bluuebunny/crossref_metadata_2025 huggingface dataset:
Depressed monocyte production of interleukin‐1 and tumor necrosis factor‐alpha in patients with alcoholic liver cirrhosis
Authors: Müzes GY. , Deák GY. , Láng I. , González‐Cabello R. , Gergely P. , Fehér J.
Published: August 2010

ABSTRACT— Interleukin‐1 and tumor necrosis factor‐alpha activity by E. coli lipopolysaccharide‐triggered monocytes were studied in patients with chronic alcoholic liver disease. Monocytes from cirrhotic patients were shown to have a significant reduction in IL‐1 and TNF‐a activity, compared with that from age‐ and sex‐matched healthy controls. These findings indicate further immunoregulatory disturbances concerning alcoholic liver cirrhosis.
Ultrastructural localization of Pre‐S2 polypeptides in the liver tissues of patients with chronic hepatitis B virus infection
Authors: Saito Takafumi , Kamimura Tomoteru , Asakura Hitoshi , Ishikawa Makoto
Published: August 2010

ABSTRACT—Pre‐S2 polypeptides in the liver tissue were investigated by immunoperoxidase staining in 26 patients with chronic hepatitis B virus (HBV) infection positive for HBeAg. Pre‐S2 polypeptides were detected in the liver of 25 patients, in 17 of whom Pre‐S2 polypeptides were localized both on the hepatocyte membrane and in the cytoplasm, and in eight of whom only in the cytoplasm. The localization pattern of Pre‐S2 polypeptides was not correlated with the histological findings but with the replicative status of HBV. In cases with a high level of DNA‐polymerase in the serum or with both nuclear and cytoplasmic expression of HBcAg in the liver, Pre‐S2 polypeptides were more frequently expressed both on the hepatocyte membrane and in the cytoplasm (P < 0.05). Membranous expression of Pre‐S2 polypeptides was speculated to be linked to active replication of HBV in the hepatocytes. Under immune electron microscopy, Pre‐S2 polypeptides were observed on the plasma membrane, membranes and cisternae of endoplasmic reticulum (ER), or perinuclear space. Moreover, Pre‐S2 polypeptides were detected on the tubular structures and the intracisternal particles about 40 nm in diameter considered to represent HBV. These findings suggest that the immunoreactions of Pre‐S2 polypeptides in the liver tissue are similar to those of HBsAg, indicating that Pre‐S2 polypeptides play an important role in the immunocharacteristics of HBsAg.
A cytokeratin‐immunohistochemical study of focal nodular hyperplasia of the liver: further evidence that ductular metaplasia of hepatocytes contributes to ductular “proliferation”
Authors: Van Eyken Peter , Sciot Raf , Callea Franscesco , Desmet Valeer J.
Published: August 2010

ABSTRACT— A cytokeratin‐immunohistochemical study was performed on eight specimens of focal nodular hyperplasia of the liver in order to determine whether hepatocytes in this lesion can express “bile duct type” cytokeratins. Serially cut cryostat sections were reacted with a panel of six monoclonal antibodies specifically reactive with cytokeratin polypeptides nr. 7, 8, 18 and 19, using a 3‐step immunoperoxidase procedure. Hepatocytes were positive for cytokeratins nr. 8 and nr. 18, whereas ductules contained in addition to cytokeratins nr. 8 and nr. 18 also polypeptides nr. 7 and nr. 19. In all cases, a variable number of hepatocytes close to fibrous septa or inside the nodules expressed cytokeratin nr. 7. In four cases, a small number of hepatocytes were also immunoreactive for cytokeratin nr. 19. Our data demonstrate that hepatocytes in focal nodular hyperplasia can express one or even two “bile duct type” cytokeratins, supporting the concept that ductular metaplasia of hepatocytes contributes to the ductular “proliferation” observed in this tumor‐like lesion.
Effect of parathyroid hormone on portal pressure in portal hypertensive rats
Authors: Yang May C. M. , Pang Peter K. T. , Lay C. S. , Wu S. L. , Jan K. M. , Tsai Y. T. , Kuo J. S.
Published: August 2010

ABSTRACT—Conflicting results have been common in the pharmacological treatments of portal hypertension. In an attempt to seek better management of portal hypertension, we studied the effect of the synthetic parathyroid hormone (PTH) fragment, [bPTH‐(1–34)], in portal hypertensive rats (partial portal vein ligation). PTH, 10 U/kg, administered via the jugular vein resulted in a reduction of both mean arterial blood pressure (MAP) and portal pressure (PP) to a similar extent (18.9% and 16.9%, respectively). A higher dose (40 U/kg) of PTH lowered the PP by 27.8% and MAP by 43.2%. Hemodynamic experiments, performed with labelled microspheres, demonstrated that PTH decreased the blood flow of the splanchnic and hepatic portal collateral vascular beds. To determine whether there is a direct vasodilatory effect on the venous vasculature, the effect of PTH on the isolated portal vein was examined. PTH was capable of inhibiting both spontaneous and drug (methacholine 10‐7mol/l or KC1 40 mmol/l‐induced contraction in a dose‐dependent manner. Therefore, it can be assumed that some of the effect of PTH on portal pressure is due to a selective effect on the portal vein.
Spontaneous reactivation of hepatitis B virus infection in patients with chronic type B hepatitis
Authors: Fattovich Giovanna , Brollo Lucio , Alberti Alfredo , Realdi Giuseppe , Pontisso Patrizia , Giustina Giuliano , Ruol Arturo
Published: August 2010

