bluuebunny/crossref_metadata_2025 huggingface dataset:
Acute biliary pancreatitis: Diagnosis and management
Authors: Liu Chi‐Leung , Lo Chung‐Mau , Fan Sheung‐Tat
Published: August 2002

Acute biliary pancreatitis is a serious complication of biliary calculous disease and is associated with significant morbidity and mortality. The role of endoscopic retrograde cholangiopancreatography (ERCP) in the management of acute biliary pancreatitis has been the focus of discussion in recent years. In addition, the exact role of laparoscopic cholecystectomy (LC) in the management of acute biliary pancreatitis has not yet been fully defined. In this report, we evaluated a protocol of emergency ERCP (within 24 hours) for predicted severe attacks, early ERCP (within 72 hours) for predicted mild attacks, and interval LC for management of acute biliary pancreatitis. Between January 1992 and June 1995 a total of 75 patients with acute biliary pancreatitis were managed according to the protocol. Bedside ultrasonography at admission diagnosed 94% of all 64 patients with gallbladder stones, but the sensitivity of visualizing choledocholithiasis was low (19%). Forty‐five (60%) of them were predicted to have a severe attack by either Ranson or glucose/urea criteria. Emergency ERCP and endoscopic sphincterotomy (ES) for identifiable common bile duct (CBD) or ampullary stones were performed on all patients predicted to have a severe attack within 24 hours from presentation. An early endoscopic procedure was performed on all patients predicted to have a mild attack within 72 hours from presentation. ERCP was successful in 95% of all patients, and CBD stones were detected in 52 (69%) of them. ES and stone clearance were successful in all of these 52 patients. The morbidity associated with the endoscopic procedure was 3%, and there were no deaths. All except one patient survived the attack of acute pancreatitis, resulting in an overall mortality of 1%. Interval LC was performed on 46 patients with a conversion rate of 4%. The median postoperative hospital stay after LC was 2 days, and there was no major intraoperative or postoperative morbidity or mortality. Our experience suggests that the policy of emergency ERCP for patients with predicted severe disease, early ERCP for patients with predicted mild disease, and interval LC are associated with favorable outcomes in patients with acute biliary pancreatitis. Acute biliary pancreatitis can be managed safely and effectively by a combined endoscopic and laparoscopic approach.
Effect of Subtotal Thyroidectomy on Natural History of Ophthalmopathy in Graves’ Disease
Authors: Abe Yoshifumi , Sato Haruhiro , Noguchi Masako , Mimura Takashi , Sugino Kiminori , Ozaki Osamu , Yoshimura Horoshi , Ito Kunihiko
Published: August 2002

Graves’ ophthalmopathy is a complex disease whose pathogenesis is thought to be autoimmune. The treatment of Graves’ disease is considered to be related to the progression of Graves’ ophthalmopathy. There have been no long‐term prospective studies on the natural course of Graves’ ophthalmopathy. In this study we assessed the influence of subtotal thyroidectomy on the outcome of Graves’ ophthalmopathy, comparing it with that of radioactive iodine treatment. Altogether 287 untreated patients with Graves’ disease referred to Ito hospital in 1989 were followed prospectively for 5 years. A group of 67 patients were treated with radioactive iodine, and 18 patients underwent surgery. Proptosis of the eyes was measured in all patients using Hertel's exophthalmometer. The mean value of proptosis in patients with untreated Graves’ disease was 14.8 ± 0.2 mm (mean ± SEM), and after 5 years it increased slightly to 15.0 ± 0.2 mm. The mean changes of proptosis in patients treated surgically and patients given radioactive iodine were −0.01 ± 0.22 and 0.93 ± 0.28 mm, respectively (p < 0.05). In patients treated by subtotal thyroidectomy, ophthalmopathy did not change in 77.8%; it progressed in 5.6% and was alleviated in 16.7%. In patients treated with radioactive iodine, the ophthalmopathy did not change in 86.6%, progressed in 10.4%, and was alleviated in 3.0%. These findings indicate that surgery can be a better treatment than radioactive iodine for Graves’ patients with ophthalmopathy.
Surgical Treatment of Spontaneous Pneumothorax: Ten‐year Experience
Authors: Athanassiadi Kalliopi , Kalavrouziotis G. , Loutsidis A. , Hatzimichalis A. , Bellenis I. , Exarchos N.
Published: August 2002

Spontaneous pneumothorax (SP) is commonly observed in young, tall, thin subjects without apparent underlying lung disease and in the elderly with chronic emphysematous lung disease. We present our experience in treating SP during the last decade. From December 1986 to November 1996 a total of 417 consecutive patients with SP were admitted to our department. There were 349 males (83.7%) and 68 females, ranging in age from 14 to 93 years. A right‐side SP was detected in 234 cases (56.1%), a left‐side SP in 175 (42.0%), and a bilateral SP in 8 (1.9%). Treatment included observation/aspiration (n= 16, 3.8%), tube thoracostomy (n= 372, 89.2%), multiple tubes (n= 29, 7.0%), blood pleurodesis (n= 13, 3.1%), midsternotomy (n= 3, 0.7%), and minithoracotomy (n= 92, 22.1%). Primary indications for operation were recurrent SP (n= 49) and persistent air leak (n= 46). Blebs or bullae were found in all patients and were ablated by stapling. Pleural abrasion was also performed. All showed good lung expansion postoperatively. Perioperative mortality was zero. The mean hospital stay was 6.5 days. Follow‐up of 89 patients who had undergone surgical treatment (93.7%) at 1 to 100 months revealed only one recurrence. Tube thoracostomy is still the treatment of choice for SP. Surgical intervention is recommended only in cases of recurrent SP or persistent air leak. Minithoracotomy is a safe surgical approach with satisfactory cosmetic results.
Short‐term Outcome and Predictors of Adverse Events following Pulmonary Thromboendarterectomy
Authors: Gilbert Timothy B. , Gaine Sean P. , Rubin Lewis J. , Sequeira Alejandro J.
Published: August 2002

