bluuebunny/crossref_metadata_2025 huggingface dataset:
Interferon–α Therapy for Hepatitis C Virus Infection After Liver Transplantation
Authors: Wright Teresa L. , Combs Connie , Kim Michael , Ferrell Linda , Bacchetti Peter , Ascher Nancy , Roberts John , Wilber Judith , Sheridan Pat , Urdea Mickey
Published: March 2007

The aims of this pilot study were to evaluate the safety and efficacy of interferon–α2b for treatment of hepatitis C virus infection in liver transplant recipients, to monitor changes in hepatitis C virus RNA levels with treatment and to determine pretreatment parameters predictive of a complete response. Eighteen patients with documented hepatitis C virus viremia and histological evidence of hepatitis after liver transplantation received 3 million units of α interferon three times weekly for at least 4 mo. Pretreatment serum aminotransferase levels were at least 1.5 times the upper limit of normal and no patient had concomitant hepatitis B virus infection. Response to therapy was defined as normalization of both aspartate and alanine aminotransferase at the end of treatment. Five patients (28%) had a complete response, whereas 13 (72%) had persistent elevation of one or both aminotransferases (nonresponders). At the end of therapy, hepatitis C virus RNA levels were reduced in both responders and nonresponders (p = 0.043 and 0.039, respectively by Wilcoxon signed rank test). After cessation of treatment, aminotransferases remained normal in four of five responders but serum hepatitis C virus RNA levels returned to pretreatment levels in responders and nonresponders. There was no significant change in histological score with therapy. Responders were more likely than nonresponders to have low pretreatment hepatitis C virus RNA levels and low serum bilirubin (p = 0.0004 and 0.0077, respectively). Responders tended to have a prolonged interval between transplantation and initiation of therapy (p = 0.10 by rank logistic regression analysis). Side effects resulted in early cessation of therapy in two patients and dose reduction in six. Histological evidence of possible rejection was seen in one patient. We conclude that normalization of liver enzymes can be seen in patients with posttransplant hepatitis C virus infection on interferon therapy and that treatment is associated with fall in hepatitis C virus RNA levels, regardless of biochemical response. However, virological changes are transient. Pretreatment variables such as hepatitis C virus RNA levels and serum bilirubin may aid in the selection of patients more likely to respond to therapy and the design of future controlled clinical trials. (Hepatology 1994;20:773-779).
Expression of Insulin–Like Growth Factor Ii in Hepatitis B, Cirrhosis and Hepatocellular Carcinoma: Its Relationship With Hepatitis B Virus Antigen Expression
Authors: Su Qin , Liu Yan-Fang , Zhang Jin-Feng , Zhang Sui-Xiang , Li De-Fang , Yang Jia-Ji
Published: March 2007

Expression of insulin–like growth factor II in two human hepatocellular carcinoma cell lines and in hepatitis B, cirrhosis and hepatocellular carcinoma in 419 cases were investigated, and its relationship with the expression of hepatitis B virus x gene was studied by means of immunohistochemical and electron microscopic techniques. The results demonstrated that hepatocellular carcinoma cells (SMMC 7721 and QGY 7703) in culture could express insulin–like growth factor II. Expression seemed to be regulated by cell density, which was suggested as the molecular basis of the contact inhibition of cell proliferation. In tissue sections, cells with high expression of insulin–like growth factor II were observed not only in hepatocellular carcinoma (93%) but also in 95% of the pericancerous liver tissues, 72% of cirrhotic livers, 64% of chronic active hepatitis and 37% of chronic persistent hepatitis. in most cases of hepatocellular carcinoma, insulin–like growth factor II was localized in the cytoplasm of the cancer cells. in the benign liver disorders, four types of cells that highly expressed insulin–like growth factor II were observed: (a) a kind of small liver cell we named the

small polygonal liver cell

; (b) multinuclear giant hepatocytes; (c) hepatocytes in most of hyperplastic and neoplastic nodules, small hepatocyte nodules and some of regenerative nodules; and (d) some proliferating ductular cells. Even more interestingly, insulin–like growth factor II expression was shown to be closely related to the expression of hepatitis B virus x gene product. We suggest that the activation of insulin–like growth factor II gene and its overexpression may be a crucial step in the processes of hepatitis B virus—associated hepatocarcinogenesis and that the x gene product may activate the insulin–like growth factor II gene through a transactivation mechanism. in addition, we studied the characteristics of

small polygonal liver cells

, and the roles they may play in the regeneration and carcinogenesis of hepatitis B virus—infected liver are discussed. (Hepatology 1994;19:788-799).
Blood and Plasma 5–Hydroxytryptamine Levels in Patients With Cirrhosis
Authors: Beaudry Patrice , Hadengue Antoine , Callebert Jacques , Gaudin Christophe , Soliman Hany , Moreau Richard , Launay Jean Marie , Lebrec Didier
Published: March 2007

