bluuebunny/crossref_metadata_2025 huggingface dataset:
Sinusoidal Ultrastructure Evaluated During the Revascularization of Regenerating Rat Liver
Authors: Wack Kathryn E. , Ross Mark A. , Zegarra Vasthy , Sysko Laura R. , Watkins Simon C. , Stolz Donna Beer
Published: August 2002

Sinusoidal endothelial cell (SEC) porosities were compared between the periportal (zone 1) and pericentral (zone 3) regions of the rat liver during regeneration following partial hepatectomy (PHx). SEC porosities and fenestration diameters were measured in control livers, as well as at 5 minutes, 24, 48, 72, 96, 120 hours, and 14 days following PHx. Bimodal maximums in both porosity and fenestration diameters were observed in both zones at 5 minutes and 5 days following PHx. SEC porosities increased significantly in both zones 1 and 3 within 5 minutes following PHx, but the increase was maintained only in zone 1 at 24 hours after resection. Following the initial rise, both zones displayed a gradual decrease to less than half their porosity values at 72 hr post–PHx. After 72 hours, porosities increased to over control levels and remained elevated until 14 days after PHx. The decrease in porosity at 72 hr post–PHx is accompanied by ultrastructural changes within the sinusoid at this time. Vascular corrosion casting and transmission electron microscopy (TEM) show sinusoid compression resulting from increased hepatic plate widths due to hepatocyte proliferation in the absence of SEC proliferation. Also at this time, we observed many SEC completely enveloped by stellate cells. The zonal variations observed for porosities throughout regeneration did not correlate with changes in laminin, collagen I and IV, or fibronectin deposition within the space of Disse. Taken together, the data reveal that SEC are dynamic regulators of porosity that respond rapidly and locally to environmental zonal stimuli during liver regeneration.
Identification of Differentially Expressed Genes in Hepatocellular Carcinoma With Cdna Microarrays
Authors: Shirota Yukihiro , Kaneko Shuichi , Honda Masao , Kawai Hiroshi F. , Kobayashi Kenichi
Published: August 2002

Genes expressed in hepatocellular carcinoma (HCC) were analyzed using cDNA microarrays to clarify gene abnormalities in HCC. mRNA was extracted from cancerous and noncancerous tissues of 10 patients with HCC, cDNA labeled with Cy5 and Cy3 fluorescence was prepared, and it was hybridized for each patient with a cDNA microarray consisting of 1,080 elements (930 unique genes). The mRNA expression rate of each element in HCC was evaluated using the level of mRNA expression in noncancerous tissue in each patient as a reference. The expression of 10 genes was enhanced 2 times or more in HCC cancerous tissue compared with noncancerous tissue in 5 or more of the 10 patients. In contrast, 9 genes were expressed at half the level or less in HCC cancerous tissue compared with noncancerous tissue. When hierarchical clustering was performed to identify genes related to clinical phenotypes of the patients, 22 genes showed changes associated with the degree of differentiation of HCC. Thirteen of these genes were transcriptional factors or tissue–specific expression proteins related to cell differentiation or development. Our present analysis clarified a number of genes that characterize HCC. This information based on examination of clinical samples is considered to be useful for clarification of the mechanism of hepatocarcinogenesis and the diagnosis and treatment of HCC.
Mitochondrial Oxidative Injury and Energy Metabolism Alteration in Rat Fatty Liver: Effect of the Nutritional Status
Authors: Vendemiale Gianluigi , Grattagliano Ignazio , Caraceni Paolo , Caraccio Giovanni , Domenicali Marco , Dall'Agata Monia , Trevisani Franco , Guerrieri Ferruccio , Bernardi Mauro , Altomare Emanuele
Published: August 2002

Hepatic steatosis is associated with mitochondrial oxidative alterations. This study aimed to characterize in a choline–deficient model of rat fatty liver whether this oxidative imbalance is related to an impairment of the capacity of ATP synthesis both under fed conditions and after starvation, which may sensitize mitochondria to oxidative injury. Mitochondria were isolated from normal and fatty livers of fed or 18–hour fasted rats. Oxidative injury was evaluated by measuring the mitochondrial content of thiobarbituric reactive substances, protein carbonyls, glutathione, and protein sulfhydryls. The mitochondrial F0F1–ATP synthase content, tissue ATP concentration, and liver histology were also determined. Compared with normal liver, under fed conditions, fatty livers showed a greater mitochondrial content of oxidized lipids and proteins together with a low concentration of sulfhydryls and glutathione. The mitochondrial catalytic β–F1 subunit of the F0F1–ATP synthase was about 35% lower in fatty livers. Hepatic ATP was also significantly reduced in fatty liver. Starvation exacerbated mitochondrial oxidative injury in both groups but to a greater extent in fatty livers. In the steatotic group, fasting induced a significant decrease of the ATP levels, which was accompanied by a 70% fall of the catalytic β–F1 subunit. These data indicate that the mitochondrial oxidative alterations in fatty livers are associated with an important reduction of the F0F1–ATP synthase. These changes, which are greatly exacerbated after starvation, may account for the reduced synthesis of the hepatic ATP observed in the presence of fatty infiltration.
Endothelin–1 Induces Vasoconstriction on Portal–Systemic Collaterals of Portal Hypertensive Rats
Authors: Chan Che-Chang , Wang Sun-Sang , Lee Fa-Yauh , Chang Full-Young , Lin Han-Chieh , Chu Chi-Jen , Chen Chien-Ting , Huang Hui-Chun , Lee Shou-Dong
Published: August 2002

