bluuebunny/crossref_metadata_2025 huggingface dataset:
Enzyme immunoassay of oestrogen receptors in needle biopsies from human liver
Authors: Becker Ulrik , Andersen Jørn , Poulsen Hans Skovgaard , Burcharth Flemming , Gluud Christian , Horn Thomas
Published: August 2010

ABSTRACT— For quantitative assessments of sex hormone receptors in liver tissue, ligand binding assays are inconvenient, as they require large biopsies (0.5–1.0 g). The present study shows that it is possible to measure oestrogen receptors (ER) quantitatively in needle biopsy specimens as small as 10 mg by modifications of a commercial enzyme immunoassay employing monoclonal antibodies. Sucrose gradient centrifugation and the dextran charcoal method served as reference methods. A consecutive series of needle biopsies from patients suspected of liver disease were investigated. The biopsies (n = 37) had a median weight of 14 mg and cytosolic protein concentrations > 1 mg/ml (median 1.28 mg/ml). The median ER concentration was 20 fmol/mg cytosolic protein (range 5 to 57 fmol/mg). The intra‐assay coefficient of variation was 8.9%, the inter‐assay 13.2%, and the detection limit 2.7 fmol/ml cytosol. Women had significantly higher ER concentrations (median 22 fmol/mg) compared to male patients (median 16 fmol/mg) (P = 0.007). The enzyme immunoassay measures ER in liver specimens as small as 10 mg, compared to the large tissue specimens necessary for the conventional DCC assay, and the method is a convenient tool for further studies of ER in routine needle biopsies from the liver.
Changes of cytokeratin filament organization in human and murine Mallory body‐containing livers as revealed by a panel of monoclonal antibodies
Authors: Preisegger Karl‐Heinz , Zatloukal Kurt , Spurej Gerlinde , Denk Helmut
Published: August 2010

ABSTRACT—Mallory bodies (MBs) are characteristic morphologic features of alcoholic hepatitis and can be produced in mouse hepatocytes by chronic griseofulvin (GF) intoxication. The formation of MBs, which share some immunological, biochemical, and ultrastructural features with cytokeratin (CK) filaments of normal liver, is accompanied by derangement and even loss of the CK cytoskeleton of hepatocytes (“empty cells”) as revealed by immunofluorescence microscopy. To clarify whether this diminution or lack of CK‐related staining of MB‐containing hepatocytes was due to loss of CK filaments or changes in antigenicity or accessibility of antigenic determinants immunohistochemical studies using a battery of monoclonal and polyclonal CK antibodies were performed. It could be shown that all these antibodies directed against different CK polypeptide components and antigenic determinants of CKs revealed a highly reduced or even undetectable cytoplasmic CK meshwork in most cells with fully developed large MBs. In the light of our present knowledge of the organization of CK intermediate filaments, these results indicate that the phenomenon of the “empty cells” reflects a diminution of CK meshwork rather than altered antigenic determinants.
Streaming liver VIII: cell production rates following partial hepatectomy
Authors: Zajicek G. , Arber N. , Schwartz‐Arad D.
Published: August 2010

ABSTRACT— Twenty‐four young, female, random‐bred rats weighing 250 g were partially hepatectomized and killed in groups of four animals at the following times: 1 h and 1, 2, 3, 7 and 14 days. One hour before killing, each rat was injected i.p. with 0.5 μCi [3H]‐thymidine, specific activity 5 Ci/mmol/g body weight. Livers were processed histologically and dipped into liquid emulsion for autoradiography. Twenty‐four hours after partial hepatectomy, hepatocyte and littoral labelling indices rose, reaching on the third day respective peak values of 3.7%, and 15.4%, whereupon they declined, remaining slightly above pre‐treatment level. Labelling indices served for cell production estimates. On day 3 the hepatocyte labelling index rose 26‐fold. At the same time hepatocytes doubled their ploidy, indicating that half of the observed L.I. increase was directed to DNA accumulation and not to cell division. The hepatocyte production rate therefore increased 13‐fold (or 1300%). The acinus diameter increased 15%, and cell density declined 5%, so that the acinus capacity to retain cells increased only 5%. Since the acinus did not enlarge proportionally to cell production, it is concluded that 95% of newly formed cells were eliminated. Partial hepatectomy thus triggers two processes: an acute process lasting about a week marked by massive and rapid cell turnover during which most newly formed cells are eliminated; and a second, more protracted process which serves for liver mass restoration. It is proposed that partial hepatectomy induces an acute shortage of a hitherto unknown metabolite that is produced by newly formed cells immediately after hepatectomy.
Measurements of hepatic malondialdehyde and conjugated diene concentrations in 24‐hour endotoxemic rats
Authors: Enochsson Lars B. , Schoenberg Michael , Ware James
Published: August 2010

Abstract: The aim of our study was to analyze if intraperitoneally administered E‐coli endotoxin (1–7 mgkg‐1) causes an increase in liver malondialdehyde (MDA) and conjugated dienes, 24 h postinjection. Blood glucose, white blood cell counts, liver edema and mortality were also measured. The results of our study show that there were no signs of increased MDA or diene production in the liver. To our surprise, blood leukocyte counts of the experimental groups were not statistically different from controls. In the endotoxemic animals there was a slight liver edema which, however, did not influence the measurements of the lipid peroxidation metabolites. Blood glucose levels were decreased in the endotoxemic animals. There was no mortality in our study, perhaps due to the fact that the rats were fed and not starved. We conclude that, in our study, a bolus dose of endotoxin i.p. does not cause any signs of increased liver lipid peroxidation 24 h postinjection. The liver seems, however, to be affected with slight edema and low blood glucose levels.
Significance of septal fibrosis for disturbance of hepatic circulation
Authors: Shibayama Yuro , Nakata Katsuji
Published: August 2010

