bluuebunny/crossref_metadata_2025 huggingface dataset:
Two-locus involvement in the association of human leukocyte antigen with the extrahepatic manifestations of autoimmune chronic active hepatitis
Authors: Makcos Yanina , Fainboim Hugo A. , Capucchio Mónica , Findor Jorge , Daruich Jorge , Reyes Beatriz , Pando Marcelo , Theiler Graciela Del C. , Méndez Nora , Satz Leonardo M. , Fainboim Leonardo
Published: March 2007

We investigated the association of human leukocyte antigen antigens and type 1 chronic active “autoimmune” hepatitis in a population of 65 white Argentinian patients, taking into account the different manifestations of the disease. Standard microlymphocytotoxicity was used for human leukocyte antigen A, B, C, DR and DQ typing. Human leukocyte antigen class 2 alleles were also typed on genomic DNA by means of polymerase chain reaction amplification and hybridization to sequence specific oligonucleotides. A primary association with human leukocyte antigen DR4 was present (human leukocyte antigen DR4: 44% in patients vs. 29% in controls; χ2, 5.6; p=0.02, relative risk, 2.1). However, a novel association was observed with human leukocyte antigen All (31% in patients vs. 6% in the controls; χ2, 25.3; corrected p=0.001; relative risk, 6.8). Moreover, of the 20 human leukocyte antigen All patients, 18 had extrahepatic manifestations associated with autoimmune chronic active hepatitis. This represented 60% of the patients bearing this form of the disease (n=30), conferring a relative risk of 22.2 (χ2, 46.3; corrected p=0.00008). In this group, human leukocyte antigen DR3 and DR4 had a weak association. When present together, human leukocyte antigen DR4 and human leukocyte antigen All had a synergistic effect, yielding an odds ratio of 357. Statistical analysis and family segregation studies suggest that the two loci products may represent independent risk factors for this form of autoimmune chronic active hepatitis. This synergistic effect was not evident with All plus DR3. In autoimmune chronic active hepatitis patients without extrahepatic manifestations, a weak association with human leukocyte antigen DR6 was found. Interestingly, in autoimmune chronic active hepatitis of childhood (in which extrahepatic manifestations are seldom observed) a strong association with human leukocyte antigen DR6 was recently observed in patients from the same ethnic group and geographic region. The clinical and genetic heterogeneity observed in this study may explain the weak human leukocyte antigen associations reported previously for autoimmune chronic active hepatitis and suggest that the extrahepatic forms in patients with autoimmune chronic active hepatitis represents a separate clinical entity. (Hepatology 1994;19:1371-1374.)
Immunohistochemical detection of chloride/bicarbonate anion exchangers in human liver
Authors: Martínez-Ansó Eduardo , Castillo José E. , Díez Javier , Medina Juan F. , Prieto Jesús
Published: March 2007

Sodium-independent Cl−/HCO3
− exchange activity has been observed in isolated rat hepatocytes and intrahepatic bile duct epithelial cells, where it is involved in intracellular pH regulation and, possibly, biliary bicarbonate secretion. Monoclonal antibodies to the membrane domain of human chloride/bicarbonate anion exchanger proteins, AE1 and AE2, were prepared so that we might determine by immunohistochemical methods the presence and location of these antiporters in the human liver. To obtain the antibody against AE1, we immunized mice with injections of washed human erythrocytes. The selected monoclonal antibody was found to be specific for the 17-kD proteolytic membrane fragment of AE1 protein. The antibody to AE2 was produced with a 14-mer synthetic peptide, whose sequence corresponds specifically to amino acid residues 871 to 884 in the deduced primary structure of human kidney AE2 protein. When the monoclonal antibody to AE2 peptide was employed for the immunohistochemical study of liver specimens (by both immunofluorescence and immunoperoxidase), a clearly defined staining was present at the canalicular membrane of hepatocytes, as well as the luminal side of the membrane of bile duct epithelial cells from small and medium-sized bile ducts. No staining was observed in the liver parenchyma with the monoclonal antibody to AE1, which instead strongly decorated the erythrocytes in liver blood vessels. We conclude that AE2 immunoreactivity is present in human liver, where it localizes very specifically to the membrane regions, which appear most probably involved in the transport of bicarbonate to bile (i.e., the canalicular membrane of hepatocytes and the apical side of epithelial cells of small and medium bile ducts). (Hepatology 1994;19:1400-1406.)
Nonsuppurative cholangitis is induced in neonatally thymectomized mice: A possible animal model for primary biliary cirrhosis
Authors: Kobashi Haruhiko , Yamamoto Kazuhide , Yoshioka Toshifumi , Tomita Minoru , Tsuji Takao
Published: March 2007