ABSTRACT— Eighty‐eight consecutive hepatitis B surface antigen (HBsAg) and hepatitis B e antigen (HBeAg) positive, heterosexual patients of Caucasian origin presenting with chronic hepatitis were followed for 1 to 15 years (mean, 5.4 years). During the study period, 45 (51%) patients cleared HBeAg and hepatitis B virus‐deoxyribonucleic acid from serum and were followed for 53 ± 29 months (mean ± SD) after seroconversion to antibody to hepatitis B e antigen. All patients manifested biochemical improvement. During follow‐up, 10 (22%) of the 45 patients experienced spontaneous reactivation of hepatitis B replication with reappearance of serum hepatitis B virus‐deoxyribonucleic acid and, in 4 patients, of hepatitis B e antigen. All patients then showed biochemical exacerbation of disease. These serologic events were transient, lasting an average of 12 months, in 8 (80%) patients. All patients were asymptomatic or minimally symptomatic. Histologic findings of liver tissue from 7 patients showed progression from chronic active hepatitis to active cirrhosis in 2 (28%) patients, while in the remaining 6 cases histology remained unchanged or improved from chronic active to chronic persistent hepatitis. These data indicate that spontaneous reactivation of hepatitis B infection occurs in heterosexual patients with chronic hepatitis B and this event is usually transient and asymptomatic, although in some patients it may be the major cause of progressive hepatic damage.
Pathomorphologic study on the extrahepatic portal vein in idiopathic portal hypertension
Authors: Kage Masayoshi , Arakawa Masahiro , Fukuda Kazunori , Kojiro Masamichi
Published: August 2010

Abstract— Patients with idiopathic portal hypertension (IPH) are known to have sclerotic changes of the intrahepatic portal vein radicles. In order to elucidate the pathological changes in the extrahepatic portal venous system in IPH, studies were carried out on the portal trunk in 12 patients with IPH, 59 patients with liver cirrhosis including some with associated hepatocellular carcinoma, and 12 normal matched control subjects. Histological examinations including histomorphometry were performed on the transverse sections of the portal trunk taken at autopsy. Most of the patients with IPH showed severe phlebosclerosis which was more pronounced than seen in liver cirrhosis. Thrombosis was also frequently observed in IPH. In IPH, the portal trunk was characterized by fibrous thickening of the intima and media with a prominent increase of elastic fibers. The mean area and thickness of the intima and media were significantly greater than in patients with liver cirrhosis. Sclerosis extensively involving both the extrahepatic and intrahepatic ramifications of the portal vein appeared to be characteristic of IPH.
Liver damage in juvenile inflammatory bowel disease
Authors: Nemeth Antal , Ejderhamn Jan , Glaumann Hans , Strandvik Birgitta
Published: August 2010

Abstract— During a 12‐year period, 46 children and adolescents with inflammatory bowel disease were followed from the time of diagnosis with regular biochemical tests of liver function. Thirty‐four patients had ulcerative colitis and 12 had Crohn's disease. Mean age at the time of diagnosis was 10.2 years (range 7 months–17 years) and the mean follow‐up period was 5.2 years (range 1–11 years). Pathological liver function tests were found in 60% of the 34 patients with ulcerative colitis: 9 of these 20 patients demonstrated more severe disturbance, usually at the time of diagnosis. Liver damage was most frequent in patients with total colitis. Liver biopsy was performed in eight patients, demonstrating “pericholangitis”, fibrosis and in one case cirrhosis. Morphometry of electron microscopical pictures revealed a significantly increased number of lysosomes and dilated cisternae of the rough endoplasmic reticulum. ERCP was performed in two patients, verifying primary sclerosing cholangitis in one. Four of the 12 patients with Crohn's disease had mildly pathological liver function tests. No correlation was found to the extent, duration or treatment of bowel disease. In our series of juvenile inflammatory bowel disease, liver damage occurred frequently, especially in ulcerative colitis. The more severe changes tended to coincide with the onset of bowel disease.
Integrity of the cytoskeletal system is important for phagocytosis by Kupffer cells
Authors: Watanabe Sumio , Hirose Miyoko , Ueno Takashi , Kominami Eiki , Namihisa Toshihiko
Published: August 2010

Abstract— The mechanism of phagocytosis by Kupffer cells is believed to involve the Ca2+‐calmodulin system. However, the role of myosin in this system is still unknown. In this study, we found that myosin light chain kinase inhibitor (ML‐9) inhibited phagocytosis by cultured Kupffer cells using polystyrene beads, a time‐lapse VTR system and fluorescent staining techniques. The inhibitory effects of ML‐9 suggest that myosin may be involved in this complex cellular function and that the integrity of the cytoskeletal system is essential for normal phagocytosis.
The metabolism of recombinant erythropoietin in the isolated perfused rat liver
Authors: NIELSEN OVE JUUL , EGFJORD MARTIN , HIRTH PETER
Published: August 2010

ABSTRACT— Indirect evidence points to extrarenal organs, presumably the liver, as the site of degradation of erythropoietin (EPO). The metabolism of both fully glycosylated and desialated intrinsically labelled 35S‐Cysteine recombinant human erythropoietin (rhEPO) was therefore studied in isolated Wistar rat livers perfused in a recirculating mode for 180 min with a hemoglobin‐free medium containing rhEPO. Perfusate and bile levels of rhEPO were measured by RIA. Total 35S‐radioactivity in liver, bile and perfusate as well as non‐acid precipitable radioactivity in perfusate were determined. In addition, detection of 35S‐radioactivity was performed after subcellular fractionation of rat livers perfused with desialo‐35S‐Cysteine rhEPO. While concentrations of fully glycosylated 35S‐Cysteine rhEPO did not exhibit any detectable decrease during perfusion, desialo‐35S‐Cysteine rhEPO was rapidly cleared from the perfusate. After 60 min of perfusion, only 32% of the initial levels of both immunoreactive rhEPO and total radioactivity remained in the perfusate. Quantitative hepatic accumulation of desialated tracer was demonstrated. Subcellular fractionation showed extensive hepatic degradation of the desialated tracer. Furthermore, during perfusion progressively larger amounts of small molecular weight degradation products of the tracer were found in the perfusate. Bile excretion of both fully glycosylated and desialated tracer was negligible. The significance of hepatic metabolism of desialo‐35S‐Cysteine rhEPO was supported by reduced removal of desialo‐35S‐Cysteine rhEPO from plasma in hepatectomized rats. It is hypothesized that continuous in vivo desialation is a crucial rate‐limiting step in the degradation of circulating EPO.
Chronic active hepatitis B. Interferon‐activated natural killer‐like cells against a hepatoma cell line transfected with the hepatitis B virus nucleic acid
Authors: Actis Giovanni C. , Ponzetto Antonio , D'Urso Nicoletta , Saracco Giorgio , Crepaldi Tiziana , Bellone Graziella , Catucci Luigia Marilena , Zarcone Daniela , Verme Giorgio
Published: August 2010