Pulmonary complications including hypoxemia, right heart failure, and prolonged ventilation may follow pulmonary thromboendarterectomy (PTE) performed via cardiopulmonary bypass (CPB) with deep hypothermic circulatory arrest. Seventeen adult patients have undergone PTE at the University of Maryland Medical System during the preceding 3 years. From these patients, clinical and hemodynamic parameters were tabulated pre‐CPB, post‐CPB, at admission to the intensive care unit (ICU), and prior to discontinuation of invasive monitoring in the ICU. Data on anthropometric variables, survival, and times of extracorporeal circulation, mechanical ventilation, and hospital stay were also collected. The mean values for pulmonary arterial systolic and diastolic pressures and pulmonary vascular resistance (PVR) decreased significantly from pre‐CPB values after PTE (all p < 0.05). Mild mixed acidosis present at ICU admission resolved prior to discharge (p= 0.002). The length of mechanical ventilation time was positively correlated with the absolute post‐CPB PVR and negatively correlated with the relative change in central venous pressure (CVP) from pre‐CPB to post‐CPB values (r= 0.75, p= 0.037). Of the pre‐CPB anthropometric variables, only body mass index was significantly higher in nonsurvivors (p= 0.037). Pulmonary artery pressures and vascular resistance fall significantly after PTE. A lower post‐CPB PVR and a relatively decreased (i.e., from pre‐CPB values) CVP predict reduced length of postoperative ventilation but not of the hospital stay. Mortality appears increased in patients with a large body habitus.
Primary Biliary Stones: Diagnosis and Management
Authors: Liu Chi‐Leung , Fan Sheung‐Tat , Wong John
Published: August 2002

To review the results of treatment of primary biliary stones, 96 consecutive patients managed from 1991 to 1996 were studied retrospectively. Acute cholangitis and abdominal pain were the presenting symptoms in 57 patients (59%) and 29 patients (30%), respectively. Fifty‐four patients (56%) had a history of biliary surgery. Endoscopic retrograde cholangiopancreatography, ultrasonography, and computed tomography were frequently employed for diagnosis of primary biliary stones and were performed on 84 patients (88%), 90 patients (94%), and 89 patients (93%), respectively. Intrahepatic stones were identified in 91 patients (95%) and biliary strictures in 34 patients (35%). Concomitant cholangiocarcinoma occurred in 15 patients (16%). Hepatic resection was required in 55 patients (57%) for removal of an atrophic liver lobe or a segment related to repeated infection, biliary strictures, liver abscesses, or cholangiocarcinoma. Intraoperative choledochoscopy was routinely performed in all patients to detect, remove, or confirm clearance of biliary stones. A hepaticocutaneous jejunostomy (HCJ) was constructed in 70 patients (73%) to facilitate postoperative choledochoscopic examination or biliary stone extraction. Twenty‐two patients (23%) had residual stones and required postoperative choledochoscopic extraction. Complete eradication of residual stones was achieved in all patients. Postoperative morbidity occurred in 28 patients (29%), and there was one hospital death (a patient with cholangiocarcinoma). With a median follow‐up of 26 months (range 2–62 months), stones recurred in three patients. In conclusion, the early results of treatment of primary biliary stones were satisfactory owing to a systematic, aggressive approach that consisted of hepatic resection, frequent construction of an HCJ, and postoperative choledochoscopy.
Concomitant Analysis of the Epidermal Growth Factor Receptor Family in Esophageal Cancer: Overexpression of Epidermal Growth Factor Receptor mRNA but Not of c‐erbB‐2 and c‐erbB‐3
Authors: Friess Helmut , Fukuda Akira , Tang Wen‐Hao , Eichenberger Adrian , Furlan Neva , Zimmermann Arthur , Korc Murray , Büchler Markus W.
Published: August 2002

The epidermal growth factor receptor family consists of four closely related transmembrane receptors: epidermal growth factor receptor (EGF‐R), c‐erbB‐2, c‐erbB‐3, and c‐erbB‐4. Overexpression of each receptor may lead to cell transformation and contributes to tumor progression in various malignancies. Although these factors have been analyzed in many cancers separately, little is known about their concomitant expression in esophageal cancer. Based on the finding that EGF‐R and c‐erbB‐2 form highly active transmembranous heterodimers that enhance cell growth and proliferation, we used Northern blot analysis and immunohistochemistry to analyze the concomitant expression of EGF‐R, c‐erbB‐2, and c‐erbB‐3 in tissue samples obtained from 39 patients undergoing esophagectomy for esophageal cancer. Northern blot analysis revealed a fourfold increase (p < 0.01) in EGF‐R mRNA levels in the esophageal cancer samples in comparison with normal tissue samples. The c‐erbB‐2 receptor was only 1.25‐fold elevated in the esophageal cancers, which failed to be statistically significant (p= 0.31). In contrast, c‐erbB‐3 mRNA levels were 3.5‐fold lower (p < 0.01) in the esophageal cancers than in the normal tissues. Immunohistochemical analysis showed weak EGF‐R, c‐erbB‐2, and c‐erbB‐3 immunostaining in the normal esophageal tissue. In esophageal cancer samples, immunoreactivity for EGF‐R, c‐erbB‐2, and c‐erbB‐3 was mainly located in the cancer cells. Strong EGF‐R, c‐erbB‐2, and c‐erbB‐3 immunoreactivity was present in 59%, 64%, and 64% of the esophageal cancer samples, respectively. In consecutive tissue sections, identical cancer cell clusters often exhibited these three closely related receptors simultaneously. However, correlation of the immunohistochemical findings with the clinicopathologic patient parameters revealed that the presence of EGF‐R, c‐erbB‐2, or c‐erbB‐3 had no influence on patient survival (p > 0.05). In addition, the simultaneous presence of these receptors did not influence survival. Our findings indicate that in esophageal cancer the presence of EGF‐R, c‐erbB‐2, and c‐erbB‐3 alone or in combination seems to have no major influence on patient prognosis and does not alter tumor growth behavior significantly.
Lung Pathology in Premature Infants with Ureaplasma urealyticum Infection
Authors: Viscardi Rose M. , Manimtim Winston M. , Sun Chen-Chih J. , Duffy Lynn , Cassell Gail H.
Published: August 2002

