaundice that developed immediately following a long distance run. Zone 3 necrosis was the most prominent histologic change of the liver during the second episode. The diagnosis of heatstroke with liver damage was based on the exclusion of known causes of acute hepatitis, the histopathologic changes and the circumstances preceding the onset of liver disease. This seems to be the first case of severe and recurrent liver impairment due to heatstroke.
Hepatitis C virus serotypes and liver pathology
Authors: Guido Maria , Rugge Massimo , Thung Swan N. , Chemello Liliana , Leandro Gioacchino , Alberti Alfredo , Cecchetto Attilio , Pontisso Patrizia , Cavalletto Luisa , Ninfo Vito
Published: August 2010

Abstract: The present study aimed to analyze the pathology of chronic hepatitis C in relation to HCV serotype, and to speculate on possible differences in the pathogenesis of liver injury. Liver biopsies were investigated from 59 consecutive patients in whom hepatitis C virus genotypes were determined by a serological genotyping assay that detects antibodies directed to epitopes encoded by the NS4 region. A morphological study was performed in each case, semiquantitatively scoring necro‐inflammatory and fibrotic liver lesions. The prevalence of HCV serotypes was as follows: 26 of the 59 patients (44%) had type 1 infection, 11 (19%) had type 2 and 20 (35%) had type 3. A significant association between intravenous drug abuse and serotype 3 infection was observed. Patients with HCV type 2 proved significantly older than patients with infection types 1 or 3, and more frequently they showed a more active liver disease, but no differences were found in the quality and acinar topographic distribution of all the morphological lesions scored. In conclusion, in chronic hepatitis C a more active liver disease can be related to HCV serotype 2 but the spectrum of liver lesions is independent of HCV types. From a morphological point of view, a different pathogenesis of liver injury related to different HCV types is unlikely.
Localization of hyaluronan in human liver sinusoids: a histochemical study using hyaluronan‐binding protein
Authors: Ichida Takafumi , Sugitani Souichi , Satoh Tomomi , Matsuda Yasunobu , Sugiyama Motoya , Yonekura Kenji , Ishikawa Tohru , Asakura Hitoshi
Published: August 2010

Abstract: Circulating hyaluronan is mostly derived from lymph, fibroblast and Ito cells in the liver, and more than 90% of hyaluronan is degraded in hepatic sinusoidal endothelial cells. Thus, elevated serum hyaluronan is regarded as an indication of hepatic fibrosis with activated Ito cells and dysfunctional sinusoidal endothelial cells. We studied the distribution of hyaluronan in human liver sinusoids to determine the influences on elevated hyaluronan levels in sera. Histochemical examination was made using hyaluronan‐binding protein (HABP) and serial sections of liver tissue for staining of alpha‐smooth muscle actin (ASMA) (an indicator of activated Ito cells) and of ulex europaeus agglutinin I lectin (UEA‐1) (closely related to hepatic sinusoidal capillarization). Positive staining, indicating the presence of hyaluronan, was noted in fibrous regions around the portal tracts, areas of focal necrosis in the liver parenchyma, and walls of the sinusoids in chronic hepatitis. In this group, hyaluronan‐positive areas corresponded to positive ASMA staining and faint staining of UEA‐1. On the contrary, in liver cirrhosis, UEA‐1‐positive areas were essentially identical to hyaluronan‐positive areas and to ASMA‐negative areas in sinusoidal walls. Hyaluronan and ASMA could be detected in the same areas of sinusoidal walls in chronic hepatitis, but not in liver cirrhosis. Hyaluronan appears to be mainly related to the staining of activated Ito cells in chronic hepatitis. Therefore, we concluded that in chronic hepatitis, the production of hyaluronan was accelerated in Ito cells; however, degradation of hyaluronan by sinusoidal endothelial cells continued. On the contrary, in liver cirrhosis, hyaluronan production decreased in Ito cells, and a marked transformation of sinusoidal endothelial cells with hepatic sinusoidal capillarization indicated loss of the ability to degrade hyaluronan. These different mechanisms in chronic hepatitis and liver cirrhosis may operate in the sinusoidal walls and may cause the elevation of hyaluronan in sera.
No significant influence of HLA determinants on susceptibility to hepatitis C virus infection in Caucasian patients with end‐stage renal disease
Authors: Chen Dong‐Feng , Entires Walter , Kliem Volker , Tillmann Hans L. , Brunkhorst Reinhard , Koch Karl M. , Manns Michael P. , Stangel Walter
Published: August 2010