ABSTRACT— In a rapid 51Cr release assay, peripheral blood mononuclear cells from 12 healthy donors did not lyse the hepatitis B virus deoxyribonucleic‐acid‐transfected human hepatoma cell line 2.2.15, but under the same experimental conditions they did lyse K562 cells. Peripheral blood mononuclear cells from 10 out of 16 patients with chronic active hepatitis B exhibited cytotoxic activity against 2.2.15 cells in the presence of a relatively reduced natural killer cell activity to the K562 cell target. Enhancement of the cytotoxic activity to 2.2.15 cells was statistically significant in the group of patients being treated with leukocyte alpha‐interferon. The activity was not influenced by the degree of human leukocyte antigen type matching between effector and target, and was enhanced by depletion of T‐cells and by in vitro interferon treatment. These results therefore support the concept of a natural killer‐like cell activated by clinical administration of interferon in chronic active hepatitis B patients. This cell effector was lytic for the virus B negative HEP‐G2 cells also. However, T‐cells purified from a few patients failed to lyse the HEP‐G2 while lysing the 2.2.15 target, thus indicating that a preferential recognition of the virus‐infected target may be exerted by certain T‐lymphocyte subsets. The use of the human leukocyte antigen type defined, highly differentiated, hepatitis B virus releasing 2.2.15 cell line as target for fresh lymphocytes in this cytolytic assay did not disclose cytolytic T‐cells in an obvious way. Further manipulation of this system perhaps using T‐cell clones may be the next step to exploit the investigative possibilities offered by the availability of the 2.2.15 cell target.
Long‐term follow‐up of chronic hepatitis non‐A, non‐B – with special reference to hepatitis C
Authors: Wejtål R. , Hermodsson S. , Norkrans G.
Published: August 2010

ABSTRACT— One hundred and twenty‐seven patients with histologically verified chronic non‐A, non‐B hepatitis were followed for up to 23 years (mean 6.3 years). Thirty‐nine were infected by blood transfusion, 58 were drug‐addicts and 30 had no obvious source of infection. Chronic persistent hepatitis (CPH) was diagnosed in 84 (66%), while 43 patients (34%) had chronic active hepatitis (CAH) with or without cirrhosis. Patients with CPH were significantly younger (29.7 years vs 46.8 years; p<0.01), irrespective of the type of virus exposure. Antibodies to hepatitis C virus (anti‐HCV) were detectable in 91 patients (72%) and 36 (28%) were anti‐HCV negative. Fifteen patients with acute onset, and negative for anti‐HCV at the start, became positive during follow‐up; 12 of them within 4.5 months. We found no differences among anti‐HCV positive and anti‐HCV negative patients in liver function tests, resolving rate, morphological progression in serial biopsies or mortality rate. Five previously anti‐HCV positive patients became negative during follow‐up and in two of them this was accompanied by decreasing hepatic inflammation.
The effect of taurine on the cholestatic potential of sulfated lithocholate and its conjugates
Authors: Belli Dominique C. , Roy Claude C. , Fournier Lyse‐Andrée , Tuchweber Beatriz , Giguère Robert , Yousef Ibrahim M.
Published: August 2010

ABSTRACT— The present study was aimed at determining whether the protection by taurine of lithocholate‐sulfate‐induced cholestasis is mediated by conjugation or by direct effect of the amino acid on bile formation. Injection of free and conjugated (glycine and taurine) sulfated lithocholate in guinea pigs significantly reduced the secretion rate of non‐sulfated bile acids in bile. There was no decrease in bile flow after the injection of taurine‐conjugated sulfated lithocholate, which was completely recovered in bile within 60 min. In contrast, injection of sulfated lithocholate and its glycine conjugate led to a marked decrease in bile flow, and neither one was significantly recovered in bile. In addition, both caused morphological changes in the liver, characterized by the accumulation of cytoplasmic vacuoles with lamellated myelin figures characteristic of phospholipidosis. Pretreatment with taurine (0.5% in drinking water for 3 days) prevented both the drop in bile flow and the histological changes in the liver, suggesting that conjugation with taurine removed the cholestatic potential of sulfated lithocholate. However, since taurine was effective not only in preventing cholestasis induced by the free form of sulfated lithocholate but also against its glycine conjugate, these results suggest that other mechanisms in addition to conjugate must be involved.
Lipid‐laden perisinusoidal cells in patients with acquired immunodeficiency syndrome
Authors: Dupon Michel , Kosaifi Tanios , Bail Brigitte Le , Lacut Yves , Balabaud Charles , Bioulac‐Sage Paulette
Published: August 2010