Abstract: To examine the significance of fibrous septa of the liver for hepatic circulatory disturbance, haemodynamic changes were investigated in rats with septal fibrosis induced with horse serum injections. The fibrotic liver showed thin fibrous bands originating in the vicinity of the peripheral branches of the hepatic vein connected with each other and partly with the portal triads, without conspicuous periportal, pericentral or perisinusoidal fibrosis. There was no evidence of hepatic cell enlargement, disarrangement of hepatic cell plates or narrowing of the sinusoids in the fibrotic livers. Portal vascular resistance, bile production and hepatic oxygen consumption, which were measured by an isolated liver perfusion method, were much the same in the normal and the fibrotic livers. Moreover, there was no significant difference in in vivo blood pressures of the portal vein, the terminal portal venule, the terminal hepatic venule and the inferior vena cava between the normal and the fibrotic rats. These data suggest that septal fibrosis in itself does not disturb hepatic circulation.
Role of septal fibrosis in development of hepatic circulatory disturbance in the presence of liver cell enlargement
Authors: Shibayama Yuro , Nakata Katsuji
Published: August 2010

Abstract: The effect of septal fibrosis on hepatic circulation was examined in rats with enlarged liver cells. Septal fibrosis was produced by horse serum injections and liver cell enlargement by a choline‐deficient diet. Septal fibrosis alone did not induce any disturbance of hepatic circulation. In fatty livers, a slight increase in sinusoidal vascular resistance and a slight elevation of portal vein pressure were found. However, in fatty livers with septal fibrosis, portal hypertension and sinusoidal vascular resistance were higher than in fatty livers without septal fibrosis. These experimental data clearly demonstrate that septal fibrosis alone has no effect on hepatic circulation, but septal fibrosis in the presence of liver cell enlargement markedly affects sinusoidal circulation and induces portal hypertension. The augmentation of sinusoidal vascular resistance by septal fibrosis in the presence of liver cell enlargement might be due to the severe deformation of the sinusoids by enlarged liver cells in the limited spaces surrounded by septal fibrous bands.
Immune response of peripheral blood mononuclear cells to antigenic determinants within hepatitis B core antigen in HB virus‐infected man
Authors: Ishikawa Tetsuya , Kakumu Shinichi , Yoshioka Kentaro , Wakita Takaji , Takayanagi Masahiro , Olido Etsuro
Published: August 2010

Abstract: Residues 72–146 within hepatitis B core Ag (HBcAg) represent T‐cell recognition site in HB‐virus‐infected man. This study was undertaken to define critical residues involved in the immunogenicity of dominant T‐cell determinants of HBcAg. For this purpose, p120–131 and its analog (p120–131 [A] containing alanine substitutions at residues 122 and 125, which were identified as epitopic residues in mice, were synthesized. These peptides and recombinant HBcAg were analyzed for their ability to stimulate peripheral blood mononuclear cells (PBMC) from 25 patients with chronic HBV infection and three patients with acute hepatitis B. PBMC from 18 out of 28 patients showed significantly increased IFN‐γ production and proliferative response in the presence of recombinant HBcAg. Eight patients responded to the two peptides, while 12 patients did not. Four patients responded only to p120–131, and four displayed a response only to p120–131 [A]. The responses to the two peptides were similar among HBeAg‐positive and anti‐HBe‐positive patients, and did not depend on disease activity, except for HBeAg‐positive asymptomatic carriers in whom there was no response to any additive. These results indicate that immune responses to p120–131 and its analog were similar in our patient groups. The dominant epitopic residues in this region of HBcAg may differ between man and mouse.
Percutaneous transluminal angioplasty of an occluded distal splenorenal shunt
Authors: Sariego J. , Wilson A. , Aoyama T. , Sano C. , Kerstein M. , Matsumoto T.
Published: August 2010

ABSTRACT— A distal splenorenal (Warren) shunt was performed on a 39‐year‐old female with bleeding esophageal varices secondary to portal hypertension and cirrhosis. On the twelfth postoperative day, however, she rebled, and angiography revealed that the shunt was occluded. Using a percutaneous approach, successful balloon angioplasty and recanalization was performed. The patient did well and was discharged without further bleeding. Percutaneous transluminal angioplasty (PTA) appears to be effective in dilating occluded splenorenal shunts, obviating a second surgical procedure in high‐risk patients.
Decreased hepatic uptake of cholesterol and retinol in the dimethylnitrosamine rat model of cirrhosis
Authors: Rogers G. W. T. , Dobbs B. R. , Fraser R.
Published: August 2010

ABSTRACT— The fenestrated endothelium of the liver sinusoids forms a sieve between the circulation and hepatocytes. Fenestrae selectively permit the entrance of relatively small chylomicron remnants into the space of Disse to contact hepatocyte receptors, but obstruct the passage of the larger parent chylomicrons. Much of dietary cholesterol and most of retinol are transported as esters in the core of chylomicrons. In the dimethyl nitrosamine rat model of cirrhosis, we have described a rapid reduction in size and number of fenestrae well before the onset of cirrhosis. Concurrent with this decreased porosity is a decreased trapping of radio‐labelled dietary cholesterol and retinol by these livers. We postulate that the less porous “liver sieve” hinders the hepatic uptake of chylomicron remnants, with consequent disturbance of cholesterol and retinol metabolism.
Immunoperoxidase localization of ursodeoxycholic acid in rat biliary epithelial cells. Evidence for a cholehepatic circulation
Authors: Lamri Yamina , Erlinger Serge , Dumont Micheline , Roda Aldo , Feldmann Gérard
Published: August 2010