Abstract: In hepatitis C, both susceptibility to infection and the course of disease may depend on differences in the immune response. As the major histocompatibility complex (MHC) plays a crucial role in antigen presentation, we investigated a possible relationship between susceptibility to hepatitis C virus (HCV) infection and human leucocyte antigen (HLA) alleles. Therefore, phenotype frequencies of HLA were compared in 186 anti‐HCV positive patients with end‐stage renal disease (ESRD) to 328 anti‐HCV negative patients with ESRD. HLA class I alleles were determined serologically and HLA class II alleles (DRB1, DQA1, DQB1) by the polymerase chain reaction sequence‐specific oligonucleotide (PCR‐SSO) technique. Additionally, in anti‐HCV positive patients we looked for a relationship between the activity of hepatitis C (indicated by elevation of transaminases or the presence of viremia) and HLA determinants. For the three criteria (antibody status, elevation of transaminases and viremia) a significant association to HLA alleles was not found in patients with ESRD. This suggests that neither susceptibility to HCV infection nor the biochemical activity of hepatitis and HCV‐RNA positivity seem to be strongly related to HLA status in Caucasian patients with end‐stage renal disease.
Stromal and blood vessel wall invasion in well‐differentiated hepatocellular carcinoma
Authors: Nakano Masayuki , Saito Akiko , Yamamoto Masakazu , Doi Mikio , Takasaki Ken
Published: August 2010

Abstract: We examined hepatocellular carcinomas (HCCs) that were smaller than 2 cm in diameter. Ninety‐nine nodules from 65 patients were removed for treatment. The nodules were divided into four types (A, B, C and D) according to the following criteria: first, the appearance of the margin of nodules, distinct or indistinct; and second, cellular atypia, uniform or multiple as “nodules in nodule”. In 45 indistinct margin nodules, 28 showed uniform atypia (type A) and 17 were of “nodules in nodule” (type B). As for 54 distinct margin nodules, 23 were “nodules in nodule” (type C) and 31 showed uniform atypia (type D). Cancer cell invasion was divided into three types: (1) stromal invasion into fibrotic tissue and/or portal tracts, (2) blood vessel wall invasion of portal veins or hepatic veins, and (3) tumor thrombus. The stromal and vessel wall invasion occurred almost at the same rate regardless of degree of atypia. This study shows that well‐differentiated HCCs in which the cancer cells show only very slight atypia have the potential for metastasis to intrahepatic and other sites.
Hepatotoxicity of ethanol: protective effect of calcium channel blockers in isolated hepatocytes
Authors: Cobreros A. , Sainz L. , Lasheras B. , Cenarruzabeitia E.
Published: August 2010

Abstract: This study examines the effects of three calcium channel blockers (verapamil, nifedipine and diltiazem) on isolated rat hepatocytes exposed to ethanol. In the first part of our study, hepatocytes were incubated with increasing concentrations of ethanol (100, 300, 500, 1000 mM) for varying times. Alanine aminotransferase (ALT), aspartate aminotransferase (AST) and lactate dehydrogenase (LDH) release were measured to evaluate the cytotoxic effects of ethanol. The concentration of 300 mM and time of incubation of 45 min were chosen for cytoprotection experiments in which calcium channel blockers, at two different concentrations, were added to the medium 30 min prior to the addition of ethanol. ALT, AST and LDH release as well as lipid peroxidation and cellular reduced gluthatione (GSH) were measured. Nifedipine and verapamil (25 μM) reduced ALT, AST and LDH activities. The highest dose of diltiazem (50 μM) was more effective than the lowest one (25 μM). Ethanol caused a significant depletion of cellular GSH content as well as a moderate enhancement of lipid peroxidation. While none of the three calcium channel blockers was able to restore the decrease in GSH levels, diltiazem (25 μM) and nifedipine (50 μM) showed the greatest effect, significantly reducing lipid peroxidation.
Interferon‐alpha does not cause significant cardiac dysfunction in patients with chronic active hepatitis
Authors: Kadayifci Abdurrahman , Aytemir Kudret , Arslan Mehmet , Aksoyek Serdar , Sivri Bulent , Kabakci Giray
Published: August 2010