Neonatally thymectomized mice are unique in that they are prone to organ-specific autoimmune diseases. We investigated whether autoimmune cholangitis could be induced in these mice when they were immunized with biliary antigens. Neonatally thymectomized A/J mice were immunized with porcine intrahepatic bile duct epithelial cells (group 1), porcine gallbladder epithelial cells (group 2), porcine splenocytes (group 3) or Freund’s adjuvant (group 4). Nonthymectomized mice were immunized with bile duct epithelial cells (group 5) or Freund’s adjuvant (group 6). The cell suspensions were injected intraperitoneally with Freund’s adjuvant once a week for 8 wk. In group 1 varying amounts of mononuclear cells infiltrated around the bile duct in 14 of 22 mice, whereas little or no accumulation was noted in other groups. Ultrastructural observations revealed that the inflammatory cells consisted of lymphocytes, plasma cells and macrophages. The bile duct showed degenerative changes and some lymphocytes infiltrated between bile duct epithelial cells. An immunohistochemical study showed that the accumulated lymphocytes consisted of CD4+ and CD8+ T cells, as well as B cells. Both major histocompatibility complex class I and class II antigens were expressed on bile duct epithelial cells. Antimitochondrial antibody was demonstrated in some mice in groups 1 (9 of 17), 2 (2 of 3) and 5 (4 of 5) by immunofluorescence; the antibody reacted with the 68, 52 and 47 kD polypeptides of the pyruvate dehydrogenase complex on Western blotting. These findings suggest that autoimmune cholangitis can be induced in neonatally thymectomized mice stimulated with biliary antigens and that these mice could be a suitable animal model for primary biliary cirrhosis. (Hepatology 1994;19:1424-1430.)
Ammonia-induced brain edema and intracranial hypertension in rats after portacaval anastomosis
Authors: Blei Andres T. , Olafsson Sigurdur , Therrien Guy , Butterworth Roger F.
Published: March 2007

Brain edema, leading to intracranial hypertension and brain herniation, is a major cause of death in fulminant liver failure. Astrocyte swelling is a prominent neuropathological feature in experimental fulminant liver failure. It has been postulated that the osmotic effects of glutamine, generated in astrocytes from ammonia and glutamate in a reaction catalyzed by glutamine synthetase, could mediate brain swelling. Normal rats and rats that received a portacaval anastomosis were infused with ammonium acetate or a sodium acetate control; brain water in cerebral cortex was measured with the gravimetry method, intracranial pressure by means of a cisterna magna catheter and cortical amino acids using high-performance liquid chromatography. Although brain edema was detected in both groups receiving ammonia, it was of a greater magnitude in portacaval anastomosis rats (80.94%+0.17% vs. 80.24%+0.09%, p<0.01), resulting in the development of intracranial hypertension. When portacaval anastomosis rats were infused with ammonium acetate and pretreated with 150 mg/kg methionine-sulfoximine, an inhibitor of glutamine synthetase activity, brain edema was ameliorated and intracranial pressure did not rise. A dose-dependent reduction in brain glutamine levels was seen with increasing doses of methionine-sulfoximine; however, brain edema did not decrease beyond the 150 mg/kg dose, suggesting that the increase in brain water was not solely a result of glutamine accumulation. We conclude that brain edema of a magnitude that results in intracranial hypertension is more likely to develop in rats after portacaval anastomosis receiving a continuous ammonia infusion. The osmotic effects of glutamine appear to mediate, but only in part, the increase in brain water seen in this preparation. Other mechanisms in addition to inhibition of glutamine synthesis may mediate the beneficial effects of methionine-sulfoximine seen in this study. (Hepatology 1994;19:1437-1444.)
Effect of irradiation on hepatic natural killer cells
Authors: Vanderkerken Karin , Bouwens Luc , Kuppen Peter , De Neve Wilfried , Van Den Berg Kit , Baekeland Marijke , Wisse Eddie
Published: March 2007

The rat liver contains a population of natural killer cells consisting of two morphologically and functionally different subsets, a low-density and a high-density fraction. In this work we describe the influence of low-dose radiation on hepatic natural killer activity. The effect on the cytotoxicity against YAC-1 lymphoma and CC531 colon adenocarcinoma tumor cells was measured in chromium-51 assays, and morphological changes were analyzed by means of electron microscopy. The low-density natural killer fraction showed increased cytotoxicity against YAC-1 which was associated with an increased binding of natural killer cells to the YAC-1 tumor cells shortly after irradiation. These phenomena were paralleled by an increased number of multivesicular bodies and cytoplasmic granules with an electron-lucent halo. In contrast, the other hepatic natural killer cell fraction, the high-density natural killer cells, did not show increased cytotoxicity, binding or morphological alterations. The radiation-stimulated lysis of YAC-1 cells was also observed when in vivo irradiated cells were isolated and tested immediately for in vitro lysis of YAC-1 cells. Sixteen hours after in vitro or in vivo irradiation, the cytotoxicity of hepatic natural killer cells against YAC-1 was no longer enhanced. The cytolysis of and binding of hepatic natural killer cells to CC531 colon adenocarcinoma cells was not stimulated by in vitro irradiation. From these experiments, we conclude that low-dose radiation stimulates the cytotoxicity of hepatic low-density natural killer cells against YAC-1 lymphoma cells immediately after irradiation as the result of enhanced binding of the cells to the tumor cells, in addition to the synthesis of new cytotoxic granules. (Hepatology 1994;19:1453-1458.)
Interleukin-6 enhances hepatic transferrin uptake and ferritin expression in rats
Authors: Kobune Masayoshi , Kohgo Yutaka , Kato Junji , Miyazaki Etsu , Niitsu Yoshiro
Published: March 2007