ABSTRACT— To explain the hypercholeretic effect of ursodeoxycholic acid, a cholehepatic shunt circulation has been postulated. This pathway includes secretion by the hepatocyte into bile and absorption by the biliary epithelial cells. To test this possibility, we have attempted to localize ursodeoxycholic acid in hepatocytes and portal bile duct cells by an indirect immunoperoxidase technique using specific polyclonal antibodies against ursodeoxycholic acid and cholic acid conjugates. Rat livers were fixed with paraformaldehyde, incubated with antibodies directed against bile acids and examined by electron microscopy. After infusion of ursodeoxycholic acid and incubation with antibodies against this bile acid, an electron‐dense staining was observed on the apical (luminal) membrane of the portal bile ductal cells. In contrast, no staining was observed when no ursodeoxycholic acid was infused, or after infusion with taurocholate and incubation with antibodies against cholic acid conjugates. These observations are consistent with absorption of ursodeoxycholic acid by the portal bile ducts and a cholehepatic circulation of this bile acid.
Liver changes in reactive haemophagocytic syndrome
Authors: Tsui W. M. S. , Wong K. F. , Tse C. C. H.
Published: August 2010

ABSTRACT— Hepatomegaly and deranged liver functions are common findings in reactive haemophagocytic syndrome (RHS). We report the findings of 12 fatal cases of RHS in which histological materials of the liver are available for study. The underlying diseases of these patients included lymphoma/leukaemia (6 cases), disseminated undifferentiated carcinoma of the ovary (1 case), disseminated nasopharyngeal carcinoma complicated by tuberculosis (1 case), adenovirus pneumonia (1 case), pneumococcal pneumonia (1 case), typhoid fever (1 case), and possible drug intoxication (1 case). Ten patients had involvement of the liver by the underlying disease process which contributed to the marked hepatic derangement. Non‐specific reactive hepatitis, sinusoidal dilatation and steatosis resulting from systemic or local effects of the associated diseases and the haemophagocytosis also added to the high incidence of liver abnormalities. A diffuse Kupffer cell hyperplasia with haemophagocytosis is characteristic of the syndrome, as all the cases showed increased numbers of bland‐looking histiocytes within the hepatic sinusoids and haemophagocytosis which was moderate to marked in 8 cases and mild in 4. Thus the finding of Kupffer cell hyperplasia with prominent haemophagocytosis in liver biopsy is indicative of an element of RHS and warrants clinical monitoring. Differential diagnoses of haemophagocytosis in liver are also discussed.
Extracellular matrix formation in piecemeal necrosis: immunoelectron microscopic study
Authors: Takahara Terumi , Nakayama Yoshihide , Itoh Hiroyuki , Miyabayashi Chiharu , Watanabe Akiharu , Sasaki Hiroshi , Inoue Kyoichi , Muragaki Yasuteru , Ooshima Akira
Published: August 2010

ABSTRACT— Immunolocalization of Type I, Type III and Type IV collagens, laminin and prolyl hydroxylase (PH), a key enzyme in collagen synthesis, was examined to clarify the fibrotic process in chronic, active liver disease. In piecemeal necrosis of chronic, active hepatitis (CAH) and active liver cirrhosis (LC), fat‐storing cells (FSCs) and transitional cells (TSCs), containing abundant rough endoplasmic reticulum (RER), were increased in number and stained intensely for PH. Immunodeposits of extracellular matrix (ECM) components were found in the RER, Golgi apparatus (GA) and vesicles of these cells, especially in cases with marked inflammation. On the other hand, in the periportal areas of chronic, persistent hepatitis (CPH) or inactive LC, immunoreaction of ECM components was seldom found in the RER of FSCs and TSCs. In the portal tract, immunodeposits of ECM components were seldom found in the organelles of fibroblasts, although ECM was increased there. These findings indicate that FSCs and TSCs in piecemeal necrosis might play a role in the production of ECM components in the progression of fibrosis during the development of chronic active liver disease. In addition, ECM component production by FSCs and TSCs is associated with marked inflammation.
Analysis of lymphoid follicles in liver of patients with chronic hepatitis C
Authors: Hino Keisuke , Okuda Michiari , Konishi Tomomi , Yamashita Aogu , Kayano Kohzoh , Kubota Masafumi , Yasunaga Mitsuru , Fukumoto Yohei , Okita Kiwamu
Published: August 2010

ABSTRACT— Using immunohistological methods, we studied the lymphoid follicles in liver biopsy specimens obtained from patients with chronic hepatitis C(CH–C) to determine whether they represented an autoimmune manifestation. In 84 specimens obtained by liver biopsy from 76 patients positive for C‐100 antibody, we assessed the presence or absence of lymphoid follicles within the portal areas, and then compared the clinical and immunohistological profiles of the two groups defined on this basis. There were no significant differences in the serological and clinical profiles of the two groups, but the T4:T8 cell ratio within the portal areas differed significantly. A T4:T8 cell ratio > 1.0 was more common in the specimens containing lymphoid follicles. This finding resembled the distribution of T cell subpopulations in the portal areas of patients with autoimmune hepatitis, and suggested that lymphoid follicles may be worth investigating to elucidate the relationship between chronic hepatitis C and autoimmune hepatitis.
Arterial blood ketone body ratio as a possible indicator for predicting fulminant hepatitis in patients with acute hepatitis
Authors: Saibara Toshiji , Onishi Saburo , Maeda Takashi , Yamamoto Yasutake
Published: August 2010

