bluuebunny/crossref_metadata_2025 huggingface dataset:
Expression of epidermal growth factor and fibroblast growth factor in human hepatocellular carcinoma: an immunohistochemical study
Authors: Motoo Yoshiharu , Sawabu Norio , Nakanuma Yasuni
Published: August 2010

ABSTRACT— Expression of epidermal growth factor (EGF) and fibroblast growth factor (FGF) was examined in 56 patients with hepatocellular carcinoma (HCC) using an immunohistochemical method. EGF and FGF were expressed on carcinoma cells in 14 (25%) and 23 cases (41%), respectively. In the 23 FGF‐positive cases, 11 cases were positive for both acidic and basic FGF, while 18 were positive for acidic FGF, and 16 were positive for basic FGF. In non‐cancerous hepatic tissues, FGF was weakly positive in macrophages, hepatocytes and vascular endothelial cells in some cases, while EGF was totally negative. There were no significant correlations between the expression of EGF or FGF on carcinoma cells and the various clinicopathologic factors examined. These data suggest that EGF and FGF are produced by human HCC cells in vivo. The roles of the expression of these growth factors in the development and progression of HCC remain only speculative.
Hepatic extraction of the aminoterminal propeptide of type III procollagen before and after bile duct ligation in pigs
Authors: Bentsen Kirsten D. , Henriksen Jens H. , Boesby Steen , Jensen Lars T. , Hørslev‐Petersen Kim , Lorenzen Ib
Published: August 2010

ABSTRACT— The aminoterminal propeptide of type III procollagen is extracted from the circulation by the liver, and PIIINP is found in bile. This study was performed in order to investigate whether biliary excretion contributes substantially to the hepatic extraction of circulating PIIINP. Hepatic extraction before and during a 4‐h period after ligation of the common bile duct was assessed from serum PIIINP concentrations in a systemic artery, the portal vein and a hepatic vein of seven healthy anaesthetized pigs. Seven sham‐operated anaesthetized pigs served as controls. Ligation of the bile duct did not cause a decrease in the hepatic extraction ratio of circulating PIIINP. The PIIINP serum levels of the cholestatic pigs and of the controls were similar throughout the investigation period. The PIIINP concentrations in bile were only 10% of the corresponding serum values. Gel filtration of sera showed that the lower PIIINP concentration in the hepatic vein, as compared to the artery and the portal vein was due to a selective decrease in the concentration of the intact propeptide. The study shows that biliary excretion does not contribute significantly to the hepatic extraction of circulating PIIINP in the normal liver. Furthermore, the hepatic extraction of circulating PIIINP preferentially affects the intact propeptide, rather than the somewhat larger PIIINP related molecule in serum.
A biliary protein identified by immunoblotting stimulates proliferation of peripheral blood T lymphocytes in primary biliary cirrhosis
Authors: Onishi Saburo , Maeda Takashi , Iwasaki Shinji , Saibara Toshiji , Miyamoto Takako , Miyazaki Masako , Yamamoto Yasutake , Enzan Hideaki
Published: August 2010

ABSTRACT— The source of activation of T lymphocytes in primary biliary cirrhosis (PBC) is undefined. Hence, T‐cell‐mediated reactivity against a biliary tract antigenic protein from human bile was studied. The bile protein was fractionated by 30–50% saturated ammonium sulphate and gel‐chromatography, and analysed by SDS‐PAGE and Western immunoblotting using rabbit antisera to the bile protein. The antisera reacted specifically with human bile duct epithelium. Western blotting of bile proteins showed two major bands, the B1 and B2 antigens. B1 stained for sialoglycoprotein but not lipid, but B2 was negative for both. Cell‐mediated reactivity was tested by proliferation of peripheral lymphocytes against B1. Taking the upper limit of the normal range for stimulation indices (S.I.) as less than 1.89 (= mean + 2 SD), a mitogenic response was detected in 14 of 16 patients with PBC (S.I.: 11.7 to 2.3), and in 4 of 15 patients with chronic active hepatitis, but in none of 12 patients with drug‐induced intrahepatic cholestasis or obstructive jaundice. The B2 protein was non‐stimulatory. Lymphocyte proliferation to B1 in PBC was confined to T cell fractions of peripheral blood leucocytes. There was no cross‐antigenicity between B1 and the M2 antigens, according to Western blotting using the rabbit antisera and PBC sera with anti‐M2 reactivity. Thus, the B1 biliary protein is a possible source of T cell activation in PBC and hence could be an immunological co‐factor in the pathogenesis of this disease.
Functional heterogeneity of rat liver macrophages: interleukin‐1 secretion and Ia antigen expression in contrast with phagocytic activity
Authors: Itoh Y. , Okanoue T. , Morimoto M. , Nagao Y. , Mori T. , Hori N. , Kagawa K. , Kashima K.
Published: August 2010

Abstract: To clarify whether immunological heterogeneity exists among liver macrophages and whether this possible heterogeneity corresponds to varying degrees of phagocytic activity, we fractionated liver macrophages according to size via centrifugal elutriation and studied their ability to ingest latex beads, to express Ia antigen and to secrete interleukin‐1 (IL‐1) in vitro. Larger liver macrophages were found to be more active in ingesting latex beads than smaller ones (p>0.01), while smaller liver macrophages were more active in Ia antigen expression and evidenced greater fluorescence at cytofluorometric examination. In addition, smaller liver macrophages elaborated more IL‐1 than the larger liver macrophages (p>0.05). These results indicate that liver macrophage heterogeneity is seen in at least two important macrophage functions: phagocytosis of large molecules, which provides an important scavenger function, and Ia antigen expression and IL‐1 secretion, important components of the immune response.
Hepatocellular carcinoma in a case of Wilson's disease
Authors: Cheng W. S. C. , Govindarajan S. , Redeker A. G.
Published: August 2010