Respiratory tract colonization with Ureaplasma urealyticum in preterm infants has been associated with a higher incidence of pneumonia, severe respiratory failure, bronchopulmonary dysplasia (BPD), and death. In this report, we characterize the lung pathology and expression of tumor necrosis factor-α (TNF-α) associated with U. urealyticum pneumonia in very low–birth weight infants (VLBW; ≤1500 g). Lung pathology of archived autopsy specimens was retrospectively reviewed in three groups of VLBW infants: 5 gestational controls who died from nonpulmonary causes, 13 infants with pneumonia who were culture and/or PCR negative for U. urealyticum, and 5 infants with pulmonary disease and positive for U. urealyticum by tracheal aspirate and/or lung tissue culture or polymerase chain reaction (PCR). Presence and extent of alveolar macrophages and neutrophils, as well as interstitial lymphocytic infiltration and fibrosis were evaluated. PCR was performed on formalin-fixed, paraffin-embedded lung sections. Additional sections were immunostained for TNF-α. The peripheral total white blood cell counts and absolute neutrophil counts were three-fold higher in infants with U. urealyticum pneumonia than cell counts in infants infected with other organisms. There was a trend toward a predominance of neutrophils in alveoli of non- Ureaplasma pneumonia infants, but a trend toward a predominance of alveolar macrophages in U. urealyticum–infected infants. The most striking finding was the presence of increased interstitial fibrosis in all Ureaplasma-inf ected infants. TNF-α immunoreactive cell density was very low in the gestational controls, but increased in both pneumonia groups. We conclude that persistent lung U. urealyticum infection may contribute to chronic inflammation and early fibrosis in the preterm lung.
Nonselective Cation Channels As Effectors of Free Radical-Induced Rat Liver Cell Necrosis
Authors: Barros Luis Felipe , Stutzin Andrés , Calixto Andrea , Catalán Marcelo , Castro Joel , Hetz Claudio , Hermosilla Tamara
Published: August 2002

Necrosis, as opposed to apoptosis, is recognized as a nonspecific cell death that induces tissue inflammation and is preceded by cell edema. In non–neuronal cells, the latter has been explained by defective outward pumping of Na+ caused by metabolic depletion or by increased Na+ influx via membrane transporters. Here we describe a novel mechanism of swelling and necrosis; namely the influx of Na+ through oxidative stress–activated nonselective cation channels. Exposure of liver epithelial Clone 9 cells to the free–radical donors calphostin C or menadione induced the rapid activation of an approximately 16–pS nonselective cation channel (NSCC). Blockage of this conductance with flufenamic acid protected the cells against swelling, calcium overload, and necrosis. Protection was also achieved by Gd3+, an inhibitor of stretch–activated cation channels, or by isosmotic replacement of extracellular Na+ with N–methyl–D–glucamine. It is proposed that NSCCs, which are ubiquitous although largely inactive in healthy cells, become activated under severe oxidative stress. The ensuing influx of Na+ initiates a positive feedback of metabolic and electrolytic disturbances leading cells to their necrotic demise.
Comparison of Two Clinical Scales for Causality Assessment in Hepatotoxicity
Authors: Lucena Ma Isabel , Camargo Raquel , Andrade Raúl J. , Perez-Sanchez Carlos J. , De La Cuesta Felipe Sanchez
Published: August 2002

This study was performed to compare the assessments of drug–induced liver injury obtained with 2 methods, the Council for International Organizations of Medical Sciences (CIOMS) scale and the recently validated Maria & Victorino (M&V) clinical scale, in cases submitted to a registry of hepatotoxicity. A total of 215 cases of hepatotoxicity reported with a structured reporting form were evaluated by 3 independent experts. Because of the use of multiple drugs, 228 ratings were generated. The probability of the diagnosis was classified as definitive, probable, possible, unlikely, or excluded, and evaluated for consistency with a weighted κ statistical test. Absolute agreement between the 2 scales was observed in 42 cases (18%, weighted κ 0.28) with disagreement of 1 level in 108 cases (47%), and of 2 levels in 70 cases (31%). The best correlation between the 2 scales was obtained for drug–induced liver injury involving a suggested immunoallergic mechanism: the disagreement was 1 level or less in 72% of the cases (34 of 48), compared with 60% of the cases (85 of 141) that involved a presumed idiosyncratic metabolic mechanism. The lowest agreement (6%) was observed in cases with evidence of cholestasis. No agreement was found in cases of fulminant hepatitis or death. The CIOMS scale showed better discriminative power and produced assessments closer to those of specialists. The performance of the M&V scale was poor in reactions with long latency periods (i.e., amoxycillin/clavulanic acid), evolution to chronicity after withdrawal (cholestatic pattern), or death.
Method to Detect Substitutions in the Interferon–Sensitivity—Determining Region of Hepatitis C Virus 1B for Prediction of Response to Interferon Therapy
Authors: Nishiguchi Shuhei , Ueda Tadashi , Itoh Teiji , Enomoto Masaru , Tanaka Motoharu , Tatsumi Nobuyuki , Fukuda Katsuhiko , Tamori Akihiro , Habu Daiki , Takeda Tadashi , Otani Shuzo , Shiomi Susumu
Published: August 2002