ABSTRACT— Encephalopathy and severe coagulopathy in patients with acute hepatitis (AH) are good markers for the diagnosis of fulminant hepatitis (FH), which occurs in only about 1% of AH patients. However, even if patients show severe coagulopathy, it is quite difficult to predict FH before the onset of encephalopathy. The ratio of acetoacetate/beta‐hydroxybutyrate in arterial blood (KBR) has been reported to reflect the cellular energy charge level in hepatocytes. In our previous report, KBR was quite low in FH patients and was an excellent marker for predicting the prognosis. KBR of normal subjects is distributed in a range of 1.0–2.1 (1.54 ± 0.26, mean ±SD). In this study, we assessed KBR serially in 15 AH patients with severe coagulopathy (hepaplastin test (HPT) < 40%), including seven patients who developed FH, to see if we could predict FH by using KBR as a marker. Seven patients with KBR < 0.6 of long duration (4 days or more) were complicated with hepatic encephalopathy (HE) and it took 3 or more days of KBR below 0.6 before HE appeared. The other eight patients with KBR < 0.6 of short duration (less than 4 days) were not complicated with HE. These data suggest that AH patients with HPT < 40% and a 3‐day duration of KBR < 0.6 are at serious risk of FH.
Peripheral blood lymphocyte subsets in patients with chronic hepatitis C – effects of interferon treatment
Authors: Wejstàl Rune , Norkrans Gunnar , Lenkei Rodiça , Weiland Ola , Schvarcz Robert , Frydén Aril
Published: August 2010

Abstract: Thirty‐three patients with chronic hepatitis non‐A, non‐B/C were included in a randomized controlled study of recombinant alpha‐2b interferon treatment 3 MU three times weekly for 36 weeks. In lysed whole blood, lymphocyte subpopulations were enumerated by flow cytometry detecting fluorescein or phycoerytrin conjugated monoclonal antibodies directed against seven different epitopes. Patients with chronic active hepatitis were significantly older than patients with chronic persistent hepatitis (p>0.05). Before treatment, the proportions of different subsets of lymphocytes were within the normal reference values and the CD4/CD8 ratio was also normal. No increased activation of T‐cells was noticed. Patients over 50 years of age, however, had a significantly increased (p>0.01) proportion of HLA‐DR + lymphocytes, mainly B‐cells. Treatment decreased the absolute number of peripheral blood leukocytes and lymphocytes. There was also a significant decline in the proportion of CD8 + lymphocytes and NK‐cells, and a significant increase in the proportion HLA‐DR + cells and of the CD4/CD8 ratio. The increased proportion of HLA‐DR + cells, however, did not reflect peripheral T‐cell activation; instead, it was due to increasing B lymphocyte numbers.
Haemodynamic alterations and their morphological basis in biliary obstruction
Authors: Shibayama Yuro , Nakata Katsuji
Published: August 2010

Abstract: The relationship between the haemodynamic alterations and morphological changes in the liver caused by biliary obstruction was investigated in rats after ligation of the common bile duct. In these rats, the portal vein pressure was markedly elevated, and the differences in blood pressure between the portal vein and the terminal portal venule and between the terminal portal venule and the terminal hepatic venule were greater than in the sham‐operated rat. The livers showed narrowing of the most peripheral branches of the portal vein due to compression by proliferated bile ductules and sinusoidal stenosis due to enlarged liver cells, but there was no perceptible change in the hepatic vein branches. These data suggest that hepatic circulatory disturbance in biliary obstruction is caused by deformation of the peripheral portal vein branches and sinusoidal stenosis.
An enzyme‐linked (alkaline phosphatase) oligonucleotide probe for the detection of serum hepatitis B virus DNA
Authors: Garcia F. G. , Quiros E. , Bernal M. C. , Salmeron J. , Maroto M. C.
Published: August 2010

Abstract: Serum HBV‐DNA detection represents a new and important diagnostic tool for the hepatologist, as well as a prognostic and therapeutic guide. Though most laboratories use genomic radioactive probes, the appearance of commercially available enzyme‐linked oligonucleotide probes, which simplify the method, represents an advance in HBV‐DNA detection. In order to verify the value of an enzyme‐linked (alkaline phosphatase) oligonucleotide probe in HBV‐DNA detection, we have first compared the results of the detection of serum HBV‐DNA in 46 patients at different serological stages of HBV infection using this probe and with those obtained using a genomic P32‐labeled probe. Fisher's test analysis of the results shows statistical significance; this indicates a high correlation between methods. Secondly, we have tested 296 sera of patients at different serological stages of HBV infection with the enzyme‐linked probe; we have found HBV‐DNA in 8 patients with acute hepatitis (61.54%), 61 HBeAg (70.12%), and 7 anti‐HBe (12.07%) chronic hepatitis. These results show the value of this probe for routine survey of serum HBV‐DNA using a simple method which avoids the use of radioactivity.
Segmental mean transit times of 99m technetium within central vascular space in liver cirrhosis
Authors: Hartleb Marek , Rudzki Krystian
Published: August 2010