ABSTRACT— Liver biopsies were taken from ten AIDS patients. Liver architecture was normal in all patients. On 1‐μm‐thick sections stained with toluidine blue, all ten cases showed lipid overload of perisinusoidal cells (1 massive, 5 moderate and 4 mild) compared to 2/8 in control patients, who had mild lipid overload. Other sinusoidal abnormalities such as hypertrophy of Kupffer cells and inclusions in endothelial cells were also noticed. Some hepatocytes presented evidence of cellular damage. Perisinusoidal cell lipid overload was not associated with hypervitaminosis A. We hypothesize that the abnormal accumulation of lipids in perisinusoidal cells (non‐induced by hypervitaminosis A) in patients with AIDS could be due to defective transport of vitamin A from perisinusoidal cells to hepatocytes, and/or from hepatocytes to blood. The cause of the defect is unknown. Since lipid overload occurs in many and diverse conditions (diabetes, cholestasis, primary biliary cirrhosis, etc.), it seems reasonable to propose that the defect is non‐specific and limited to functional or structural damage of the liver whether induced by drugs, liver or systemic diseases.
Enzyme‐histochemical studies of griseofulvin‐intoxicated mouse livers
Authors: Woltsche Michael , Zatloukal Kurt , Denk Helmut
Published: August 2010

ABSTRACT— Enzyme‐histochemical studies were conducted on livers of mice chronically fed griseofulvin (GF) in order to produce Mallory bodies (MBs) in hepatocytes. The development of MBs is associated with derangement of the immunohistochemically detectable intermediate filament (IF) cytoskeleton of the cytokeratin (CK) type, although no strict correlation between appearance or involution of MBs and the cytoskeletal alterations exists. Since the function of the IF cytoskeleton and the relationship of its disturbance to cell injury is unknown, the aim of the present study was to correlate the activities of several key enzymes of cellular metabolic pathways with the disturbance of the cytoskeleton architecture. For that purpose enzyme‐histochemistry in combination with immunohistochemical CK‐IF stainings were performed on identical sections. In GF‐intoxicated mouse livers the normal topography of enzyme activities was disturbed, but no strict colocalization of enzymatic and cytoskeletal changes was found. Glucose‐6‐phosphatase, a microsomal enzyme involved in glucose output and gluconeogenesis, showed elevated activity in MB‐free hepatocytes with diminished immunostainable CK‐IF cytoskeleton refuting the concept of a disability of those cells to export glucose. It could indeed indicate that those cells without MBs are in the state of recovery. However, these cells could also resemble “hyperactive foci“. Glycogen was decreased in MB‐containing hepatocytes with disturbed cytoskeleton, and this feature favours the assumption of cell degeneration. On the other hand, the mitochondrial marker enzymes, i.e. succinate dehydrogenase, cytochrome‐c‐oxidase and 3‐hydroxybutyrate dehydrogenase, remained unchanged in altered hepatocytes. Alkaline phosphatase activity at the canalicular pole of GF‐intoxicated hepatocytes was elevated, indicating cholestatic features associated with this disorder. However, since altered hepatocytes did not show impairment of oxido‐reductase activities, a severe impairment of bile secretion as a consequence of cell damage is unlikely. Unchanged or even increased ATPase activity of altered hepatocytes also indicated their sustained metabolic abilities. The results presented provide indirect evidence that hepatocytes with disturbed IF cytoskeleton do not significantly differ from normal cells with respect to oxidative metabolism, fatty acid synthesis and gluconeogenesis. This suggests that alterations of the IF cytoskeleton associated with GF intoxication and MB formation have no significant adverse influence on the metabolic functions of liver cells, as far as can be assessed by evaluation by enzyme‐histochemical staining of several key enzymes.
Incidence of spontaneous bacterial peritonitis in patients with ascites. Diagnostic value of white blood cell count and pH measurement in ascitic fluid
Authors: Storgaard John Skaaning , Svendsen Jesper Hastrup , Hegnhøj Jens , Krintel Jens Jacob , Nielsen Peder Bo
Published: August 2010

ABSTRACT—During a 21‐month period, 65 consecutive patients admitted with ascites were included in a prospective study of the incidence of spontaneous bacterial peritonitis, and paracentesis was performed on admission. The ascitic fluid was cultured, ascitic leucocytes were counted and pH was measured. Bacterial growth was found in five patients with chronic liver disease, who were diagnosed as having spontaneous bacterial peritonitis (SBP), since no intra‐abdominal focus could be demonstrated. Thus, the incidence of SBP in this material was 7.7% (95% confidence limits: 2.5–17%). SBP was caused byEscherichia coli(n = 3), coagulase negative staphylococcus (n = 1), andBacteroidesspecies (n = 1). Abdominal tenderness, abnormal intestinal sounds, fever and hepatic encephalopathy were equally frequent in the group with SBP and in patients with sterile ascites. Infection was not anticipated in any of the patients with SBP. In contrast to several previous studies, neither ascites pH nor ascites leucocyte counts were any help in obtaining a rapid diagnosis. Survival time of patients with SBP was significantly shorter than of patients without SBP.
Liver pathology associated with the use of anabolic‐androgenic steroids
Authors: See Katrine Lydolph , See Morten , Gluud Christian
Published: August 2010

Abstract: This review examines the liver‐damaging side effects of anabolic‐androgenic steroids (AAS). It seems that AAS can cause development of peliosis hepatis, subcellular changes of hepatocytes, hepatocellular hyperplasia and hepatocellular adenomas. On the other hand, it has not been convincingly proved that AAS can cause development of hepatocellular carcinomas when used in the usual therapeutic doses. Tumours reported as hepatocellular carcinomas caused by AAS seem to be hyperplastic lesions of a benign nature able to regress with withdrawal of the putative agent. The effects of untraditional combinations and high‐dose AAS are not yet known, leaving the possibility of a carcinogenic effect in those cases.
Glutathione treatment of hepatocellular carcinoma
Authors: Dalhoff Kim , Ranek Leo , Mantoni Margit , Poulsen Henrik E.
Published: August 2010

