bluuebunny/crossref_metadata_2025 huggingface dataset:
Chronic ethanol consumption increases hepatic sinusoidal contractile response to endothelin-1 in the rat
Authors: Bauer Michael , Paquette Nicole C. , Zhang Jian X. , Bauer Inge , Pannen Benedikt H. J. , Kleeberger Steven R. , Clemens Mark G.
Published: March 2007

Recent evidence suggests that hepatic stellate cells function as liver-specific pericytes that are highly contractile in response to endothelin-1 (ET-1). Liver injury has been shown to lead to “activation” of stellate cells producing a phenotypic change to a more myofibroblastic cell type including loss of vitamin A and increased contractility. The present study was undertaken to test the effects of short-term chronic ethanol consumption (36% of total calories for 5 weeks according to the Lieber-DeCarli protocol) on hepatic vitamin A storage, expression of smooth muscle α-actin, and sinusoidal contractility in Sprague-Dawley rats. Using in vivo epifluorescence video microscopy, we quantified the number of sites of vitamin A fluorescence (purportedly stellate cells) and assessed sinusoidal microhemodynamics at baseline and during a 20-minute infusion period of ET-1 (1 pmol* 100 g body weight [bw]−1*min−1). Retinol and retinyl palmitate were measured after the experiment by means of high-pressure liquid chromatography (HPLC). A highly significant decrease in liver retinyl palmitate level (control: 622.5 ± 50.9; ethanol: 273.0 ± 38.0 μg/g liver; P < .001) was found that correlated with a decrease in sites of vitamin A fluorescence (control: 531.4 ± 76.1; ethanol: 141.1 ± 30.2* mm−2; r = .82, P < .001). Concomitantly scattered expression of smooth muscle α-actin in sinusoids was observed. Although sinusoidal hemodynamics were not affected at baseline, a significant increase in sinusoidal contractility on endothelin-1 infusion (e.g., sinusoidal resistance [% of baseline value]: control: 10 minutes: 288.7 ± 71.7, 20 minutes: 200.5 ± 46.9; ethanol: 10 minutes: 1,916.0 ± 701.7, 20 minutes: 656.8 ± 103.3; P < .05 and .01, respectively) was observed. These data indicate that chronic ethanol consumption in this moderate model initiates stellate cell activation. Increased sinusoidal responsiveness to the vasoconstrictor ET-1 in vivo may contribute to the increased susceptibility of ethanol-fed rats to secondary stresses that increase ET-1 expression, such as endotoxemia. (Hepatology 1995; 22:1565-1576).
Feedback-inhibition of glucagon-stimulated glycogenolysis in hepatocyte/kupffer cell cocultures by glucagon-elicited prostaglandin production in kupffer cells
Authors: Hespeling Ursula , Jungermann Kurt , Püschel Gerhard P.
Published: March 2007

Prostaglandins, released from Kupffer cells, have been shown to mediate the increase in hepatic glycogenolysis by various stimuli such as zymosan, endotoxin, immune complexes, and anaphylotoxin C3a involving prostaglandin (PG) receptors coupled to phospholipase C via a G0 protein. PGs also decreased glucagon-stimulated glycogenolysis in hepatocytes by a different signal chain involving PGE2 receptors coupled to adenylate cyclase via a Gi protein (EP3 receptors). The source of the prostaglandins for this latter glucagon-antagonistic action is so far unknown. This study provides evidence that Kupffer cells may be one source: in Kupffer cells, maintained in primary culture for 72 hours, glucagon (0.1 to 10 nmol/L) increased PGE2, PGF2α, and PGD2 synthesis rapidly and transiently. Maximal prostaglandin concentrations were reached after 5 minutes. Glucagon (1 nmol/L) elevated the cyclic adenosine monophosphate (cAMP) and inositol triphosphate (InsP3) levels in Kupffer cells about fivefold and twofold, respectively. The increase in glycogen phosphorylase activity elicited by 1 nmol/L glucagon was about twice as large in monocultures of hepatocytes than in cocultures of hepatocytes and Kupffer cells with the same hepatocyte density. Treatment of cocultures with 500 μmol/L acetylsalicylic acid (ASA) to irreversibly inhibit cyclooxygenase (PGH-synthase) 30 minutes before addition of glucagon abolished this difference. These data support the hypothesis that PGs produced by Kupffer cells in response to glucagon might participate in a feedback loop inhibiting glucagon-stimulated glycogenolysis in hepatocytes. (Hepatology 1995; 22:1577-1583).
Recombinant interferon-alfa therapy in children with chronic hepatitis C
Authors: Bortolotti Flavia , Giacchino Raffaella , Vajro Pietro , Barbera Cristiana , Crivellaro Carlo , Alberti Alfredo , Nebbia Gabriella , Zancan Lucia , de Moliner Lorena , Bertolini Adalberto , Balli Fiorella , Callea Francesco
Published: March 2007