Abstract: Mean transit time (MTT) through a given vascular space is closely related to the effective blood volume in this compartment. Central vascular blood volume in liver cirrhosis is believed to be reduced, but more precise data on the location of the underfilled vascular area are lacking. 99mTechnetium first‐pass angiography was performed in 15 cirrhotic patients and in 10 age‐matched normals. The method of segmental analysis of MTT was validated by performing an analog study in a plastic tubing/chamber system. The mean tracer sojourn in the central circulation was described in cirrhotic patients (10.25 ± 2.17 s vs 12.92±2.88 s; p>0.05); however, this finding was not observed beyond cardiopulmonary circulation (MTT between right and left heart chambers). Segmental comparative analysis between cirrhotics and normals revealed significant MTT differences in two vascular subcompartments, i.e. right chamber – pulmonary artery, and within the lung vascular bed. Analysis of time activity lung curves in patients with cirrhosis disclosed a shorter time to peak and more rapid washout of the tracer from this area, without any change in curve symmetry as compared to normals. Ascites had no apparent impact on MTT rate, and cirrhotics with most advanced disease (grade C Child‐Pugh) had longer MTT through the cardiopulmonary circulation as compared to combined groups A & B (7.32 ± 0.13 s vs 6.03 ± 1.23 s; p > 0.05). Our data provide further evidence for contraction of the cardiopulmonary vascular space in liver cirrhosis.
Quantitative analysis of activated Kupffer cells in viral hepatitis: application of computer image analysis for lectin histochemistry
Authors: Yamamoto Kazuhide , Makino Yasuhiro , Yoshioka Toshifumi , Kobashi Haruhiko , Tomita Minoru , Tsuji Takao
Published: August 2010

Abstract: Lectin histochemistry revealed that Kupffer cells in the normal liver bound lectins such as Concanavalin A (Con A), Ricinus communis agglutinin (RCA) and Wheat germ agglutinin (WGA), but did not bind Peanut agglutinin (PNA), Dolichos biflorus agglutinin (DBA), Ulex europaeus agglutinin I (UEA‐I) or Soybean agglutinin (SBA). Kupffer cells in viral liver diseases, however, bound the PNA lectin and the binding was specific to Kupffer cells in liver parenchyma. Computer image analysis was performed using light micrographs of sections stained with immuno‐peroxidase and diaminobenzidine (DAB). The dark brown area of reaction products was detected by analyzing each color component (red, green and blue) in the picture and was expressed as the percent area in the parenchyma. Quantitative analysis revealed the percent area occupied by Kupffer cells positive for the PNA lectin was as follows: acute hepatitis, 2.83 ± 0.74; chronic persistent hepatitis, 2.51 ±0.88; chronic aggressive hepatitis, activity moderate and severe, 4.71 ± 2.23 and 3.45 ± 1.84; and liver cirrhosis, 1.96 ± 0.99. The percent area of Kupffer cells was significantly higher in CAH2A than that in chronic persistent hepatitis or in liver cirrhosis. These results suggest that the PNA lectin could be used as a marker for activated Kupffer cells and that activated Kupffer cells were increased in volume in chronic aggressive hepatitis.
In situ hybridizaiton in Viral hepatitis
Authors: Nagro F. , Pacchiionl D. , Meadardini A. , Bussolati G. , Bonine F.
Published: August 2010

Abstract: In situ hybridizaiton (ISH) is a sensitive and specific technique for detecting nucleic acids in intact cells. Visualization of the target sequences by autoradiography or immunohistochemistry allows their precise subcellular localization and quantitation. The application of ISH techniques has contributed to the understanding of the complex replicative cycle of hepatitis B virus. More recently, hepatitis delta and C virus replication has also been studied by this technique. ISH‐based assays have finally been used to follow the replication of cytomegalovirus within the transplanted liver. Although ISH is a powerful tool for the molecular biologist, its clinical significance in the diagnosis and prognosis of human I hepatitis virus infections has yet to be fully evaluated.
Use of sulphonated probes for detecting human immunodeficiency virus‐1 trascripts by in situ hybridizaation
Authors: Pezzella Marle , Vonesch Nicoletta , Sturchie Elena
Published: August 2010

Abstract: A detailed procedure is described that allows detection of the presence of human immunodeficiency virus‐1 (HIV‐1) transcripts within both acetone‐fixed tisseues and peripheral blood mononuclear cells. This assay uses cDNA probes labelled by a non‐procedure that results in the modification of cytosine residues trhough covalent linkage to a sulophone group. In sit hybridized probe is than detected by an alkaline phostphatase‐conjugated antibody specifically directed againsts the sulphone hapten. This procedure is specific, rapid and safe and can be applied in the research as well as in the clinical pathology settings.
Localization of cloned human DNA sequences and analysis of chromosomal alteration by in situ hybridization
Authors: Patracchini P. , Aiello V. , Marchetti G. , Palazzi P. , Ferrati M. , Calzolari E. , Bernardi F.
Published: August 2010

Abstract: The in situ hybridization technique was used for the localization on human chromosomes of single‐copy and repeated sequences and, in addition, for the characterization of altered human chromosomes. Two anonymous clones, single or low‐copy, obtained from a human X chromosome library were localized on the distal part of the long arm and in the paracentromeric region of X chromosome, respectively. A genomic fragment of the single‐copy thyroglobulin (TG) gene was used to confirm the localization on the distal part of the long arm of chromosome 8. The localization and distribution on human chromosomes of the glyceraldehyde‐3‐phosphate dehydrogenase (GAPD) multigene family obtained by in situ hybridization and by somatic cell hybrids were compared. A phosphoglycerate kinase (PGK) c‐DNA clone, which detects genic and pseudogenic sequences on the X chromosome, was used for the characterization of three small ring markers present in unrelated female patients.
Genetic variants of alpha‐1‐antitrypsin (AAT)
Authors: Fabbretti Glovanna , Sergl Censolate , Consales Glacomo , Faa Gavino , Brisigotti Massimo , Romeo Glovannl , Callea Francesco
Published: August 2010