Portal hypertension is associated with increased hepatic and collateral resistance to an increased portal blood flow. Endothelin–1 (ET–1) can induce intrahepatic vasoconstriction and consequently increase portal pressure. It is unknown if ET–1 also modulates portal pressure by a direct vasoconstrictive effect on collaterals. This study investigated the collateral vascular responses to ET–1, the receptors in mediation, and the regulation of ET–1 action by nitric oxide and prostaglandin. The portal–systemic collaterals of partially portal vein—ligated rats were tested by in situ perfusion. The concentration–response curves of collaterals to graded concentrations of ET–1 (10-10–10-7 mol/L) with or without BQ–123 (ETA receptor antagonist, 2 × 10-6 mol/L), BQ–788 (ETB receptor antagonist, 10-7 mol/L) or both were recorded. In addition, the collateral responses to ET–1 with preincubation of nω–nitro–L–arginine (NNA; 100 mol/L), indomethacin (INDO; 10 mol/L), or in combination were performed. ET–1 increased the perfusion pressure of collaterals and its effect was significantly suppressed by BQ–123 alone and BQ–123 plus BQ–788, but not BQ–788 alone (P < .05). Incubation with NNA, INDO, or both significantly enhanced the response of collaterals to ET–1 (P < .05). These results show that ET–1 produces a direct vasoconstrictive effect on the collateral vessels of portal hypertensive rats. This effect is mediated by ETA, but not ETB, receptors. Both nitric oxide and prostaglandin modulate the collateral vascular response to ET–1 and may therefore participate in the development and maintenance of portal hypertension.
A Critical Role of T–Cell Receptor γ/δ Cells in Antibacterial Protection in Mice Early in Life
Authors: Emoto Masashi , Miyamoto Mamiko , Emoto Yoshiko , Zerrahn Jens , Kaufmann Stefan H.E.
Published: August 2002

Although it is generally assumed that T–cell receptor (TCR) γ/δ cells participate in protection against intracellular microbial pathogens, their impact remains controversial. In our study, young (14–day–old) mice lacking TCRγ/δ cells were far more susceptible to Listeria monocytogenes than wild–type (WT) mice of the same age. The number of interferon gamma (IFN–γ) producers responsible for antilisterial resistance was significantly higher among natural killer (NK)1+ TCRγ/δ cells than among NK1- TCRγ/δ cells. Endogenous IFN–γ neutralization increased susceptibility of young WT mice to L. monocytogenes infection. Liver was a major residence of peripheral NK1+ TCRγ/δ cells, whereas NK1- TCR γ/δ cells were broadly distributed in various lymphoid organs. Numbers of both NK1+ and NK1- TCRγ/δ cells increased in the liver of WT mice prior to TCRα/β cells and represented a substantial population in early life (14 days after birth). Virtually all NK1+ TCRγ/δ cells expressed activation markers, whereas substantial numbers of NK1- TCRγ/δ cells showed a naive phenotype. We conclude that TCRγ/δ cells play a critical role in protection against L. monocytogenes in the early life of mice, probably because their TCRα/β cell compartment is not fully competent. For this antibacterial function, we assign NK1+ TCRγ/δ cells a more important role than their NK1- cognates.
A Randomized Trial of Amantadine and Interferon Versus Interferon Alone As Initial Treatment for Chronic Hepatitis C
Authors: Mangia Alessandra , Minerva Nicola , Annese Mauro , Leandro Gioacchino , Villani Maria Rosaria , Santoro Rosanna , Carretta Vito , Bacca Donato , Giangaspero Antonio , Bisceglia Michele , Ventrella Francesco , Dell'Erba G. , Andriulli Angelo
Published: August 2002

The aim of this study was to compare, in an open–label study, the efficacy and safety of a combination of interferon (IFN) and amantadine (AMA) with that of IFN alone in previously untreated patients with chronic hepatitis C. A total of 200 patients were randomized to 6 MU of IFN–α2a 3 times per week, with 200 mg of AMA daily (n = 99) or to an identical dose of interferon α2a (n = 101). Patients were treated for 12 months and observed for 6 months' posttreatment. At the completion of treatment, 28.7% of patients in the monotherapy group and 45.5% in the combination group had a virologic response (P = .014). At 6 months' posttreatment, a sustained virologic response was observed in 16.8% (95% CI: 9–23) of patients with IFN alone versus 29.3% (95% CI: 19–37) of patients who were treated with combination therapy (P = .036). In each of the 2 treatments, genotype was the only predictive parameter for a sustained response. At the logistic regression analysis, therapy and genotype were the only 2 parameters with an independent predictive value. In the combination group, at examination of month 3, hepatitis C virus (HCV)–RNA status had a 97.6% (95% CI: 93–102) positive predictive value and a 50% (95% CI: 37–63) negative predictive value for a sustained virologic clearance. A substantial proportion of naïve patients with chronic hepatitis C have an end–of–treatment and end–of–follow–up virologic and biochemical response to a combination of IFN and AMA. This new treatment appears safe and well tolerated. (Hepatology 2001;33:989–993.)
Vascular Invasion and Histopathologic Grading Determine Outcome After Liver Transplantation for Hepatocellular Carcinoma in Cirrhosis
Authors: Jonas Sven , Bechstein Wolf O. , Steinmüller Thomas , Herrmann Martin , Radke Cornelia , Berg Thomas , Settmacher Utz , Neuhaus Peter
Published: August 2002