The efficacy of recombinant interferon-alfa therapy in children with chronic hepatitis C has been evaluated in a randomized, controlled pilot study including 27 patients, aged 2 to 14 years, without underlying systemic diseases. On entry, all patients had abnormal alanine transaminase (ALT) levels, 22 were hepatitis C virus (HCV) RNA positive, 19 had mild chronic active hepatitis, and 8 had chronic persistent hepatitis on liver biopsy. Fourteen children received 5 MU/m2 of recombinant interferon-alfa2b thrice weekly for 4 months. If at this time ALT had been reduced to at least 50% the baseline level, treatment was continued up to 12 months. The other 13 children remained untreated. The whole follow-up period lasted 24 months. Interferon was stopped at 4 months in 4 children because of an ALT increase (2 cases), unchanged ALT and febrile convulsions (1 case), and slight ALT decrease (1 case). This latter patient, however, had normal ALT at 6 months and throughout further follow-up, and cleared HCV RNA, thus behaving as a sustained responder. All 10 children treated for 12 months had normal levels of ALT, and 9 were HCV RNA negative at the end of treatment. Of the 9 children who could be followed to 24 months, 4 relapsed soon after therapy withdrawal and 5 maintained a sustained biochemical and virologic response. Overall, 6 (43%) of 14 treated children had a sustained ALT normalization associated with HCV RNA clearance as compared with only 1 (7.5%) untreated child who had a sustained ALT normalization but did not clear HCV RNA. These results suggest that recombinant interferon therapy, at the regimen used in this study, may induce sustained ALT normalization and HCV clearance in a significant proportion of children with chronic hepatitis C. (Hepatology 1995;22:1623-1627).
Fulminant hepatitis associated with hepatitis B virus e antigen-negative infection: Importance of host factors
Authors: Karayiannis Peter , Alexopoulou Alexandra , Hadziyannis Stephanos , Thursz Mark , Watts Richard , Seito Seiji , Thomas Howard C.
Published: March 2007

The precore stop-codon variant of hepatitis B virus (HBV) has been implicated in fulminant hepatitis. The precore/core regions of such variants from two sets of patients with interpartner transmission resulting in fulminant hepatitis in the contact, were sequenced to establish whether further sequence variations in the core region are specifically associated with the fulminant disease. In both sets of patients, there was sequence diversity of the precore/core region from the wild type, leading to numerous amino acid substitutions in the core region. Between the infecting source and the contact, there was only one amino acid change in one set of patients and none in the other. In addition, in the second set of patients, serum samples from four different time points were investigated. Sequence data showed no variation in each patient at the nucleotide level in the core region, even in the case of the source, who was followed for 3 years. In this same pair of subjects, the remainder of the genome was sequenced and was identical at the nucleotide level. Therefore, it appears that, at least in some cases of fulminant hepatitis caused by infection with the precore variant, the nucleotide sequence of the patient with fulminant hepatitis is identical to that observed in the asymptomatic source of infection. These data indicate that the severity and outcome of infection in such cases are unrelated to any additional variation in the entire HBV genome, and that the changed clinical picture is dependent on host factors, possibly the HLA environment. HLA typing of both the contact and source in the two sets of patients revealed that the contacts with fulminant hepatitis had one or more alleles at the HLA class II locus, previously shown to be associated with an acute hepatitis sufficient to clear the virus. (Hepatology 1995; 22:1628-1634).
Immunoglobulin M antibody to hepatitis C virus core antigen: Correlations with viral replication, histological activity, and liver disease outcome
Authors: Quiroga Juan Antonio , van Binsbergen Jan , Wang Chang Yi , Pardo Margarita , Navas Sonia , Trines Chantal , Herrero Montserrat , Carreno Vicente
Published: March 2007

Immunoglobulin M (IgM) antibody to hepatitis C core antigen (anti-HCV-core) was tested by enzyme immunoassay against a synthetic peptide representing amino acids 1 to 62 of the core protein. Of 214 patients with different categories of histological activity, 193 (90%) showed positive results for IgM anti-HCV-core, and 207 (97%) had HCV RNA; most cases (186, 87%) had both markers detectable simultaneously. No differences in the frequency of IgM anti-HCV-core were observed with respect to epidemiological, biochemical, or histological parameters. In 175 interferon alfa (IFN-α) recipients, and in 39 untreated controls, pretreatment IgM anti-HCV-core frequencies were similar: 28 of 32 (88%) in sustained responders; 55 of 61 (90%) in responders with relapse; 72 of 82 (88%) in nonresponders; and 38 of 39 (97%) in untreated controls. After IFN-α therapy, IgM anti-HCV-core levels became undetectable with significantly greater frequency in sustained responders (P = .014); a similar trend was observed for HCV RNA (P < .0001). IgM anti-HCV-core levels decreased after therapy in responders (P < .001) but increased in nonresponders. Fifty-one cases were longitudinally tested in relation to long-term disease outcome. Both markers remained detectable in most nonresponders with persistent liver disease, in most responders before relapse, and in all but one case at the time of biochemical relapse. IgM anti-HCV-core and HCV RNA became undetectable in most sustained responders, but reappeared despite a long-lasting transaminase normalization, behaving as asymptomatic HCV carriers; the possibility that disease reactivation may take place years afterwards cannot be excluded. These data indicate that IgM anti-HCV-core may be useful in the assessment of HCV replication and in monitoring biochemical and virological responses to IFN-α treatment. (Hepatology 1995; 22:1635-1640).
Proteolytic activity of NS3 serine proteinase of hepatitis C virus efficiently expressed in escherichia coli
Authors: Shoji Ikuo , Suzuki Tetsuro , Chieda Shinya , Sato Mitsuru , Harada Takashi , Chiba Tsutomu , Matsuura Yoshiharu , Miyamura Tatsuo
Published: March 2007