To explore a mechanism of interleukin (IL)-6-induced hypoferremia in rats, iron metabolism was investigated both in vivo and in vitro. Recombinant IL-6 was intraperitoneally administered to male Wistar rats and the serial change of parameters related to iron metabolism was examined. After administration of IL-6, plasma IL-6 concentration increased rapidly, reached its maximum in 1 hr and thereafter decreased quickly. Plasma IL-6 3 hr after IL-6 injection (50 μg/kg) was 3 units/ml, which is a concentration capable of inducing hepatic 125I-labeled transferrin uptake in vitro using isolated hepatocytes. Plasma iron concentration and transferrin saturation had decreased to approximately one third of the initial level within 3 hr and then recovered. Total iron binding capacity remained unchanged for 6 hr, then began to decrease. Red blood cell count and hemoglobin concentration showed no remarkable changes during this period. By ferrokinetic study with plasma that contained iron 59-labeled transferrin, the plasma iron disappearance half time, calculated from the disappearance curve, was significantly shortened from 55 min to 22 min by IL-6 treatment (p<0.01). The ferritin concentration in the liver was increased significantly after the administration of IL-6 (p<0.001), but transiently decreased in the spleen. The plasma ferritin showed a gradual increase during the 6-hr period after IL-6 injection. The uptake of 125I-labeled diferric transferrin by isolated hepatocytes was increased by IL-6 treatment and this increment was inhibited by addition of 100-fold excess unlabeled transferrin. On the other hand, no significant increment of 125I-labeled differic transferrin uptake was observed in Kupffer cells. These results confirmed that in rats, IL-6 produced a rapid increase of hepatic uptake of serum iron by means of transferrin receptor and resulted in hypoferremia. (Hepatology 1994;19:1468-1475.)
Conversion of xanthine dehydrogenase into xanthine oxidase in rat liver and plasma at the onset of reperfusion after ischemia
Authors: Kooij Arnold , Schiller Henry J. , Schijns Martin , Van Noorden Cornelis J. F. , Frederiks Wilma M.
Published: March 2007

The aim of this study was to test whether conversion of xanthine dehydrogenase into xanthine oxidase as induced by fasting, ischemia of the liver or both is an in vivo process or only occurs in vitro in homogenates. For this purpose, the conversion rate of xanthine dehydrogenase into xanthine oxidase was studied in liver homogenates obtained from rats after normal feeding or 24 hr of fasting followed or not by 2 hr of ischemia of the liver. In fed rats, the conversion rate of xanthine dehydrogenase into xanthine oxidase was studied as well in liver homogenates after different periods of reperfusion after 2 hr of ischemia. Homogenization was carried out under strictly controlled conditions, after which the supernatants were incubated at 37° C in buffer for 0 to 5 hr. Enzyme activities were assayed spectrophotometrically by measuring urate production at 295 nm. Conversion started only after 2 to 3 hr of incubation of supernatants of control fed livers, whereas conversion started immediately after 24 hr of fasting. The percentage oxidase activity of total xanthine oxidoreductase activity in ischemic livers from fed animals was slightly higher (26.7% ± 1.7%; p<0.05) than in control livers (19.3% ± 1.4%), whereas the percent oxidase activity in ischemic livers from fasted animals (16.7% ± 1.0%) was not different from that in control animals (16.8% ± 1.1%). Ischemia for 2 hr caused in vitro a substantial increase in the conversion rate in supernatants of livers of fed and fasted rats as compared with their controls. Furthermore, the appearance of xanthine dehydrogenase and xanthine oxidase in the blood during reperfusion up to 60 min after 2 hr ischemia of the liver was studied. The enzyme started to appear in the blood after 5 min of reperfusion, predominantly as xanthine dehydrogenase. Very rapid conversion of xanthine dehydrogenase into xanthine oxidase was observed in the plasma. We conclude that fasting and 2-hr in vivo ischemia of rat liver can affect the conversion rate of xanthine dehydrogenase into xanthine oxidase during homogenization but not in vivo. Therefore initial tissue injury due to in vivo ischemia and reperfusion in liver is not caused by conversion of xanthine dehydrogenase into xanthine oxidase. It cannot be ruled out that, after leakage of the enzyme to the circulation, additional damage is induced. (Hepatology 1994;19:1488-1495.)
Hepatobiliary function in senescent male sprague-dawley rats
Authors: Handler Jeffrey A. , Genell Caroline A. , Goldstein Robin S.
Published: March 2007

The purpose of these studies was to investigate intrahepatic changes underlying age-related decreases in bile flow by evaluating the effects of aging on bile acid-dependent and -independent flow, canalicular versus ductular flow and hepatic tight junction permeability. The isolated perfused liver was used to assess age-related changes in intrinsic hepatobiliary function without the complications of extrahepatic factors such as circulating hormones or hemodynamics. Livers from young adults (3 to 6 mo old) or senescent (22 to 26 mo old) male Sprague-Dawley rats were isolated and perfused in a nonrecirculating, hemoglobin-free system to assess oxygen uptake, bile acid-dependent and -independent bile flow, bile acid uptake, carbon 14-labeled erythritol clearance as a measure of canalicular flow, tight junction permeability and transcellular transport into bile. Rates of oxygen uptake by livers from senescent rats were significantly lower than those of young adults (75 ± 8 μmol/gm/hr vs. 121 ± 5 μmol/gm/hr). Age-related decreases in total bile flow were observed and were associated with similar reductions in 14C-erythritol clearance suggestive of decreased canalicular bile flow. Bile aciddependent and -independent flow was decreased by 50% and 60%, respectively, in isolated perfused livers from senescent rats. Hepatocellular uptake of taurocholate and rates of bile acid excretion also were about 50% lower in senescent than in young adult rats. Tight junction permeability and transcellular transport were assessed by monitoring appearance of tritiated inulin and horseradish peroxidase in bile after bolus injections of these compounds through the portal vein. Tritiated inulin appearance in bile was decreased slightly in senescent compared with young rats. No age-related differences in tight junction permeability to horseradish peroxidase were observed. In addition, biliary excretion of horseradish peroxidase in livers from senescent rats was not affected by colchicine, suggesting that microtubule-related transport was minimal or absent in the senescent rats. Taken together, these data indicate that decreased rates of bile flow in senescent compared with young rats are caused by age-related effects on intrinsic hepatobiliary function, specifically decreases in canalicular bile flow associated with decreased bile acid-dependent and -independent bile flow. The decreased canalicular bile flow appears to be a result in part of impairment of energy-dependent transport processes and microtubule-dependent transcellular transport across cells. These findings suggest that changes in intrinsic hepatobiliary function occur in aged populations and are not caused exclusively by extrahepatic effects such as altered blood flow or changes in hormonal status. Further, age-related decreases in bile flow may be caused at least in part by decreased active transport of osmotically active solutes into bile. (Hepatology 1994;19:1496-1503.)
Ontogeny of hepatobiliary secretion: Role of glutathione
Authors: Mohan Parvathi , Ling Saidee C. , Watkins John B.
Published: March 2007