Selection of patients suffering from hepatocellular carcinoma (HCC) in cirrhosis for liver transplantation follows limits of number and diameter of tumor nodules. It has not been investigated whether there is a correlation of these parameters with vascular invasion. From 1989 to 2000, 1,188 liver transplantations were performed in 1,087 patients, including 120 patients (11%) with an HCC in cirrhosis. Selection criteria were a maximal diameter of up to 5 cm if the tumor appeared to be uninodular or of up to 3 cm in the case of 2 or 3 nodules. The postoperative mortality rate was 1.7%. One–, 5–, and 10–year survival was 90%, 71%, and 60%, respectively. In 940 transplantation patients without an HCC, these rates were 93%, 87%, and 83% (P < .0001). Vascular invasion and histopathologic grading were identified as prognostic parameters by multivariate analysis. In a logistic regression analysis, histopathologic grading and maximal diameter showed a significant correlation with a vascular invasion. Analyzing tumors larger than 5 cm, i.e., tumors not fulfilling the selection criteria as a result of diagnostic inaccuracy or progression thereafter, the rates of vascular invasion were significantly (P < .01) lower in patients suffering from well–differentiated tumors (25%) when compared with moderately and poorly differentiated tumors (100%). Liver transplantation is a safe and effective long–term treatment for small HCC in cirrhosis. Tumor diameter and number of nodules in correlation with the histopathologic grading were predictive of a vascular invasion only in HCC larger than 5 cm.
Intracellular Signaling Pathways Involved in Acetaldehyde–Induced Collagen and Fibronectin Gene Expression in Human Hepatic Stellate Cells
Authors: Svegliati-Baroni Gianluca , Ridolfi Francesco , Di Sario Antonio , Saccomanno Stefania , Bendia Emanuele , Benedetti Antonio , Greenwel Patricia
Published: August 2002

Ethanol induces liver fibrosis by several means that include, among others, the direct fibrogenic action of acetaldehyde on hepatic stellate cells (HSC). However the mechanisms responsible for this effect are not well understood. In this communication we investigated signal transduction pathways triggered by acetaldehyde leading to upregulation of α2(I) collagen and fibronectin gene expression in human HSC. Run–on assays showed that acetaldehyde–enhanced transcription of these 2 genes as early as 2 hours, via de novo protein synthesis–independent and –dependent mechanisms. It also stimulated a time–dependent induction in phosphorylation of pp70S6K and extracellular–regulated kinase ½ (ERK½). These effects were completely prevented by calphostin C, a protein kinase C inhibitor. As expected, acetaldehyde–elicited ERK½ phosphorylation was inhibited by PD98059, a MEK inhibitor, but not by wortmannin, a PI3K inhibitor. On the other hand, both of these inhibitors partially inhibited phosphorylation of pp70S6K induced by acetaldehyde suggesting that its activation is ERK½– and PI3K–dependent. Acetaldehyde–elicited fibronectin and α2(I) collagen upregulation was inhibited by calphostin C. However, while PD98059, wortmannin and rapamycin (a pp70S6K inhibitor) completely abrogated α2(I) collagen upregulation, they had no effect on fibronectin expression. Overall, these data suggest that protein kinase C is an upstream component from which acetaldehyde signals are transduced to other pathways such as PI3K and ERK½. In addition, differential activation of these pathways is needed for the increase in fibronectin and α2(I) collagen gene expression induced by acetaldehyde in human HSC.
Effects of Treatment With Deoxycholic Acid and Chenodeoxycholic Acid on the Hepatic Synthesis of Cholesterol and Bile Acids in Healthy Subjects
Authors: Einarsson Curt , Hillebrant Carl-Gustaf , Axelson Magnus
Published: August 2002

The degradation of cholesterol to bile acids is regulated by a negative–feedback mechanism by the bile acids, especially the hydrophobic bile acids, returning to the liver via the portal vein. Chenodeoxycholic acid (CDCA) is a potent suppressor of the cholesterol 7α–hydroxylase, the rate–determining enzyme in bile acid formation. CDCA may also suppress hepatic 3–hydroxy–3–methyl glutaryl coenzyme A (HMG CoA) reductase, the rate–limiting enzyme in cholesterol synthesis. Conflicting reports have appeared regarding the suppression on bile acid synthesis by the most hydrophobic bile acid of human bile, deoxycholic acid (DCA). To study the suppressive effects of CDCA and DCA on hepatic cholesterol and bile acid synthesis in humans, 10 healthy subjects were treated with CDCA or DCA for 3 weeks in a randomized cross–over study with a washout period of 4 weeks in between. Serum levels of 7α–hydroxy–4–cholesten–3–one, reflecting cholesterol 7α–hydroxylase activity, and 7–dehydrocholesterol, reflecting HMG CoA reductase activity, and bile acids were repeatedly measured during the study periods. After 3 weeks of treatment with CDCA or DCA, CDCA constituted 70% and DCA 74% of the total serum bile acids, respectively. CDCA and DCA decreased the serum levels of 7α–hydroxy–4–cholesten–3–one by 80% and 75%, respectively. Negative correlations between the percentages of CDCA and DCA and the serum concentration of 7α–hydroxy–4–cholesten–3–one were obtained. CDCA reduced the serum level of 7–dehydrocholesterol by 29%, whereas treatment with DCA tended to increase the level of 7–dehydrocholesterol. Treatment of healthy subjects with CDCA and DCA reduces bile acid synthesis. CDCA also inhibits cholesterol synthesis, whereas DCA does not.
Etiologic Significance of Defects in Cholesterol, Phospholipid, and Bile Acid Metabolism in the Liver of Patients With Intrahepatic Calculi
Authors: Shoda Junichi , Oda Koji , Suzuki Hiroshi , Sugiyama Yuichi , Ito Kosei , Cohen David E. , Feng Li , Kamiya Junichi , Nimura Yuji , Miyazaki Hiroshi , Kano Masahito , Matsuzaki Yasushi , Tanaka Naomi
Published: August 2002