Abstract: The cardiac adverse effects of interferon (INF) treatment have been reported recently in various clinical trials of INF. In this study, the cardiac effects of recombinant INF‐alpha treatment were evaluated prospectively in a group of patients with chronic active viral hepatitis (CAH). Sixteen patients with CAH type B, 14 patients with CAH type C and one patients with CAH type D were included in this study, and 4.5, 3 and 9 MU of recombinant INF‐alpha‐2a was administrated three times a week to these patients, respectively. The durations of treatment were 6 months for CAH type B and C, and 12 months for type D. The cardiac status of all patients was evaluated and monitored with a detailed medical history, physical examination, electrocardiography (ECG), telecardiography, echocardiography and heart rate variability tests at the beginning of the study and at the first and sixth months of INF therapy and also 6 months after ceasing the therapy. The clinical evaluation of patients before the treatment revealed that three had hypertension, one had a past medical history of myocardial infarction, one had a prosthetic mitral valve replacement and another had left hemiblock in her ECG record. No significant changes and adverse effects were detected in clinical examination and cardiovascular tests of all patients, either in preexisting cardiovascular diseases, during therapy and after stopping the treatment. The cardiac adverse effects of INF reported in previous studies are questionable and we conclude that it can be used safely in CAH patients. Therefore, it should be kept in mind that fever and tachycardia may occur during the first and second weeks of INF therapy and patients with high risk for cardiac disease should be monitored closely in this period of treatment.
Preventive effect of cyclosporin A on experimentally induced acute liver injury in rats
Authors: Wasaki Shuji , Sakaida Isao , Uchida Koichi , Kimura Teruaki , Kayano Kozo , Okita Kiwamu
Published: August 2010

Abstract: We examined the effect of cyclosporin A (CsA) on the pathogenesis of acute experimental liver injury in rats induced by injection of heat‐killed Propionibacterium acnes (P. acnes) and subsequent injection of lipopolysaccharide (LPS). Pretreatment with CsA significantly reduced serum alanine aminotransferase (ALT), serum tumor necrosis factor‐α (TNF‐α) production, without changing the TNF‐α mRNA level in the liver, and plasma interferon‐γ (IFN‐γ), following LPS injection in this model. Twenty‐four‐hour mortality was also markedly improved, from 100% in the P. acnes plus LPS group to 0% in the CsA‐pretreated group. Although direct addition of CsA to isolated hepatic macrophages from P. acnes‐pretreated rats did not prevent the production of TNF‐α and active oxygen species, isolated hepatic macrophages from P. acnes plus CsA‐pretreated rats significantly reduced their production in response to the addition of LPS. These results suggest that CsA protects against P. acnes plus LPS‐induced acute liver injury, not by direct inhibition of hepatic macrophage activation, but by indirect prevention of hepatic macrophage activation, presumably related to the reduction in plasma IFN‐γ levels.
The relationship of histology to genotype in chronic HCV infection
Authors: Booth Jonathan C. L. , Foster Graham R. , Levine Tanya , Thomas Howard C. , Goldin Robert D.
Published: August 2010

Abstract: The histological description of chronic hepatitis is undergoing considerable change at present. It has become important to define chronic hepatitis aetiologically and then define levels of necro‐inflammatory change (grade) and fibrosis (stage). The aim of this study was to compare the ability of different histological scoring systems to detect differences in the pathological changes associated with infection with the different HCV genotypes that are known to have different natural histories. The histological appearances of liver biopsies from 29 HCV infected patients were compared by the Knodell histological activity index (HAI), modified histological activity index and the Scheuer histological scoring system. HCV genotyping was performed for each patient by sequence analysis of the 5′ non‐coding region. The histological appearances from HCV 1 infected patients showed a tendency towards more active necro‐inflammatory changes when compared with those from HCV 2 or 3 infected patients. The levels of fibrosis were similar for all genotypes. The modified HAI and Scheuer scoring systems detected differences, not revealed by the Knodell system, in the types of inflammatory pathology produced by the different genotypes of HCV In particular these scoring systems noted significant differences in the component scores of inflammation, in addition to the total inflammatory scores. In conclusion, the recently introduced scoring systems were able to detect differences in liver pathology produced by infection of similar duration with different viral genotypes. As genotype is considered an important determinant of disease progression and response to anti‐viral therapy, it is likely that those scoring systems correlating with genotype will yield more useful histological information than those that do not.
The importance of cofactors in the histologic progression of minimal and mild chronic hepatitis C
Authors: Shev Steven , Dhillon Amar P. , Lindh Magnus , Serléus Zita , Wejstål Rune , Widell Anders , Norkrans Gunnar
Published: August 2010