Substitutions deduced by direct sequencing in the interferon–sensitivity—determining region (ISDR) of hepatitis C virus (HCV) are related to patients' responses to interferon (IFN), but sequencing is time consuming and results are only for the dominant virus. We developed a rapid method to detect such changes. With serum from 50 patients with chronic hepatitis C (genotype 1b) given IFN–α, a way to detect changes in ISDR by hybridization with oligonucleotide probes that had a prototype nucleotide sequence of HCV–J was established. Hybridization intensity was expressed as optical density (ODNS5A). The method was checked with serum from 100 more patients. In the study of 50 patients, all 21 with the prototype sequences had a high ODNS5A (≥0.4), and all 8 patients with a mutant–type sequence had low values (≤0.2). Twelve (95% confidence interval, 36–81%) of 20 patients with ODNS5A of <0.4 and 2 (1%–22%) of 30 patients with ODNS5A ≥0.4 had complete responses (CR). All nine (66%–100%) patients with ODNS5A <0.4 and little HCV RNA (<100 kIU/mL) had CR, but none (0%–14%) of the 24 patients with high values from both predictors had CR. In the study of 100 patients, ODNS5A and the HCV RNA level were independent predictors of the effects of IFN. By multivariate analysis, the odds ratio for a CR in patients with ODNS5A of ≥0.4 was 0.015 (0.001–0.190) compared with the other patients (P = .001). In conclusion, our method should be useful in identification of prototype strains, which generally resist IFN therapy.
Signal Pathway Involved in the Development of Hypoxic Preconditioning in Rat Hepatocytes
Authors: Carini Rita , De Cesaris Maria Grazia , Splendore Roberta , Vay Daria , Domenicotti Cinzia , Nitti Maria Paola , Paola Dimitri , Pronzato Maria Adelaide , Albano Emanuele
Published: August 2002

Ischemic preconditioning improves liver resistance to hypoxia and reduces reperfusion injury following transplantation. However, the intracellular signals that mediate the development of liver hypoxic preconditioning are largely unknown. We have investigated the signal pathway leading to preconditioning in freshly isolated rat hepatocytes. Hepatocytes were preconditioned by 10–minute incubation under hypoxic conditions followed by 10 minutes of reoxygenation and subsequently exposed to 90 minutes of hypoxia. Preconditioning reduced hepatocyte killing by hypoxia by about 35%. A similar protection was also obtained by preincubation with chloro–adenosine or with A2A–adenosine receptor agonist CGS21680, whereas A1–adenosine receptor agonist N–phenyl–isopropyladenosine (R–PIA) was inactive. Conversely, the development of preconditioning was blocked by A2–receptor antagonist 3,7–dimethyl–1–propargylxanthine (DMPX), but not by A1–receptor antagonist 8–cyclopenthyl–1,3–dipropylxanthine (DPCPX). In either preconditioned or CGS21680–treated hepatocytes a selective activation of δ and ε protein kinase C (PKC) isoforms was also evident. Inhibition of heterotrimeric Gi protein or of phospholypase C by, respectively, pertussis toxin or U73122, prevented PKC activation as well as the development of preconditioning. MEK inhibitor PD98509 did not interfere with preconditioning that was instead blocked by p38 MAP kinase inhibitor SB203580. The direct activation of p38 MAPK by anisomycin A mimicked the protection against hypoxic injury given by preconditioning. Consistently, an increased phosphorylation of p38 MAPK was observed in preconditioned or CGS21680–treated hepatocytes, and this effect was abolished by PKC–blocker, chelerythrine. We propose that a signal pathway involving A2A–adenosine receptors, Gi–proteins, phospholypase C, δ– and ε–PKCs, and p38 MAPK, is responsible for the development of liver ischemic preconditioning.
Activation of Retinoic X Receptor and Peroxisome Proliferator-Activated Receptor–γ Inhibits Nitric Oxide and Tumor Necrosis Factor–α Production in Rat Kupffer Cells
Authors: Uchimura Koutaro , Nakamuta Makoto , Enjoji Munechika , Irie Takashi , Sugimoto Rie , Muta Tatsushi , Iwamoto Hiroaki , Nawata Hajime
Published: August 2002

Activators of peroxisome proliferator–activated receptor γ (PPARγ), which forms a heterodimer with retinoic X receptor (RXR), inhibit the production of certain inflammatory mediators. To clarify the role of the PPARγ:RXR signaling pathway in Kupffer cells, we studied the effect of an RXR agonist and PPARγ agonist on LPS–induced nitric oxide (NO) and tumor necrosis factor–α (TNF–α) production. An RXR–specific agonist, Ro47–5944, and a PPARγ–specific agonist, AD4833 (pioglitazone hydrochloride), each inhibited LPS–induced NO and TNF–α production. The combined treatment of Ro47–5944 and AD4833 resulted in enhanced inhibition, and suppressed the mRNA levels of NO and TNF–α. PPARγ:RXR activation did not affect the level of LPS–induced phosphorylation of c–jun N–terminal kinase and p38 mitogen–activated protein kinase. PPARγ:RXR activation also did not affect nuclear factor kappa B (NF–κB) nuclear translocation nor NF–κB and activator protein 1 (AP–1) activation in the electrophoretic mobility–shift assay. Finally, PPARγ:RXR activation suppressed the LPS–induced promoter activity of the NF–κB–luciferase reporter gene in RAW 264.7 cells. These data imply that PPARγ:RXR activation suppresses LPS–induced NO and TNF–α production in Kupffer cells, and that this inhibition occurred at the transcriptional level. Although no consensus PPARγ:RXR–responsive element in the promoter regions of the inducible isoform of nitric oxide synthase (iNOS) and TNF–α genes was found, PPARγ:RXR may interfere with NF–κB and AP–1 transcriptional activity. Our data also suggest a potential therapeutic approach for moderating hepatic injury such as endotoxin shock in which Kupffer cell activation has been implicated.
Estimation of Early Hepatitis C Viral Clearance in Patients Receiving Daily Interferon and Ribavirin Therapy Using A Mathematical Model
Authors: Bekkering Frank C. , Stalgis Carlos , McHutchison John G. , Brouwer Johannes T. , Perelson Alan S.
Published: August 2002