Intrahepatic calculi, highly prevalent in the Far East, including Japan, are characterized clinically by chronic proliferative cholangitis with frequent stone recurrences. Intrahepatic calculi consist of 2 groups, i.e., brown pigment stones, including a high cholesterol content, and cholesterol stones, with the former predominating. To gain insights into the pathogenesis of intrahepatic calculi, cholesterol and bile acid biosynthesis, as well as alterations in intracellular transport and/or canalicular secretion of phospholipid and bile acid were investigated in liver of patients with intrahepatic calculi. Enzyme activities of 3–hydroxy–3–methylglutaryl-coenzyme A (HMG–CoA) reductase were increased (12.8 ± 1.9 pmol/min/mg protein, mean ± SEM vs. 5.5 ± 0.4 in controls; P < .01) and cholesterol 7 α–hydroxylase activities were decreased (1.3 ± 0.4 vs. 4.9 ± 0.6; P < .01) in liver specimens of patients with brown pigment stones. In addition, messenger RNA (mRNA) levels of multidrug resistance P–glycoprotein 3 (MDR3 Pgp) and phosphatidylcholine transfer protein (PCTP) were markedly low in the liver specimens compared with the levels in specimens of control subjects, gallbladder stone patients, and patients with obstructive cholestasis. The protein levels and the immunohistochemical staining were decreased for MDR3 Pgp and PCTP in the liver. Consistently, the concentrations of phospholipid were markedly reduced in the hepatic bile from both affected and unaffected hepatic segments. In patients with intrahepatic calculi, biliary cholesterol supersaturation and the formation of cholesterol–rich brown pigment as well as cholesterol stones may be attributed to decreased hepatic transport and biliary secretion of phospholipids, in the setting of increased cholesterogenesis and decreased bile acid synthesis. (Hepatology 2001;33:1194–1205.)
Contact Sensitization Pretransplantation Predicts Acute Hepatic Allograft Rejection
Authors: Bathgate Andrew , Dollinger Matthias , Plevris John , Bellamy Christopher , Hayes Peter
Published: August 2002

The loss of hepatic allografts to the rejection processes is now relatively rare, and the reduction of adverse effects related to immunosuppressive therapy is becoming more important as patients survive longer after transplantation. We therefore investigated the response to a contact neoantigen before liver transplantation as a predictor of acute rejection after transplantation. Forty–one patients with chronic liver disease were sensitized with 0.1% diphenylcyclopropenone while on the waiting list for orthotopic liver transplantation. Fourteen days later an elicitation reaction was performed with 5 different concentrations of diphenylcyclopropenone. Nineteen responded to diphenylcyclopropenone (score range, 1–9). Twenty–two patients had no response. Three patients died before transplantation (all nonresponders). Twelve (63%) of 19 responders had treatment for acute rejection compared with 1 of 19 nonresponders (P < .0001). In addition univariate analysis revealed recipient age, donor age, Child–Pugh class, and immunosuppressive agent to be associated with acute rejection. On multivariate analysis only skin test response was a significant predictor of acute rejection (P = .02). All nonresponders had no or only mild rejection on biopsy, but 12 of 19 responders had moderate or severe acute rejection on biopsy. All patients requiring additional therapy to a single course of corticosteroids for acute rejection had skin test scores greater than 1. We concluded that patients who do not respond to diphenylcyclopropenone sensitization before transplantation develop at most mild acute rejection and that skin test scores identify patients with troublesome rejection. Evaluation of skin test responses to a contact neoantigen may facilitate tailoring of immunosuppressive therapy.
Human Reg I Gene Is Up–Regulated in Intrahepatic Cholangiocarcinoma and Its Precursor Lesions
Authors: Harada Kenichi , Zen Yoh , Kanemori Yoshiko , Chen Tse-Ching , Chen Miin-Fu , Yeh Ta-Sen , Jan Yi-Yin , Masuda Shinji , Nimura Yuji , Takasawa Shin , Okamoto Hiroshi , Nakanuma Yasuni
Published: August 2002

The Reg I gene (regenerating gene) and its product (Reg protein) are a regenerating and/or proliferating factor(s) of pancreatic islet cells. The ectopic expression of REG Iα was shown in colorectal carcinomas, suggesting that REG Iα is related to their carcinogenesis. In this study, we examined the expression of REG I in intrahepatic cholangiocarcinoma (ICC) and its precursor lesion (biliary dysplasia). By polymerase chain reaction and in situ hybridization (ISH) studies using a total of 16 fresh liver specimens, REG Iα mRNA was demonstrated in 6 of 11 (55%) ICC cases, but in 0 of 5 (0%) normal livers. Immunohistochemistry for REG I protein was performed in 100 formalin–fixed, paraffin–embedded sections obtained from the 18 cases of ICC alone, 45 hepatolithiasis with ICC (n = 19) or biliary dysplasia (n = 26), 21 hepatolithiasis alone (all with hyperplasia), and 16 normal livers. In ICC, the expression of REG I protein was significantly dependent on the histologic differentiation; 12 of 13 (92%) cases in papillary and well–differentiated, 6 of 16 (38%) cases in moderately differentiated, and 0 of 8 (0%) cases in poorly differentiated types. Moreover, in the lesions of hyperplasia, low–grade dysplasia, and high–grade dysplasia in hepatolithiasis, REG I protein was expressed in 4 of 21 (19%), 7 of 12 (58%), and 13 of 14 (93%) cases, respectively. In normal liver, intrahepatic bile ducts were constantly negative for REG I protein. These findings suggest that neoexpression of REG I is a good marker for biliary mucosa at risk for development of ICC, and also that REG I plays a role in the early stages of biliary carcinogenesis, probably via a cell–proliferative effect. (Hepatology 2001;33:1036–1042.)
Promoter Polymorphism of the Cd14 Endotoxin Receptor Gene As A Risk Factor for Alcoholic Liver Disease
Authors: Järveläinen Harri A. , Orpana Arto , Perola Markus , Savolainen Vesa T. , Karhunen Pekka J. , Lindros Kai O.
Published: August 2002