The ontogeny of hepatic synthesis and biliary secretion of glutathione was characterized and correlated with hepatic gamma glutamyl transpeptidase, bile flow rate, biliary bile acids and amino acids in Sprague-Dawley rats at 18 days of gestation and postnatally at ages 7, 14, 21, 28 and 54 days. Bile was collected by bile duct cannulation under intraperitoneal anesthesia with nembutal/ketamine. Glutathione, gamma glutamyl transpeptidase and bile acids were analyzed enzymatically and amino acids by high-performance liquid chromatography using established methods. Hepatic glutathione was low in the fetus, but increased to approximate adult levels by 7 days postnatally (2.77 μmol/gm liver). Hepatic gamma glutamyl transpeptidase activity, high in the fetus, declined to adult levels by day 7 (0.03 μmol/mg protein). In contrast, significant efflux of glutathione and its constituent amino acids into bile did not occur until weaning (21 days of age). During weaning, there was a fivefold increase in the biliary glutathione and with a twofold increase in bile flow rate. Biliary bile acids concentration remained constant throughout development, with only a 30% to 50% increase in its secretion rate. The data suggest that the developmentally related increase in bile flow rate was principally a result of the increase in bile acid independent flow from 1.1 μl/(min · 100 gm body wt) in the suckling to 4.3 μl/(min · 100 gm body wt) in the post-weanling animal. Conclusions: (1) bile formation in the suckling is largely dependent on bile acids secretion, in the absence of a significant bile acids independent component; (2) glutathione and amino acids play a major role in promoting bile acid independent flow; and (3) the development of mechanisms for biliary glutathione secretion succeeds those of hepatic glutathione synthesis and provides a potential model for investigating the ontogeny of canalicular transport processes and the role of glutathione in bile formation. (Hepatology 1994;19:1504-1512.)
Serum ALT: Tarnished gold standard for interferon response in hepatitis C virus infection
Authors: Yee Hal F. , Wright Teresa L.
Published: March 2007

Serial serum samples and pretreatment and post-treatment liver tissue from patients with chronic hepatitis C virus (HCV) infection were tested for HCV RNA by reverse-transcription polymerase chain reaction and branched DNA signal amplification assays. At the end of treatment with interferon-α (IFNα), 4 of 5 patients showing no biochemical response (in alanine aminotransferase activity), 4 of 5 with transient responses, and 1 of 5 showing complete and sustained responses had HCV RNA detectable in serum. The corresponding numbers for liver tissue were 5, 5, and 0 (of 4). However, all 5 complete responders had virological relapses wdthin 6 months. Biochemical response may not reflect virological profile during IFNα treatment of HCV.
Effects of Anisotonic Exposure on Duck Hepatitis B Virus Replication
Authors: Offensperger Wolf-Bernhard , Offensperger Silke , Stoll Barbara , Gerok Wolfgang , Häussinger Dieter
Published: March 2007

ABSTRACT

In cultured hepatocytes from

in vivo

duck hepatitis B virus–infected ducks the effect of medium osmolarity on viral replication was studied. A 10–day exposure to hypotonic media (277 mOsm/L due to removal of 26 mmol/L NaCl) lowered the duck hepatitis B virus DNA content of cells and of the medium by about 50%, whereas hyperosmotic exposure (421 mOsm/L by addition of 46 mmol/L NaCl) increased it about four–fold compared with normotonic standard incubation medium (329 mOsm/L). The tissue levels of viral RNA transcripts increased during the 10 days of hypertonic exposure but decreased only slightly after hypoosmotic treatment. Western–blot analysis for the production of viral pre–S/S proteins revealed a marked stimulation of viral protein synthesis in hypertonic media, whereas hypotonic exposure inhibited it. Conversely, total cellular protein synthesis as assessed from [3H]leucine incorporation into acid–precipitable material decreased during hyperosmotic exposure but increased during hypoosmotic exposure. We noted a comparable increase of duck hepatitis B virus DNA when raffinose (80 mmol/L) was added to hypotonic or normotonic media, without change in the NaCl concentrations. This suggests that the effects of anisotonicity on viral replication were not due to alterations of Na+ or Cl- activity in the incubation media, but might reflect changes of cellular volume. The effects of anisotonicity on viral replication were only seen after exposure of more than 8 hr of the cells to anisotonicity. The findings suggest that the cellular volume is an important determinant for duck hepatitis B virus replication, yet the underlying molecular mechanisms remain elusive. (Hepatology 1994;20:1-7.)
Intraobserver and Interobserver Variations in Liver Biopsy Interpretation in Patients With Chronic Hepatitis C
Authors: Bedossa P.
Published: March 2007