The serine proteinase of hepatitis C virus (HCV) nonstructural protein NS3 was efficiently expressed in an active form as a fused protein with oligohistidine in Escherichia coli. The recombinant fusion protein was purified to near homogeneity by affinity chromatography on a metal chelation column. Trans-cleavage activity of this protein was investigated by using the substrate NS5 protein expressed in insect cells. The purified serine proteinase trans-cleaved the partially purified NS5 protein. In contrast, the NS3 proteins with mutations at the proposed catalytic site, Ser1165 or His1083, lost the trans-cleavage activity. Analysis of the authentic enzyme and variants with site-directed mutations provides a useful tool for understanding the structure-function relationship of the NS3 serine proteinase. We then developed an in vivo trans-cleavage assay system by coexpression of the NS3 proteinase and the NS5 substrate in E coli, and examined the effect of known inhibitors of serine proteinase. Inhibition of its proteolytic activity by N-p-tosyl-l-lysine chloromethyl ketone (TLCK) was observed, but only at high concentrations. The in vitro and in vivo trans-cleavage assays for NS3 serine proteinase will facilitate efficient testing for inhibitors of the replication of HCV and specific treatment for hepatitis C. (Hepatology 1995; 22:1648-1655).
Increased production of nitric oxide by neutrophils and monocytes from cirrhotic patients with ascites and hyperdynamic circulation
Authors: Laffi Giacomo , Foschi Marco , Masini Emanuela , Simoni Antonella , Mugnai Laura , la Villa Giorgio , Barletta Giuseppe , Mannaioni Pier Francesco , Gentilini Paolo
Published: March 2007

An increased release of nitric oxide (NO), a powerful vasodilating agent, has been proposed to play a role in the pathogenesis of vasodilation and hyperdynamic circulation associated with advanced cirrhosis. We evaluated NO synthase (NOS) activity in peripheral leukocytes of 12 cirrhotic patients and 9 healthy subjects together with plasma endotoxin levels and systemic hemodynamic (by a noninvasive echocardiographic method). NOS activity was evaluated by (1) measuring the capacity of isolated polymorphonuclear cells (PMNs) and monocytes to convert [3H]arginine to [3H]citrulline; (2) measuring the ability of neutrophils and monocytes to inhibit thrombin-induced platelet aggregation and to increase guanosine 3′-5′-cyclic monophosphate content in coincubated platelets, an expression of NO release from these cells. Both neutrophils and monocytes from cirrhotic patients produced significantly higher amounts of [3H]citrulline than cells obtained from healthy subjects (P < .001 and P < .02 for neutrophils and monocytes, respectively) and were more effective than control cells in inhibiting platelet aggregation (P < .05 and P < .001, respectively for 2 × 106 cells) and in increasing guanosine 3′-5′-cyclic monophosphate content in coincubated platelets (P < .05 and P < .001, respectively). The anti-aggregating activity expressed by leukocytes has a pharmacological profile similar to that described for NO, because it increased after addition of superoxide dismutase, a superoxide anion scavenger, and markedly decreased after inhibition of nitric oxide synthesis with N
G-monomethyl-l-arginine (L-NMMA) and N
G-nitro-l-arginine-methyl ester (L-NAME). Cirrhotic patients had significantly higher plasma endotoxin levels (P < .001) and cardiac index (P < .01) when compared with controls. These data support the contention that the NO synthetic pathway is activated in cirrhotic patients with ascites, hyperdynamic circulation, and endotoxemia. (Hepatology 1995; 22:1666-1673).
Changes in platelet kinetics after a partial splenic arterial embolization in cirrhotic patients with hypersplenism
Authors: Noguchi Hideya , Hirai Kenji , Aoki Yoshinori , Sakata Kenji , Tanikawa Kyuichi
Published: March 2007

We performed a partial splenic arterial embolization in 22 patients with cirrhosis associated with thrombocy-topenia and then evaluated the changes in platelet kinetics after undergoing the procedure using 111In-tropolone-labeled platelets. The controls consisted of eight chronic hepatitis patients who showed a normal platelet count and normal spleen size. The mean splenic infarction ratio after the procedure was 54.9%. A platelet kinetics study was performed before and 2 months after the procedure. Before the procedure, the cirrhotic patients showed increases in the splenic volume and the spleen/liver uptake ratio of the 111In-labeled platelets on both the third and seventh days, and a decrease in the platelet recovery compared with the controls, which suggested an increased platelet pool in the spleen. In addition, the platelet survival time in cirrhotic patients was shortened, whereas the platelet-associated immuno-globulin G (PA-IgG) was higher than that of the controls, which suggested the involvement of immunologic mechanisms in the thrombocytopenia. With an increase of the platelet count after a partial splenic arterial embolization, the spleen/liver uptake ratio of the 111In-labeled platelets decreased, whereas the platelet recovery increased. Furthermore, the platelet survival time was prolonged, whereas the PA-IgG decreased. The platelet count showed a positive correlation with the platelet survival time and a negative correlation with PA-IgG before and after the procedure. These results suggest that a transcatheter splenic arterial embolization not only may reduce the increased platelet pool in the spleen but also may improve the thrombocytopenia induced by immunologic mechanisms in patients with cirrhosis. (Hepatology 1995; 22:1682-1688).
Paraumbilical vein patency in cirrhosis: Effects on hepatic hemodynamics evaluated by doppler sonography
Authors: Sacerdoti David , Bolognesi Massimo , Bombonato Giancarlo , Gatta Angelo
Published: March 2007