ABSTRACT— This prospective study was undertaken to substantiate observations that glutathione (GSH) inhibits or reverses tumor growth in humans with hepatocellular carcinoma (HCC), a neoplasm with an extremely poor prognosis. Eight patients with biopsy‐proven HCC not amenable to surgery were given 5 g of GSH daily from the time of diagnosis. Two patients withdrew shortly after receiving GSH due to intolerable side‐effects. Of the six eligible patients, two had mildly advanced tumors and four moderately advanced tumors. At 1–2‐month intervals the liver was CT and ultra‐sound scanned to assess the growth status of the tumor (progression, stagnation or regression). All the patients, except a male with a fibrolamellar type of HCC, died within 1 year after diagnosis. Two women with moderately advanced tumors survived almost 1 year, tumor growth stopped or regressed and in one of the women an initially abnormal alfa‐1‐fetoprotein (AFP) returned to normal after GSH treatment. AFP remained normal throughout the treatment period in the other woman. These observations indicate that GSH may have a sex‐dependent effect on HCC. However, further studies involving more patients are required to pursue this hypothesis.
Immunoquantification of cytochrome P‐450 3A on rat paraffin‐embedded liver tissue
Authors: Horsmans Yves , Stevens Martine , Geubel André , Harvengt Carl , Rahier Jacques
Published: August 2010

ABSTRACT—The cytochrome P‐450 3A family is involved in the metabolism of several drugs, including nifedipine, cyclosporine, quinidine and erythromycin. The purpose of this study was to develop a reliable method to obtain a relative quantification of cytochrome P‐450 3A apoproteins in rat liver specimens by immunocytochemistry and to correlate such quantification to erythromycin N‐demethylase activity, a biochemical pathway sustained by that enzymatic system. Thirty‐six male Wistar rats were treated with an injection of either saline or dexamethasone phosphate (10, 30 or 50 mg/kg), a potent inducer of cytochrome P‐450 3A. Specimens taken from the same lobe were processed for immunocytochemistry and determination of erythromycin N‐demethylase activity. Paraffin sections were treated with a polyclonal antiserum directed against cytochrome P‐450 3A. The density of cytochrome P‐450 3A immunostaining measured by an automatic image analyzer increased with the dose of dexamethasone pretreatment, and with the erythromycin N‐demethylase activity, both parameters being closely correlated. Our data indicate that in rats, when cytochrome P‐450 3A is concerned, there is a close correlation between the results of immunoquantitation and biochemical activity. This suggests that such a method of investigation might be used on small paraffin‐embedded liver specimens obtained by needle biopsy.
In situ hybridization: optimization of the techniques for collecting and fixing the specimens
Authors: Biffo Stefane
Published: August 2010

Abstract: An overview is presented concerning the different protocols for collecting and fixing the specimens for in situ hybridization analysis. Particular attention is devoted to the problem of avoiding the tissue RNA degradation due to the ubiquitous and rather resistant RNases. The choice of the final protocol largely depends on a compromise between the need to maintain morphological details, the care to avoid loss of the target nucleic acids and the capability of the probe to diffuse within the specimen.
Autoradiographic detection of radiolabelled probes by in situ hybridizaiton
Authors: Gowans Eric J.
Published: August 2010

Abstract: In the context of in situ hybridization techniques, autoradiography is used to localize radiolabelled probes within intact tissue sections or cell preparations. The present report describes in detail a standardized autoradiographic procedure. Topics include the choice of the emulsion, its preparation and layering onto the slides, the development procedure and the analysis of the results. Special attention is devoted to the practical aspects, so that the technique will be generally applicable, according to the reader's needs.
Non‐readioactive nucleiec acid probes: labelling and detection procedures
Authors: Vale Gluseppe , Dell'Orto Patrizia
Published: August 2010

Abstract: A comprehensive overview is given concerning the in situ hybridizaiton tecnhiques using non‐radioactively labelled probes. Details are given on the preparation of the probes with different reporter moelecules and on the corresponding revealing assays. The development of non‐isotopic labeeling methods for nucleic acid probes has facilitated the diffusion of the in situ hybridizaiton techniques to the research as well as to the clinical pathology laboratory. Non‐radioactive probes have serveral advantages over the radiolabelled ones, including cost, shelf‐life, safety and, last but not least. resolutions of the signal.
Intercellular adhesion molecule‐1 expression on the hepatocyte membrane of patients with chronic hepatitis B and C
Authors: Horiike Norio , Onji Morikazu , Kumon Izumi , Kanaoka Mitsuo , Michitaka Kojiro , Ohta Yasuyuki
Published: August 2010

ABSTRACT— The expression of intercellular adhesion molecule‐1 (ICAM‐1) on the hepatocyte membrane was studied in 27 patients with chronic hepatitis B and C (CHB, CHC) by immunostaining using a monoclonal antibody. ICAM‐1 was expressed focally in a honeycomb‐like pattern by hepatocytes in livers of 26/27 patients. The degree of ICAM‐1 expression was closely related to the ALT level and the histological grade of liver damage. Abundant cytotoxic T cells (CD8 +, CD11b –) were found in ICAM‐1‐positive areas of the liver. Zones of focal necrosis contained both ICAM‐1‐positive hepatocytes and cytotoxic T cells. The expression of ICAM‐1 was decreased in 4/6 CHB patients after interferon‐α therapy. No relationship between the degree of hepatocyte ICAM‐1 expression and viral replication markers (DNA polymerase activity and the presence of HBcAg in the liver) was observed in patients with CHB. In addition, no positive correlation was found between the distribution of ICAM‐1‐positive hepatocytes and HBcAg‐positive hepatocytes. These results suggest that ICAM‐1 may play an important role in the pathogenesis of hepatocellular injury mediated by cytotoxic T cells in CHB and CHC.
Treatment of decompensated viral hepatitis B‐induced cirrhosis with low doses of interferon alpha
Authors: Nevens F. , Goubau P. , Van Eyken P. , Desmyter J. , Desmet V. , Fevery J.
Published: August 2010