Twin concordance studies indicate that genetic factors influence the individual susceptibility for alcoholic liver disease (ALD). Both clinical and experimental data suggest that Kupffer cell activation by gut–derived endotoxins and other bacterial products is an important pathogenic factor. Activated Kupffer cells release proinflammatory cytokines, a process that is regulated by the CD14 endotoxin receptor (CD14). Recently, a C→T (-159) polymorphism in the promoter region of the CD14 gene was detected and found to confer increased CD14 expression. In the present study, the association of CD14 promoter polymorphism with different forms of ALD was examined in 3 separate autopsy series. Among 442 men with valid alcohol–consumption data, 381 men had been moderate or heavy alcohol consumers. The allele frequency of the CD14 promoter genotype, determined by a modified cycle minisequencing technique, was 0.34 (CC), 0.51 (CT), and 0.16 (TT). The T allele was found to be associated with advanced ALD, i.e., with alcoholic hepatitis (odds ratio [OR]: 2.48; P = .018), and especially with cirrhosis (OR: 3.45; P = .004), but not with fatty liver, periportal fibrosis, or bridging fibrosis. The overall age–adjusted risk for cirrhosis was 3.08 (P = .01) for the carriers of the CT genotype, and 4.17 (P = .005) for the homozygous TT genotype. These results suggest that in the relatively isolated Finnish population, the T allele confers increased risk of alcoholic liver damage. In particular, TT homozygotes are at a high risk to develop cirrhosis.
Detoxification of Reactive Oxygen Species by A Nonpeptidyl Mimic of Superoxide Dismutase Cures Acetaminophen–Induced Acute Liver Failure in the Mouse
Authors: Ferret Pierre-Jacques , Hammoud Renaud , Tulliez Micheline , Tran Agnès , Trébéden Hélène , Jaffray Patrick , Malassagne Benoît , Calmus Yvon , Weill Bernard , Batteux Frédéric
Published: August 2002

Drug–induced acute liver failure (ALF) is a devastating and often fatal disease mainly caused by poisoning by acetaminophen (APAP). The toxic metabolite, N–acetyl–p–benzoquinone–imine (NAPQI), that leads to gluthatione depletion has been suspected to be the main effector of hepatocyte apoptosis during APAP–induced ALF. We have investigated whether reactive oxygen species (ROS) also play a role in APAP–induced ALF, and whether manganese III tetrakis (5,10,15,20 benzoic acid) (MnTBAP), a mimic of superoxide dismutase (SOD) with catalase–like activity, can treat the disease in mice. The effects of MnTBAP were tested on APAP–intoxicated mice and on isolated hepatocytes incubated with APAP. MnTBAP preventively and curatively administered significantly improved survival times, and dramatically reduced serum transaminase activity levels and parenchymal lesions in APAP–intoxicated mice. Whereas pretreatment with N–acetyl–l–cysteine (NAC) prevented ALF in a dose–dependent manner, the molecule was ineffective when curatively administered. The significant increase in glutathione peroxidase (Gpx) activity following APAP administration, and the beneficial effects of MnTBAP suggested that ROS were produced during APAP–induced ALF. A direct evidence of ROS generation was provided by flow cytometry of isolated hepatocytes incubated with APAP. In vitro, ROS production was associated with mitochondrial damage characterized by the collapse of transmembrane potential and the loss of cardiolipin content. In livers of intoxicated mice, ALF was associated with cytochrome c release that led to the activation of caspases–9 and –3. The capacity of MnTBAP to abrogate all those alterations suggests that ROS play a role in APAP–induced apoptosis of hepatocytes, and explains the beneficial effects of MnTBAP, which could be of interest in APAP–induced ALF in humans.
Global Changes in Interleukin–6-Dependent Gene Expression Patterns in Mouse Livers After Partial Hepatectomy
Authors: Li Wei , Liang Xianping , Leu Julie I. , Kovalovich Kellen , Ciliberto Gennaro , Taub Rebecca
Published: August 2002

Liver regeneration following 70% partial hepatectomy leads to rapid activation of genes in the remnant liver. Interleukin–6 deficient (IL–6 -/-) mice have impaired liver regeneration and abnormalities in immediate early gene expression. In this study, the gene expression program in the IL–6 +/+ and -/- livers at 2 hours posthepatectomy was examined with a cDNA array representing 588 highly regulated mouse genes. Thirty–six percent of the 103 immediate early genes were induced differently in IL–6 +/+ compared with IL–6 -/- livers, implying regulation by IL–6. IL–6 treatment of the IL–6 -/- mice in the absence of hepatectomy induced a much smaller set of genes in the liver, suggesting that IL–6 cooperates with other hepatectomy–induced factors to activate the large number of genes. Northern blot analyses were used to verify gene expression data obtained from the arrays. The expression of urokinase type plasminogen activator receptor (uPAR) and plasminogen activator inhibitor–1 (PAI–1), critical components of the urokinase plasminogen activator (uPA) system, was lower and delayed in IL–6 -/- livers. Despite the fact that active uPAR/uPA complex is critical for hepatocyte growth factor (HGF) activation, no differences were detected between the IL–6 +/+ and -/- livers in HGF activation as measured by receptor phosphorylation. On the contrary, the mitogen–activated protein kinase (MAPK) pathway was activated in IL–6 +/+ livers early during regeneration but remarkably delayed in IL–6 -/- livers. Defective liver regeneration may be explained by the large number of gene activation pathways altered in IL–6 -/- livers and further supports the finding that IL–6 is necessary for normal liver regeneration.
Liver Transplantation for Hepatocellular Carcinoma: Expansion of the Tumor Size Limits Does Not Adversely Impact Survival
Authors: Yao Francis Y. , Ferrell Linda , Bass Nathan M. , Watson Jessica J. , Bacchetti Peter , Venook Alan , Ascher Nancy L. , Roberts John P.
Published: August 2002