Serotoninergic mechanisms are thought to play a role in portal hypertension. Because this biomine is metabolized by the liver, peripheral blood and plasma levels of 5–hydroxytryptamine and 5–hydroxyindole acetic acid (the main metabolite of 5–hydroxytryptamine) were measured in 30 patients with cirrhosis. Whole–blood 5–hydroxytryptamine levels were significantly lower in patients with cirrhosis (158 ± 28 nM) than in age–matched controls (332 ± 19 nM), and no correlation was found between these levels and the severity of cirrhosis. Unconjugated plasma 5–hydroxytryptamine levels, an indication of the active form of 5–hydroxytryptamine, were significantly higher in patients with cirrhosis than in controls (6.8 ± 1.7 nM and 3.4 ± 0.5 nM, respectively), and in patients with cirrhosis these levels were higher in Pugh grade A than in Pugh grade C patients. Conjugated–plasma 5–hydroxytryptamine levels were not significantly different between patients with cirrhosis (32.2 ± 8.1 nmol/L) and controls (16.4 ± 1.4 nmol/L). Plasma 5–hydroxyindole acetic acid was significantly lower in patients with cirrhosis than in controls (1.5 ± 0.1 nmol/L and 2.3 ± 0.1 nmol/L, respectively). in conclusion, this study shows that serotoninergic mechanisms are altered in patients with cirrhosis. (Hepatology 1994;20:800-803).
Cysteinyl Leukotrienes in the Urine of Patients With Liver Diseases
Authors: Uemura Masahito , Buchholz Ulrike , Kojima Hideyuki , Keppler Andrea , Hafkemeyer Peter , Fukui Hiroshi , Tsujii Tadasu , Keppler Dietrich
Published: March 2007

The significance of cysteinyl leukotrienes was investigated in patients with liver diseases by measurements of leukotriene E4 and

N

–acetyl–leukotriene E4 in urine. A marked increase of renal cysteinyl leukotriene excretion was observed in patients with cirrhosis without and with ascites, intrahepatic cholestasis, and obstructive jaundice as compared with healthy subjects (leukotriene E4: means 82, 264, 221 and 142 versus 40 nmol/mol creatinine, respectively;

N

–acetyl–leukotriene E4: means 25, 64, 61 and 47 versus 13 nmol/mol creatinine, respectively). The urinary concentration of leukotriene E4 was positively correlated with the one of

N

–acetyl–leukotriene E4 (r = 0.81, p < 0.001). In patients with cirrhosis, the excretion of cysteinyl leukotrienes was strongly increased in patients in Child–Turcotte stage C as compared with those in Child–Turcotte stages A and B. In patients with intrahepatic cholestasis and in those with obstructive jaundice, the excretion of leukotriene E4 plus

N

–acetyl–leukotriene E4 was positively correlated with total serum bilirubin. In patients with cirrhosis and in those with obstructive jaundice, the cysteinyl leukotrienes in urine were negatively correlated with creatinine clearance. The elevated renal excretion of cysteinyl leukotrienes decreased after biliary drainage in patients with obstructive jaundice. These data support the concept that increased urinary excretion of cysteinyl leukotrienes in patients with cirrhosis is due to a reduced functional liver mass and that in patients with cholestasis it is mainly due to an impaired elimination into the biliary tract that results in a diversion to renal excretion. The increased concentrations of biologically active cysteinyl leukotrienes may play an important role in the renal circulatory disturbance in patients with cirrhosis and obstructive jaundice. (Hepatology 1994;20:804-812).
Aplastic Anemia After Liver Transplantation for Fulminant Liver Failure
Authors: Cattral Mark S. , Langnas Alan N. , Markin Rodney S. , Antonson Dean L. , Heffron Thomas G. , Fox Ira J. , Sorrell Michael F. , Shaw Byers W.
Published: March 2007

We determined the incidence and outcome of aplastic anemia among 56 patients who underwent liver transplantation for fulminant liver failure at the University of Nebraska Medical Center between July 1985 and December 1993. Aplastic anemia developed in 6 of 18 (33%) children and 1 of 19 (5%) adults who had fulminant non–A, non–B hepatitis; no cases of aplastic anemia occurred among patients with other causes of fulminant liver failure. None of these patients had evidence of a preexisting hematological disorder or in fection with hepatitis C virus (as determined with a second–generation ELISA). Aplastic anemia was diagnosed at a median of 4 wk after the onset of hepatitis, with five cases seen before transplantation. Six patients received antithymocyte globulin to promote remission of aplastic anemia. Three children died (fungal infection in two, intracranial hemorrhage in one)—one at 43, one at 108 and one at 119 days after transplantation—without remission of aplastic anemia. Among the four surviving patients, with median follow–up of 25 mo, complete and partial remission of aplastic anemia have occurred in three and one, respectively. Liver allograft function is stable in all surviving patients. The data demonstrate that aplastic anemia is a common complication among children who undergo liver transplantation for fulminant non–A, non–B hepatitis. It is associated with a high rate of mortality, although most survivors appear to have full hematological recovery. (Hepatology 1994;20:813–818).
Fetal Bile Acid Metabolism During Infancy: Analysis of 1β–Hydroxylated Bile Acids in Urine, Meconium and Feces
Authors: Kimura Akihiko , Yamakawa Ryoichi , Ushijima Kosuke , Fujisawa Takuji , Kuriya Norikazu , Kato Hirohisa , Inokuchi Takahiro , Mahara Reijiro , Kurosawa Takao , Tohma Masahiko
Published: March 2007

Fetal bile acids (1β–hydroxylated, 6α–hydroxylated and unsaturated bile acids), especially 1β,3α,7α, 12α–tetrahydroxy–5β–cholan–24–oic acid (CA–1β–ol), have been detected in urine and feces early in life. To investigate whether a fetal pathway of bile acid synthesis exists in infancy, we measured the concentrations of bile acids in the urine, meconium and feces from normal newborns and infants by means of gas chromatography—mass spectrometry. The mean ratio of total bile acids to creatinine in urine increased between birth and 7 days and then gradually decreased; however, the concentration of total bile acids in urine remained significantly higher than that in adult urine until 3 mo of age. The main urinary bile acid was CA–1β–ol, and substantial amounts of fetal bile acids were detected in urine until 3 mo of age. The ratio of cholic acid to chenodeoxycholic acid was abnormally low in meconium (mean, 0.44; range, 0.19 to 0.74), and hyocholic acid constituted 19.3% of total bile acids. The mean total bile acid content of feces decreased between birth and 7 days of age and thereafter increased. The mean percentage of fetal bile acids in feces decreased after birth, but substantial amounts were present in feces until 1 mo of age. Our data support the notion that a primitive pathway for the synthesis of fetal bile acids—such as CA–1β–ol, hyocholic acid and 3β–hydroxy–5–cholen–24–oic acid—may be responsible for excretion mechanisms of cholestatic bile acids during immaturity of hepatic excretory function in infants from birth until 3 mo of age. (Hepatology 1994;20:819-824).
Effects of Large–Volume Paracentesis on Pulmonary Function in Patients With Tense Cirrhotic Ascites
Authors: Angueira Carlos E. , Kadakia Shailesh C.
Published: March 2007