Abstract: This paper reviews the genetic variants of alpha‐1‐antitrypsin (AAT) which have been sequenced with special emphasis on the s.c. deficiency variants. These result in AAT low plasma levels via three main mechanisms: 1) intracellular storage; 2) intracellular degradation; 3) lack of synthesis. Intracellular storage occurs with the classical Z variant and with a few variants called M‐like, because of their isoelectric focusing (IF) pattern. The storage phenomen causes liver damage and can be demonstrated at both light and electron microscopic level with the help of immuno‐histochemistry. We report a new deficiency variant of AAT (M‐Cagliari) characterized by very low plasma levels, massive storage of AAT and liver cirrhosis. By using immunohistochemical techniques and DNA analysis we could demonstrate that M‐Cagliari has antigenic and genetic properties other than the Z AAT.
Hepatic arterial and portal venous oxygen content and extraction in liver cirrhosis
Authors: Sezai Shuichi , Sakurabayashi Shin , Yamamoto Yoshihire , Morita Toshikazu , Hirano Masanori , Oka Hiroshi
Published: August 2010

ABSTRACT— We examined the oxygen content in the hepatic arterial, hepatic venous and portal venous blood to evaluate the oxygen supply to the liver and hepatic oxygen extraction in cirrhosis. The arterial‐portal venous difference of the oxygen content was within the normal range in cirrhosis patients, although the oxygen content of the hepatic artery and portal vein was lower than in the control patients. The hepatic venous oxygen content was normal in the cirrhosis patients. The oxygen tension and saturation were always higher in the splenic vein than in the other branches of the portal system. Oxygen was supplied chiefly by the hepatic artery, and arterial oxygen extraction was normal in cirrhosis. In addition, there was no change in arterial extraction during oxygen inhalation by cirrhosis patients. Portal venous oxygen extraction was decreased in cirrhosis and was increased by oxygen inhalation. These findings indicate the autoregulation of hepatic oxygen through a mutual relationship between the hepatic arterial and the portovenous oxygen supply.
Effects of prednisolone pretreatment in interferon alfa therapy for patients with chronic non‐A, non‐B (C) hepatitis
Authors: Liaw Yun‐Fan , Sheen I‐Shyan , Lin Shi‐Ming , Chen Tong‐Jong , Chu Chia‐Ming
Published: August 2010

ABSTRACT— The effect of corticosteroid pretreatment on interferon alfa treatment of chronic non‐A, non‐B or C hepatitis was evaluated in an open, randomized, controlled study. Normalization of the serum alanine aminotransferase level, which suggested complete response, was recorded in 14 (56%) of 25 patients at the end of a 6‐month treatment with 3 million units of interferon alfa three times weekly alone. Prednisolone withdrawal followed by the same interferon alfa treatment resulted in complete response in 14 (61%) of another 23 patients. Both of these rates are significantly higher than the spontaneous ALT normalization rate of 9% observed in 2 of 22 patients during the same period (p<0.01). ALT normalized within 2 months of interferon therapy in most (89%) of the complete responders. Of the patients pretreated with prednisolone, 43% showed a “rebound” following prednisolone withdrawal and 89% of the patients with “rebound” responded to subsequent interferon therapy. The cumulative probability of sustained remission in patients with and without prednisolone pretreatment in a 24‐month period of follow‐up after the end of therapy was 79% and 30%, respectively (p<0.05). Clinical relapse mostly (85%) occurred within 3 months after the end of therapy. No obvious factor could be identified as a predictor of response or relapse. The results of this study suggest that prednisolone withdrawal tends to enhance the durability of the beneficial effects of interferon alfa therapy in patients with chronic non‐A, non‐B or C hepatitis.
Neutralizing antibodies to recombinant alpha‐interferon and response to therapy in chronic hepatitis C virus infection
Authors: Milella Michele , Antoneill Guido , Santantonio Teresa , Currenti Maurizio , Monno Laura , Mariano Nicola , Angarano Gioacchino , Dianzanl Ferdinando , Pastors Giuseppe
Published: August 2010

ABSTRACT— Forty‐seven patients with chronic hepatitis C were treated with recombinant interferon alpha‐2a (rIFNα2a) given subcutaneously in a standard dose of 3 MU thrice weekly for 12 months. Stored baseline sera and monthly samples during treatment were assayed for anti‐interferon neutralizing antibodies using the antiviral neutralization bioassay against 5 IU of rIFNα2a. During therapy, 15 of 47 patients (31.9%) developed detectable levels of neutralizing antibodies within 2–8 months after starting treatment. After 12 months of therapy, 26 of 32 antibody‐negative patients (81.3%) showed normalization or marked reduction of ALT levels compared to 4 of 15 (26.6%) who developed anti‐IFN neutralizing antibodies (p = 0.0009). Four patients demonstrated antiviral response during treatment even in the presence of low levels or late occurrence of neutralizing antibodies. Six of the seven patients who had disease reactivation after an initial response developed high titers of neutralizing antibodies. Our results suggest that reactivation of chronic hepatitis C before completion of therapy seems to be an obvious consequence of anti‐IFN neutralizing antibody formation.
Histological changes of the liver in experimental graft‐versus‐host disease across minor histocompatibility barriers. VII. A light and electron microscopic study of the large bile duct
Authors: Nonomura Akitaka , Kono Naoko , Mizukami Yuji , Nakanuma Yasunl
Published: August 2010