Abstract: Hepatocellular carcinoma has rarely been reported in Wilson's disease, particularly in women. We describe the case of a female patient who was diagnosed with Wilson's disease at the age of 39 years, after presenting with severe neurological symptoms. She had significant neurological improvement following penicillamine therapy and succumbed to hepatocellular carcinoma at the age of 72 years, following 33 years of penicillamine therapy. The patient described here was the oldest and only the third female patient with hepatocellular carcinoma complicating Wilson's disease to be reported in the literature.
Primary hepatocellular carcinoma with severe hypoglycemia; involvement of insulin‐like growth factors
Authors: Yonei Yoshikazu , Tanaka Masato , Ozawa Yukako , Miyazaki Koji , Tsukada Nobuhiro , Inada Shinichi , Inagaki Yasutaka , Miyamota Kei , Suzuki Osamu , Okawa Hideo , Kiryu Yasuyoshi
Published: August 2010

Abstract: We report a case of severe hypoglycemia and hepatic masses suspected to be an insulin‐like growth factor‐II (IGF‐II)‐producing hepatocellular carcinoma. A 62‐year‐old man presented with mental disorder in the night and early morning associated with extremely low blood sugar levels (> 21 mg/dl). Computerized axial tomography and ultrasonography revealed a massive tumor in the right lobe of the liver with multiple secondary nodules, and a tumor thrombus in the portal vein. At autopsy 107 days after admission, the liver weighed 3070 g, histologically showing an Edmondson type II tumor with liver cirrhosis. IGF‐II in plasma (899 ng/ml) and tumor tissue (2.4 μg/g) was higher than that in normal plasma (374–804 ng/ml) and non‐tumor liver tissue (0.2 μg/ml), while IGF‐I (14 ng/ml) was significantly reduced. IGF‐II, probably produced by the liver tumor, appeared to be involved in the mechanism of hypoglycemia.
Estradiol‐17β modifies the induction of spermidine/spermine N1‐acetyltransferase activity in the liver of lipopolysaccharide‐treated mice
Authors: Sugimoto Hiroyuki , Matsuzaki Yutaka , Yuasa Keilchiro , Kaneko Mieko , Yamada Shoji , Hamana Koei , Matsuzaki Shigeru
Published: August 2010

ABSTRACT— In an attempt to elucidate the effects of estrogen on polyamine metabolism in lipopolysaccharide (LPS)‐treated mice, we assayed polyamine content and the activity of spermidine/spermine N1‐acetyltransferase (SAT) and ornithine decarboxylase (ODC) in some organs. LPS elevated N'‐acetylspermidine levels in the liver and lung and putrescine levels in the liver, lung and spleen. LPS increased the activity of ODC at 6 h and that of SAT at 12 h in the liver. When estradiol‐17β was simultaneously administered with LPS, the maximum increase in hepatic N1‐acetylspermidine levels was found 6 h earlier than in the LPS control. Likewise, the peak of the hepatic SAT activity after LPS‐treatment was observed 6 h earlier in the estradiol‐17β‐treated mice than in the LPS control. No such effect of estradiol‐17β was found in the lung and spleen. The LPS‐induced ODC activity was not affected by estradiol‐17β in the liver, lung or spleen. Estrone and 16β‐ethylestradiol (an anti‐estrogen) were also effective in enhancing the LPS‐induced elevation of N1‐acetyl‐spermidine and putrescine in the liver, while both diethylstilbestrol, which has a potent estrogenic activity without steroid structure and estradiol‐17α (a non‐estrogenic isomer of estradiol‐17β) were without effect. Tamoxifen (an estrogen receptor antagonist) did not suppress the estrogen‐induced increase in hepatic N1‐acetylspermidine levels.
Liver function after bilateral nephrectomy
Authors: Fleck C. , Börner A. , Kretzschmar M. , Machnik G. , Sprott H. , Zimmermann T. , Kell E. , Bräunlich H.
Published: August 2010

ABSTRACT—Consequences of bilateral nephrectomy (NX) for liver functions and for hepatic excretion of various endogenous substances were characterized in rats 24 h after NX. Plasma concentrations of urea, creatinine, fibrinogen, and glutathione increased significantly after NX, whereas the concentrations of total protein, albumin, and lipids decreased. The hepatic excretion of urea, creatinine, phospholipids, cholesterol, and aldosterone significantly increased in uremia, and excretions of protein and glutathione diminished. Active biliary transport can be diminished after NX by the effects of uremic toxins on the liver cells or by the competition phenomena between endogenous substances, which are normally excreted in urine, at the hepatocellular level. Reduced glutathione content and increased lipid peroxidation in hepatocytes have been found. Changes in lipid and protein metabolism after NX can be proved.
Modulation of thioacetamide‐induced hepatocellular necrosis by prostaglandins is associated with novel histologic changes
Authors: Bergasa Nora V. , Borque Maria J. , Wahl Larry M. , Rabin Lionel , Jones E. Anthony
Published: August 2010

Abstract:Cytoprotective effects of the prostaglandins 16, 16‐dimethyl PGE2(dmPGE2) and PGF2α tromethamine (PGF2α) were evaluated in the rat model of acute hepatocellular necrosis induced by thioacetamide (TAA). dmPGE2(100 μg/kg SC 8 hourly) did not induce a significant increase in survival when started after the onset of TAA‐induced fulminant hepatic failure. However, priming with dmPGE2(100 μg/kg SC 30 min before TAA) reduced TAA‐induced elevations in serum ALT (684 ± 68 (SEM) vs 274 ± 135 IU/1, p>0.01). This phenomenon did not occur if dmPGE2was administered after TAA or by the IP route. Modulation of TAA‐induced centrizonal hepatocellular necrosis by dmPGE2was associated with a striking increase in centrizonal ballooning of hepatocytes (p>0.01), and, as assessed by stereology, less hepatocellular necrosis and degenerative changes. PGF2α, which in contrast to dmPGE2does not act via cAMP, had no effect on TAA‐induced changes in serum ALT or hepatic histology. These findings suggest that dmPGE2decreases hepatocellular necrosis by activating surface membrane adenylate cyclase and consequently stimulating cAMP. Ballooning of hepatocytes could occur secondary to these membrane events and appears to be a marker of dmPGE2‐induced cytoprotection in this model.