Large–volume paracentesis is an accepted therapeutic modality for the treatment of tense ascites in patients with cirrhosis. Whereas the effects of large–volume paracentesis on the cardiovascular system have been studied in great detail, the effects of tense ascites and large–volume paracentesis on the respiratory system have undergone only limited evaluation. Most patients report symptomatic improvement in breathing after large–volume paracentesis. The aim of our study was to prospectively evaluate the effects of large–volume paracentesis on the pulmonary function of patients with tense ascites. Nine patients with alcoholic cirrhosis and tense ascites underwent large–volume paracentesis (mean, 3,500 ml of ascites removed) followed by intravenous albumin infusion (10 gm/L ascites removed). Pulmonary function tests were performed just before and 2 hr after large–volume paracentesis. The functional residual capacity increased from 2.46 ± 0.92 to 2.99 ± 0.97 L (p = 0.01) and total lung capacity from 5.08 ± 1.18 to 5.72 ± 1.32 L (p = 0.03). The forced vital capacity, expiratory reserve volume and residual volume also increased but were not statistically significant (p = 0.07 for all). We noted no significant improvement in forced expiratory volume in 1 sec, forced expiratory effort 25% to 75%, vital capacity or diffusion capacity. There was marked symptomatic improvement in all patients. We conclude that large–volume paracentesis improves measured lung volumes within hours in patients with tense ascites. (Hepatology 1994;20:825-828).
Selecting the Donor Liver: Risk Factors for Poor Function After Orthotopic Liver Transplantation
Authors: Strasberg Steven M. , Howard Todd K. , Molmenti Ernesto P. , Hertl Martin
Published: March 2007

Initial poor function and primary nonfunction are important problems in clinical transplantation. The incidence of primary nonfunction is about 6% and that of initial poor function is about 15%. Grafts with initial poor function have a higher graft failure rate in the first 3 mo after transplantation. Severe steatosis and cold preservation in University of Wisconsin solution for over 30 hr will alone cause primary nonfunction. However, primary nonfunction is probably most often caused by the presence of multiple relative risk factors. The major donor–relative risk factors are moderate steatosis, cold preservation over 12 hr and donor age over 50 yr, whereas retransplantation, high (United Network of Organ Sharing class 4) medical status and kidney failure are recipient relative risk factors. The most important perioperative risk factor is warm ischemia time. Rates of primary nonfunction and initial poor function might be reduced by avoidance of combinations of risk factors. Several tests have been developed to predict primary nonfunction and initial poor function, but none is yet clinically efficient. (Hepatology 1994;20:829-838).
Diagnosis of Functional Kidney Failure of Cirrhosis With Doppler Sonography: Prognostic Value of Resistive Index
Authors: Maroto Albert , Ginès Angels , Saló Joan , Clària Joan , Ginès Pere , Anibarro Luis , Jiménez Wladimiro , Arroyo Vicente , Rodés Juan
Published: March 2007

Time–velocity wave–form analysis of Doppler signals from small intrarenal arteries allows estimation of intrarenal arteriolar vascular resistance. Among the various indexes proposed, the resistive index is the most widely used for this estimation. To investigate whether the resistive index is useful in the diagnosis of functional kidney failure and prediction of survival in cirrhotic patients with ascites, we measured resistive index, kidney and liver function and plasma levels of renin, aldosterone and antidiuretic hormone in 10 healthy subjects, 12 patients with compensated cirrhosis and 32 patients with cirrhosis and ascites (17 with kidney failure). A total of 28 clinical and laboratory variables were analyzed for prognostic value. Resistive index was significantly increased in patients with kidney failure (0.74 ± 0.01) compared with those in the other three groups (0.64 ± 0.01, 0.64 ± 0.02 and 0.67 ± 0.01) and correlated significantly with glomerular filtration rate, arterial pressure, plasma renin activity and free water clearance in the cirrhotic patients. The sensitivity and specificity of the resistive index in detecting kidney failure in patients with ascites were 71% and 80%, respectively. Nine variables were correlated with survival in the univariate analysis, including resistive index, age, hepatomegaly, blood urea nitrogen, serum creatinine, plasma sodium concentration, glomerular filtration rate, plasma renin activity and plasma concentration of antidiuretic hormone. Multivariate analysis disclosed only three independent predictors of survival: plasma renin activity, plasma concentration of antidiuretic hormone and serum sodium concentration. In conclusion, resistive index is a sensitive method to assess intrarenal hemodynamics in patients with cirrhosis and ascites. It also has predictive value for survival in these patients. (Hepatology 1994;20:839-844).
The Major Metabolites of Ursodeoxycholic Acid in Human Urine Are Conjugated With N –Acetylglucosamine
Authors: Marschall Hanns-Ulrich , Griffiths William James , Götze Ulrich , Zhang Jie , Wietholtz Hubertus , Busch Norbert , Sjövall Jan , Matern Siegfried
Published: March 2007