Patients with hepatitis C virus (HCV) genotype 1 infection are resistant to standard interferon (IFN) therapy. We used a mathematical model to estimate the duration of daily therapy necessary to maximize the number of patients achieving viral negativity before 12 weeks of therapy. Patients from a study to determine HCV RNA reduction over 4 weeks using 3 million units (MU), 5 MU, or 10 MU of IFN alfa daily plus Ribavirin were compared with a group receiving IFN alfa 3 MU three times a week. By extending the linear regression and prediction interval lines, the estimated time to negativity was greater than 12 weeks for the standard IFN group, 42 to greater than 84 days for the 3 MU IFN daily plus Ribavirin, 39 to 60 days for 5 MU IFN daily plus Ribavirin and 25 to 45 days for the 10 MU IFN daily and Ribavirin group, respectively. Thus, the use of a predictive model based on log transformation and linear regression of the early HCV RNA response suggests daily doses of 5 or 10 million units of IFN plus Ribavirin will be theoretically necessary for longer than 4 weeks to maximize the number of patients who clear virus by 12 weeks of therapy. This model may be useful in predicting response in groups of patients receiving other therapies.
Expression and Matrix Deposition of Latent Transforming Growth Factor β Binding Proteins in Normal and Fibrotic Rat Liver and Transdifferentiating Hepatic Stellate Cells in Culture
Authors: Breitkopf Katja , Lahme Birgit , Tag Carmen G. , Gressner Axel M.
Published: August 2002

Latent transforming growth factor β binding protein (LTBP), a high–molecular–weight glycoprotein of the large latent TGF–β complex is suggested to serve as an anchor for latent TGF–β in the extracellular matrix and as a component of microfibrillar structures. Proteolytic cleavage of LTBP is supposed to be a prerequisite for the release and generation of bioactive (mature) TGF–β. We investigated the expression of LTBP isoforms in normal and fibrotic rat liver and in cultured rat hepatic stellate cells (HSC) transdifferentiating to myofibroblasts (MFB). We further determined their interaction with the matrix and some of their basic functions. Immunostainings of normal and fibrotic livers demonstrate intense signals for LTBP–1 and –2, preferably in parenchymal, but also nonparenchymal, cells and in fibrotic extracellular matrix. However, in situ hybridization points to a restriction of transcripts to nonparenchymal cells from fibrotic livers, whereas hepatocytes were always devoid of LTBP transcripts. The findings were confirmed by real–time quantitative reverse–transcription polymerase chain reaction (RT–PCR), which showed isoform–specific increases of LTBP transcripts in cultured stellate cells transdifferentiating to MFB and by Northern blot analyses showing the absence of LTBP–1 mRNA in freshly isolated hepatocytes. Using a cell enzyme–linked immunosorbent assay (ELISA), a differential increase of partly deoxycholate (DOC)–resistant, matrix–bound LTBP–1 and –2 was measured in cultured stellate cells. Treatment with plasmin generated soluble LTBP–1 and bioactive TGF–β, which was able to induce Smad7 expression in an autocrine fashion. Our data propose (transdifferentiating) stellate cells, respectively MFB, as the major source of LTBP in normal and fibrotic liver, which here probably fulfills structural and TGF–β–regulating functions as suggested for nonhepatic tissues.
Hepatitis B Infection in Patients With Acute Liver Failure in the United States
Authors: Teo Eng-Kiong , Ostapowicz George , Hussain Munira , Lee William M. , Fontana Robert J. , Lok Anna S.F.
Published: August 2002

Occult hepatitis B virus (HBV) infection has been reported in 30% to 50% of patients with acute liver failure (ALF) in small case series. The aim of this study was to determine the prevalence of occult HBV infection in a large series of ALF patients in the United States and the prevalence of precore and core promoter variants in patients with ALF caused by hepatitis B. Sera from patients in the US ALF study and liver, when available, were tested using nested polymerase chain reaction (PCR) with primers in the HBV S and precore regions. PCR–positive samples were sequenced. Sera and/or liver from 139 patients (39 males, 100 females; mean age, 42 years) enrolled between January 1998 and December 1999 were studied. Twelve patients were diagnosed with hepatitis B, 1 with hepatitis B+C+D coinfection, and 22 had indeterminate etiology. HBV DNA was detected in the sera of 9 (6%) patients; all 9 had ALF caused by hepatitis B. HBV genotypes A, B, C, and D were present in 4, 3, 1, and 1 patients, respectively. Seven of these 9 patients had precore and/or core promoter variants. Liver from 19 patients were examined. HBV DNA was detected in the liver of 3 patients with ALF caused by hepatitis B, but in none of the remaining 16 patients with non–B ALF. Contrary to earlier reports, occult HBV infection was not present in this large series of ALF patients in the United States. HBV precore and/or core promoter variants were common among US patients with ALF caused by hepatitis B. (Hepatology 2001;33:972–976.)
Detection of Hepatitis C Virus in the Bile and Bile Duct Epithelial Cells of Hepatitis C Virus-Infected Patients
Authors: Haruna Yoshimichi , Kanda Tsutomu , Honda Masaharu , Takao Tetsuto , Hayashi Norio
Published: August 2002