The precise staging of hepatocellular carcinoma (HCC) based on the size and number of lesions that predict recurrence after orthotopic liver transplantation (OLT) has not been clearly established. We therefore analyzed the outcome of 70 consecutive patients with cirrhosis and HCC who underwent OLT over a 12–year period at our institution. Pathologic tumor staging of the explanted liver was based on the American Tumor Study Group modified Tumor–Node–Metastases (TNM) Staging Classification. Tumor recurrence occurred in 11.4% of patients after OLT. The Kaplan–Meier survival rates at 1 and 5 years were 91.3% and 72.4%, respectively, for patients with pT1 or pT2 HCC; and 82.4% and 74.1%, respectively, for pT3 tumors (P = .87). Patients with pT4 tumors, however, had a significantly worse 1–year survival of 33.3% (P = .0001). An α–fetoprotein (AFP) level > 1,000 ng/mL, total tumor diameter > 8 cm, age ≥ 55 years and poorly differentiated histologic grade were also significant predictors for reduced survival in univariate analysis. Only pT4 stage and total tumor diameter remained statistically significant in multivariate analysis. Patients with HCC meeting the following criteria: solitary tumor ≤ 6.5 cm, or ≤ 3 nodules with the largest lesion ≤ 4.5 cm and total tumor diameter ≤ 8 cm, had survival rates of 90% and 75.2%, at 1 and 5 years, respectively, after OLT versus a 50% 1–year survival for patients with tumors exceeding these limits (P = .0005). We conclude that the current criteria for OLT based on tumor size may be modestly expanded while still preserving excellent survival after OLT.
Partially Reversible Renal Tubular Damage in Patients With Obstructive Jaundice
Authors: Bairaktari Eleni , Liamis George , Tsolas Orestes , Elisaf Moses
Published: August 2002

Decreased serum uric acid levels resulting from renal urate wasting have been occasionally encountered in jaundiced patients. However, in these cases, there are no data concerning the underlying renal tubular defects. In the present study, we investigated the renal tubular function in 35 patients with obstructive jaundice of various severity and causes (11 with lithiasis, 17 with carcinoma, and 7 with intrahepatic cholestasis). A detailed study of the renal tubular function was performed. Beyond the conventional methods, 1H–NMR spectroscopy of urine was used to evaluate noninvasively renal damage by the characteristic perturbation in the excretion pattern of low–molecular weight endogenous metabolites. On admission, patients with obstructive jaundice had significantly lower serum uric acid and phosphate levels and higher bile acid concentrations compared with 40 age– and sex–matched controls. Serum uric acid levels presented a negative correlation with the total and direct bilirubin as well as the fractional excretion of uric acid. Furthermore, a great number of the patients studied developed one or more proximal tubular dysfunction manifestations beyond uricosuria, such as renal glucosuria, phosphaturia, and increased excretion of α1–microglobulin. 1H–NMR spectroscopy of the urine showed decreased levels of citrate and hippurate and increased levels of 3–hydroxybutyrate and acetate. In 12 patients partial or complete remission of jaundice was followed by an improvement of the proximal renal tubular damage. In conclusion, obstructive jaundice can cause a partially reversible generalized proximal tubular dysfunction.
Inhibition of Hepatitis B Virus Dna Replication by Imino Sugars Without the Inhibition of the Dna Polymerase: Therapeutic Implications
Authors: Mehta Anand , Carrouée Sandra , Conyers Bertha , Jordan Robert , Butters Terry , Dwek Raymond A. , Block Timothy M.
Published: August 2002

Previously we have shown that the imino sugar inhibitor of N–linked glycan processing, N–nonyl–deoxynojirimycin (N–nonyl–DNJ), had antiviral activity in the woodchuck model of chronic hepatitis B virus (HBV) infection. In studying the mechanism of action of this compound, it was discovered that imino sugars could inhibit HBV secretion without inhibiting N–linked glycoprocessing. Although N–nonyl–DNJ is an inhibitor of the endoplasmic reticulum (ER) glucosidase, here it is shown that N–nonyl–DNJ retained antiviral activity at concentrations that had no significant impact on ER glucosidase function. Taken together, these results suggested that N–nonyl–DNJ possessed an antiviral activity attributable to a function other than an impact on glycoprocessing. This hypothesis was confirmed by experiments showing that N–nonyl–deoxygalactojirimycin (N–nonyl–DGJ), an alkyl derivative of galactose with no impact on glycoprocessing, retains anti–HBV activity. The data suggest that N–nonyl–DGJ exerts its antiviral action at a point before viral envelopment and may prevent the proper encapsidation of the HBV pregenomic RNA.
Reduced Mitochondrial Adenosine Triphosphate Synthesis in Skeletal Muscle in Patients With Child–Pugh Class B and C Cirrhosis
Authors: Jacobsen Esben Bjørn , Hamberg Ole , Quistorff Bjørn , Ott Peter
Published: August 2002

Patients with cirrhosis of the liver often complain of tiredness and a lack of strength at physical exercise. Other investigators have found that muscle strength, work capacity, and maximal oxygen consumption are reduced in cirrhosis. We hypothesized that mitochondrial maximal rate of ATP synthesis in skeletal muscle may be impaired in these patients. This was tested with 31P nuclear magnetic resonance spectroscopy in anterior tibial muscle of cirrhotic patients and healthy controls at rest, during exercise, and subsequent recovery. In patients with Child–Pugh class B and C cirrhosis resting PCr/Pi ratio (8.3 ± 1.0; n = 7) was lower than in patients with Child–Pugh class A cirrhosis (12.1 ± 2.1; n = 7) and controls (11. 7 ± 1.1; n = 6; P = .03), while the resting Pi/γATP ratio was higher in Child–Pugh class B and C patients (0.43, 0.30, and 0.27, respectively; P = .03). Maximal rate of mitochondrial adenosine triphosphate (ATP) synthesis (V
max) as calculated from the initial rate of phosphocreatine (PCr) recovery after work was lower in Child–Pugh class B and C cirrhosis (0.189 mmol/L/s ± 0.034) than in both Child–Pugh class A patients (0.402 mmol/L/s ± 0.103) and controls (0.425 mmol/L/s ± 0.064; P = .01). V
max was significantly correlated to intracellular free [Mg2+] obtained from the 31P nuclear magnetic resonance (NMR) spectra (P = .003). Insufficient oxygen delivery did not seem a likely cause of reduced ATP synthesis in the patients. These findings suggest either a decreased number of mitochondria in skeletal muscle of the cirrhotic patient in Child–Pugh class B and C or a defective mitochondrial function that could be related to low intracellular free [Mg2+].
Increased Extracellular Matrix Remodeling Is Associated With Tumor Progression in Human Hepatocellular Carcinomas
Authors: Théret Nathalie , Musso Orlando , Turlin Bruno , Lotrian Dominique , Bioulac-Sage Paulette , Campion Jean-Pierre , Boudjéma Karim , Clément Bruno
Published: August 2002