ABSTRACT— Although morphologic changes of the intrahepatic bile ducts in graft‐versus‐host disease (GVHD) have been well studied, those of the large extrahepatic bile ducts in the porta hepatis or common bile ducts have not been so well elucidated. In the present study, pathologic changes of the extrahepatic bile ducts in experimental mouse GVHD across minor histocompatibility barriers were examined up to 14 months after transplantation. Mononuclear cell infiltration was most striking around 2 weeks after transplantation. Although it gradually decreased, infiltration persisted during the entire period of observation. Fibrous thickening and sclerosis of the bile duct wall continued over time following transplantation, especially 3 months after transplantation. The appearance was similar to sclerosing cholangitis, but obliteration of the lumen was not demonstrated. Electron microscopically, the bile duct epithelial layer was frequently infiltrated by lymphocytes, and often accompanied by polymorphonuclear leukocytes, monocytes, and rarely by plasma cells. The epithelial cells in close contact with and in the vicinity of these cells showed a variety of degenerative changes. These results suggest that not only the interlobular and/or small bile ducts but also the large hilar and extrahepatic bile ducts are involved in hepatic GVHD, and thus bile duct injury in GVHD may occur along the full length of the biliary tree.
Cytotoxic activity of spleen‐derived T lymphocytes against autologous biliary epithelial cells in autopsy patients with primary biliary cirrhosis
Authors: Onishi Saburo , Saibara Toshiji , Nakata Sbusaku , Maeda Takashi , Iwasaki Shlnji , Iwamura Shinichi , Miyazaki Masako , Yamamoto Yasutake , Enzan Hideaki
Published: August 2010

ABSTRACT— Autoimmunity against biliary epithelial cells is considered to be involved in the pathogenesis of primary biliary cirrhosis (PBC). However, cytotoxic activity of T lymphocytes against biliary epithelial cells has not previously been examined. This study has demonstrated that spleen‐derived T lymphocytes were cytotoxic for autologous biliary epithelial cells in all of five patients with PBC, even though it was only detectable at high effector to target ratios. Such cytotoxicity was not found in non‐PBC patients. CD8‐positive T lymphocytes were shown to be responsible for the cytotoxicity by negative selection, and its inhibition was dependent on the ratio of cold to hot target cells. These observations may support a current hypothesis that the pathogenesis of PBC is partly due to T cell autoimmunity directed against the bile duct epithelium.
Bile and plasma lipid composition in non‐obese normo‐lipidemic subjects with and without cholesterol gallstones
Authors: Halpern Zamir , Rubin Moshe , Harach Gideon , Grotto Ittamar , Mosor Asher , Dvir Alisa , Lichtenberg Dov , Gilat Tuvia
Published: August 2010

ABSTRACT— A two‐stage study was carried out to characterize the bile and plasma lipid composition in normolipidemic non‐obese patients with and without cholesterol gallstones. The first stage involved 11 patients with cholesterol gallstones admitted for elective cholecystectomy and a control group of 16 patients without cholesterol gallstones undergoing elective laparotomy. Bile samples were obtained intraoperatively by aspiration from the gallbladder. The bile of all the gallstone patients was supersaturated with cholesterol and its nucleation time was much shorter than that of bile in the control group (2.5 days vs 22.5 days, respectively, P<0.001). The biliary fatty acid profile of phosphatidylcholine (PC) and free fatty acids (FFA) of gallstone patients was similar to that of the control group. C‐22 fatty acids were found in a higher concentration in the FFA than in the PC fatty acids (P<0.05) in both groups of patients. Plasma triglyceride levels in the gallstone patients were significantly higher than those in the control group and the biliary cholesterol level correlated with that of plasma triglycerides. In the second stage of the study, plasma lipid profiles were obtained in two additional groups of patients, 20 patients with and 24 patients without cholesterol gallstones, for an in‐depth characterization of the differences in plasma lipid profiles. The gallstone patients were found to have not only significantly higher concentrations of plasma triglycerides but increased cholesterol and phospholipid level as well. These differences were essentially due to a higher lipid content of the plasma VLDL fraction, similar to the pattern of patients with type IV hyperlipoproteinemia.
Taurine conjugate of ursodeoxycholate plays a major role in the hepatoprotective effect against cholestasis induced by taurocheno‐deoxycholate in rats
Authors: Tsukahara Kosho , Kanal Setsuko , Ohta Minora , Kitani Kenichi
Published: August 2010

ABSTRACT— Rats which were taurine‐deprived through ß‐alanine administration and untreated rats were used to elucidate the mechanism of hepatoprotective effects of ursodeoxycholate (UDC). Animals were infused with taurochenodeoxycholate (TCDC, 0.4 μmol · min‐1 · 100 g‐1) alone or in combination with tauroursodeoxycholate (TUDC), or with UDC (both 0.6 μmol · min‐1 · 100 g‐1) for 2 h. Ursodeoxycholate as well as TUDC prevented severe cholestasis and liver damage induced by TCDC infusion in both untreated and taurine‐deprived rat groups. In untreated rats, however, UDC was less effective in hepatoprotection than TUDC as indicated by sequential changes in biliary LDH output during the period of 30 to 120 min (P<0.05). In rats receiving UDC and TCDC, total biliary output of LDH for 2 h was significantly higher in taurine‐deprived rats than that in the control (73.40±10.10 vs 41.14±12.56: P < 0.05), suggesting that the difference became greater upon taurine deprivation. In contrast, in rats receiving TUDC and TCDC, the protective effect was comparable for the taurine‐deprived and untreated rats. When the animals were infused with UDC and TCDC, taurine‐deprived rats exhibited a biliary excretion rate for TUDC half that of control rats (P<0.05). Furthermore, a highly significant correlation was observed between the biliary excretion rate of TUDC and biliary output of LDH (r = –0.886, P<0.0001). These results suggest that UDC conjugates, especially TUDC, and not UDC may play a major role in the prevention of cholestasis and liver cell damage caused by TCDC infusion.
Fine‐needle liver biopsy in patients with severely impaired coagulation
Authors: Caturelli Eugenio , Squillante Maria Maddalena , Andriulli Angelo , Siena Domenico Angelo , Cellerino Caterina , De Luca Francescantonio , Marzano Maria Adelaide , Pompili Maurizio , Rapacclni Gian Ludovico
Published: August 2010