ABSTRACT

Liver biopsy is used as the “gold standard” for the assessment of the stage and degree of activity in chronic hepatitis C and is of major importance in evaluating the effects of treatment. Because numerous therapeutic trials are undertaken with histological control, the reproducibility of liver biopsy interpretation appears essential. Therefore the aim of this study was to estimate intraobserver and interobserver variations in the assessment of features, classification, and numerical scoring of chronic viral hepatitis C among 10 pathologists specializing in liver diseases. These pathologists independently reviewed 30 liver biopsy specimens of viral hepatitis C and completed a histological form for each of the specimens. Five pairs of pathologists then were randomly designated. They independently reviewed the biopsy specimens and filled out a new coding form. The interobserver variation was calculated for each item among the 10 individuals and then among the five pairs with the intraclass correlation coefficient or κ statistics. Five features showed an almost perfect or a substantial degree of concordance among the 10 observers (i.e., cirrhosis, fibrosis, fibrosis grading of Knodell index, steatosis, portal lymphoid aggregates). The 17 other indicators showed a weaker concordance with, for instance, a moderate degree of concordance for piecemeal necrosis, disease activity, Knodell index, a fair degree of concordance for lobular necrosis, and only a slight degree of concordance for six items. Five items had a higher concordance when viewed by a pair of pathologists than when studied by only one pathologist (i.e., steatosis, periportal necrosis grading of Knodell index, lobular necrosis grading of Knodell index, centrilobular fibrosis, and ductular proliferation). Two showed a significant decrease, and the others were unchanged. This study reveals that certain features of major importance in assessing disease activity show significant observer variation. The acceptable degree of concordance of the Knodell index was related mainly to the substantial degree of concordance of the fibrosis score, whereas other numerical items displayed substantial observer variations. Simultaneous observation by two pathologists increased the reproducibility of numerical scoring and of certain viral hepatitis C lesions. A classification of chronic hepatitis C based on dissociated semiquantitative assessment of necroinflammatory lesions and fibrosis offered more reproducibility than the use of a global numerical index. (Hepatology 1994;20:15-20.)
Infantile Hemangioendothelioma of the Liver
Authors: Selby Dena M. , Stocker Thomas J. , Waclawiw Myron A. , Hitchcock Charles L. , Ishak Kamal G.
Published: March 2007

ABSTRACT

Clinical and morphological findings in 91 patients with infantile hemangioendothelioma of the liver are reported. The study comprised 56 girls and 35 boys ranging in age from premature infant to 3 yr; one outlier patient was 18 yr old. Most patients with infantile hemangioendothelioma (87%) were first seen before the age of 6 mo. Congestive heart failure was evident in 15%. Skin hemangiomas were noted in 11%. Anemia, hyperbilirubinemia and increased AST level were present. Solitary lesions were more common than multiple ones (3:2). Immunohistochemical staining of tumor cells for factor VIII was positive in 20 of 21 cases tested; testing for blood group antigen was positive in 8 of 28 cases. Cytokeratin staining verified the presence of bile ducts, some of which appeared to be the result of transformation of injured liver. No pericytes were identified on electron microscopy. The 6–mo survival rate, based on 71 cases, was 70%. Average time of follow–up for the survivors was 7.7 yr. All deaths occurred during the initial presentation/hospitalization of infants, with the exception of two patients who died 3 mo and 7 mo after diagnosis. More recent analytic methods, including immunohistochemical stains and flow cytometric studies, do not contribute to the practical assessment of this tumor. Covariates with significant value in predicting death 6 mo after diagnosis included presence of congestive heart failure, jaundice, multiple tumor nodules and absence of cavernous differentiation. (Hepatology 1994;20:39-45.)
Nodular Regenerative Hyperplasia of the Liver Graft After Liver Transplantation
Authors: Gane Edward , Portmann Bernard , Saxena Romil , Wong Philip , Ramage John , Williams Roger
Published: March 2007

ABSTRACT

Nodular regenerative hyperplasia has not previously been reported in patients who have undergone orthotopic liver transplantation. In this report we describe the clinical, biochemical and histologic findings in nine liver transplant recipients in whom nodular regenerative hyperplasia developed between 6 and 144 mo (median, 64 mo) after transplantation. Six of the patients manifested features of portal hypertension (variceal bleeding, ascites, severe intractable peripheral edema), whereas three patients were asymptomatic at the time of diagnosis. All patients had elevated serum alkaline phosphatase (median, 269 IU; range, 159 to 1182 IU) and gamma–glutamyl transferase levels (median, 247 IU; range 104 to 1513 IU). Other liver function tests remained normal. In five patients, earlier biopsies had identified histological features of venous outflow block, consistent with azathioprine hepatotoxicity. All nine patients had been taking azathioprine up to the time of presentation. After withdrawal of azathioprine, liver function tests improved appreciably in five patients and histologic improvement was documented in four of these who underwent follow–up liver biopsies. In the remaining four patients progressive graft failure ensued and all subsequently underwent repeat liver retransplantation. At a certain stage, nodular regenerative hyperplasia, in the context of a liver graft, becomes an irreversible lesion, emphasizing the need for early identification and withdrawal of the agent. (Hepatology 1994;20:88-94.)
Cancer Risk in Primary Biliary Cirrhosis: A Population–Based Study From Sweden
Authors: Lööf Lars , Adami Hans-Olov , Sparén Pär , Danielsson Åke , Eriksson Ljusk Siw , Hultcrantz Rolf , Lindgren Stefan , Olsson Rolf , Prytz Hanne , Ryden Bengt-Olof , Sandberg-Gertzen Hanna , Wallerstedt Sven
Published: March 2007