Matrix metalloproteinase–2 (MMP2) is a key enzyme in the process of extracellular matrix remodeling involved in tumor invasion and metastasis. The activation of MMP2 involves interplay with the membrane type–matrix metalloproteinase–1 (MT1–MMP) and the tissue inhibitor of metalloproteinase–2 (TIMP2). In vitro, activated hepatic stellate cells are a main source of MMP2 and collagen I induces MMP2 activation. The steady–state mRNA levels of MMP2, MT1–MMP, TIMP2, collagen I, collagen IV, and laminin γ1 were compared with MMP2 activity in 55 hepatocellular carcinomas, 47 matching nontumor biopsies and 19 histologically normal livers. In hepatocellular carcinomas, increased collagen I mRNA levels were strongly associated with those of MMP2 (Spearman R = .74, P < .001), MT1–MMP (R = .65, P < .001) and TIMP2 (R = 0.61, P < .001). MMP2 activity was correlated with the mRNA expression of collagen I (R = .45 P < .01), collagen IV (R = .40, P < .01) and laminin γ1 (R = .33, P < .05). Unlike collagen IV and laminin γ1 mRNAs, MMP2, MT1–MMP, TIMP2, collagen I mRNA levels were increased in nonencapsulated compared with encapsulated tumors (P < .05). In addition, MMP2 activity was fourfold higher (P < .01) in tumors arising in cirrhotic livers than in those arising in noncirrhotic livers. Moreover, tumor recurrence was associated with 4.6– and 2.8–fold (P < .05) higher collagen I and MMP2 mRNA levels, respectively, in hepatocellular carcinomas arising in cirrhotic livers. Thus, a high extracellular matrix remodeling favors tumor progression in hepatocellular carcinomas.
Protein Kinase C-Dependent Distribution of the Multidrug Resistance Protein 2 From the Canalicular to the Basolateral Membrane in Human Hepg2 Cells
Authors: Kubitz Ralf , Huth Clemens , Schmitt Marcus , Horbach Axel , Kullak-Ublick Gerd , Häussinger Dieter
Published: August 2002

The subcellular localization of hepatobiliary transport proteins directly affects the rate of bile formation, e.g., the conjugate export pump multidrug resistance protein 2 (MRP2) is regulated on a short–term scale by retrieval from and insertion into the canalicular membrane in the liver. This study reports on the effects of protein kinase C on MRP2 localization and activity in human hepatoblastoma HepG2 cells. MRP2 was detected in HepG2 cells by immunocytochemistry and Western blot analysis. Functional activity was assessed by confocal laser scanning microscopy using fluorescent MRP2 substrates. In untreated HepG2 cells MRP2 was almost exclusively localized at the apical membrane. Treatment of HepG2 cells with phorbol–12–myristate–13–acetate (PMA) resulted in a rapid decrease of apically localized MRP2 and a loss of more than 90% of pseudocanaliculi within 4 hours. This was accompanied by a reduced pseudocanalicular secretion of the MRP2 substrate glutathione–methylfluorescein. Interestingly, PMA treatment (1–100 nmol/L) led to the appearance of immunoreactive MRP2 at the basolateral membrane within 30 minutes. This was shown by its colocalization with MRP1, human dipeptidylpeptidase IV (DPPIV), and transfected rat Ntcp. The effects of PMA on MRP2 localization were sensitive to the protein kinase C (PKC) inhibitor Gö6850 but insensitive to inhibition of MEK by PD098059. Basolateral MRP2–appearance was not inhibited by cycloheximide or by disruption of microtubules or microfilaments. In rat livers cholestasis was induced by PMA (100 nmol) and MRP2 was detected at the basolateral membrane in some areas, colocalizing with Ntcp. The data suggest that retargeting of canalicular MRP2 to the basolateral membrane due to PKC activation may represent a novel mechanism that may contribute to cholestasis.
Detection of Antimitochondrial Autoantibodies in Immunofluorescent Ama–Negative Patients With Primary Biliary Cirrhosis Using Recombinant Autoantigens
Authors: Miyakawa Hiroshi , Tanaka Atsushi , Kikuchi Kentaro , Matsushita Masanao , Kitazawa Eriko , Kawaguchi Naomi , Fujikawa Hirotoshi , Gershwin Eric M.
Published: August 2002

Antimitochondrial antibodies (AMA) are the serologic hallmark of primary biliary cirrhosis (PBC). However, depending on the clinical laboratory, from 5% to 17% of PBC patients are consistently AMA–negative, using native mitochondrial antigens and a variety of conventional assays including immunofluorescence (IMF) and enzyme–linked immunosorbent assay (ELISA). The major immunoreactive mitochondrial autoantigens are the E2 members of the 2–oxo–acid dehydrogenase complex family, including pyruvate dehydrogenase complex–E2 (PDC–E2), branched chain 2–oxo acid dehydrogenase complex–E2 (BCOADC–E2), and oxo–glutarate dehydrogenase complex–E2 (OGDC–E2); cDNAs of these proteins have now been cloned, sequenced, and their B–cell epitopes defined. In the present study, we cloned cDNAs encoding these proteins from human, not bovine, sources, and expressed the recombinant proteins in a newly developed ELISA that employs a unique Escherichia coli buffer, and compared the data with previous assays using both AMA–positive and –negative patients. Using this new assay and our criteria for positive as an optical density (OD) greater than 10 SD above the mean of control sera, the AMA–positive rate of 191 PBC sera was 94% (179 of 191) compared with 84% (161 of 191) by IMF. None of the 316 control sera were reactive. Using our recombinant assays, we focused attention on the 30 IMF–AMA-negative patients. Twenty–two of 30 (73%) of these patients were positive using this new ELISA. The group of 30 IMF–AMA-negative/ELISA–positive patients did not differ significantly from a comparable population of IMF–AMA-positive patients with respect to age, sex distribution, liver function tests, elevation of serum IgM, or pathologic stage.
Intrahepatic Amino Acid and Glucose Metabolism in A D–Galactosamine-Induced Rat Liver Failure Model
Authors: Arai Kosuke , Lee Kyongbum , Berthiaume François , Tompkins Ronald G. , Yarmush Martin L.
Published: August 2002