Abstract: A follow‐up liver biopsy was done 9–16 years (mean 12 years) after initial biopsy in 20 untreated Swedish patients infected with hepatitis C (8 men, 12 women; mean age 30 years at initial biopsy) in whom first biopsy had been classified as chronic persistent hepatitis. A significant progression of liver damage was found when using Histology Activity Index (HAI) scoring according to Knodell (p=0.006 for total HAI score; p=0.03 for grading, i.e., sum of HAI components 1, 2, and 3; p=0.01 for staging, i.e., HAI component 4, fibrosis). Fourteen of 20 (70%) patients had increased while 6 had decreased or unchanged HAI scores on follow‐up biopsy. Occasional heavy alcohol drinkers (n=6) had an increased follow‐up HAI score as compared with nondrinkers (p<0.05). Eight of 14 who deteriorated on follow‐up versus 0 of 6 with improved or unchanged liver histology were anti‐HBc positive (p=0.04). There was no significant correlation between HCV genotype and prognosis; however, the only two patients with liver cirrhosis on follow‐up had genotype lb. In conclusion, most patients with minimal or mild chronic hepatitis C in the present study had histologic progression on the latest biopsy. Cofactors such as alcohol abuse and exposure to hepatitis B may have a greater influence than HCV alone in determining the rate of deterioration of liver disease.
Platelet‐associated IgM elevated in patients with chronic hepatitis C contains no anti‐platelet autoantibodies
Authors: Kosugi S. , Imai Y. , Kurata Y. , Tomiyama Y. , Shiraga M. , Honda S. , Nishikawa M. , Yonezawa T. , Kanakura Y. , Matsuzawa Y.
Published: August 2010

Abstract: We investigated whether thrombocytopenia in patients with chronic hepatitis C is due to anti‐platelet autoantibodies. Platelet‐associated IgG (PAIgG) and platelet‐associated IgM (PAIgM) were measured by direct immunofluorescent flow cytometric analysis. Elevation of PAIgM level was detected in 70% of chronic hepatitis C patients, while only a mild elevation of PAIgG level was detected in 32% of the cases. The elevation of PAIgM values in these patients was comparable to that in patients with chronic immune thrombocytopenic purpura (ITP). However, elevated PAIgM was also found in both patients with and without thrombocytopenia, and no correlation was found between PAIgM and platelet count. Eluted PAIgM did not react with normal platelets in all cases with a positive PAIgM value, indicating that eluted PAIgM contained no detectable anti‐platelet antibodies. During alpha‐interferon therapy, the level of PAIgM increased in association with the decrease in platelet counts in 75% of the cases; however, eluted PAIgM at any day point never reacted with platelets from normal donors. PAIgM was elevated in patients with chronic hepatitis C, but contained no detectable anti‐platelet autoantibodies. Thrombocytopenia in these patients is not due to anti‐platelet autoantibodies.
Serum levels of tissue inhibitor of metalloproteinases‐2 and of precursor form of matrix metalloproteinase‐2 in patients with liver disease
Authors: Ebata Mieko , Fukuda Yoshihide , Nakano Isao , Katano Yoshiaki , Fujimoto Noboru , Hayakawa Tetsuo
Published: August 2010