The opinions or assertions contained herein are the private views of the authors and are not to be construed as official or as reflecting the views of the Department of the Army or the Department of Defense.
Intracellular distribution and effect of the antimalarial drug mefloquine on lysosomes of rat liver
Authors: Glaumann Hans , Motakefi Anne‐Marie , Jansson Helena
Published: August 2010

Abstract: Mefloquine was administered in a single dose (1–30 mg/100 g) to rats in order to study its subcellular distribution and effects on rat liver lysosomal structure and function. Subcellular fractionation showed a significant enrichment of mefloquine in lysosomes. Even repeated administration of mefloquine did not affect the levels of cytochrome‐P‐450 or its reductase, indicating, although not proving, that it is not metabolized by this mono‐oxygenase system. Mefloquine caused an expansion of the lysosomal apparatus, earliest seen by 24 h and lasting for some 7 days. Initially, cytoplasmic constituents were seen inside the lysosomes. Later, the lysosomes harboured myelin‐like figures (multilamellar bodies) disappearing after 7–10 days. The proteolytic and lipolytic capacity was assessed in isolated lysosomes. Mefloquine caused increased protein degradation but decreased breakdown of lipids. Concomitantly, all five major phospholipids (phosphatidyl‐choline, ‐ethanolamine, ‐inositol, ‐serine and sphingomyelin) increased in the lysosomes. It is concluded that: (1) mefloquine is a lysosomotropic drug that accumulates in lysosomes; (2) mefloquine impairs lipid degradation with ensuing accumulation of lipids in lysosomes; and (3) lysosomal trapping explains the high volume distribution of mefloquine.
Enhanced tumor necrosis factor alpha in coronavirus but not in paracetamol‐induced acute hepatic necrosis in mice
Authors: Devictor D. , Decimo D. , Sebire G. , Tardieu M. , Hadchouel M.
Published: August 2010

Abstract:Previous reports have demonstrated that tumor necrosis factor alpha (TNF‐α) plays an important role in the pathogenesis of fulminant hepatic necrosis. The purpose of this experimental study was to measure TNF‐α blood activity in paracetamol‐induced liver necrosis and in coronavirus (MHV3)‐induced fulminant hepatitis in mice. No elevation of TNF‐α activity was found in hepatic failure complicating paracetamol poisoning. In contrast, TNF‐α activity significantly increased in response to MHV3, reaching 16.3 ± 5.5 U/ml from 24 h post infection (P>0.01). This augmentation was observed even though the virus was not detectable in the liver. Serum alanine aminotransferase levels were low and no histological lesion was observed. In conclusion, our study further supports the implication of TNF‐α in virus‐induced hepatitis failure and confirms that paracetamol poisoning does not cause increased TNF‐α activity in the circulation.
Detection of cytomegalovirus by in situ hybridization using a digoxigenin‐tailed oligonucleotiede
Authors: Pacchlenl Benatella , Papettl Mauro , Benlne Ferrucclo , Bussolati Glanni , Negro Francesco
Published: August 2010

Abstract: A non‐isotopic in situ hybridizaiton procedure was used to detect cytomeglavirus (CMV sequences within routinely fixed tissue. A digoxigenin‐tailed oligonucletoide was hybridized to sections of specimens obtianed at autopsy from 2 patients with CMV infection. Hybrids wer revealed by an alkaline phosphatase‐conjugated anti‐digoxigenin antibody. Serila sections were also assayed for the presence of CMV by in situ hybridizaiotn with a biotin‐labelled cDNA probe and by immunohistochemistry and routinenly stained for morphological evaluation. Results show that the two in situ hybridization procedures are equally sensitive but superior to the immunohistochemical detection of the viral antigen. Most cells positive for CMV DNA had the cytopathological features characteristic of CMV infection. A minor population of infected cells lacking morphological changes was also found. We recommend the routine application of the oligoncucleotied‐based assay because it is specific, easy and less expensive than other similar procedures.
From two dimensional (2D) to three dimensional (3D) analysis by confocal microscopy
Authors: Neri L. M. , Martelli A. M. , Previati M. , Capitani S.
Published: August 2010

Abstract: The confocal microscope is becoming increasingly important as an apparatus to analyze the 3‐D topography of the cell. Main reasons are the high resolution optical sectioning capacity, the non‐invasiveness which leaves the object intact, and the imaging capabilities. This chapter introduces a description of the confocal principle, the basic concepts of confocal fluorescence microscopy and some criteria for cell preservation. Optimization of in situ immunofluorescence, hybridization and detection procedures in combination with new digital microscope techniques can fully express their capacities only if the preparation of biological specimens is accurate for 3‐D analysis. Some applications of confocal microscopy to the study of intranucleolar antigens, enzyme translocations and fluorescence in situ hybridization, are described in association with 3‐D software image processing, as a useful framework for the study of the 3‐D visualization of proteins and chromatin domains.
Arterial and mixed venous acid‐base status in patients with cirrhosis. Influence of liver failure
Authors: Moreau Richard , Hadengue Antoine , Soupison Thierry , Mamzer Marie‐France , Kirstetter Philippe , Saraux Jean‐Luc , Assous Michel , Roche‐Sicot Jacqueline , Sicot Christian
Published: August 2010