ABSTRACT

A cohort of 559 patients in Sweden who satisfied predetermined criteria for the diagnosis of primary biliary cirrhosis was followed with respect to the incidence of cancer during the period of 1958 to 1988. The mean follow–up time from the time of primary biliary cirrhosis diagnosis was 9.0 ± 54 yr. During the follow–up period, 148 patients died and the primary cause of death was liver insufficiency. An overall excess risk for cancer, standardized incidence ratio 1.6; 95% confidence interval, 1.1 to 2.2, was found in the cohort. In contrast to previous reports, we found no excess risk for breast cancer (standardized incidence ratio, 0.9; 95% confidence interval, 0.3 to 2.1). The number of hepatocellular cancers in the primary biliary cirrhosis cohort did not significantly differ from expected (standardized incidence ratio, 2.91; 95% confidence interval, 0.4 to 10.5). (Hepatology 1994;20:101-104.)
Vasopressin and Phorbol–12, 13–Dibutyrate Inhibit Glucagon– or Cyclic Amp–Stimulated Taurocholate Uptake in Isolated Rat Hepatocytes
Authors: Divald Andras , Simpser Edwin , Fisher Stanley E. , Karl Peter I.
Published: March 2007

ABSTRACT

Bile salt uptake by hepatocytes is modulated in part by changes in intracellular cyclic AMP. We studied the effect of activation of protein kinase C on cyclic AMP–mediated taurocholate uptake in isolated rat hepatocytes. Both dibutyryl cyclic AMP (2 × 10-6 mol/L) and glucagon (10-6 mol/L), which increase intracellular cyclic AMP, enhanced the initial uptake rate of taurocholate into hepatocytes, with maximal increases of 45% to 50% over the basal uptake rate. Vasopressin (10-9 mol/L), a hormone known to activate protein kinase C, and phorbol–12, 13–dibutyrate (10-5 mol/L) significantly inhibited the glucagon–stimulated increase in taurocholate uptake rate (72% ± 10% and 105% ± 13% inhibition, respectively). Basal (unstimulated) taurocholate uptake rate was not affected by vasopressin or phorbol–12, 13–dibutyrate. Down–regulation of the glucagon–stimulated transport was rapid and persisted during the 20–min experimental period. Angiotensin II had a similar but more transient inhibitory effect. Vasopressin and phorbol–12, 13–dibutyrate suppression of glucagon–stimulated taurocholate uptake rate was not accompanied by diminished cyclic AMP levels. Moreover, vasopressin and phorbol–12, 13–dibutyrate inhibited dibutyryl cyclic AMP–stimulated taurocholate uptake rate can be dissociated from alterations in the cyclic AMP levels. (Hepatology 1994;20:159-165.)
Bile Acid Inhibition of P–Glycoprotein-Mediated Transport in Multidrug–Resistant Cells and Rat Liver Canalicular Membrane Vesicles
Authors: Mazzanti Roberto , Fantappié Ornella , Kamimoto Yukkio , Gatmaitan Zenaida , Gentilini Paolo , Arias Irwin M.
Published: March 2007

ABSTRACT

To study the effect of bile acids on P–glycoprotein-mediated drug transport, we performed experiments using multidrug resistant cells and rat canalicular membrane vesicles. Cellular accumulation and efflux of rhodamine 123 were measured in drug–resistant cells by means of computerized quantitative image analysis and fluorescence microscopy. ATP–dependent [3H]daunomycin transport was studied by means of rapid filtration in canalicular membrane vesicles prepared from normal rats. Doxorubicin–sensitive (PSI–2) and –resistant (PN1A) 3T3 cells and human–derived hepatocellular carcinoma doxorubicin–sensitive and –resistant cells were used. Taurochenodeoxycholate and glycochenodeoxycholate, taurolithocholate and ursodeoxycholate (50 to 200 μmol/L) inhibited rhodamine 123 and [3H]daunomycin transport in multidrug–resistant cells and canalicular membrane vesicles, respectively, whereas taurocholate, taurode–oxycholate and tauroursodeoxycholate did not. Primary and secondary unconjugated bile acids had no effect. These results reveal that taurolithocholate, taurochenodeoxycholate and glycochenodeoxycholate and ursodeoxycholate inhibit P–glycoprotein-mediated drug transport function in multidrug resistant cell lines and in canalicular membrane vesicles. These results suggest possible interaction between P–glycoprotein function and bile acids in cholestasis and after treatment of patients with ursodeoxycholic or chenodeoxycholic acid. (Hepatology 1994;20:170-176.)
The Molecular Genetics of Autoimmune Liver Disease
Authors: Donaldson Peter , Doherty Derek , Underhill James , Williams Roger
Published: March 2007

ABSTRACT

The dual observations that human leukocyte antigens have an antigen–binding groove and that the polymorphism we study as human leukocyte antigen types is largely related to amino acid substitutions in and around that groove have provided a new focus for immunogenetic studies. In autoimmune liver disease, recent studies have described specific amino acid substitutions in the antigen–binding groove of human leukocyte antigen DR molecules that may determine both disease susceptibility, through their direct influence on antigen binding, and the severity of the disease. In autoimmune hepatitis, lysine residues at DRβ position 71 in European subjects and arginine or histidine residues at DRβ position 13 in Japanese subjects may be responsible for much human leukocyte antigen—encoded disease susceptibility. Similar claims have been made for leucine residues at DRβ 38 in primary sclerosing cholangitis and for leucine residues at DPβ 35 in Japanese patients with primary biliary cirrhosis. To date, our knowledge of genetic susceptibility to autoimmune liver disease is incomplete. Other genes may contribute to susceptibility to autoimmune liver disease—for example the contribution of TAP genes, upstream promoter sequences and class III genes on chromosome 6 and the T–cell receptor genes and complement genes elsewhere in the human genome is currently unclear. Additional information concerning the immunogenetic contribution to disease severity is needed to complete the picture. (Hepatology 1994;20:225-239.)
Diagnostic and Clinical Implications of the Different Genotypes of Hepatitis C Virus
Authors: Bukh Jens , Miller Roger H.
Published: March 2007