Abstract: Serum levels of tissue inhibitor of metalloproteinases‐2 (TIMP2) and of precursor form of matrix metalloproteinase‐2 (proMMP2) were determined in patients with chronic hepatitis and hepatocellular carcinoma by a one‐step sandwich enzyme immunoassay. Serum levels of TIMP2 and proMMP2 were significantly higher in patients with chronic liver disease, than in normal controls. Serum levels of TIMP2 showed a weak negative correlation with the serum albumin level and prothrombin time (PT). Serum levels of proMMP2 in patients with chronic hepatitis were strongly correlated with those of type IV collagen and were negatively correlated with PT and serum albumin levels. Serum proMMP2 levels were also significantly correlated with histological stages. These data indicate that serum levels of proMMP2 might be useful in the follow‐up of patients with chronic hepatitis.
Hepatitis C virus genotypes: epidemiological and clinical associations
Authors: Kleter Bernhard , Brouwer Johannes T. , Nevens Frederik , Van Doorn Leen‐Jan , Elewaut Andre , Versieck Jacques , Michielsen Peter P. , Hautekeete Marc L. , Chamuleau Rob A. F. M. , Brénard Réginald , Bourgeois Nadine , Adler Michael , Quint Wim G. V. , Bronkhorst Carolien M. , Heijtink Rudolf A. , Hop Wim J. C. , Fevery Johan , Schalm Solko W. ,   The Benelux Study Group on Treatment of Chronic Hepatitis C
Published: August 2010

ABSTRACT— In a cohort of 292 chronic hepatitis C patients living in the Benelux countries the relationship between viral genotype and geographical origin, route of transmission, clinical characteristics and severity of liver disease was analyzed. HCV‐RNA isolates could be classified by the Line Probe Assay (LiPA) as 1a, 1b, 2, 3, 4 or 5 in 286 (98%) cases. Patients of European origin were predominantly infected with HCV subtype 1b (164/254, 65%, CI 58–70%), as were patients of Asian origin (7/13, 54%). Patients originating from Surinam (South America) had predominantly type 2 (9/10, 90%), whereas Africans were mainly infected with type 4 (7/9, 77%). Blood transfusion was the mode of transmission in 142 (50%) patients, intravenous drug abuse (IVDA) in 40 (14%), occupational needle accident or tattoo in 11 (4%); no obvious source of infection was found in 93 (33%). In patients infected by blood transfusion, subtype 1b was predominant (70%, CI 61–77%), whereas subtypes 1a and 3 were predominant in those infected by IVDA (25% and 45%, respectively, p<0.001). Cirrhosis was observed in 68 (24%) patients; in multi‐variate analysis, factors independently related to cirrhosis were: the duration of infection, age and prior hepatitis B. No significant relationship was found between the severity of fibrosis or liver inflammation and the HCV (sub)types. In summary, in this large cohort of patients in the Benelux countries the hepatitis C virus (sub)type present was clearly related to the country of origin and the route of transmission, but not to the severity of liver disease.
An endothelin receptor antagonist TAK‐044 ameliorates carbon tetrachloride‐induced acute liver injury and portal hypertension in rats
Authors: Gandhi Chandrashekhar R. , Nemoto Edwin M. , Watkins Simon C. , Subbotin Vladimir M.
Published: August 2010

ABSTRACT—Hepatic levels of a powerful vasoconstrictor endothelin‐1 (ET‐1) and its receptors increase in human and carbon tetrachloride (CCl4)‐induced liver cirrhosis. The aim of this study was to determine whether antagonism of hepatic ET‐1 receptors ameliorates CCl4‐induced hepatic injury and portal hypertension in rats. Acute liver injury was induced by a single intraperitoneal injection of CCl4(0.3 ml/kg), whereas cirrhosis and portal hypertension were induced by CCl4treatment (0.15 ml/kg twice a week) for 8 weeks. Hepatic morphology, ET‐1 and its receptors, and portal venous pressures were determined. Increases in ET‐1 and its receptors occurred within 24 h of CCl4administration, and progressively thereafter during the development of cirrhosis. The acute CCl4‐induced hepatic injury was characterized by significant increases in portal pressure (from 8.7 ± 1.8 to 17.6 ± 3.3 mmHg;p<0.01) and serum levels of liver enzymes, as well as massive hepatocellular necrosis (62±8%). Intravenous administration of an ET‐1 receptor antagonist TAK‐044 reduced portal pressure to 13.6±2.8 mmHg (p<0.05), and ameliorated hepatocellular necrosis by about 35% (p<0.001). TAK‐044 treatment also produced significant reduction in serum levels of liver enzymes. In cirrhotic rats, portal venous infusion of TAK‐044 reduced portal hypertension by about 40% (p<0.05). In conclusion, these results indicate involvement of ET‐1 in acute liver injury as well as portal hypertension associated with hepatic cirrhosis, and a potential for ET‐1 receptor antagonists in the treatment of these pathologic conditions.
Fulminant hepatic failure associated with dicoumarol therapy
Authors: Castedal Maria , Aldenborg Frank , Olsson Rolf
Published: August 2010