Ursodeoxycholic acid (750 mg/day) was administered orally to ten healthy subjects over a period of 10 days; 24 hr urine samples were collected the day before and on the last day of the study. Urinary bile acids were extracted, separated into groups of conjugates and analyzed by gas chromatography–mass spectrometry and fast atom bombardment mass spectrometry. Excretion of ursodeoxycholic acid rose from 70 to 2,915 μg/24 h. The highest increase was observed among

N

–acetylglucosamine conjugates, 90% of which constituted the previously unknown double conjugate of ursodeoxycholic acid with

N

–acetylglucosamine and glycine. Excretion of isoursodeoxycholic acid increased from 50 to 738 μg/24 h. This isomerization product of ursodeoxycholic acid was excreted almost exclusively as

N

–acetylglucosamine conjugate. In total,

N

–acetylglucosamine conjugates constituted 50% of urinary metabolites of ursodeoxycholic acid. In addition, metabolites of ursodeoxycholic acid hydroxlated at carbon atoms 1, 6, 22 and possibly 21 were observed. These compounds were also found as conjugates with

N

–acetylglucosamine. Their formation from ursodeoxycholic acid was definitely demonstrated by 13C–labeling after giving [24–13C]ursodeoxycholic acid to one of the healthy subjects and to a patient with extrahepatic cholestasis in whom hydroxylation of ursodeoxycholic acid at C–23 was also observed. The patient was also found to excrete the double conjugate of ursodeoxycholic acid with

N

–acetylglucosamine and taurine. The

N

–acetylglucosaminidation of ursodeoxycholic acid

in vivo

was shown to occur at C–7. This was established by collision–induced dissociation of the pseudomolecular anions generated by fast atom bombardment ionization of samples from pooled urine collected from the healthy subjects during administration of ursodeoxycholic acid and from the patient given [24–13C] ursodeoxycholic acid. The study shows that

N

–acetylglucosaminides are major urinary metabolites of ursodeoxycholic acid and that the selective

N

–acetylglucosaminidation of 7β–hydroxy bile acids occurs

in vivo

at the 7β–position. (Hepatology 1994;20:845-853).
Prospective Randomized Trial of Emergency Portacaval Shunt and Emergency Medical Therapy in Unselected Cirrhotic Patients With Bleeding Varices
Authors: Orloff Marshall J. , Bell Richard H. , Orloff Mark S. , Hardison William G.M. , Greenburg Gerson A.
Published: March 2007

A prospective randomized trial was conducted in unselected, consecutive patients with bleeding esophageal varices resulting from cirrhosis comparing (1) emergency portacaval shunt performed within 8 hr of initial contact (21 patients) with (2) emergency medical therapy (intravenous vasopressin and esophageal balloon tamponade) followed in 9 to 30 days by elective portacaval shunt in survivors (22 patients). All patients underwent the same diagnostic workup within 3 to 6 hr of initial contact, and received identical supportive therapy initially. All patients were followed up for at least 10 yr. The protocol contained no escape or crossover provisions. There were no statistically significant differences between the two treatment groups in the incidence of any of the clinical variables, results of laboratory tests or degree of portal hypertension. Child's risk classes in the shunt group were A—2 patients, B—8 patients and C—11 patients, whereas in the medical group they were A—10 patients, B—5 patients, and C—7 patients, a significant difference (p < 0.01) that might have favored emergency medical treatment. Bleeding was controlled initially and permanently by emergency shunt in every patient, but by medical therapy in only 45% (p < 0.001). Mean requirement for blood transfusion was 7.1 ± 2.6 units in the shunt group and 21.4 ± 2.6 units in the medical group (p < 0.001). Eighty–one percent of the patients in the shunt group were discharged alive compared with 45% in the medical group (p = 0.027). Five– and 10–yr observed survival rates were 67% and 57%, respectively, after emergency shunt compared with 18% and 18%, respectively, after the combination of emergency medical therapy and elective shunt (p < 0.01). These survival rates produced by emergency shunt performed within 8 hr of initial contact confirm the effectiveness of this procedure observed in our previous unrandomized studies. (Hepatology 1994;20:863-872).
Soluble Intercellular Adhesion Molecule–1 in Primary Biliary Cirrhosis: Relationship With Disease Stage, Immune Activity and Cholestasis
Authors: Lim Guan A. , Jazrawi Riadh P. , Ahmed Hafez A. , Levy Joanne H. , Zuin Massimo , Douds Andrew C. , Douglas J. , Northfield Tim C.
Published: March 2007

Intercellular adhesion molecule—1, strongly expressed on the interlobular and proliferating bile ducts in primary biliary cirrhosis, is important in the migration and adhesion of inflammatory cells from the circulation to these structures. A soluble form has been found to be elevated in serum in primary biliary cirrhosis. Our aim was to check on the role of soluble intercellular adhesion molecule—1 in primary biliary cirrhosis with particular reference to its specificity by comparison with other disease control groups and to assess its relationship with stage of disease activity, circulating lymphocyte activation and cholestasis. Soluble intercellular adhesion molecule—1 (enzymelinked immunosorbent assay) and liver biochemistry were measured in 41 patients with primary biliary cir rhosis, 9 with primary sclerosing cholangitis, 12 with alcoholic liver disease and 17 healthy controls. In subgroups of patients with primary biliary cirrhosis, lymphocyte activation and hepatic bile acid uptake and excretory rates were determined. Soluble intercellular adhesion molecule—1 was significantly higher in all three disease groups. Levels in primary biliary cirrhosis and primary sclerosing cholangitis were similar and significantly higher than alcoholic liver disease. Soluble intercellular adhesion molecule—1 expression was greater in late primary biliary cirrhosis than early disease and correlated with histological progression. Correlations were also found with alkaline phosphatase, γ–glutamyl transpeptidase and conjugated bilirubin. A trend toward an inverse correlation with hepatic excretory rate was found, but no correlation was detected with circulating lymphocyte interleukin–2 receptor expression. We conclude that serum soluble intercellular adhesion molecule—1 is markedly elevated in immune liver diseases characterized by chronic cholestasis. In primary biliary cirrhosis, it is a marker of stage of disease activity and cholestasis. Measurements of soluble intercellular adhesion molecule—1 may be useful in monitoring disease progression and in assessing the effects of therapy. (Hepatology 1994;20:882-888).
Systemic Release of Interleukin–10 During Orthotopic Liver Transplantation
Authors: Moine Olivier Le , Marchhant Arnaud , Durand François , Ickx Brigitte , Pradier Olrvier , Belghiti Jacques , Abramowicz Daniel , Gelin Michel , Goldman Michel , Devière Jacques
Published: March 2007