The pathobiology of hepatitis C virus (HCV) in the biliary system has not been clarified yet, although bile duct damage is a histological finding characteristic of chronic hepatitis C. In this study, we examined whether HCV infects bile ducts and is released into the bile. Twelve patients positive for serum HCV antibody were examined in this study, and eight were seropositive for HCV RNA by polymerase chain reaction (PCR). For those who underwent abdominal surgery, the bile was aspirated from the gall bladders by fine needle puncture. Five underwent wedge liver biopsy. Series of saline–diluted bile were assayed for HCV RNA by PCR to determine the HCV RNA titers. To examine HCV expression in the biliary system, the liver specimens were immunostained using monoclonal antibodies to the HCV proteins. HCV RNA was detected in the bile of 5 patients with high serum HCV RNA load (≥2.5 Meq/mL). Comparison of viral loads between serum and bile revealed that the HCV RNA level in the bile was as high as that in serum. Furthermore, immunohistological study showed that bile duct epithelial cells were infected with HCV. In contrast, HCV was not found in either the bile or bile duct of patients seronegative for HCV RNA or with low serum HCV load (≤1.1 Meq/mL). These findings suggest that the biliary system is involved in the pathobiology of HCV and that the bile is as highly infectious as the serum. (Hepatology 2001;33;977–980.)
Effects of Blood Volume Restitution Following A Portal Hypertensive-Related Bleeding in Anesthetized Cirrhotic Rats
Authors: Castañeda Beatriz , Morales Josephine , Lionetti Raffaella , Moitinho Eduardo , Andreu Victoria , Pérez-del-Pulgar Sofía , Pizcueta Pilar , Rodés Juan , Bosch Jaime
Published: August 2002

The aim of this study was to investigate the influence of different strategies of blood volume restitution in the outcome of portal hypertension-related bleeding in anesthetized cirrhotic rats. Gastrointestinal hemorrhage was induced by sectioning a first order branch of the ileocolic vein in 38 cirrhotic rats (common bile duct ligation and occlusion). The subsequent hypovolemic shock was treated with no transfusion (n = 17), moderate transfusion (50% of expected blood loss, 5 mL, n = 11), and total transfusion (100% of expected blood loss, 10 mL, n = 10). At the end of the blood transfusion period (minute 15), mean arterial pressure (MAP) partially recovered in rats receiving moderate transfusion or no transfusion but decreased in the 10–mL transfusion group (↓12 ± 43%, P < .05 vs. no transfusion and 5 mL transfusion). After transfusion, groups given no or 5 mL transfusion remained hemodynamically stable. However, rats receiving 10 mL transfusion continued to deteriorate with persistent bleeding and progressive fall in MAP (↓65 ± 12%; P < .05 vs. no transfusion and 5 mL transfusion). Collected blood loss was significantly greater in the 10–mL group (20.0 ± 1.5 g) than in groups given 5 mL (15.9 ± 2.8 g; P < .05) or no transfusion (13.2 ± 2.1 g; P < .05 vs. 10 mL and 5 mL transfusion). Survival in the no transfusion group was 47%. Rats given 5–mL transfusion had 64% survival. The worst survival was observed in the 10–mL transfusion group (0% survival; P < .05). We concluded that a transfusion policy aimed at completely replacing blood loss worsens the magnitude of bleeding and mortality from portal hypertensive–related bleeding in cirrhotic rats. On the contrary, moderate blood transfusion allowed hemodynamic stabilization and increased survival.
Cellular Localization and Up–Regulation of Multidrug Resistance-Associated Protein 3 in Hepatocytes and Cholangiocytes During Obstructive Cholestasis in Rat Liver
Authors: Soroka Carol J. , Lee John M. , Azzaroli Francesco , Boyer James L.
Published: August 2002

The hepatic expression of the ATP–dependent conjugate export pump multidrug resistance-associated protein 2 (Mrp2) is diminished in experimentally induced models of cholestasis. In this study we have examined the localization and expression of Mrp3, another member of the multidrug resistance-associated protein family, in normal liver and after obstructive cholestasis in the rat. Indirect immunofluorescence and confocal microscopy were used to determine the tissue localization and Western blot analysis was performed to quantitate the expression. In normal rat liver Mrp3 was found on the basolateral membrane of cholangiocytes and a single layer of hepatocytes surrounding the central vein. Three and 7 days after bile duct ligation Mrp3 expression was significantly increased, predominantly in hepatocytes in the pericentral region. By 14 days all hepatocytes showed basolateral membrane labeling for Mrp3 at a time when apical Mrp2 staining was significantly diminished. Proliferating bile ducts continued to stain positive, although the intensity of staining did not seem to vary. After 14 days Western blot quantitation showed that Mrp3 had increased approximately 30–fold in total liver membranes. Quantitation of Mrp3 in membranes from isolated hepatocytes of livers of sham and common bile duct-ligated (CBDL) animals showed a significant up–regulation beginning at 1 day and continuing to increase through 14 days postligation. This was in contrast to the progressive decrease in Mrp2 protein. Because Mrp3 is capable of transporting toxic bile acids, up–regulation of Mrp3 may compensate for the down–regulation of Mrp2 in obstructive cholestasis.
Expression of Hepatitis C Virus Proteins Does Not Interfere With Major Histocompatibility Complex Class I Processing and Presentation In Vitro
Authors: Moradpour Darius , Grabscheid Benno , Kammer Andreas R. , Schmidtke Gunter , Groettrup Marcus , Blum Hubert E. , Cerny Andreas
Published: August 2002