ABSTRACT

Hepatitis C virus (HCV) samples in 155 sera, from patients with chronic non–A, non–B liver disease and blood donors, were grouped into four genotypes (I, II, III, and IV) by amplification of core–gene sequences by polymerase chain reaction with type–specific primers. HCV genotypes were compared with various HCV–associated antibodies detectable by the first–generation ELISA (ELISA–1) with C100–3 protein and a second–generation immunoblot assay with four recombinant HCV proteins. Antibodies to C100–3 protein and those to its subsequence (5-1-1) were detected in 13 (93%) and 12 (86%), respectively, of 14 sera with genotype I HCV; 56 (79%) and 58 (82%) of 71 sera with genotype II; 13 (34%) and 6 (16%) of 38 sera with genotype III; and 11 (34%) and 4 (13%) of 32 sera with genotype IV. Amino acid sequences of C100–3 of genotype I HCV are conserved by ˜90% in genotype II, but only by ˜75% in genotypes III and IV. The sensitivity of ELSA–1, therefore, would be influenced by heterogeneity in C100–3 sequences of different genotypes.
Heterogeneity of Combinatorial Human Autoantibodies Against Pdc–E2 and Biliary Epithelial Cells in Patients With Primary Biliary Cirrhosis
Authors: Cha Sanghoon , Leung Patrick S.C. , Coppel Ross L. , Van De Water Judy , Ansari Aftab A. , Gershwin Eric M.
Published: March 2007

The polyclonal nature of antimitochondrial autoantibodies and the limited success of generating human monoclonal antibodies have made analysis of fine specificity and antibody heterogeneity difficult to define. The major autoantigen of primary biliary cirrhosis is the E2 component of the pyruvate dehydrogenase pathway (PDC–E2). To address the relative importance of the region(s) in the PDC–E2 inner lipoyl domain to antibody binding, we report herein detailed profiles of 12 PDC–E2—specific antigen–binding fragments, SP1 through SP12, derived by screening of a combinatorial immunoglobulin library (derived from a primary biliary cirrhosis patient) with full–length native PDC–E2. All antigen–binding fragments are IgG isotypes and include a similar number of λ– and κ–chains. The antigen–binding fragments react specifically to PDC–E2 with high affinity (Ka = 10-7 to 10-10 mol/L-1) and recognize a conformational epitope in the inner lipoyl domain of PDC–E2. Furthermore, the antibodies demonstrate substantial heterogeneity in recognition of different recombinant PDC–E2 fragments and differential recognition patterns against mutant constructs of the human PDC–E2 inner lipoyl domain (amino acid residues 91 to 227). In addition, five of the antigen–binding fragment clones (SP1, 3, 4, 8 and 12) demonstrate different staining patterns on biliary epithelial cells of patients with primary biliary cirrhosis but not control liver disease; some antigenbinding fragments specifically stained the apical region of biliary epithelium, a pattern distinct from that of typical mitochondrial staining. The response to the inner lipoyl domain is not, however, monospecific, and there is much more heterogeneity in fine specificity than could be accounted for by arbitrary reshuffling of variable immunoglobulin heavy and light chains into unnatural combinations. (Hepatology 1994;20:574–;583).
Acetylsalicylic Acid in the Prevention of Early Stenosis and Occlusion of Transjugular Intrahepatic Portal–Systemic Stent Shunts: A Controlled Study
Authors: Theilmann Lorenz , Sauer Peter , Roeren Thomas , Otto Gerd , Arnold Joachim C. , Noeldge Gerd , Richter Goetz , Stiehl Adolf
Published: March 2007

Stenosis or occlusion of the transjugular intrahepatic portal–systemic stent shunt may be initiated by aggregation and activation of thrombocytes on the surface of the metallic stent material. To find effective prevention of this event, we conducted a controlled trial administering acetylsalicylic acid for 3 mo. Forty–four patients (8 women and 36 men) with portal hypertension were included in this study. The patients were randomized into a group receiving 100 mg acetylsalicylic acid/day (n = 21) or into a control group (n = 23). Treatment was started immediately after transjugular intrahepatic portal–systemic stent shunt. Three months after transjugular intrahepatic portal–systemic stent shunt, 15 patients in the acetylsalicyclic acid group and 19 patients in the control group underwent clinical reevaluation, gastroscopy and recatheterization with determination of the portal–systemic pressure gradient. No variceal bleeding occurred in any patients. In four patients in the acetylsalicylic acid group, erosive gastritis was observed in gastroscopy in contrast to only one patient in the control group. Complete patency of the stent was noted in 10 of 15 patients in the acetylsalicylic acid group and in 14 of 19 patients in the control group. Transjugular intrahepatic portal–systemic stent shunt restenosis associated with a significant increase of the portal–systemic gradient occurred in five patients in the acetylsalicylic acid group, which required redilation in all and additional stent placement for expansion of the stented tract in two patients. In the control group, redilation was necessary in five patients with additional stent extension in two patients. Transjugular intrahepatic portal–systemic stent shunt effectively prevented recurrence of variceal bleeding in the first 3 mo despite significant stent restenosis. The latter could not be prevented by acetylsalicylic acid treatment. (Hepatology 1994;20:592-597).
Effect of Orthotopic Liver Transplantation on Bone Mineral Content and Serum Vitamin D Metabolites in Infants and Children With Chronic Cholestasis
Authors: Argao Eric A. , Balistreri William F. , Hollis Bruce W. , Ryckman Frederick C. , Heubi James E.
Published: March 2007