Experimental and clinical observations indicate that the liver allograft is less immunogenic than other organ transplants and can promote immune tolerance. Because interleukin–10 recently emerged as a macrophage and T–cell–derived cytokine with potent immunosuppressive properties, we studied its production in 28 patients undergoing orthotopic liver transplantation. Plasma levels of immunoreactive interleukin–10 dramatically increased within 2 hr after liver allograft reperfusion, with peak levels ranging between 214 and 4998 pglml (median = 677 pglml). This systemic release of interleukin–10 was transient because it returned to low levels by 48 hr (range = 26 to 51 pg/ml). The higher interleukin–10 levels measured in right atrial blood as compared with portal blood indicated that interleukin–10 was most likely synthesized within the liver graft. To get insight into the cellular origin of interleukin–10, we also measured serum levels of interleukin–4 and interferon–γ, both produced by T cells, and interleukin–8, a cytokine secreted by macrophages, in eight patients. Interleukin–4 and interferon–γ levels remained undetectable in most of the patients, whereas interleukin–8 levels paralleled those of interleukin–10. Portal endotoxemia was probably not involved in interleukin–10 production because endotoxin levels remained low (<20 pg/ml) before and after liver allograft reperfusion. Interleukin–10 plasma levels did not correlate either with cold ischemia time or with the occurrence of rejection episodes. We conclude that orthotopic liver transplantation is associated with a massive release of interleukin–10 and interleukin–8, most likely produced by allograft macrophages. (HEPATOLOGY 1994;20:889-892).
Epstein–Barr Virus in Inflammatory Diseases of the Liver and Liver Allografts: An In Situ Hybridization Study
Authors: Hubscher Stefan G. , Williams Ann , Davison Suzanne M. , Young Lawrence S. , Niedobitek Gerald
Published: March 2007

The Epstein–Barr virus is associated with a broad spectrum of lymphoproliferative diseases in liver allograft recipients. To investigate the effects of primary infection in children following liver transplantation and the possible role of Epstein–Barr virus in the pathogenesis of unexplained chronic hepatitis after transplantation, we used an in situ hybridization technique to detect small virus encoded nuclear RNAs in posttransplant specimens. For comparison, other posttransplant complications and diseases occurring in the nontransplanted liver were studied with the same method. We examined 114 specimens in the following categories: (a) children with primary Epstein–Barr virus infection after transplant (n = 25), (b) chronic hepatitis more than 12 mo after transplant (n = 14), (c) rejection (n = 15), (d) normahear–normal histology more than 12 mo after transplant (n = 13) (e) end–stage acute liver disease (n = 9) and (f) end–stage cirrhosis (n = 38). Thirty–three of 114 specimens had labeling of occasional portal and parenchymal lymphoid cells. These were present in each of the six main diagnostic categories listed above: 2 of 25, pediatric; 6 of 14, chronic hepatitis; 4 of 15, rejection; 1 of 13, normal; 4 of 9, acute; and 16 of 38, cirrhosis. In none of the patients with Epstein–Barr virus–infected cells did lymphoproliferative disease subsequently develop. In conclusion, we have been unable to show an obvious association between Epstein–Barr virus infection and graft dysfunction after liver transplantation. Our results suggest that immunological response to primary infection in children after liver transplantation may be adequate. Small numbers of Epstein–Barr virus–infected cells are present in a wide range of liver diseases. The finding of occasional infected cells in posttransplant biopsy tissue does not predict progression to lymphoproliferative disease. (HEPATOLOGY 1994;20:899-907).
Undulin Rna and Protein Expression in Normal and Fibrotic Human Liver
Authors: Milani Stefano , Grappone Cecilia , Pellegrini Giulia , Schuppan Detlef , Herbst Hermann , Calabró Antonio , Casini Alessandro , Pinzani Massimo , Surrenti Calogero
Published: March 2007

We have analyzed the distribution, gene expression and cellular origin of undulin, a large extracellular matrix glycoprotein associated with mature collagen fibrils, in human liver by immunohistochemistry, Northern–blot analysis and

in situ

hybridization. In normal liver, undulin was distributed as densely packed fibers in portal tract stroma, and as fine fibers along sinusoids, and around central veins. Undulin ribonucleic acid expression was low in normal liver, and confined to mesenchymal cells of portal tract stroma, vessel walls and perisinusoidal space. In fibrotic liver, undulin deposition and gene expression were enhanced in fibrotic stroma and areas of fibrogenesis identified by the presence of active septa and inflammatory infiltrate. Undulin gene expression in fibrotic liver was exclusively localized in mesenchymal cells that could be identified by staining for vimentin, and partially for α–smooth muscle actin as (myo)fibroblasts, and possibly fat–storing cells. These data suggest that undulin is a constituent of the hepatic extracellular matrix of normal human liver, and that it participates in the rearrangement of connective tissue occurring in hepatic fibrosis. (Hepatology 1994;20:908-916).
Effect of Different Sepsis–Related Cytokines on Lipid Synthesis by Isolated Hepatocytes
Authors: Vara Elena , Arias-Díaz Javier , Torres-Melero Juan , García Cruz , Rodríguez José M. , Balibrea José L.
Published: March 2007