Hepatitis C virus (HCV) infection takes a chronic course in the majority of patients. The mechanisms underlying the evasion of the host immune response and viral persistence are poorly understood. In this context, we investigated interactions of HCV proteins with major histocompatibility complex (MHC) class I processing and presentation pathways using cell lines that allow the tetracycline–regulated expression of viral structural and nonstructural proteins. These well–characterized inducible cell lines were found to efficiently process and present endogenously synthesized HCV proteins via MHC class I. Functional MHC class I cell–surface expression and intracellular proteasome activity were not affected by the expression of HCV proteins. These results suggest that viral evasion of the host immune response does not involve interactions of HCV with MHC class I processing and presentation. Other mechanisms, such as interference with the interferon system, may be operative in HCV infection, leading to viral persistence. (Hepatology 2001;33:1282–1287.)
Positron Emission Tomography With [18F]Fluoro–2–Deoxy–D–Glucose for Diagnosis and Staging of Bile Duct Cancer
Authors: Kluge Regine , Schmidt Frank , Caca Karel , Barthel Henryk , Hesse Swen , Georgi Peter , Seese Anita , Huster Dominik , Berr Frieder
Published: August 2002

Malignant tumors with high glucose metabolic rates accumulate [18F]–fluorodeoxyglucose (FDG), a positron emitting tracer. The aim of this study was to evaluate FDG positron emission tomography (PET) for detection and staging of human cholangiocarcinoma (CC). Patients with adenocarcinoma of the biliary tree (n = 26), with benign lesions of the bile ducts (n = 8), and 20 control patients underwent FDG–PET (370 MBq [18F]–FDG, Siemens ECAT EXACT HR+). In a blinded fashion, 4 independent experts evaluated the PET scans visually and semiquantitatively using the standardized uptake value and a tumor/non–tumor ratio. All adenocarcinomas and benign lesions (sclerosing cholangitis, bile duct adenoma, Caroli's disease) were histologically proven and imaged by magnetic resonance imaging and endoscopic retrograde cholangioscopy. True–positive PET scans were obtained in 24 of 26 CC and false–negative scans in the other 2 (sensitivity 92.3%). The PET scan was true–negative in 18 of 20 controls and in all 8 benign biliary lesions (specificity 92.9%). Visual and semiquantitative evaluation using tumor/non–tumor ratios were equally accurate (accuracy 92.6%) whereas evaluation by standardized uptake value revealed lower accuracy (P < .05). Regional or hepatoduodenal lymph node metastases were detected with PET in only 2 of 15 cases whereas distant metastases (peritoneal carcinomatosis, pulmonary metastases) were diagnosed in 7 of 10 cases. In conclusion, PET is highly sensitive and specific for the detection and localization of CC. It can be helpful for diagnosis of distant metastases but is not suitable for detection of regional lymph node metastases. (Hepatology 2001;33:1029–1035.)
Central Venulitis in Pediatric Liver Allografts
Authors: Krasinskas Alyssa M. , Ruchelli Eduardo D. , Rand Elizabeth B. , Chittams Jesse L. , Furth Emma E.
Published: August 2002

Central venulitis (CV), a distinct histologic lesion described in adult liver transplants, can occur with acute portal tract rejection or in isolation (ICV). Possible etiologies include immunosuppressive drug toxicity, acute cellular rejection, viral hepatitis, ischemic injury, and recurrent disease. This study was designed to characterize ICV and to assess its potential etiology in pediatric liver recipients because this population generally does not develop recurrent disease or viral hepatitis. All posttransplantation liver biopsy specimens that were obtained from children who received liver allografts over a 4–year period were reviewed. ICV was identified in 12 of 127 posttransplantation biopsies and in 7 of 45 allograft recipients. Only 4 liver transplantations were performed for potentially recurrent diseases (primary sclerosing cholangitis). ICV first appeared in posttransplantation biopsy specimens significantly later than did portal rejection alone. The finding of CV was not significantly correlated with elevation of Tacrolimus levels, reason for transplantation, donor/recipient cytomegalovirus (CMV) status or blood type, cold ischemic times, or the incidence of outflow obstruction. The responses of CV to therapy were variable and, although the majority of cases resolved, several episodes persisted or recurred. In conclusion, ICV occurs in 16% of pediatric liver allograft recipients and does not appear to be related to recurrent disease, viral hepatitis, drug toxicity, or graft ischemia. CV may be a variant of acute rejection, but longer follow–up is required to determine the most adequate therapy for this entity.
A Molecular Comparison of T Lymphocyte Populations Infiltrating the Liver and Circulating in the Blood of Patients With Chronic Hepatitis B: Evidence for Antigen–Driven Selection of A Public Complementarity–Determining Region 3 (Cdr3) Motif
Authors: Sing Garwin K. , Li Dili , Chen Xiaohua , Macnaughton Tom , Lichanska Agnieszka M. , Butterworth Lesley , Ladhams Andrew , Cooksley Graham
Published: August 2002

Despite a large number of T cells infiltrating the liver of patients with chronic hepatitis B, little is known about their complexity or specificity. To characterize the composition of these T cells involved with the pathogenesis of chronic hepatitis B (CHB), we have studied the clonality of VβT cell receptor (TCR)–bearing populations in liver tissue by size spectratyping the complementarity–determining region (CDR3) lengths of TCR transcripts. We have also compared the CDR3 profiles of the lymphocytes infiltrating the liver with those circulating in the blood to see whether identical clonotypes may be detected that would indicate a virus–induced expansion in both compartments. Our studies show that in most of the patients examined, the T cell composition of liver infiltrating lymphocytes is highly restricted, with evidence of clonotypic expansions in 4 to 9 TCR Vβ subfamilies. In contrast, the blood compartment contains an average of 1 to 3 expansions. This pattern is seen irrespective of the patient's viral load or degree of liver pathology. Although the TCR repertoire profiles between the 2 compartments are generally distinct, there is evidence of some T cell subsets being equally distributed between the blood and the liver. Finally, we provide evidence for a putative public binding motif within the CDR3 region with the sequence G–X–S, which may be involved with hepatitis B virus recognition. (Hepatology 2001;33:1288–1298.)
The Role of P27kip1 Protein Expression on the Biological Behavior of Intrahepatic Cholangiocarcinoma
Authors: Taguchi Ken-Ichi , Aishima Shin-Ichi , Asayama Yoshiki , Kajiyama Kiyoshi , Kinukawa Naoko , Shimada Mitsuo , Sugimachi Keizo , Tsuneyoshi Masazumi
Published: August 2002