ABSTRACT—Seven patients with decompensated posthepatitis B cirrhosis were treated with low doses of interferon alpha. The initial plasma level of HBV‐DNA ranged from 3.0 to 189.3 pg/ml, and that of ALT from 37 to 156 IU/1. Liver biopsies demonstrated ongoing piecemeal necrosis. In sera of all but one patient, HBV‐DNA became undetectable by hybridisation techniques within 10 to 28 weeks. Plasma HBeAg became negative in four and HBe‐antibodies positive in three patients. Serum transaminase levels showed a marked initial rise 3 to 13 weeks after onset of therapy; they dropped to normal values later in all except one patient. Therapy was initiated at 1 MU (million units) three times a week for 2 weeks and was increased to 2.5 MU for 16 weeks. Later, this dosage was raised to 5 MU three times a week in some patients. Complications included variceal haemorrhage, aggravation of ascites or of encephalopathy, development of pneumonia, recurrence of spontaneous bacterial peritonitis or of gastric ulcer bleeding. One year after stopping the therapy, three patients are well and without any feature of liver decompensation. Three patients died before they could undergo a liver transplantation. In one patient treatment was interrupted because of marked exacerbation of liver cell necrosis. It thus seems possible to suppress HBeAg and HBV‐DNA in patients with decompensated cirrhosis. This is important to prepare them for possible liver transplantation. Interferon should be initiated at low doses and the patients be very carefully monitored. Prophylactic therapy for bacterial peritonitis and for variceal haemorrhage is warranted.
Effect of recombinant human transforming growth factor β1 on immune responses in patients with chronic hepatitis B
Authors: Kakumu Shinichi , Ito Yuji , Takayanag Masahiro , Yoshioka Kentaro , Wakita Takaji , Ishikawa Tetsuya , Higashi Yasuyuki , Yang Zhi‐Qi
Published: August 2010

ABSTRACT— Studies were undertaken to examine the effect of recombinant human transforming growth factor beta 1 (rTGF‐β1) on cellular and humoral immune responses of peripheral blood mononuclear cells (PBMC) from patients with chronic hepatitis B. The addition of TGF‐β1 caused a significant dose‐dependent inhibition of hepatitis B (HB) core Ag‐stimulated interferon‐γ and antibody to HB core Ag production and proliferation of PBMC from chronic hepatitis patients and HB‐immune donors. TGF‐β1 also induced a significant reduction in pokeweed mitogen‐stimulated IgG and IgM production, as well as phytohemagglutinin p‐stimulated proliferative response of PBMC. The degree of inhibition of TGF‐β1 did not differ between antigen‐specific and ‐nonspecific cellular and humoral immune responses, and between control individuals and patients. Pretreatment study with TGF‐β1 showed that the activities of T cells, B cells and monocytes were similarly inhibited. Further, TGF‐β1 inhibited activities of HLA class I antigen‐matched cytotoxic T cells from patients with chronic hepatitis B for HBV DNA‐transfected HepG2 cells in a 51Cr release assay. The results suggest that TGF‐β1 may play a role in the regulation of antigen‐dependent and ‐independent immune responses in patients with chronic hepatitis B.
Effects of interferon‐α treatment on hepatitis B virus antigen‐specific immunologic responses in patients with chronic hepatitis B
Authors: Ishikawa Tetsuya , Kakumu Shinichi , Yoshioka Kentaro , Kurokawa Susumu , Kusakabe Atsuhiko , Tahara Hirofumi , Hirofuji Hideo , Kawabe Masami
Published: August 2010

ABSTRACT— Studies were undertaken to evaluate the relationship between the immune responses and the effectiveness of interferon‐α treatment in 21 patients with HBeAg‐positive chronic active hepatitis. Peripheral blood mononuclear cells (PBMC), obtained on four occasions during an 8‐week course of IFN‐α therapy, were cultured with recombinant HBcAg, purified HBeAg or pokeweed mitogen (PWM). During follow‐up for 6 months after therapy, clearance of serum HBeAg was observed in eight patients designated as responders. Immunological responses of PBMC obtained before treatment did not differ between responders and non‐responders. In responders, IFN‐γ and anti‐HBc production was depressed during therapy, but recovered to above the pretreatment level at the end of and/or after cessation of therapy, while lymphocyte proliferation was enhanced during therapy with a subsequent decline to baseline value. In non‐responders, such changes were modest throughout the study, and anti‐HBc response remained decreased even after cessation of therapy. These results indicate that PBMC of responders have immunologically different responses to IFN‐α therapy when compared with non‐responders.
Expression of transforming growth factor alpha in human hepatocellular carcinoma
Authors: Collier J. D. , Guo K. , Gullick W. J. , Bassendine M. F. , Burt A. D.
Published: August 2010