ABSTRACT— Although it has been established that liver failure is associated with arterial hypocapnia and alkalaemia (i.e., respiratory alkalosis), the influence of liver failure on mixed venous acid‐base status has not yet been studied. Thus, arterial and mixed venous acid‐base status were simultaneously measured in controls and in a large series of patients with cirrhosis. Grade B patients (n = 28) or Grade C patients (n = 21) had significantly lower arterial and mixed venous carbon dioxide tensions than controls (n = 29). Grade B or Grade C patients also had significantly higher arterial, mixed venous pH, and lower mixed venous bicarbonate concentrations than controls. Among Grade A patients (n = 27), those with the lowest Pugh's score (i.e., equal to five) had significantly lower mixed venous carbon dioxide tension than controls. The other arterial and mixed venous acid‐base values did not differ significantly between Grade A patients with the lowest Pugh's score and controls. Grade A patients with a Pugh's score equal to six and Grade B patients had similar acid‐base disorders. No significant differences were found between groups concerning the anion gap and plasma chloride concentrations. In conclusion, this study shows that in Grade B or C patients, respiratory alkalosis was responsible for mixed venous hypocapnia, alkalaemia and hypobicar‐bonataemia. In addition, in Grade A patients with the lowest Pugh's score (equal to five), analysis of arterial and mixed venous blood revealed that mixed venous hypocapnia was the sole anomaly of the acid‐base status. This last finding suggests that mixed venous hypocapnia might be an early event preceding the onset of arterial hypocapnia.
Antibody to two forms of dihydrolipoamide acetyltransferase (PDC‐E2) in primary biliary cirrhosis
Authors: Chen Qlao‐YI , Rowley Merrill J. , Mackay Ian R.
Published: August 2010

ABSTRACT— Dihydrolipoamide acetyltransferase, the E2 subunit of the pyruvate dehydrogenase complex (PDC‐E2), is the major autoantigen in primary biliary cirrhosis. By immunoblotting with sera from patients with primary biliary cirrhosis, we observed a double band, of molecular weight 70 and 74 kD for PDC‐E2, when a preparation of bovine heart mitochondria was not boiled prior to electrophoresis. This double band could also be detected using antisera raised in rats or rabbits against intact PDC or PDC‐E2, but not in antisera raised against a synthetic decamer representing the lipoic acid binding sequence of PDC‐E2; the latter reacted only with the 74 kD component. Antibody eluted from either the 70 or 74 kD component reacted with both 70 and 74 kD components. By ELISA, sera from patients with primary biliary cirrhosis reacted more strongly with a non‐boiled than a boiled PDC‐E2, whereas immune animal sera reacted equally with both preparations. Thus, according to whether preparations of PDC are boiled or not, two conformationally alternative forms of the PDC‐E2 protein can be revealed by immunoblotting. The two forms in non‐boiled preparations migrate at molecular weights corresponding to 70 and 74 kD.
Disappearance of serum hepatitis B virus DNA by polymerase chain reaction after adenine arabinoside 5‘‐monophosphate therapy in chronic hepatitis B
Authors: Buti M. , Rodriguez‐Frlas F. , Genesca J. , Arranz A. , Jardi R. , Esteban R. , Guardia J.
Published: August 2010

ABSTRACT— The aim of antiviral therapy in chronic hepatitis B is the cessation of viral replication, which may be demonstrated by the loss of hepatitis B “e” antigen (HBeAg) and serum hepatitis B virus DNA (HBV‐DNA) detected by dot‐blot hybridization. With the development of the sensitive polymerase chain reaction (PCR) technique for detecting HBV‐DNA, it has become apparent that many HBeAg negative patients may still have small amounts of circulating viral DNA. We assessed 19 of 25 patients with chronic hepatitis B who seroconverted from HBeAg to anti‐HBe after adenine arabinoside 5′‐monophosphate therapy (5 mg ·kg‐1 · day‐1 for 7 weeks) to determine whether serum HBV‐DNA became undetectable. Sixteen of the 19 HBeAg negative patients remained hepatitis B surface antigen (HBsAg) positive, and the other three lost HBsAg during follow‐up. All of them were HBV‐DNA negative by dot‐blot hybridization. Using the PCR technique, HBV‐DNA became negative in 13 (81.2%) of the 16 patients who seroconverted to anti‐HBe without losing HBsAg, and in all the patients who lost HBsAg. These data suggest that the majority of patients who respond to adenine arabinoside 5′‐monophosphate show a complete inhibition of hepatitis B virus replication, as demonstrated by the absence of viral DNA by PCR. This inhibition was present in all patients who, at the same time, lost HBsAg.
Efficacy and safety of the stepped care medical treatment of ascites in liver cirrhosis: a randomized controlled clinical trial comparing two diets with different sodium content
Authors: Bernardi Mauro , Laffi Glacomo , Salvagnini Mario , Azzena Giuseppe , Bonato Stefano , Marra Fabio , Trevlsani Franco , Gasbarrini Giovannl , Naccarato Remo , Gentillni Paolo
Published: August 2010

ABSTRACT— In order to clarify debated issues of the medical treatment of ascites in cirrhosis – the usefulness of a low sodium diet and washout period preceding diuretic administration, maximal dosage of antimineralocorticoid to be reached before the addition of a loop diuretic, identifications of factors influencing treatment efficacy – 115 hospitalized patients with non‐azotemic cirrhosis and ascites were recruited and randomized to receive a diet providing either 40 or 120 mmol of sodium daily. After a washout period from the outpatient diuretic regimen for 7 days (Step 1), increasing dosages of K‐canrenoate (200 mg/day every 4th day up to 600 mg) were administered to patients not undergoing spontaneous diuresis (Step 2). Upon the failure of Step 2, K‐canrenoate (400 mg/day) and furosemide at increasing dosage (25–50–100 mg every other day) were given (Step 3). Nine percent of patients underwent spontaneous diuresis, and 77% developed a negative sodium balance by the end of Step 2 (69% with a dosage of K‐canrenoate ≤400 mg/day) and 93% by the end of Step 3. Two patients were withdrawn from the protocol due to diuretic side‐effects. Univariate analysis showed that the type of diet did not influence the response to treatment. The washout period led to a significant increase in endogenous creatinine clearance; natremia significantly rose in hyponatremic patients. Multivariate analysis showed that creatinine clearance and plasma aldosterone were independent predictive factors of the response to treatment. The use of low‐sodium diet did not enhance the efficacy of the medical treatment of non‐azotemic cirrhotic ascites and a washout period from diuretics improved renal function and natremia. Antimineralocorticoids up to 400 mg/day were successful in over 2/3 of cases and the improvement in efficacy obtained by a further increase in dose was small. Endogenous creatinine clearance and plasma aldosterone significantly influenced treatment efficacy. The therapeutic regimen employed in this study was highly effective and had a very low incidence of side effects.
Serum alpha‐fetoprotein and lens culinaris agglutinin‐reactive fraction of alpha‐fetoprotein in patients with hepatocellular carcinoma
Authors: Kuromatsu Ryoko , Tanaka Masatoshi , Tanikawa Kyuichi
Published: August 2010