Doppler sonographic portal vein parameters are used for the noninvasive evaluation of portal hypertension in cirrhosis. The patency of a paraumbilical vein is a rather frequent finding in cirrhosis, which may affect hepatic hemodynamics and function. We evaluated portal and hepatic arterial parameters in 184 cirrhotic patients with and without a patent paraumbilical vein and the relationships with paraumbilical blood flow. A patent paraumbilical vein was found in 33.7% of patients. The prevalence was higher (56.8%) in Child C patients. Portal blood flow velocity (PBV) (10.8 ± 2.2 vs. 9.8 ± 2.4 cm/sec; P < .01) and volume (PBF) (995.0 ± 383.8 vs. 811.6 ± 318.7 mL/min; P < .001) were significantly higher, and effective portal liver perfusion (PLP) (portal blood flow - paraumbilical blood flow) (621,3 ± 420.8 vs. 811.6 ± 318.7 mL/min; P < .001) was significantly lower in patients with a patent paraumbilical vein than in those without. These differences were more evident in Child C patients (10.7 ± 2.0 vs. 8.3 ± 2.3 cm/sec; 935.7 ± 378.3 vs. 680.6 ± 239.4 mL/min; 369.0 ± 282.0 vs. 680.6 ± 239.4 mL/min). Portal vein diameter, the congestion index (CI) of the portal vein, hepatic arterial resistance indexes, and the severity of esophageal varices did not differ between the two groups. In patients with a patent paraumbilical vein, the Child-Pugh score and the prevalence of ascites were significantly higher than in those without. In conclusion, the evaluation of PBV and PBF in cirrhotic patients can provide misleading results if a paraumbilical vein is patent, underestimating the degree of portal hypertension. The CI and hepatic arterial resistance indexes are not influenced by the paraumbilical vein patency. (Hepatology 1995; 22:1689-1694).
Renal effects of dietary supplementation with fish oil in cyclosporine-treated liver transplant recipients
Authors: Badalamenti Salvatore , Salerno Francesco , Lorenzano Elettra , Paone Gianni , Como Giovanna , Finazzi Silvia , Sacchetta Anna Cristina , Rimola Antoni , Graziani Giorgio , Galmarini Dinangelo , Ponticelli Claudio
Published: March 2007

Nephrotoxicity is the main untoward effect of cyclosporine (CsA) treatment. Experimental and clinical data suggest that dietary supplementation with fish oil may lessen cyclosporine nephrotoxicity, possibly by lowering renal thromboxane (Tx) production. We have studied the renal effects of a daily supplementation for 2 months of 12 g fish oil (18% C20:5 n-3 eicosapentaenoic acid [EPA] and 12% C22:6 n-3 docosahexanoic acid [DHA]) in a placebo-controlled (12 g corn oil), prospective, randomized, double-blind study of stable CsA-treated liver transplant recipients. Thirteen patients ingested corn oil capsules and 13 fish oil. Compliance with dietary regimen was confirmed by fatty acid chromatography that showed increased plasma concentrations of EPA (from 0.4 ± 0.02% to 4.6 ± 0.5%, P < .0001) and DHA (from 1.8 ± 0.2% to 3.9 ± 0.1%, P < .0001) in the fish oil group and increased plasma concentration of linoleic acid (C18:2 n-6) in the corn oil group (from 25 ± 2% to 28.4 ± 2%, P < .001). At the end of the 2 months of the study, in the fish oil group the effective renal plasma flow increased by 22% (P = .012), the glomerular filtration rate increased by 33% (P = .057), the renal blood flow increased by 17% (P = .024), and the calculated total renal vascular resistances decreased by 20% (P = .034). In contrast, none of these parameters changed in the corn oil group. The renal functional reserve determined during l-arginine infusion, plasma renin activity (PRA), and plasma aldosterone (PA) remained unchanged during the study in either group. The urine excretion of Tx B2 decreased from 707 ± 192 to 276 ± 76 pg/hr (P = .013) in fish oil group, whereas it increased from 428 ± 195 to 870 ± 310 pg/hr (P = .016) in corn oil group. In conclusion, 2 months of diet supplementation with fish oil in stable CsA-treated liver transplant recipients was associated with significant improvement of renal hemodynamics and with significant reduction of the renal production of TxB2. (Hepatology 1995; 22:1695-1701).
The clinical significance of p53 gene mutation in hepatocellular carcinomas from japan
Authors: Hayashi Hiroshi , Sugio Kenji , Matsumata Takashi , Adachi Eisuke , Takenaka Kenji , Sugimachi Keizo
Published: March 2007

To clarify the clinical significance of the mutation of p53 gene in hepatocellular carcinoma (HCC), 90 resected specimens from Japanese patients were assayed using a polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) analysis. p53 mutations were detected in 25 cases (27.8%) at exons 4, 5, 6, 7, and 8, and the most frequent region of the mutation was at exons 5 and 7. No statistically significant correlation was observed between the p53 mutations and the clinical features except for the preoperative alpha-fetoprotein (AFP) level (P < .05). According to the pathological features, prognostic factors, such as size of the tumor, vascular invasion, fibrous capsule infiltration, and intrahepatic metastasis, showed no relationship to the existence of the mutation. However, p53 mutations were significantly associated with the degree of differentiation of HCC; that is, the mutation was found in 19 cases of 53 poorly differentiated HCCs (35.9%) and 2 of 3 cases of anaplastic HCCs (66.7%). The presence of p53 mutations was associated with a shortened cancer-free survival (P < .001, by log rank test) and a shortened survival (P < .05). A multivariate analysis by the Cox regression analysis showed that the p53 mutations were an unfavorable prognostic factor related to recurrence (P < .005), which is especially significant within the first postoperative year. These results suggest that the mutations of p53 gene of HCC might be an independent prognostic predictor to help in the selection of candidates who should undergo more intensive postoperative treatment. (Hepatology 1995; 22:1702-1707).
Expression of intercellular adhesion molecule 1 in human hepatocellular carcinoma
Authors: Momosaki Seiya , Yano Hirohisa , Ogasawara Sachiko , Higaki Koichi , Hisaka Tohru , Kojiro Masamichi
Published: March 2007