Recent studies have reported that a decreased p27kip1 protein (p27) expression is associated with aggressive tumor behavior in several human malignancies. In this study, we examined the role of p27 expression in intrahepatic cholangiocarcinoma (ICC) and the noncancerous intrahepatic bile duct epithelium and assessed its association with clinicopathologic features, especially the influence of decreased p27 expression by the tumor on its prognosis as determined by immunohistochemistry. Almost all noncancerous bile duct epithelia show positive nuclear staining and reveal a faintly positive reaction in the cytoplasm. In 62 surgically resected ICCs, a variable degree of positive p27 nuclear staining is recognized. A subset (13 of 62 cases; 21.0%) of the carcinomas displayed cytoplasmic staining. We classified 62 cases into 2 groups, a low expression group (<50% of p27 nuclear positive cells) and a high expression group (≥50% of p27 nuclear positive cells), and tested for the association between positive expression and clinicopathologic features. We found that a low expression of nuclear p27 was correlated with positive vascular invasion (P < .05). The survival curve of the group with low p27 nuclear expression was significantly lower than that of the group with high p27 nuclear expression (P < .005). According to the multivariate analyses, low p27 expression can be considered an independent prognostic factor whose occurrence indicates a worse prognosis. We conclude that a decreased p27 expression adds an independent parameter that can be used when determining the prognosis of patients with ICC.
Fas–Induced Apoptosis in Mouse Hepatocytes Is Dependent on C/Ebpβ
Authors: Mukherjee Debjani , Kaestner Klaus H. , Kovalovich Kellen K. , Greenbaum Linda E.
Published: August 2002

Apoptotic cell death in the liver in response to activation of the Fas pathway has been implicated in human disease states as well as liver remodeling and tissue repair. C/EBPβ, a member of the CCAAT enhancer binding protein family of bZIP transcription factors has been linked to both growth response and apoptotic targets in the liver, and, therefore, is a likely candidate for the regulation of apoptotic liver injury. We investigated differences in apoptotic cell death in the livers of C/EBPβ–null mice using the Jo–2 agonistic anti–Fas antibody. Apoptotic injury was dramatically reduced in C/EBPβ -/- livers as shown by a nearly 20–fold reduction in apoptotic hepatocytes 6 hours post–Jo–2 treatment in C/EBPβ -/- hepatocytes compared with controls (P < .04) and reduced activation of caspase 3. Bid cleavage occurred in Jo–2 treated C/EBPβ -/- livers indicating a block of Fas–induced injury distal to the death–inducing signaling complex. The level of the antiapoptotic protein bcl–xL was increased greater than tenfold in the mutant animals (P < .04), which can, at least in part, account for the protection from Fas–mediated apoptosis. In contrast, bcl–xL mRNA levels were unchanged. These observations link C/EBPβ to Fas–induced hepatocyte apoptosis through a mechanism that likely involves translational or posttranslational regulation of bcl–xL.
Tauroursodeoxycholic Acid Inserts the Apical Conjugate Export Pump, Mrp2, Into Canalicular Membranes and Stimulates Organic Anion Secretion by Protein Kinase C-Dependent Mechanisms in Cholestatic Rat Liver
Authors: Beuers Ulrich , Bilzer Manfred , Chittattu Anila , Kullak-Ublick Gerd A. , Keppler Dietrich , Paumgartner Gustav , Dombrowski Frank
Published: August 2002

Ursodeoxycholic acid (UDCA) exerts anticholestatic effects by undefined mechanisms. Previous work suggested that UDCA stimulates biliary exocytosis via Ca++– and protein kinase C (PKC)–dependent mechanisms. Therefore, the effect of taurine–conjugated UDCA (TUDCA) was studied in the experimental model of taurolithocholic acid (TLCA)–induced cholestasis on bile flow, hepatobiliary exocytosis, distribution of PKC isoforms, and density of the apical conjugate export pump, Mrp2, in canalicular membranes. Isolated perfused rat livers were preloaded with horseradish peroxidase (HRP), a marker of vesicular exocytosis, and were perfused with bile acids or dimethylsulfoxide (control) only. PKC isoform distribution and membrane density of Mrp2 were studied using immunoblotting and immunoelectron–microscopic techniques. Biliary secretion of the Mrp2 substrate, 2,4–dinitrophenyl–S–glutathione (GS–DNP), was studied in the presence or absence of the PKC inhibitor, bisindolylmaleimide I (BIM–I; 1 μmol/L). TLCA (10 μmol/L) impaired bile flow by 51%; biliary secretion of HRP and GS–DNP by 46% and 95%, respectively; membrane binding of the Ca++–sensitive α–isoform of PKC by 32%; and density of Mrp2 in the canalicular membrane by 79%. TUDCA (25 μmol/L) reversed the effects of TLCA on bile flow, secretion of HRP and GS–DNP, and distribution of α–PKC. TUDCA reduced membrane binding of ε–PKC and increased Mrp2 density 4–fold in canalicular membranes of cholestatic hepatocytes. BIM–I inhibited the effect of TUDCA on GS–DNP secretion in cholestatic livers by 49% without affecting secretion in controls. In conclusion, TUDCA may enhance the secretory capacity of cholestatic hepatocytes by stimulation of exocytosis and insertion of transport proteins into apical membranes via PKC–dependent mechanisms. (Hepatology 2001;33:1206–1216.)