ABSTRACT— A fraction of serum α‐fetoprotein (AFP) reactive with lens culinaris agglutinin (LCA) was measured by affinity chromatography in serum samples from 102 patients with hepatocellular carcinoma (HCC) and 48 patients with chronic liver diseases without HCC. Its usefulness as a marker of HCC was evaluated. The mean ± SD percentage of this fraction in total AFP was 3.10 ± 3.17% in 48 patients with chronic liver diseases without HCC. When the cut‐off level was set at 12.6% (mean + 3 SD), the sensitivity was 36.3%, the specificity was 100%, and the accuracy was 56.7% in the 102 patients with HCC. This lentil lectin‐reactive AFP was positive in 7 of 25 patients (28%) who had single small liver cancer (Ψ < 20 mm), suggesting its clinical usefulness as a tumor marker. The lentil lectin‐reactive AFP showed no correlation with the serum concentration of AFP or des‐γ‐carboxy prothrombin (DCP). In patients with HCC showing an AFP level of 20 ng/ml or above, the lentil lectin‐reactive fraction is a highly specific tumor marker. We consider it to be useful as an adjunct in the diagnosis of HCC.
Influence of oral treatment with ursodeoxycholic and tauroursodeoxycholic acids on estrogen‐induced cholestasis in rats: effects on bile formation and liver plasma membranes
Authors: Bouchard Guylaine , Yousef Ibrahim M. , Tuchweber Beatrix
Published: August 2010

ABSTRACT— In this study, we examined whether ursodeoxycholic acid (UDC) and its taurine conjugate, tauroursodeoxycholic acid (TUDC), given per os, can prevent the cholestasis induced in rats by 17‐α‐ethynyl estradiol (EE) and whether this protection is mediated by choleretic activity or altered plasma membrane composition. EE (5 mg/kg body weight/day for 5 days) markedly reduced bile flow and bile salt secretion without significantly affecting plasma membrane composition and function. Treatment with UDC or TUDC (100, 150 or 200 (TUDC only) μmol/100 g body weight/day for 5 days) did not significantly modify bile flow, but the bile salt secretion rate increased in a dose‐dependent manner. UDC was the main biliary bile acid secreted in groups given higher doses of UDC or TUDC. At these dose levels, bile acid treatment did not affect plasma membrane fluidity as assessed by fluorescence anisotropy, the cholesterol/phospholipid molar ratio as well as Na+K+‐ and Mg++‐ ATPase activities. The highest dose of UDC and TUDC prevented the reduction of both bile flow and bile salt secretion induced by EE, reestablishing these parameters to the values of the corresponding control for the UDC group. In conclusion, UDC and TUDC, given per os, improve EE‐induced cholestasis, an effect that cannot be attributed to choleretic activity or altered plasma membrane composition.
Effect of dobutamine on the arterial ketone body ratio and portal blood flow velocity in cirrhosis
Authors: Kajlmura Kozo , Morlyasu Fuminori , Kimura Tohru , Okuma Minora , Mori Keiichiro , Ozawa Kazuo
Published: August 2010

ABSTRACT—We studied the relationship between the portal blood flow velocity and the arterial ketone body ratio in patients with chronic liver disease receiving a dobutamine infusion. We used an ultrasonic Doppler duplex system to evaluate the portal blood flow velocity. Dobutamine was given intravenously at 5 μg/kg/min for 20 min. Dobutamine infusion induced smaller changes in the portal blood flow velocity and ketone body ratio in liver cirrhosis than in chronic hepatitis. The existence of shunts and the poor increase of the cardiac index in response to dobutamine explained the limited improvement of portal blood flow velocity in cirrhosis patients. The ketone body ratio was improved by dobutamine in cirrhosis patients whose portal blood flow velocity was increased by more than 10%, while this ratio decreased when the increase of it was less than 10%. There was no change in portal oxygen extraction in the cirrhosis group, and portal oxygen uptake only increased when the portal blood flow velocity rose by more than 10%. Dobutamine should only be used to treat liver failure if the portal blood flow velocity is increased by more than 10% or the arterial ketone body ratio is improved by a test infusion.
The mutation of codon 249 in the p53 gene is not specific in Japanese hepatocellular carcinoma
Authors: Hayashi Hiroshi , Sugio Kenji , Matsumata Takashi , Adachi Eisuke , Urata Keiko , Tanaka Shlnji , Sugimachi Keizo
Published: August 2010