Cytokines seem to play an important role in the metabolic disturbances that are commonly associated with sepsis. In this study, we analyzed the effect of tumor necrosis factor, interleukin–1 and interleukin–6, as well as that of tumor necrosis factor in combination with interleukin–1 or interleukin–6, both on free fatty acids and on phospholipid synthesis by isolated rat hepatocytes. All three cytokines and combinations caused inhibited D–[U–14C]glucose incorporation into phosphatidylcholine (tumor necrosis factor = 6.39 ± 1.13 pmol/μg protein vs. control = 12.90 ± 0.98 pmol/μg protein, n = 7; p < 0.001). However, when [U–14C]palmitate was used as radioactive precursor, tumor necrosis factor, either alone or in the presence of the other cytokines, stimulated phosphatidylcholine synthesis. D–[U–14C]glucose incorporation into free fatty acids and triacylglycerol was also significantly stimulated, whereas phosphatidylinositol labeling was found inhibited by the assayed cytokines. Our results demonstrate an effect of sepsis–related cytokines, more evident for tumor necrosis factor, on hepatocyte lipid synthesis either from glucose or palmitate. Also, the findings support the hypothesis that cytokine–induced changes in hepatocyte lipid synthesis can contribute to the impairment in lipidic metabolism seen in patients with sepsis. (Hepatology 1994;20:924-931).
In Vivo and in Vitro Uptake of An Anti–Cd30/Saporin Immunotoxin by Rat Liver Parenchymal and Nonparenchymal Cells
Authors: Battelli Maria Giulia , Buonamici Laura , Bolognesi Andrea , Stirpe Fiorenzo
Published: March 2007

A Ber–H2/saporin immunotoxin, consisting of the single–chain ribosome–inactivating protein saporin–S6 and the anti–CD30 monoclonal antibody Ber–H2, gave encouraging results in the treatment of refractory Hodgkin's disease but caused a transient hepatotoxicity. The accumulation of Ber–H2/saporin conjugate and of its components by rat liver parenchymal and nonparenchymal cells was studied. The

in vivo

concentration of intravenously injected Ber–H2/saporin, saporin or Ber–H2 in nonparenchymal cells was 4–, 25–and 11–fold higher, respectively, than that in parenchymal cells. Adherent

in vitro

cultured nonparenchymal cells, mostly Kupffer cells, accumulated the proteins approximately 10 times more than parenchymal cells; traces of free saporin were taken up by both types of cells.

In vitro

protein synthesis by both cell types was inhibited by 50% at nanomolar concentrations of saporin. Nonparenchymal cells were sensitive to Ber–H2/saporin at picomolar concentrations, whereas parenchymal cells were unaffected by the immunotoxin up to 100 pmol/L. The results of the uptake of, and the sensitivity to, the immunotoxin suggest that the sensitivity of liver cells is proportional to the uptake and that the

in vivo

damage to parenchymal cells is at least in part mediated by the toxicity to nonparenchymal liver cells. (Hepatology 1994;20:940-947).
Renal Effects of Natriuretic Peptide Receptor Blockade in Cirrhotic Rats With Ascites
Authors: Angeli Paolo , Jiménez Wladimiro , Arroyo Vicente , Mackenzie Harald S. , Zhang Ping L. , Clária Joan , Rivera Francisca , Brenner Barry M. , Rodés Joan
Published: March 2007

The aim of this study was to assess the effect of HS–142–1, a recently discovered specific antagonist of endogenous natriuretic peptides, on systemic hemodynamics, renal function, and the renin–aldosterone system in rats with cirrhosis and ascites. The study consisted of three protocols, each including 10 conscious control rats and 10 conscious rats with carbontetrachloride–induced cirrhosis with ascites. In protocol 1, HS–142–1 administration (by intravenous bolus of 20 mg · kg-1 body weight in all protocols) was not associated with significant changes in mean arterial pressure, heart rate, cardiac output or total peripheral resistance in the two groups of animals. In protocol 2, HS–142–1 induced a significant reduction in glomerular filtration rate (from 4.2 ± 0.5 to 2.6 ± 0.3 ml/min, p < 0.025) in control animals. A decrease in renal plasma flow and an increase in renal vascular resistance also occurred, but these changes were not statistically significant. In cirrhotic rats, HS–142–1 resulted in a significant decrease in renal plasma flow (from 10.9 ± 0.7 to 4.3 ± 0.6 ml/min, p < 0.001) and a significant increase in renal vascular resistance (from 6.0 ± 0.6 to 16.3 ± 2.7 mm Hg · min · ml-1, p < 0.025). Glomerular filtration rate decreased more in cirrhotic rats with ascites than in control rats (from 3.8 ± 0.3 to 1.3 ± 0.2 ml/min, p < 0.001). Changes in urine flow rate and urinary sodium excretion rate paralleled those of glomerular filtration rate in both groups of animals. In protocol 3, HS–142–1 administration was associated with a marked increase in plasma renin activity (from 17 ± 3 to 39 ± 9 ng · ml-1 ± hr-1, p < 0.01) and plasma aldosterone concentration (from 130 ± 12 to 268 ± 48 pg · ml-1, p < 0.025) in cirrhotic rats with ascites. This association was not observed in control animals (plasma renin activity from 1.6 ± 0.3 to 1.6 ± 0.5 ng ± ml·1 · hr-1, aldosterone from 45 ± 15 to 44 ± 16 pg · ml-1). These findings indicate that endogenous natriuretic peptides play a critical role in the maintenance of renal function and in the regulation of the renin–aldosterone system in cirrhosis with ascites. These peptides, however, are not critical to arterial pressure homeostasis under the conditions studied. (Hepatology 1994;20:948-954).
Rapid Induction of Mrnas for Liver Regeneration Factor and Insulin–Like Growth Factor Binding Protein–1 in Primary Cultures of Rat Hepatocytes by Hepatocyte Growth Factor and Epidermal Growth Factor
Authors: Weir Edward , Chen Qiuyan , Defrances Marie C. , Bell Aaron , Taub Rebecca , Zarnegar Reza
Published: March 2007