We investigated the expression of intercellular adhesion molecule 1 (ICAM-1) in ex vivo human hepatocellular carcinoma (HCC) cells and in vitro in eight liver cancer cell lines, including six HCC cell lines and two combined hepatocholangiocarcinoma (CHC) cell lines. Immunohistochemistry showed the expression of ICAM-1 on the HCC cell surface with honeycomblike appearance in most cases (96.2%). On the other hand, hepatocytes in noncancerous areas did not express ICAM-1, except those hepatocytes in the periportal and intra-acinar areas with inflammation. Immunohistochemical study on cultured cells revealed that four cultured HCC cell lines and one CHC cell line constitutively expressed ICAM-1 on the cell surface and in the cytoplasm. Flow cytometric analysis revealed that immunostain-positive cells expressed surface ICAM-1 with more than a 90% positive cell rate, and their expressions were upregulated by incubation of cells with inflammatory cytokines, such as interferon alfa, interferon gamma, tumor necrosis factor-α, and interleukin 1β. Soluble ICAM-1 was detected in supernatants of cell lines expressing cell surface ICAM-1, in addition to one HCC cell line without surface ICAM-1 expression, and was increased in amounts 2- to 20-fold by inflammatory cytokines. These findings suggest that liver cancer cells in ex vivo may express not only surface but also a soluble form of ICAM-1, differently from normal hepatocytes, and that both expressions are upregulated by inflammatory cytokines. (Hepatology 1995; 22:1708-1713).
Screening for hemochromatosis in children of homozygotes: Prevalence and cost-effectiveness
Authors: Adams Paul C. , Kertesz Ann E. , Valberg Leslie S.
Published: March 2007

Although hereditary hemochromatosis is an autosomal recessive disease, most homozygotes are concerned with the genetic implications for their children. The optimal age for testing children and the cost implications of screening their children have not been clearly established. A clinical database consisting of 255 children from families with at least one homozygote is used to assess the prevalence of homozygotes among children of homozygous parents and to review the biochemical abnormalities and life-threatening symptoms in these young adults. Decision analysis is used to estimate the cost and utility of screening children of a homozygous parent. Eleven homozygotes were discovered among children of homozygotes. Only one male had a life-threatening event, cirrhosis. Decision analysis estimated cost saving of $12 per child screened ($ net present value) and a saving of 10 quality-adjusted days per child screened at age 10 years compared with not screening. If screening began at age 20 years, there is a cost saving of $65 per child screened. Sensitivity analysis showed that the major factors influencing cost savings were the cost of venesections, sensitivity and specificity of the screening tests, and prevalence of disease. Because the prevalence of hemochromatosis is higher in children of homozygotes than in the general population, screening with transferrin saturation and ferritin as early as age 10 years is recommended. Savings are augmented if the cost per venesection is eliminated by allowing hemochromatosis patients to become voluntary blood donors. (Hepatology 1995;22:1720-1727).
Evidence for the presence of an inactive precursor of human hepatocyte growth factor in plasma and sera of patients with liver diseases
Authors: Arakaki Naokatu , Kawakami Shuichi , Nakamura Osamu , Ohnishi Tomokazu , Miyazaki Hiroomi , Ishii Takehisa , Tsubouchi Hirohito , Daikuhara Yasushi
Published: March 2007

Human hepatocyte growth factor (hHGF), which is now known to be the same protein as the scatter factor and the tumor cytotoxic factor, is a heterodimeric protein with one heavy chain and one light chain linked together by a disulfide bond, and is thought to be involved in liver regeneration. Using an enzyme-linked immunosorbent assay (ELISA) and Western blot analysis, we found that a significant amount of single chain precursor of hHGF (pro-hHGF) was present in plasma of patients with fulminant hepatic failure (FHF) and that normal human serum contained a protease or proteases that convert pro-hHGF to a heterodimeric (mature) form of hHGF. We also showed that the processing protease activity for hHGF was suppressed by such serine protease inhibitors as leupeptin, antipain, and aprotinin, and that sera of patients with liver diseases such as fulminant hepatic failure, acute hepatitis, chronic hepatitis, and cirrhosis contained not only pro-hHGF but also the protease. This is the first report showing the presence of pro-hHGF in human blood, and our observations suggest that hHGF is synthesized and secreted from the hHGF-producing cells as an inactive pro-hHGF after hepatic injuries, and the pro-hHGF is then converted to an active heterodimeric form of hHGF in the blood. It is also suggested that plasma of patients with liver diseases contains an active protease or proteases that convert pro-hHGF to a mature form of hHGF. (Hepatology 1995; 22:1728-1734).
The integrity of the cholecystokinin receptor gene in gallbladder disease and obesity
Authors: Nardone Gerardo , Ferber Irene A. , Miller Laurence J.
Published: March 2007

Cholesterol gallstone disease and obesity are common and often associated disorders that could be affected by dysfunction of the receptor for cholecystokinin (CCK). Extending earlier studies that identified a defect at the level of receptor-G protein coupling in cholesterol gallstone disease, we characterized the primary structure of the gallbladder CCK receptor in patients undergoing a cholecystectomy. Represented were patients with cholesterol gallstones, as well as controls with pigment gallstones or without gallbladder disease. Both groups were composed of the range of body habitus from lean to morbidly obese. No evidence of any sequence mutation or polymorphism in the CCK receptor gene was found in any patient. This should lead future investigations of the pathogenesis of these problems toward the possible contribution of the plasmalemmal environment in affecting the association between normal receptors and G proteins. (Hepatology 1995; 22:1751-1753).
Effect of donor age and sex on the outcome of liver transplantation
Authors: Marino Ignazio Roberto , Doyle Howard R. , Aldrighetti Luca , Doria Cataldo , McMichael John , Gayowski Timothy , Fung John J. , Tzakis Andreas G. , Starzl Thomas E.
Published: March 2007