ABSTRACT— Hepatocellular carcinoma samples obtained from 59 patients at surgical resection were examined for mutations of the third base at codon 249 of the p53 gene, using the polymerase chain reaction and oligonucleotide hybridization techniques. This point mutation, which is frequently observed in HCC cases from Southern Africa and Quidong in China, was not recognized in either 60 hepatocellular carcinomas or 53 noncancerous liver tissue samples from Japan. Thirty‐four of 45 patients (75.6%) were positive for the hepatitis C virus, which was a higher rate than that for hepatitis B virus infection (9 of 55; 16.4%). The exposure to aflatoxin B1 was not considered to be remarkable. These results suggest that the point mutation of the third base at codon 249 is not common in Japanese patients, and it is suggested that numerous other factors affect the mutation of the p53 gene and the development of hepatocellular carcinoma.
Hypercalcemia and parathyroid hormone‐related protein in hepatocellular carcinoma
Authors: Yen Tzu‐Chen , Hwang Shinn‐Jang , Wang Ching‐Chen , Lee Shou‐Dong , Yen Shin‐Hwa
Published: August 2010

ABSTRACT— A two‐site immunoradiometric assay (IRMA) of parathyroid hormone‐related protein (PTHrP) was employed to react with circulating concentrations of PTHrP in 14 patients with hepatocellular carcinoma (HCC) and hypercalcemia (> 10.6 mg/dl). Eleven of them had unresectable lesions and three received transcatheter arterial chemo‐embolization (TACE) treatment. Patients had no evidence of bony metastases and only one had evidence of a parathyroid lesion (by bone scan and serum parathyroid hormone level, respectively). The urinary cAMP level was increased in all patients, but the serum 1,25‐dihydroxyvitamin D and plasma cAMP levels varied. Twelve patients had elevated alpha‐fetoprotein (AFP) (> 400 ng/ml) and two of them had mildly elevated AFP levels (11 and 147 ng/ml). Their PTHrP concentrations were elevated (7.1 to 33.2 pmol/l), compared with normal levels obtained in our laboratory (< 3.5 pmol/l). A significant decrease in plasma PTHrP (from 27.4 to 5.2 pmol/1), serum calcium concentrations (from 16.3 to 9.4 mg/dl) and AFP levels (from 64 787 to 3129 ng/ml) was observed on the day following TACE treatment. These results, by using an improved technique, extend the findings that hypercalcemia in patients with HCC is associated with increased renal reabsorption of calcium and increased bone resorption of PTHrP generated by HCC.
Manifestations of Epstein‐Barr virus‐associated disorders in liver
Authors: Markin Rodney S.
Published: August 2010

Abstract: Epstein‐Barr virus is a ubiquitous virus associated with a variety of different diseases and disorders. The manifestations of Epstein‐Barr virus‐associated diseases or disorders within the liver, which involve a broad spectrum of histologic and clinical features, ranging from hepatitis through lymphoproliferative disorders to lymphoma, are presented. An important aspect of Epstein‐Barr virus expression and infection is the biology of the Epstein‐Barr virus. Documentation of infection can be performed using serology to detect the interaction of Epstein‐Barr virus with the immune system, and the detection of EBV proteins and use of molecular biologic techniques to identify the presence of EBV RNA, and DNA sequences. Of particular utility are in situ hybridization, Southern blot analysis, and polymerase chain reaction as diagnostic methods to identify specific RNA or DNA sequences. Epstein‐Barr virus‐associated diseases and disorders including infectious mononucleosis, sporadic fatal infectious mononucleosis, X‐linked proliferative disorder (Duncan's disease), post‐transplant lymphoproliferative disorders, lymphoma, and AIDS are discussed. The histopathologic findings present in liver associated with each disease are presented with illustrative examples. Handling the tissue and interaction with clinical services are also discussed as a method for appropriate diagnosis of Epstein‐Barr virus‐driven processes affecting the liver.
Immunohistochemical analysis of S‐phase cells in normal human and rat liver by PC10 monoclonal antibody
Authors: Mancini Raniero , Marucci Luca , Benedetti Antonio , Jezequel Anne‐Marie , Orlandi Francesco
Published: August 2010

The expression of proliferating cell nuclear antigen (PCNA) was examined in normal human and rat liver fixed in either formaldehyde or methanol, and was compared with the incorporation of bromo‐deoxyuridine (BrdU) in S‐phase cells. Codistribution of PCNA and BrdU was assessed in rat liver by double immunohistochemical staining using PC10 and anti‐BrdU monoclonal antibodies to identify labelled nuclei of parenchymal and sinusoidal cells. In formaldehyde‐fixed human biopsies (n = 13) PCNA‐labelling index (PCNA LI) was 0.43 ± 0.24% (mean ± SEM) for hepatocytes and 0.09 ± 0.03% for sinusoidal cells. A great inter‐specimen variability was observed and a preferential lobular distribution was not evident. In methanol‐fixed human liver (n = 8) the immunostaining was strong. PCNA LI was 0.05 ± 0.01%) for hepatocytes and 0.14 ± 0.01% for sinusoidal cells. 75% of labelled hepatocytes and 60% of labelled sinusoidal cells were found in acinar zone 1. In formaldehyde‐fixed rat liver (n = 10) a weak nuclear staining and a great interspecimen variability were evident. LI was 0.13 ± 0.07%) for hepatocytes and 0.40 ± 0.21% for sinusoidal cells without preferential acinar distribution. In methanol‐fixed rat liver (n = 10), PCNA LI was 0.14 ± 0.02% for hepatocytes and 0.40 ± 0.04% for sinusoidal cells. 64% of labelled hepatocytes and 50% of labelled sinusoidal cells were found in zone 1. Only on methanol‐fixed material did double immunohistochemistry show an almost complete overlap of BrdU and PCNA labelling. The PCNA LIs and the zonal distribution of labelled nuclei as obtained in methanol‐fixed material are in keeping with previous reports using 3H‐thymidine (3H‐Thy) incorporation, suggesting that PCNA immunostaining represents a valid alternative to 3H‐Thy. In addition, the present data support the hypothesis that S‐phase associated PCNA is more selectively retained in methanol‐fixed liver tissue.
Diclofenac‐induced hepatitis: case report and literature review
Authors: Ramakrishna Banumathi , Viswanath N.
Published: August 2010