Liver regeneration factor belongs to the leucinezipper family of transcription factors. It was originally cloned and characterized through differential screening of a regenerating rat liver cDNA library. The mRNA for liver regeneration factor–1 is barely detectable in normal rat liver but is dramatically induced after two–thirds hepatectomy, with a peak 1 to 3 hr after surgery. The nature of the signaling molecule(s) for this rapid induction is not known. It has been suggested that the liver regeneration factor–1 protein product, through complex interactions with other transcription factors such as c–Jun and Jun–B, controls expression of genes that are required during the G1 phase of hepatic growth. Hepatocyte growth factor has been shown to be the most potent mitogen for hepatocytes

in vitro

and

in vivo.

Plasma levels of hepatocyte growth factor rapidly (within 30 min) increase after loss of hepatic parenchyma induced by partial hepatectomy or carbon tetrachloride treatment. It has been postulated that hepatocyte growth factor plays a crucial role in stimulating the hepatocyte to enter the cell cycle. In this communication, we report that addition of pure hepatocyte growth factor to primary cultures of rat hepatocytes in the absence of serum and insulin results in rapid and transient induction of liver regeneration factor–1 mRNA (more than 20–fold) with a peak of expression 1 hr after treatment. The levels of

jun

–B and

c

–

fos

mRNAs, which are also known to be induced during the early hours of liver regeneration, were also increased after treatment of isolated hepatocytes with hepatocyte growth factor. Epidermal growth factor, another potent hepatomitogen, induced liver regeneration factor–1 mRNA with time kinetics similar to those of hepatocyte growth factor; however, the magnitude of induction by epidermal growth factor was much lower than that of hepatocyte growth factor. Hepatocyte growth factor and epidermal growth factor quickly (1 to 2 hr) increased the levels of mRNAs for two other immediate early genes, namely, early growth response gene–1 and insulin–like growth factor binding protein–1 in cultured hepatocytes. These two genes are known to be up–regulated during the early hours of liver regeneration. Taken together, our results support the notion that,

in vivo

, hepatocyte growth factor and epidermal growth factor play important functions in triggering and initiating the early events required for subsequent hepatocyte growth and liver regeneration. (Hepatology 1994;20:955-960).
Characterization of Signaling Pathways to Na+/H+ Exchanger Activation With Epidermal Growth Factor in Hepatocytes
Authors: Tanaka Yuji , Hayashi Norio , Kaneko Akira , Ito Toshifumi , Horimoto Masayoshi , Sasaki Yutaka , Kasahara Akinori , Fusamoto Hideyuki , Kamada Takenobu
Published: March 2007

To investigate the signaling pathways to Na+/H+ exchanger activation with epidermal growth factor in hepatocytes, we measured changes in cytosolic free calcium and intracellular pH levels at the single–cell level using digital imaging fluorescence microscopy of fura–2—or BCECF–loaded hepatocytes in primary culture. Epidermal growth factor induced cytosolic free calcium oscillations consisting of periodic trains of spikes with a latency period of up to several minutes. These calcium responses were inhibited by tyrosine kinase inhibitor genistein (100 μmol/L) and abolished by emptying of intracellular Ca2+ pools with 3 μmol/L thapsigargin, an inhibitor of Ca2+–ATPase on the endoplasmic reticulum. Epidermal growth factor (1 nmol/L) induced an intracellular pH increase of 0.12 ± 0.07 units from the basal level of 7.25 ± 0.09 units after several minutes of latency. This effect was completely abolished by 1 mmol/L amiloride, an inhibitor of the Na+/H+ exchanger. The epidermal growth factor—induced intracellular pH increase was inhibited by pretreatment of hepatocytes with genistein (100 μmol/L), thapsigargin (3 μmol/L) or calmodulin inhibitor W–7 (25 μmol/L), but not with protein kinase C inhibitor H–7 (50 μmol/L) or with cyclic AMP—dependent kinase inhibitor H–8 (60 μmol/L). Phorbol ester PMA (phorbol 12–myristate 13–acetate), a potent activator of protein kinase C, induced a slight intracellular pH increase significantly smaller than that with epidermal growth factor, whereas this effect was completely blocked by pretreatment with H–7, indicating that PMA–induced intracellular pH increase is mediated by protein kinase C pathways, unlike epidermal growth factor. These data indicate that epidermal growth factor—induced intracellular pH increase mediated by activation of Na+/H+ exchanger in primary cultured hepatocytes depends on tyrosine kinase and is downstream from Ca2+ mobilization and Ca2+–calmodulin—dependent pathways. (Hepatology 1994;20:966-974).
Heat Shock Response in the Liver: Expression and Regulation of the Hsp70 Gene Family and Early Response Genes After In Vivo Hyperthermia
Authors: Schiaffonati Luisa , Tacchini Lorenza , Pappalardo Carmela
Published: March 2007