ABSTRACT—Transforming growth factor alpha (TGF‐α) is a mitogenic polypeptide which acts on the epidermal growth factor receptor (EGFR). The aim of this study was to examine the expression of TGF‐α in human hepatocellular carcinoma (HCC) and surrounding cirrhotic tissue, and to compare it with normal liver. Immunoreactive TGF‐α was detected using two antibodies raised against its C terminus, a polyclonal antibody 26T and a monoclonal antibody Ab‐2. In normal liver immunoreactive TGF‐α was localised strongly to bile duct epithelium and weakly in occasional parenchymal cells but was notably absent from perisinusoidal and Kupffer cells. Eight out of twenty‐eight (28%) cases of HCC expressed TGF‐α as demonstrated by cytoplasmic staining with both antibodies and in four cases additional membrane immunoreactivity was demonstrated using 26T. However, where cirrhotic tissue surrounding TGF‐α positive tumours was available for analysis immunoreactive TGF‐α was detected in only 1/7 cases. TGF‐α synthesis by malignant hepatocytes was supported by the detection of specific RNA by Northern blotting from two cases with TGF‐α immunoreactivity. These results implicate bile duct epithelium as an important source of TGF‐α in human liver. Furthermore, in HCC the expression of TGF‐α in some cases, together with paucity of TGF‐α immunoreactivity in surrounding cirrhotic tissue, suggests that TGF‐α may play a role in continued cell proliferation in human hepatocarcinogenesis.
Total therapeutic paracentesis (TTP) with and without intravenous albumin in the treatment of cirrhotic tense ascites: a randomized controlled trial
Authors: Garcia‐Compeán Diego , Villarreal Jesüs Zacarias , Cuevas Horacio Bahena , Cantü Dora Alicia Garcia , Estrella Miguel , Tamez Eduardo Garza , Castillo Ricardo Valadez , Barragán Rodrigo F.
Published: August 2010

ABSTRACT— We studied 35 cirrhotic patients with tense ascites assigned at random into two groups: Group I consisted of 17 patients treated by total therapeutic paracentesis (TTP) (6–15 1) plus i.v. albumin (5 g/l of fluid) and Group II consisted of 18 patients treated by TTP (5.5–15.5 1) without albumin. On 19 patients we performed a sequential assessment of cardiac output (CO), plasma renin activity (PRA) and plasma aldosterone (PA). Both groups were similar in age, sex, and etiology of cirrhosis. CO, PRA and PA values were expressed as mean changes occurring in relation to their respective baseline values. CO changes after TTP (1/min): Group I: 2.5 after 6 h and 2.2 after 12 h; Group II: 2.2 after 6 h and –0.4 after 12 h, (p<0.05 comparing values after 12 h between the two groups). PRA changes after TTP (ng/dl/h): Group I: –7.4 after 1 h, –7.8 after 6 h and –3.2 after 24 h; Group II: –2.4 after 1 h, –0.8 after 6 h and 3.9 after 24 h (p<0.05 comparing values between both groups after 6 and 24 h). PA changes after TTP (ng/dl): Group I: –50.5 after 1 h, –36.7 after 6 h and –34.6 after 24 h; Group II: –18.2 after 1 h, –2.2 after 6 h and 20 after 24 h, (p < 0.05 comparing values between both groups after 1 and 6 h). Complications were minimal in both groups. In conclusion, there was an increase of CO and a decrease of PRA and PA in all patients early after TTP. However, after 6 h there was a decrease of CO and an increase of PRA and PA, suggesting hypovolemia, only in patients without albumin. The frequency of complications after 24 h was similar in both groups. Since we do not exclude the possibility of complications over a longer period of time, we recommend the use of plasma expanders until their impact on morbidity and mortality in the long term is well established. The best time to infuse them is about 6 h after the procedure.
Outcome of acute symptomatic non‐A, non‐B hepatitis: a 13‐year follow‐up study of hepatitis C virus markers
Authors: Mattsson Lars , Sönnerborg Anders , Weiland Ola
Published: August 2010

ABSTRACT— Thirty‐nine of 61 prospectively followed patients who had had acute non‐A, non‐B hepatitis in 1978 were clinically reexamined in 1991 and tested for antibodies to hepatitis C virus (anti‐HCV) with a second generation ELISA and RIBA and for HCV RNA by PCR. Acute hepatitis C was diagnosed in stored sera from 1978 in 24 patients, who were found still to be anti‐HCV positive in 1991, and 16 of them were also HCV RNA positive. The majority of anti‐HCV positive patients with or without HCV RNA had elevated serum ALT levels 13 years after onset of their acute hepatitis C. After 13 years follow‐up, 1.6% of the patients had died of end‐stage liver disease, 8% of anti‐HCV positive patients had histologically confirmed liver cirrhosis, 79% of anti‐HCV positive patients were judged to have chronic infection, whereas 21% seemed to have recovered. To conclude, we found that a majority of our patients with acute symptomatic hepatitis C continued to be viraemic 13 years after onset of hepatitis C, and that all continued to be anti‐HCV positive by second‐generation ELISA.
Pregnancy in patients with primary biliary cirrhosis — a case for dissuasion?
Authors: Olsson Rolf , Lööf Lars , Wallerstedt Sven ,   The Swedish Internal Medicine Liver Club
Published: August 2010

ABSTRACT— The medical literature on pregnancy in primary biliary cirrhosis is restricted; it mostly dates from several decades ago and mostly reports a bad prognosis for the fetus as well as for the liver disease. We report experiences based on four pregnancies in three patients with primary biliary cirrhosis which are at variance with those reported earlier.
Encephalopathy from acute liver failure and from acute hyperammonemia in the rabbit. A clinical and biochemical study
Authors: de Knegt Robert J. , Groeneweg Michael , Schalm Solko W. , Hekking‐Weijma Ineke
Published: August 2010