A case of diclofenac‐induced hepatitis, in a 53‐year‐old Indian woman with recurrent jaundice, is presented. The diagnosis was suspected from the appearance of the liver biopsy and was confirmed by the temporal association with drug usage and cure on withdrawal of the drug.
Flucloxacillin‐associated hepatic injury
Authors: Koek G. H. , Stricker B. H. Ch. , Blok A. P. R. , Schalm S. W. , Desmet V. J.
Published: August 2010

Eleven cases of hepatic injury attributed to the intake of flucloxacillin were reported to the Netherlands Center for Monitoring of Adverse Reactions to Drugs between 1982 and 1992. They concerned four men and seven women, with a mean age of 57 years, treated for 2–28 days with an oral dose varying from 1500–4000 mg per day. Symptoms mostly appeared 10 to 30 days after starting treatment with flucloxacillin. Biochemically, the pattern was compatible with cholestatic hepatitis in seven cases, with a mixed cholestatic‐hepatocellular type of injury in one case, a hepatocellular pattern in two cases, and mild liver enzyme elevations in one patient. Two patients died, one due to fatal bleeding from the liver after biopsy, and the second patient to a combination of hepatic and cardiac failure. The other patients recovered, on average 72 days after peaking of serum aminotransferase values. Histology in seven cases showed cholestatic hepatitis in five, with cholangitis or cholangiolitis in four of these patients. In the other two patients, there was centrilobular cholestasis with extensive bridging fibrosis and portal‐central bridging necrosis, respectively.
Immunological functions of mouse liver resident high density lymphocytes
Authors: Ueta Hiroshi , Saibara Toshiji , Onishi Saburo , Yamamoto Yasutake
Published: August 2010

Liver‐derived high density lymphocytes (Matsunaga cells) have been detected as members of resident T cells in the mouse liver. In this study, we assessed the immunological functions of liver‐derived high density lymphocytes of BALB/c mice in comparison with those derived from spleen and peripheral blood. Liver‐derived high density lymphocytes proliferated in response to the syngeneic and allogeneic mixed lymphocyte reaction, as well as those derived from spleen and peripheral blood. The allo‐activated cytotoxic T lymphocyte activity of liver‐derived high density lymphocytes against PHA‐blasts of C57BL/6 mice was lower than that of spleen‐ and peripheral blood‐derived lymphocytes. The suppressor activity of syngeneic‐ or allo‐activated high density lymphocytes of the liver, spleen, and the peripheral blood was assessed by measuring their suppressive effect on the proliferation or on the generation of allo‐specific cytotoxic activity in allogeneic mixed lymphocyte reaction. The suppression was concentration‐dependent and strongest in liver‐derived lymphocytes.
Pathology of livers infected with “silent” hepatitis B virus mutant
Authors: Uchida Toshikazu , Shimojima Seiichi , Gotoh Kenichiro , Shikata Toshio , Mima Satoaki
Published: August 2010

We have discovered that non‐A, non‐B, non‐C, non‐D, non‐E (so‐called type F) acute and chronic hepatitis is caused by a hepatitis B virus (HBV) variant with mutations in the X open reading frame. This silent HBV mutant does not induce immunoserological markers. In the present investigation we attempted to elucidate the putative mechanism of hepatocellular necrosis and expression patterns of hepatitis B surface antigen (HBsAg) and core antigen (HBcAg) in biopsied liver tissue. The subjects consisted of 14 patients with acute hepatitis, 11 with chronic hepatitis and eight with liver cirrhosis, all of whom had been previously diagnosed as having so‐called hepatitis F. Nine of the 14, 10 of the 11 and all eight, respectively, of the above patients exhibited significant positive immunostaining for HBsAg within their hepatocellular cytoplasm, diffusely or focally. HBcAg stained in a few hepatocellular nuclei in 24.2% of the patients. Histological features were characterized by necroinflammation, indicating immune‐mediated hepatocellular necrosis. Despite the serological‐marker negativity, the results of immunostaining for HBsAg and HBcAg support replication and expression of HBV DNA, though weak.
Kinetics of hepatic amino‐nitrogen conversion in ageing man
Authors: Fabbri Andrea , Marchesini Giulio , Bianchi Giampaolo , Bugianesi Elisabetta , Zoli Marco , Pisi Emilio
Published: August 2010

ABSTRACT: Previous studies have shown that hepatic function, quantitatively measured by dynamic liver function tests, progressively declines with ageing. Urea synthesis is a specific process taking place in the liver; a reduced urea synthesis in response to a protein‐rich metal has previously been demonstrated in the elderly, but the process has never been standardized in relation to amino acid supply. We measured the hepatic conversion of α‐amino nitrogen into urea nitrogen in response to alanine infusion in 32 subjects, with normal routine liver and renal function tests and without evidence of previous hepatic disorders, belonging to three different age‐groups (≤55 years, 56–70, ≥71). The functional hepatic nitrogen clearance was reduced on average by 20% in subjects aged 56–70 years, and by 30% in subjects over 70 years old in comparison to the age‐group under 55 years (ANOVA: P=0.0001), and significantly correlated with age (r=‐0.684). No sex differences were observed on the effects of age on hepatic clearance. Also, liver volume, measured by ultrasonography, was reduced with advancing age, but the age‐related decrease in hepatic nitrogen conversion was not primarily dependent on decreased liver volume. The measurement of functional hepatic nitrogen clearance has already been validated as a quantitative liver function test in clinical hepatology. In keeping with previous studies, the age‐related deceline in hepatic nitrogen conversion points to a decreased functional capacity of the ageing hepatic parenchyma.
Predictive factors in the response to interferon therapy in chronic hepatitis C
Authors: Yoshioka Kentaro , Higashi Yasuyuki , Yamada Masaki , Aiyama Toshiyuki , Takayanagi Masahiro , Tanaka Kazuma , Okumura Akihiko , Iwata Kazuo , Kakumu Shinichi
Published: August 2010