We correlated donor and recipient factors with graft outcome in 436 adult patients who underwent 462 liver transplants. Donor variables analyzed were age, gender, ABO blood group, cause of death, length of stay in the intensive care unit, use of pressors or pitressin, need for cardiopulmonary resuscitation, terminal serum transaminases, and ischemia time. Recipient variables analyzed were age, gender, primary diagnosis, history of previous liver transplant, ABO blood group, cytotoxic antibody crossmatch, United Network for Organ Sharing (UNOS) status, and waiting time (except for the cross-match results, they were all known at the time of the operation). The endpoint of the analysis was graft failure, defined as patient death or retransplantation. Using multivariate analysis, graft failure was significantly associated with donor age, donor gender, previous liver transplantation, and UNOS 4 status of the recipient. The effect of donor age became evident only when they were older than 45 years. Livers from female donors yielded significantly poorer results, with 2-year graft survival of female to male 55% (95% CI, 45% to 67%); female to female, 64% (95% CI, 54% to 77%); male to male, 72% (95% CI, 66% to 78%); and male to female, 78% (95% CI, 70% to 88%). The only donors identified as questionable for liver procurement were old (±60 years) women in whom the adverse age and gender factors were at least additive. However, rather than discard even these livers, in the face of an organ shortage crisis, their individualized use is suggested with case reporting in a special category. (Hepatology 1995; 22:1754-1762).
Disruption of circadian locomotor activity in rats after portacaval anastomosis is not gender dependent
Authors: Steindl Petra E. , Gottstein Jeanne , Blei Andres T.
Published: March 2007

A recent study suggested that female rats are less affected by a portacaval anastomosis (PCA) than their male counterparts, as measured by body weight and changes in locomotor activity. In this study, we evaluated the entrainment of locomotor activity to the light/dark (LD) cycle, a consistent abnormality in the portacaval shunted rat. The degree of entrainment was measured in male and female rats before and after PCA or sham operation. All four groups of animals showed strong entrainment to an LD cycle before surgery. After portacaval anastomosis, male and female rats exhibited a highly significant decrease in overall motor activity as compared with the preoperative period and as compared with sham-operated animals of the same gender. The percentage of total activity during daytime was significantly increased after portacaval anastomosis. The reduction in parameters of entrainment indicates a disruption of circadian function in both portacavalshunted groups. Portal pressure measurements confirmed the patency of the shunts. Cortical brain glutamine levels were similarly increased in male and female shunted rats. The loss of body weight was slightly, but not significantly, more pronounced in male animals after shunt surgery. In conclusion, our results do not support a role for gender in the disruption of circadian function in rats after PCA. (Hepatology 1995; 22:1763-1768).
Regulation of gene expression by interleukin-6 in fetal rat hepatocyte primary cultures: Role of epidermal growth factor and dexamethasone
Authors: Roncero Cesar , Fabregat Isabel , Benito Manuel
Published: March 2007

Fetal rat hepatocytes incubated in the absence of hormonal signals, or under proliferative (presence of epidermal growth factor [EGF]) or differentiative (presence of dexamethasone) culture conditions, showed responsiveness to interleukin-6 (IL-6). Northern blotting analysis for some typical acute phase genes such as haptoglobin and other proteins not previously identified as acute-phase reactants, such as α-fetoprotein, β2-microglobulin, and fibronectin, showed a positive modulation by IL-6, in a dose-dependent manner. However, a wellcharacterized negative acute-phase reactant such as albumin was not responsive to IL-6. The well-established synergism between glucocorticoids and IL-6 on inducing transcription is absent in fetal hepatocytes. Conversely, the combination of IL-6 and EGF produced different patterns of expression, depending on the messenger RNA (mRNA) analyzed. Thus, EGF abolished the increased mRNA levels of haptoglobin caused by IL-6 but had no effect on other genes such as α-fetoprotein and fibronectin. (Hepatology 1995; 22:1769-1775).
Acetaminophen-induced microvascular injury in the rat liver: Protection with misoprostol
Authors: Lim Sook Ping , Andrews Fiona Jane , O’Brien Paul Edmond
Published: March 2007

Studies into the mechanism of acetaminophen (APAP)-induced hepatotoxicity have focused mainly at the hepatocellular level. This study aimed to investigate the effect of acetaminophen on the hepatic microvasculature using a vascular casting technique. Acetaminophen was administered at a dose of 650 mg/kg body weight (intraperitoneally) to fasted male Long Evans rats. Microvascular casting was performed at various points after drug administration. Liver casts from control rats showed good patency with normal hepatic microvasculature. Thirty-six hours after overdose with acetaminophen, liver casts showed rounded centrilobular cavities of various sizes, representing regions in which cast-filled sinusoids were absent with relatively normal microvasculature within periportal regions. Evidence of microvascular injury occurred as early as 5 hours after acetaminophen overdose. This injury consisted of changes to centrilobular sinusoids including areas of incomplete filling and dilated centrilobular sinusoids. Misoprostol (a prostaglandin E1 analog) treatment (6 × 25 μ/kg) given before and after acetaminophen administration markedly reduced the extent of microvascular injury with only small focal unfilled areas in the casts and a generally intact microvasculature. In conclusion, this study shows that overdosage with APAP resulted in an extensive, characteristic pattern of hepatic microvascular injury in the centrilobular region. The results also suggest that microvascular injury is an early event in the pathogenesis of acetaminophen hepatotoxicity. Misoprostol was found to protect against injury occurring at the microvascular level. (Hepatology 1995; 22:1776-1781).
An array of mitochondrial alterations in the hepatocytes of long-evans cinnamon rats
Authors: Sternlieb Irmin , Quintana Nelson , Volenberg Irene , Schilsky Michael L.
Published: March 2007