ABSTRACT— Coumarins have been associated with non‐predictable hepatic injury. In the case of dicoumarol, there is no hard evidence in the literature of a causal connection with liver damage. We report the case of a 73‐year‐old woman who developed a fatal liver disease of a mixed hepatocellular‐cholestatic type after 3 months of treatment with dicoumarol. A thorough diagnostic work‐up did not reveal any other possible cause of the liver disease.
The Fas system is not significantly involved in apoptosis in human hepatocellular carcinoma
Authors: Kubo Keiko , Matsuzaki Yuko , Okazaki Muneko , Kato Akira , Kobayashi Nobuyuki , Okita Kiwamu
Published: August 2010

ABSTRACT— We investigated the role of apoptosis in relation to proliferative activity in hepatocellular carcinoma (HCC) using in situ DNA nick end labeling (ISNEL) and immunostaining for the Ki‐67 antigen in 35 patients with HCC. We also performed immunostaining for Fas and Fas ligand (Fas L) to determine the relationship between the Fas system and apoptosis. The ratio of the ISNEL labeling index (LI) to the Ki‐67 LI was significantly lower in HCC than in surrounding nontumorous liver tissue (p<0.0001), suggesting that a decrease in apoptosis relative to cell proliferation is important in the pathogenesis of HCC. Fas and Fas L were expressed in both HCC and nontumorous tissue, but Fas and Fas L LIs were significantly lower in HCC (p<0.0001). Fas expression by cells near ISNEL‐positive cells tended to be increased in nontumorous tissue in mirror‐image sections, suggesting that apoptosis is related to Fas expression. However, this pattern was rarely observed in HCC. These findings indicate that the Fas system may not play a major role in apoptosis in HCC.
Absence of human herpesvirus 8 DNA sequences in vascular tumors of the liver*
Authors: Ishak Kamal G. , Bijwaard Karen E. , Makhlouf Hala R. , Taubenberger Jeffery K. , Lichy Jack H. , Goodman Zachary D.
Published: August 2010

ABSTRACT— Kaposi's sarcoma‐associated herpes virus (KSHV), also designated human herpesvirus 8 (HHV8), has been detected consistently in Kaposi's sarcoma, body cavity lymphoma and multicentric Castleman's disease, both in human immunodeficiency virus (HIV)‐positive and ‐negative patients. Identification of KSHV/HHV8 DNA sequences in various benign and malignant vascular tumors in HIV‐negative patients was reported in one study, but was not confirmed in several other studies. The vascular lesions, other than Kaposi's sarcoma, in which sequences could not be detected have included malignant vascular tumors of serous membranes, infantile capillary hemangiomas, and several benign and malignant vascular tumors of the spleen. We studied 30 primary benign and malignant vascular tumors of the liver; KSHV/HHV8 DNA sequences could not be detected in any. We conclude that this virus plays no role in the etiology of vascular tumors of the liver.
Microsatellite instability in human hepatocellular carcinoma: relationship to p53 abnormalities
Authors: Kazachkov Y. , Yoffe B. , Khaoustov V. I. , Solomon H. , Klintmalm G. B. G. , Tabor E.
Published: August 2010