Factors predicting the efficacy of interferon therapy were statistically analyzed on 111 patients with chronic hepatitis C. Of the treated patients (total doses of interferon; 96–468 MU), 35 (31.5%) had a long‐term remission. On multivariate analysis, hepatitis C virus genotype (p<0.0001), histological diagnosis (p<0.05), fibrosis score of histological activity index (p<0.01) and source of infection (p<0.05) were found useful for predicting the response to interferon therapy. Our findings suggest that the outcome of interferon therapy can be predicted to some degree from pretreatment data, and that a new therapeutic strategy is necessary for the group of patients who are predicted to be nonresponders.
Histochemical and electron microscopic characterization of hepatic macrophage subfractions isolated from normal and liposomal muramyl dipeptide treated rats
Authors: Hoedemakers Rein M. J. , Atmosoerodjo‐Briggs Jane E. , Morselt Henriëtte W. M. , Daemen Toos , Scherphof Gerrit L. , Hardonk Machiel J.
Published: August 2010

Abstract: Subfractions of the hepatic macrophage population, differing in cell size, were isolated from normal rats and rats treated with liposomal muramyl dipeptide (lipMDP) and analyzed histochemically and by ultra‐structural peroxidase cytochemistry. The majority of cells in all subfractions of control rats displayed the ultrastructural endogenous peroxidase pattern of resident liver macrophages and showed positive staining with the general macrophage markers nonspecific esterase (NSE) and monoclonal antibody ED1. Heterogeneity in intensity of NSE and ED1 staining was observed among macrophages of different size. Generally, the intensity of NSE and ED1 staining decreased with decreasing cell size. After injection of lipMDP, we observed the appearance of a discrete subpopulation of cells in the liver in addition to the resident macrophages. These cells, containing a nucleus with a characteristic shape, were predominantly recovered in the small‐sized fractions and were characterized by an immature ultrastructural macrophage morphology (no or only a few lyso‐somes and phagosomes) and a lack of ED1 reactivity, NSE, and endogenous peroxidase. We suggest an important role for these cells in lipMDP induced antitumor capacity of the liver.
High interleukin‐6 production within the peritoneal cavity in decompensated cirrhosis and malignancy‐related ascites
Authors: Bac D. J. , Pruimboom W. M. , Mulder P. G. H. , Zijlstra F. J. , Wilson J. H. P.
Published: August 2010

Abstract: To assess the diagnostic and prognostic value of interleukin‐6, interleukin 1β, and tumor necrosis factor‐α assays in plasma and ascites, we measured these cytokines in eight patients with malignancy‐related ascites and 32 patients with decompensated cirrhosis. Five patients had an episode of bacterial peritonitis, during which one or more ascitic fluid samples were analyzed. Interleukin‐6 and tumor necrosis factor‐α were not significantly different between the cirrhotic and the malignant groups: ascitic interleukin‐6 13 816±15 314 vs 28 138±23 403 pg/ml, plasma interleukin‐6 542±719 vs 559±604 pg/ml; ascitic tumor necrosis factor‐α 19±50 vs 12±31 pg/ml, plasma tumor necrosis factor‐α 3.4±8.2 vs 6.1±13.8 pg/ml. During an episode of bacterial peritonitis there was a significant increase only in ascitic interleukin‐6 (133 268±99 743 pg/ml), which declined after antibiotic treatment. None of the parameters was associated with 6‐month survival (11 of the 40 patients died within 6 months). There was a correlation (r=0.675; p=0.002) between plasma interleukin‐6 levels and the Child‐Pugh score in patients with cirrhosis, but not with the etiology of the liver disorder. Plasma interleukin‐6 levels correlated with IgA levels (r=0.649; p=0.004) but not with C reactive protein, sedimentation rate, fibrinogen, IgM or IgG. These results do suggest that interleukin‐6 is produced within the peritoneal cavity in hepatic and malignant ascites. There is a sharp increase in the local production of interleukin‐6 during an episode of bacterial peritonitis. This increase was not detectable for tumor necrosis factor‐α. The physiological meaning of this over‐production of locally generated cytokines and whether it relates to the poor prognosis of patients with cirrhosis and infectious disorders remain to be assessed.
Heterogeneity in secretory responses of rat liver macrophages of different size
Authors: Hoedemakers Rein M. J. , Morselt Henriëtte W.M. , Scherphof Gerrit L. , Daemen Toos
Published: August 2010

Abstract: Four subpopulations of hepatic macrophages, differing in size, were isolated from rat liver. The secretion of nitric oxide (NO), tumor necrosis factor‐α (TNF‐α), prostaglandin E (PGE) and interleukin‐1 (IL‐1) by freshly isolated as well as cultured cells was studied after in vitro stimulation with the immunomodulator muramyl dipeptide (MDP) in free or liposome‐encapsulated form. Freshly isolated liver macrophages could be induced to secrete significant levels of NO, TNF‐α, PGE and IL‐1. The extent of secretion, however, varied substantially between macrophages of different size. The highest levels of secretion of TNF‐α, PGE and IL‐1 were observed in the fraction containing the large‐size macrophages, while progressively lower levels of secretion were observed with decreasing size. In constrast, the highest levels of NO secretion were observed by small macrophages and steadily decreased with increasing size. Hepatic macrophages of different size displayed differences in secretory potential during in vitro culture. The ability of small liver macrophages to secrete NO, TNF‐α, or PGE, following activation with MDP, gradually increased with time in culture. In contrast, large liver macrophages gradually lost their secretory ability after 1–2 days and the intermedicate‐size cells after 2–3 days in culture. This functional heterogeneity in secretory properties among rat liver macrophages of different size is discussed with reference to their potential role and significance in host defense against metastatic tumor growth in the liver.