Abstract:To study the molecular basis of ammonia toxicity, highly reproducible models of acute liver failure and acute hyperammonemia in the rabbit were developed. Acute liver failure was induced by two‐stage liver devascularization, and acute hyperammonemia by prolonged ammonia infusion such that the plasma ammonia pattern found in acute liver failure was simulated. Clinical symptoms, spectral analysis of the EEG, biochemistry (blood gasses, renal function, electrolytes and markers of hepatic injury) and the presence of cerebral edema were studied. During acute liver failure severe encephalopathy developed after 10.2 ± 1.9 h (n = 6, mean ± SEM). Other liver‐failure‐associated abnormalities were cerebral edema, lactic acidosis, renal dysfunction, hypothermia and septicemia. During acute hyperammonemia, severe encephalopathy developed after 18.2 ± 0.4 h (n = 6, mean ± SEM). Other abnormalities found were cerebral edema and lactic acidosis. In both animal models comparable EEG changes were observed (a decrease in mean dominant frequency and theta‐activity, and an increase in delta activity). However, these changes were not statistically significant, and non‐specific as they also occurred in control rabbits despite their clinical wellbeing. This study demonstrates in the rabbit the similarity between encephalopathy due to acute ischemic liver failure and that due to hyperammonemia. An observed difference in hyperammonemia‐induced encephalopathy was pronounced ataxia, which did not occur during acute liver failure, whereas hypothermia, sepsis and renal failure occurred exclusively in acute liver failure. Our models appear satisfactory for the study of hepatic encephalopathy and ammonia toxicity.
Phenotypic change in portal fibroblasts in biliary fibrosis
Authors: Tang Liang , Tanaka Yujiro , Marumo Fumiaki , Sato Chifumi
Published: August 2010

Portal fibroblasts have been considered responsible for biliary fibrosis. Since lipocytes show differentiation toward myofibroblast‐like cells in hepatic fibrogenesis, we studied whether similar differentiation of portal fibroblasts could be observed in biliary fibrosis. We examined rat livers after bile duct ligation by double immunofluorescent staining of α‐smooth‐muscle actin (α‐smA) and desmin and also by electronmicroscopy. In the portal tract of normal livers, α‐smA‐positive cells were noted only in the vessel wall, whereas desmin‐positive cells were occasionally seen in the connective tissue as well. With the development of biliary fibrosis, α‐smA was remarkably expressed in the portal connective tissue, while desmin was seen in a small portion of α‐smA‐positive cells around proliferating bile ducts. In normal livers, portal fibroblasts presented quiescent features, such as a small Golgi complex and a few cisternal profiles of endoplasmic reticulum under electron microscopy. After 7 days of bile duct ligation, portal fibroblasts proliferated, were arrayed in multilayers, and were associated with collagen bundles. Some of these fibroblasts had numerous cytoskeletal components, and developed rough endoplasmic reticulum and a dense body. These data suggest that portal fibroblasts appear to differentiate toward myofibroblasts in biliary fibrosis.
Serum tenascin levels in chronic liver disease
Authors: Yamauchi Masayoshi , Mizuhara Yuji , Maezawa Yoshihiko , Toda Gotaro
Published: August 2010

To evaluate the diagnostic significance of tenascin, the extracellular matrix glycoprotein in chronic liver disease, serum tenascin levels were measured by a newly developed ELISA in 21 patients with chronic persistent hepatitis, in 55 with chronic active hepatitis, in 59 with liver cirrhosis, in 31 with hepatocellular carcinoma, in 26 with acute hepatitis and in 66 healthy subjects. The serum tenascin level was significantly elevated in the patients with chronic active hepatitis, liver cirrhosis, hepatocellular carcinoma, and acute hepatitis when compared with the healthy subjects (P<0.001). The serum tenascin level also increased with increasing severity of chronic liver diseases. A significant correlation was observed between the serum tenascin levels and serum levels of various extracellular matrix proteins such as type III procollagen N‐aminoterminal peptide (PIIIP), laminin and the 7S domain of type IV collagen (P<0.001). A strong positive correlation was observed between the serum tenascin levels and histologic findings, particularly in the degree of hepatic fibrosis. This is the first report documenting serum tenascin level increases in patients with various chronic liver diseases. The measurement of the serum tenascin levels may provide additional information relevant to the study of connective tissue.
Endotoxin‐induced defenestration of the hepatic sinusoidal endothelium: a factor in the pathogenesis of cirrhosis?
Authors: Dobbs B. R. , Rogers G. W. T. , Xing H. Y. , Fraser R.
Published: August 2010

Defenestration and capillarisation of the hepatic sinusoidal endothelium occurs early in the pathogenesis of cirrhosis, both in patients suffering from alcohol abuse and in animal models. It it possible also that alcohol abuse promotes the absorption of bacterial endotoxins from the gastrointestinal tract. In this study we have investigated the effects of a single intravenous injection of endotoxin on the hepatic sinusoidal endothelium of rats. Seven days after the dose of endotoxin, the porosity of the sinusoidal endothelium was reduced to 40% of that of controls, due to a decrease in both diameter and number of fenestrae. The livers examined 14 days after dosing exhibited normal porosity. We postulate that bacterial endotoxins play a role in the pathogenesis of cirrhosis by modulating the fenestrated sinusoidal endothelium (liver sieve).
Oxidative stress in hepatocytes and stimulatory state of Kupffer cells after reperfusion differ between warm and cold ischemia in rats
Authors: Mochida Satoshi , Aral Masahiro , Ohno Akihiko , Masaki Naohiko , Ogata Itsuro , Fujiwara Kenji
Published: August 2010

Rat liver was kept at 4°C or 37°C in MEM, and reperfused through a closed circulation from the hepatic vein to the portal vein at 37°C with the same solution. Although purine nucleoside phosphorylase and ALT activities were increased in the perfusate, depending on the duration of ischemia at both 4°C and 37°C, the ratio of the latter to the former was significantly higher after 37°C‐ischemia than after 4°C‐ischemia. The stimulation stage of Kupffer cells evaluated in situ by formazan deposition after liver perfusion with nitro blue tetrazolium and phorbol myristate acetate was elevated after 4°C‐ischemia longer than 1 h, but not after 37°C‐ischemia. In contrast, the degree of oxidative stress in hepatocytes assessed by formazan deposition after liver perfusion with nitro blue tetrazolium alone was greater after 37°C‐ischemia than after 4°C‐ischemia. These results suggest that oxidative stress in hepatocytes and the stimulatory state of Kupffer cells after ischemia‐reperfusion may differ between 4°C‐ischemia and 37°C‐ischemia, probably leading to different development of liver damage.