A better understanding of the hepatic metabolic pathways affected by fulminant hepatic failure (FHF) would help develop nutritional support and other nonsurgical medical therapies for FHF. We used an isolated perfused liver system in combination with a mass–balance model of hepatic intermediary metabolism to generate a comprehensive map of metabolic alterations in the liver in FHF. To induce FHF, rats were fasted for 36 hours, during which they received 2 d–galactosamine injections. The livers were then perfused for 60 minutes via the portal vein with amino acid-supplemented Eagle minimal essential medium containing 3% wt/vol bovine serum albumin and oxygenated with 95% O2/5% CO2. Control rats were fasted for 36 hours with no other treatment before perfusion. FHF rat livers exhibited reduced amino acid uptake, a switch from gluconeogenesis to glycolysis, and a decrease in urea synthesis, but no change in ammonia consumption compared with normal fasted rat livers. Mass–balance analysis showed that hepatic glucose synthesis was inhibited as a result of a reduction in amino acid entry into the tricarboxylic acid cycle by anaplerosis. Furthermore, FHF inhibited intrahepatic aspartate synthesis, which resulted in a 50% reduction in urea cycle flux. Urea synthesis by conversion of exogenous arginine to ornithine was unchanged. Ammonia removal was quantitatively maintained by glutamine synthesis from glutamate and a decrease in the conversion of glutamate to α–ketoglutarate. Mass–balance analysis of hepatic metabolism will be useful in characterizing changes during FHF, and in elucidating the effects of nutritional supplements and other treatments on hepatic function.
Safety and efficacy of oral entecavir given for 28 days in patients with chronic hepatitis B virus infection
Authors: de Man Robert A. , Wolters Leonieke M. M. , Nevens Frederik , Chua David , Sherman Morris , Lai Cing L. , Gadano Adrian , Lee Youngmee , Mazzotta Fransesco , Thomas Neil , DeHertogh Deborah
Published: August 2002

Entecavir is an oral antiviral drug with selective activity against hepatitis B virus (HBV). We conducted a randomized, placebo-controlled, dose-escalating study in patients with chronic hepatitis B infection in which we evaluated the efficacy and safety of entecavir given for 28 days. Follow-up was 24 weeks. All doses of entecavir (0.05 mg, 0.1 mg, 0.5 mg, and 1.0 mg) showed a pronounced suppression of replication of the HBV with a 2.21, 2.29, 2.81, and 2.55 mean log10 reduction of viral load, respectively. Approximately 25% of patients on entecavir showed a decline of HBV DNA below the limit of detection of the Chiron HBV-DNA assay (<0.7 MEq/mL). In the postdosing follow-up period patients who were treated with 0.5 and 1.0 mg of entecavir showed a considerably slower return in their HBV DNA levels to baseline compared with those patients treated with lower dosages (P < .05). All doses of entecavir were well tolerated with no significant difference between treated patients and those receiving placebo. No significant changes in alanine transaminase (ALT) levels within the dose groups and the placebo group between baseline and the end of treatment were observed. Three patients (9%) (1 each in the 0.05-, 0.1-, and 0.5-mg groups) experienced asymptomatic hepatitis flares 16 weeks (2 patients) and 24 weeks (1 patient) after withdrawal of entecavir. In conclusion, in this 28-day study of entecavir a pronounced decrease of HBV DNA was observed and there were no significant side effects in entecavir patients in comparison with placebo-treated patients. (Hepatology 2001;34:578-582.)
Psychosocial Practice Trends in Pediatric Oncology
Authors: Hicks Melissa Deifer , Lavender Rebecca
Published: August 2002

Children with cancer are now living longer lives. As a result, the focus on care has shifted to include psychosocial components to help promote healthy adjustment to the diagnosis of cancer. In addition, the focus extends to the systems that encompass the patient and family. This ranges from cultural needs to comfort care and how the medical team and institutions address these issues. Through well-defined psychosocial practices, children and families are better prepared to cope with cancer and become well-adjusted survivors. However, regardless of the eventual outcome of the disease, providing psychosocial services can add to the quality of life. Many common practices have emerged over the last 25 years, and many more will continue to arise as outcome studies continue to address the impact of psychosocial services and interventions. This report explores some current trends and common psychosocial practices in working with children with cancer and their families.
A Living Legend in Pediatric Oncology Nursing: Jean Fergusson
Authors: Ruccione Kathy , Fergusson Jean
Published: August 2002

Jean Fergusson is a true pioneer in pediatric oncology nursing. Her many professional accomplishments include working alongside Dr. Sidney Farber and others in the first pediatric Tumor Therapy Clinic in the United States, establishing a model pediatric nurse practitioner program that graduated an influential cadre of pediatric oncology nurse practitioners, publishing landmark papers about late sequelae of childhood cancer treatment, and serving as a role model and mentor to countless nurses over the past 50 years. Jean has brought all she is to her life as a nurse—she is gentle, funny, wise, easily moved, curious, generous, and, most of all, she truly loves children. An eyewitness to the dramatic changes in pediatric oncology over the decades, she herself is a survivor, having overcome dyslexia and other daunting life challenges. In this interview, Jean responds to questions about what shaped her interest in nursing and how she chose pediatric oncology, her impressions and recollections of the early days in pediatric oncology nursing, and her vision for the future of our specialty.