In an attempt to identify the cellular targets of copper toxicity, we studied the ultrastructure of hepatocytes in the livers of 23 Long-Evans Cinnamon (LEC) rats ranging in age from 10 to 89 weeks. The hepatic copper concentration ranged from 325 to 2,126 (mean, 930) μg/g dry weight. Thirteen rats displayed varying degrees of jaundice at the time of killing. Numerous nuclei were indented by cytoplasmic invaginations. Conspicuous abnormalities were displayed by mitochondria. These included marked pleomorphism; increased or diminished matrical density; rearrangement, elongation, dilatation, stacking, or disappearance of cristae; absence of matrical granules; presence of matrical inclusions of flocculent electron-dense deposits; and degenerative changes. The severity of the ultrastructural changes did not correlate with the hepatic copper concentration but did correlate with the degree of the icterus displayed by the rats. Certain mitochondrial changes resembled those encountered in the hepatocytes of patients with Wilson’s disease, confirming that these organelles are important targets of copper toxicity. (Hepatology 1995; 22:1782-1787).
Hepatoma cell-specific expression of a retrovirally transferred gene is achieved by α-fetoprotein but not insulinlike growth factor II regulatory sequences
Authors: Arbuthnot Patrick , Bralet Marie-Pierre , Thomassin Helene , Danan Jean-Louis , Bréchot Christian , Ferry Nicolas
Published: March 2007

To target gene expression to malignant hepatic cells, we have constructed recombinant retroviral vectors containing a reporter gene encoding nuclear β-galactosidase (nls-LacZ) under transcriptional control of regulatory sequences from the rat α-fetoprotein (AFP) or human insulinlike growth factor II (IGFII) genes. The AFP and IGFII P3 promoters activate transcription during fetal development and are often reactivated in hepatocellular carcinoma (HCC). Infection of several cultured cell types with the retroviral vector containing the IGFII P3 sequence resulted in expression of the reporter gene in all cell lines tested, including those that do not produce IGFII. In contrast, selective expression was achieved by vectors containing the AFP transcriptional regulatory sequence. Nuclear β-galactosidase activity was detectable in cells from lines that produce AFP, and not in cells that do not express the AFP, gene. In most infected cell lines, retroviral RNA synthesis from the 5′ LTR was inhibited, and deletion of the retroviral LTR enhancer did not change expression from either the IGFII P3-nls-LacZ or the AFP-nls-LacZ cassettes. After treatment of cells with 12-O-tetradecanoylphorbol-13-acetate and epidermal growth factor (EGF), the decrease in concentrations of endogenous AFP messenger RNA (mRNA) and nls-LacZ mRNA transcribed from the transferred AFP regulatory sequence were similar. In the context of an integrated provirus, the AFP transcriptional regulatory sequence is therefore subject to similar regulatory control as that of the endogenous gene. These data show that the AFP sequence, and not the IGFII P3 promoter we used, is suitable for targeting gene expression to malignant hepatic cells. (Hepatology 1995; 22:1788-1796).
Ciprofloxacin prevents the inhibitory effects of acute ethanol exposure on hepatic regeneration in the rat
Authors: Minuk Gerald Y. , Gauthier Tony , Zhang Xin K. , Wang Gu Qi , Pettigrew Norman M. , Burczynski Frank J.
Published: March 2007

To determine whether the inhibitory effects of ethanol on hepatic regeneration could be prevented by ciprofloxacin, a fluroquinolone antibiotic with gammaaminobutyric acid (GABAA), receptor antagonist properties, adult, male Sprague-Dawley rats (n = 6-8/group) received intraperitoneal injections of saline, putrescine (a hepatic growth promotor, 50 mg/kg), or ciprofloxacin (100 mg/kg), followed 1 hour later by gastric gavage with saline or ethanol (5 g/kg). One hour post-gavage, all rats underwent a 70% partial hepatectomy (PHx). Hepatic regenerative activity was documented 24 hours post-PHx by 3H-thymidine incorporation into hepatic DNA (DNA synthesis), proliferating cell nuclear antigen staining, and hepatic tissue putrescine levels. Compared with healthy controls, DNA synthesis rates were significantly lower in ethanol-gavaged/saline-treated rats (22.7 ± 4.4 × 103 vs. 12.3 ± 6.9 × 103 DPM/mg DNA, respectively, P < .001) but unaltered in putrescine-(18.8 ± 3.4 × 103 DPM/ mg DNA) and ciprofloxacin-treated (18.3 ± 2.6 × 103 DPM/mg DNA) rats. Hepatic proliferating cell nuclear antigen staining supported these findings. Hepatic putrescine levels also correlated with DNA synthesis data, being decreased in ethanol-gavaged/saline-treated rats (86 ± 14 pmoles/mg tissue) compared with healthy controls (120 ± 12 pmoles/mg, P < .01), ethanol-gavaged/putrescine-treated (112 ± 14 pmoles/mg, P < .05) and ethanol-gavaged/ciprofloxacin-treated (125 ± 17 pmoles/mg, P < .05) rats. To determine whether these effects resulted from GABAA receptor-mediated changes in liver membrane potentials, intracellular membrane potentials were recorded before and 1 hour after PHx in healthy control, ethanol-gavaged/saline-treated and ethanol-gavaged/ciprofloxacin-treated rats. In these studies, ciprofloxacin prevented ethanol-induced depolarization of the liver (change in membrane potential of healthy controls, ethanol-gavaged/saline-treated, and ethanol-gavaged/ciprofloxacin-treated rats were −9 ± 1, −15 ± 2, and −3 ± 1 mV, respectively). In conclusion, the results of this study indicate that the inhibitory effects of acute ethanol exposure on hepatic regenerative activity in rats can be prevented by exogenous ciprofloxacin. (Hepatology 1995; 22:1797-1800).
Portal vein embolization with steel coils and absolute ethanol: A comparative experimental study with canine liver
Authors: Yamakado Koichiro , Takeda Kan , Nishide Yoshiya , Jin Jugangu , Matsumura Kaname , Nakatsuka Atsuhiro , Hirano Tadanori , Kato Noriyuki , Nakagawa Tsuyoshi
Published: March 2007