ABSTRACT— Aims/Background: Microsatellite instability was sought in 10 human hepatocellular carcinomas (HCCs) to determine whether defective DNA mismatch repair might be implicated in the multiple genetic alterations observed in the p53 tumor suppressor gene in some of these patients' tumors. Methods: Genomic DNA from HCCs and adjacent non‐tumorous livers was subjected to PCR with primers for nine microsatellites, and PCR products were resolved in a denaturing gel. Microsatellite instability was defined as the presence of band shifts or additional bands for at least two microsatellite sequences in an HCC compared to the nontumorous liver tissue from the same patient. Results: Microsatellite instability was detected in four of ten HCCs. Three of these four HCCs did not have p53 exon mutations. However, one HCC had microsatellite instability as well as multiple p53 exon mutations and multiple intron alterations. Four other patients with multiple p53 intron alterations in HCC (compared to their own nontumorous liver), three of whom also had a mutation in the exons, had no microsatellite instability. Conclusions: Defective DNA mismatch repair, as indicated by microsatellite instability, might have played a role in hepatocarcinogenesis in four of the ten patients, but in general it was not associated with p53 alterations. In one of the ten patients, defective DNA mismatch repair might have been the cause of multiple mutations in both the coding and intron sequences of the p53 gene.
Plasma from patients with cirrhosis increases tissue plasminogen activator release from vascular endothelial cells in vitro
Authors: Hayashi Takeshi , Kamogawa Asahi , Ro Shinsei , Yamaguchi Kai , Kobayashi Yutaro , Takahashi Yasuhito , Murayama Masahiro
Published: August 2010

ABSTRACT— Aims/Background: In patients with severe liver disease, blood levels of many coagulation and fibrinolytic factors are lowered due to a diminished synthetic capability in the liver. Tissue plasminogen activator (t‐PA) is synthesized by the vascular endothelial cells, however, and is increased in such patients. Methods: Amounts of t‐PA secreted were determined by immunosorbent assay after plasma from patients with cirrhosis was added to cultured human umbilical vein endothelial cells to determine whether a plasma factor directly enhanced t‐PA secretion from vascular endothelial cells. Results: Release of t‐PA was significantly higher with exposure to plasma from patients with decompensated cirrhosis than when plasma from patients with compensated cirrhosis or normal subjects was used (p<0.01 and p<0.05, respectively). Plasminogen activator inhibitor 1(PAI‐1) concentrations were measured similarly but did not differ among the three groups. Conclusions: Our results indicate that factors in plasma from patients with decompensated cirrhosis directly stimulate t‐PA release from the vascular endothelial cells, while any increased PAI‐1 release observed in comparable in vivo situations is probably an indirect response to an increase of t‐PA or a result of impaired hepatic clearance.
Viral kinetics can predict early response to alpha‐interferon in chronic hepatitis C
Authors: Walsh K. M. , Good T. , Cameron S. , Thorburn D. , McCruden E. A. B. , Mills P. R. , Morris A. J.
Published: August 2010

ABSTRACT— Aims/Background: Response rates to alpha‐interferon therapy for chronic hepatitis C are poor. An early indication of efficacy would reduce the need for prolonged therapy, leading to significant cost savings. It was established that a change in quantitative hepatitis C virus RNA (HCV‐RNA) titre at 4 weeks could predict the outcome of alpha‐interferon therapy in chronic hepatitis C. Methods: Serum HCV‐RNA titres were quantified using branched chain DNA (bDNA) assay in 26 patients who responded to alpha‐interferon (serum HCV‐RNA negative after 12 weeks therapy) and 11 age and sex matched non‐responders. Quantitative bDNA and qualitative RT‐PCR assays for HCV‐RNA were measured pre‐treatment and at 4 weeks. The change in quantitative HCV‐RNA titre between pre‐therapy and after 4 weeks was compared in the two groups. Results: Seventeen of the 37 patients had become RT‐PCR negative at 4 weeks (early responders) and had an undetectable HCV‐RNA titre on bDNA assay. Nine patients were RT‐PCR positive at 4 weeks but negative by 12 weeks (delayed responders), and of these, 8 had an undetectable viral titre at 4 weeks on bDNA assay. The patient with a detectable HCV titre did become RT‐PCR negative after 12 weeks, but subsequently became RT‐PCR positive again at 24 weeks. All the non‐responders had a detectable bDNA titre (>0.2 Meq/ml) at 4 weeks. Conclusions: Change in quantitative HCV‐RNA titre measured by bDNA assay at 4 weeks predicts response to alpha interferon. If HCV‐RNA remains detectable on bDNA assay at 4 weeks, no sustained response to treatment is found and alpha‐interferon can be discontinued.