Heat shock response in cultured cells has been studied extensively; however few data are available on heat shock response in an intact organ of a living animal. In this study we analyzed the kinetics of expression of the heat shock protein 70 gene family (heat shock protein 70, heat shock cognate protein 73 and glucose–regulated protein 78) in the liver of the thermally stressed rat. New synthesis of heat shock protein 70 and heat shock cognate protein 73 was shown in liver slices pulse labeled

in vitro

with 35S–methionine. Accumulation of heat shock protein 70 and heat shock cognate protein 73 proteins was shown in total cellular extracts. 32P–labeled complementary DNA probes encoding heat shock protein 70, heat shock cognate protein 73 and glucose–regulated protein 78 were used to show that the levels of the corresponding messenger RNAs increase as a fraction of total RNA and in polysomes at different extents and with different kinetics. The induction of heat shock protein 70 and heat shock cognate protein 73 messenger RNAs reflected the increase in the synthesis of the corresponding proteins. Run–on transcription analysis indicated that the expression of heat shock protein 70 and heat shock cognate protein 73 genes was mainly regulated at the transcriptional level. On the contrary, both transcriptional and posttranscriptional regulatory mechanisms can explain the induction of the glucose–regulated protein 78 gene. The binding of a transiently activated heat shock factor to the heat shock element was detected by gel retardation assays and was directly correlated to the increase in the transcription of the heat shock protein 70 gene. In hyperthermic livers the increase in the expression of the heat shock protein 70 gene family was associated with an increase in the expression of the early response genes c–

fos

and c–

jun.

The observed differences in timing of expression and in regulatory mechanisms of different members of the heat shock protein 70 family are probably related to the specific liver response to an increase in body temperature. (Hepatology 1994;20:975-983).
Cocaine–Induced Liver Injury in Mice Elicits Specific Changes in Dna Ploidy and Induces Programmed Death of Hepatocytes
Authors: Cascales María , Alvarez Alberto , Gascó Pilar , Fernández-Simón Lourdes , Sanz Nuria , Boscá Lisardo
Published: March 2007

Liver injury was induced by a single dose (60 mg/kg) of cocaine in male albino Swiss mice untreated or pretreated with phenobarbital (in drinking water 1 gm/L), for 5 days before cocaine administration. One parameter of liver injury, serum isocitrate dehydrogenase activity, showed sharp increases at 24 hr of cocaine treatment; we also noted decreased hepatic levels of ATP, GSH, cytochrome P–450 and NADPH/NADP+ ratio and increases in malondialdehyde concentration. Histopathological study of liver slices showed perivenous and periportal necrosis induced by cocaine in untreated mice and mice pretreated with phenobarbital, respectively. A regenerative postnecrotic response, which peaked at 48 hr, was demonstrated by the appearance of mitotic cells. Mitotic index analysis showed that proliferative cells appear to be unevenly distributed in the hepatic acinus and were mainly located in the vicinity of the damaged acinar region. Genomic DNA ploidy and the distribution of DNA in the phases of the cell cycle were studied in nuclei of isolated hepatocytes. At 12 hr of cocaine administration, both in untreated and phenobarbital–pretreated mice, the following changes were observed: a sharp decrease in tetraploid (4N) cells (40% to 17% and 25% to 6%, respectively) and octoploid (8N) cells (5% to 2% and 2% to 1%, respectively), together with the appearance of a hypodiploid population (13% and 31%, respectively). Hypodiploid population was characterized as apoptotic cells by detection of DNA fragmentation in agarose gel. These results suggest that a significant percentage of cell death induced by cocaine occurs by means of the apoptosis death program. Comparison of the initial values of DNA ploidy with those obtained at 7 days of cocaine administration showed remarkable increases in polyploid populations (4N and 8N) and a decrease in diploid cells (2N), indicating that the process of differentiation occurs when liver restores its functionality. (Hepatology 1994;20:992-1001).
Calmodulin Expression During Rat Liver Regeneration
Authors: Agell Neus , Pujol Maria Jesus , López-Girona Antonia , Bosch Marta , Rosa Jose Luis , Bachs Oriol
Published: March 2007

We have investigated the messenger RNAs expressed from the three calmodulin genes during rat liver regeneration. The results revealed that all the calmodulin transcripts increased from 8 hr after a partial hepatectomy, although differences in the timing and the level of expression from the three genes were observed. Calmodulin I transcripts peaked at 16 hr, whereas calmodulin II and calmodulin III progressively increased from 8 to 24 hr. At 24 hr after surgery, calmodulin I, calmodulin II and the 2.3 kb calmodulin III transcripts reached values of a 6–fold increase, whereas the 0.8 kb product of calmodulin III increased 25–fold. At 30 hr the levels of all the calmodulin transcripts were similar to those observed at 24 hr. The transcription rates of the three calmodulin genes augmented after hepatectomy (calmodulin I and calmodulin II twofold and calmodulin III fourfold), indicating that the elevation of the calmodulin transcripts could be, at least partially, the result of this increase in the transcription rates. The total calmodulin concentration also increased twofold at 24 hr after hepatectomy. We also report that the administration of the β–adrenergic blocker, D, L–propranolol inhibited the accumulation of calmodulin protein without significantly affecting the increase of the messenger RNAs. These results indicate that the expression of calmodulin observed during liver regeneration could be regulated by cyclic AMP at the translational or post–translational level. (Hepatology 1994;20:1002-1008).