Portal vein embolization (PVE) is expected to have an adjuvant effect on the treatment of human liver tumors with transcatheter hepatic arterial embolization (TAE). In this experimental study, PVE was tried using two kinds of embolic materials, steel coils (coils) and absolute ethanol (ethanol). The aim of this article is to compare the effectiveness between the two methods. Five dogs and 12 dogs were treated with coils and ethanol, respectively. The second order portal branches were embolized. Embolization effects of each embolic material on the hepatic vessels and the liver parenchyma were studied angiographically and histologically. PVE with coils achieved immediate occlusion of the embolized vessels in all dogs. Unexpectedly, unembolized vessels were also constricted and/or obstructed in 4 of 5 dogs during a 4-week follow-up period. Histological examination showed no hepatic necrosis in all dogs. By contrast, in the dogs treated with ethanol, venous occlusion was achieved immediately after embolization with ethanol of 0.4 mL/kg or more. The embolized vessels remained occluded and no obstruction of unembolized vessels was observed until 4 weeks after embolization. Hepatic necrosis was observed in the segments where portal veins were occluded. The degree of parenchymal damage was proportional to the injected ethanol dose. Judging from these results, ethanol is likely to be more suitable than coils as an embolic material for PVE. PVE with coils is often dangerous because unembolized vessels might be occluded unexpectedly. For the clinical use of PVE with thanol, the appropriate dose of injection should be investigated. (Hepatology 1995; 22:1812-1818).
Independent modulation of galactose-specific receptor expression in rat liver cells
Authors: Massimi Mara , Devirgiliis Laura Conti , Kolb-Bachofen Victoria , Dini Luciana
Published: March 2007

The expression of galactose-specific receptors on liver cells from rats at the end of pregnancy and from estrogen-treated animals was studied. The number and distribution of binding sites were estimated on hepatocytes and Kupffer and endothelial cells in vitro as well as in situ by means of protein-gold complexes. Hepatocytes and endothelial cells from pregnant rats showed an increased binding activity of at least three times for hepatocytes and one and a half times for endothelial cells with respect to normal rat livers. The increase in the hepatocyte receptor expression was paralleled by an increase in the level of its specific messenger RNA (mRNA). On Kupffer cells, a decreased number of binding sites, at least three times less than control values, was measured. The correlation between the altered hormonal level during pregnancy and the expression of galactose binding sites was examined in hepatocytes and Kupffer cells isolated from virgin rats treated with the synthetic estrogen diethylstilbesterol. In estrogen-treated rats both the binding sites and the specific mRNA of hepatocytes increased as compared with vehicle-treated or untreated animals. In contrast, in Kupffer cells both the estrogen treatment as well as vehicle-only injection led to a significant reduction in the expression of binding sites as compared with virgin untreated animals. To establish whether the decrease of galactose binding sites in Kupffer cells was related to the activation of macrophages or to the removal of plasma membrane caused by enhanced nonspecific phagocytosis, in situ binding experiments were performed after lipopolysaccharide (LPS)-stimulation or latex-bead phagocytosis. Nonspecific phagocytosis does not affect the binding activity, which instead appears strongly reduced after LPS injection. These findings suggest an independent response of galactose-specific receptor expression systems in the different types of liver cells to modulating agents. (Hepatology 1995; 22:1819-1828).
Interleukin-1 and nitric oxide protect against tumor necrosis factor α-induced liver injury through distinct pathways
Authors: Bohlinger Ines , Leist Marcel , Barsig Johannes , Uhlig Tefan , Tiegs Gisa , Wendel Albrecht
Published: March 2007

Mice sensitized with d-galactosamine (GalN) and challenged with recombinant murine tumor necrosis factor α (TNFα) developed severe apoptotic and secondary necrotic liver injury as assessed by histology, measurement of cytosolic DNA fragments, and determination of liver specific enzymes in plasma. Pretreatment with recombinant human interleukin-1β (IL-1) rendered mice insensitive to this TNFα toxicity. Coadministration of the liver-specific transcriptional inhibitor GalN with IL-1 prevented the development of tolerance, implicating de novo synthesis of liver specific proteins in the induction of tolerance. Pretreatment of mice with IL-1 resulted in elevated levels of nitrite/nitrate in serum and in enhanced nitric oxide synthase (NOS) activity in liver cells isolated from these animals. In addition, pharmacological doses of the nitric oxide (NO) donor sodium nitroprusside conferred complete protection against TNFα-induced liver injury in galactosamine-sensitized mice, suggesting a possible link between IL-1- and NO-in-duced protection. However, prevention of NO-synthesis by N
G-monomethyl-l-arginine (NMMA) did not abolish IL-1-induced tolerance to TNFα in vivo. Cytotoxicity of TNFα to isolated hepatocytes sensitized with actinomycin D (ActD) was not significantly altered by inhibition of endogenous nitrite release. Also, enhanced NO production elicited in vitro by glycerol trinitrate or ex vivo by pretreatment with IL-1 had no significant effect in this system. We conclude that IL-1- and NO-induced protection of mice against TNFα-mediated liver damage follow distinct pathways. (Hepatology 1995; 22:1829-1837).