Almost all infants and children with chronic cholestasis have osteopenia. We evaluated the effect of orthotopic liver transplantation on bone mineral content and serum 25(OH)–vitamin D-[25(OH)D]-of nine infants and children (five girls; age, 6 to 21 mo at the time of orthotopic liver transplantation) with end–stage liver disease resulting from chronic cholestasis. We hypothesized that after orthotopic liver transplantation, decreased bone mineral content will recover and the serum 25(OH)D level will either normalize or remain normal in those who were previously vitamin D deficient or sufficient, respectively. All had subnormal bone mineral content before transplant. On long–term follow–up (>4 mo) of seven patients, bone mineral content normalized in all between 6.5 and 19 mo after transplant, with a mean of 11.2 + 4.5 mo. In six patients with normal serum 25(OH)D levels before orthotopic liver transplantation, the serum 25(OH)D levels had declined markedly 1 to 2 mo after transplant, followed by return to normal by 3 to 6 mo. Low serum 25(OH)D levels (<15 ng/ml) in three patients before orthotopic liver transplantation normalized after transplant. Although there was no correlation between bone mineral content and serum 25(OH)D level before transplant, sustained normal serum 25(OH)D and 1,25(OH)2D levels preceded or accompanied normalization of bone mineral content in the seven patients available for long–term follow–up. We conclude that (a) in infants and children younger than 2 yr with chronic cholestasis, bone mineral content normalizes approximately 11 mo after orthotopic liver transplantation. This normalization is preceded by a sustained period of normal serum 25(OH)D levels. (b) Normal baseline serum 25(OH)D levels decrease 1 to 2 mo after orthotopic liver transplantation, with subsequent normalization, whereas initially low serum 25(OH)D levels gradually normalize within 2 to 4 mo. We speculate that maintenance of normal serum 25(OH)D levels with oral vitamin D supplementation may enhance recovery of bone disease after orthotopic liver transplantation. (Hepatology 1994;20:598-603).
Sclerotherapy for Male Alcoholic Cirrhotic Patients Who Have Bled From Esophageal Varices: Results of A Randomized, Multicenter Clinical Trial
Authors: Hartigan Pamela
Published: March 2007

Sclerotherapy and sham–sclerotherapy were compared in male alcoholic patients with cirrhosis and bleeding esophageal varices. The prospective, singleblind, randomized clinical trial of 5 yr duration entered 253 male alcoholic patients at 12 Veterans Affairs medical centers. Patients were either actively bleeding from esophageal varices at randomization or they had a history of such bleeding. They were treated by endoscopy with sham–sclerotherapy or endoscopy with sclerotherapy at randomization, 4 to 6 days, 9 to 11 days, 1 mo, 3 mo and every 3 mo for 2 yr and followed for the remainder of the study period. of the patients randomized, 131 were assigned to sham–therapy and 122 were assigned to sclerotherapy. At entry the two patient groups were comparable. The upper gastrointestinal rebleeding rates during the study were 101 and 66 per 100 person years of follow–up in sham–therapy and sclerotherapy, respectively (rate ratio, 1.54; 95% confidence interval, 1.06 to 2.22; p = 0.01). A significantly higher number of episodes were attributable to esophageal varices in the shamtherapy group (112 vs. 52; p = 0.005). Seventy–four sham–therapy patients (56%) and 77 sclerotherapy patients (63%) died (p = 0.54; relative risk, 0.91; 95% confidence interval, 0.66 to 1.25). The mean transfusion requirement was higher in the sham–therapy group (16.0 units vs. 9.4 units; p = 0.002) and more patients in this group required shunt surgery (18 vs. 5; p = 0.005). We conclude that sclerotherapy reduces the rate of rebleeding from varices, the transfusion requirement, and the need for shunt surgery, but has no long–term effect on survival. (Hepatology 1994;20:618-625).
Immunoglobulins and α1–Acid Glycoprotein Do Not Contribute to the Cholesterol Crystallization—Promoting Effect of Concanavalin A—Binding Biliary Protein
Authors: De Bruijn Marianne A.C. , Mok Kam S. , Out Tiny , Tytgat Guido N.J. , Groen Albert K.
Published: March 2007

Human bile contains cholesterol crystallization—stimulating proteins that can be isolated by concanavalin A—Sepharose chromatography. In the past few years an increasing number of different pronucleating proteins have been identified in the concanavalin A—binding fraction. In this study we attempted to estimate the relative contribution of a number of these proteins to total concanavalin A—binding pronucleating activity. For this purpose, concanavalin A—binding glycoproteins were isolated from gallbladder bile samples from 12 patients with gallstones. The role of IgA, IgG and IgM and α1–acid glycoprotein was investigated by means of immunoextraction. No decrease in crystallization–promoting activity was observed after precipitation of more than 98% of the different immunoglobulins. In addition, removal of more than 95% of α1–acid glycoprotein from different concanavalin A—binding fractions had no significant effect on cholesterol crystallization—promoting activity. The influence of fibronectin was estimated by addition of physiological concentrations to a model bile system. At these concentrations fibronectin did not promote crystallization. From these data we conclude that immunoglobulins, α1–acid glycoprotein and probably also fibronectin do not significantly contribute to total concanavalin A—binding activity. (Hepatology 1994;20:626-632).