ABSTRACT— Severe coagulation defects, as reflected by platelet count and prothrombin time, have always been considered a contraindication to needle biopsy of the liver, but there are very limited data on the actual rate of bleeding in patients with such severe alterations and none whatsoever on the bleeding risk associated with newer, fine‐gauge needles that produce less trauma to the liver tissue. In addition, there has never been any evidence that platelet count and/or prothrombin time are the most sensitive indices of bleeding risk. This retrospective study of 85 patients, with platelet counts less than 50 000/mm3 and/or prothrombin times less than 50% of controls, subjected to ultrasound‐guided fine‐needle liver punctures for diagnostic or therapeutic (percutaneous ethanol injection) purposes showed no bleeding episodes after any of the 229 punctures performed. No type of replacement therapy was administered to correct clotting defects prior to the procedure. Correct pathologic diagnoses were obtained in 81.2% of all patients. Ultrasound‐guided fine needle puncture appears to be safer than currently believed in patients with severe clotting defects and deserves further evaluation as an alternative to surgical procedures to diagnose and treat liver lesions, even when severe coagulation impairment is present.
Liver function and morphology during long‐term fatty acid supplementation in cystic fibrosis
Authors: Strandvik Birgitta , Hultcrantz Rolf
Published: August 2010

Abstract: Liver function tests and liver morphology were studied in 18 patients with cystic fibrosis (CF) for a period of 3 years. The patients were 5–19 years of age at the start of the study. Nine patients received regular supplementation of essential fatty acids (Intralipid®) and nine sex‐ and age‐matched patients, with as similar clinical status as possible but without fat emulsion treatment, were followed as controls. There were no significant differences between the two groups with regard to the parameters of liver function, including determinations of bile acids in serum and urine. Histological examinations of liver biopsies suggested less progression of liver disease in the fatty acid supplemented group, including fatty infiltration of the hepatocytes, evaluated morphometrically. The results show that regular supplementation of fat emulsions has no negative effects in the liver of patients with CF, and the lessening of progression of liver damage suggests that essential fatty acid deficiency might contribute to the liver damage in this disease.
Effects of vasopressin on haemodynamics in portal hypertensive rats receiving clonidine
Authors: Moreau Richard , Cailmail Stéphane , Lebrec Didier
Published: August 2010

Abstract: The effects of clonidine, vasopressin or a combination of both substances on splanchnic and systemic haemodynamics were measured in conscious rats with portal vein stenosis. Clonidine alone significantly decreased portal pressure, portal tributary blood flow and cardiac index, but did not change arterial pressure. Vasopressin alone significantly increased arterial pressure and significantly decreased portal pressure, portal tributary blood flow and cardiac index. Changes in portal tributary blood flow, arterial pressure and cardiac index were significantly higher with vasopressin than with clonidine alone. Vasopressin infusion in rats pretreated with clonidine significantly increased arterial pressure and significantly decreased portal pressure, portal tributary blood flow and cardiac index. Changes in arterial and portal pressures and portal tributary blood flow were significantly higher with combined therapy than with clonidine alone. Changes in arterial and portal pressures and portal tributary blood flow did not differ between combined therapy and vasopressin alone. Changes in cardiac index were significantly higher with combined therapy than with clonidine or vasopressin alone. Hepatic artery blood flow was not affected by either clonidine or vasopressin but significantly declined with combined therapy. In conclusion, this study suggests that a combination of vasopressin and clonidine accentuates the portal hypotensive action of clonidine but not the action of vasopressin. Moreover, this study suggests that a combination of vasopressin and clonidine may have deleterious effects on systemic and hepatic artery vascular beds.
Preservation of human liver grafts in UW solution. Ultrastructural evidence for endothelial and Kupffer cell activation during cold ischemia and after ischemia‐reperfusion
Authors: Carles Jacques , Fawaz Rabia , Hamoudi Nour Eddine , Neaud Veronique , Balabaud Charles , Bioulac‐Sage Paulette
Published: August 2010

Abstract: Biopsies taken from 13 human liver grafts at different stages of the transplantation process were used for study of the morphology of sinusoidal cells prior to harvesting (5 biopsies), after preservation in UW solution (10 biopsies), and after complete revascularization (13 biopsies). The mean cold ischemic period was 12 h 30. Immediate follow up was uneventful and the mean peak of post‐operative transaminases below 1300 IU/1. Biopsies were perfusion‐fixed by the transparenchymal route to ensure satisfactory ultrastructural results. There were no loose sinusoidal endothelial cells in the lumen and no signs of cellular death. Some endothelial cells presented signs of activation at the end of the preservation period, and even more after revascularization, with numerous lucent vacuoles resembling endosomes in the cytoplasm. Kupffer cells also presented signs of activation, particularly after reperfusion. The retraction of endothelial cell processes which formed large gaps during cold ischemia proved to be partly reversible after reperfusion. Signs of endothelial cell damage with gaps and partial rupture of the plasmic membrane were also observed, particularly after revascularization, in areas which contained numerous inflammatory cells adhering to the wall. The Disse space was not generally enlarged and contained no inflammatory cells. The sinusoidal pole of hepatocytes was occasionally damaged with the formation of blebs. These results strongly suggest that any drug or perservation solution that will inhibit endothelial and Kupffer cell activation could be beneficial in the prevention of preservation and reperfusion injury.