bluuebunny/crossref_metadata_2025 huggingface dataset:
Low frequency of precore hepatitis B virus mutants in anti-hepatitis B e—positive reactivation after loss of hepatitis B e antigen in patients with chronic hepatitis B
Authors: Loriot Marie Anne , Marcellin Patrick , Talbodec Nathalie , Guigonis Véronique , Gigou Michèle , Boyer Nathalie , Bezeaud Annie , Erlinger Serge , Benhamou Jean Pierre
Published: March 2007

The objective of this study was to evaluate the role of hepatitis B virus (HBV) precore mutations in patients with anti-HBe—positive chronic hepatitis B with or without previous known HBe antigen (HBeAg) viremic phase, and to assess the potential implication of precore mutants in HBeAg—negative reactivation after loss of HBeAg. Nineteen patients were studied: 7 had a previous HBeAg-positive phase and had spontaneous or therapeutically induced loss of HBeAg (group A); 12 had no previous HBeAg-positive phase (group B). Direct sequencing of PCR products was performed on serum collected during the anti-HBe—positive phase in the two groups. In group A, precore sequencing showed that 5 patients were infected by wild-type virus, 1 patient was infected with a precore mutant, and 1 patient was found to be infected by a mixture of wild-type and precore mutant viruses. In group B, precore sequencing showed that only 1 patient was infected with wild-type virus and that 11 were infected with precore mutants. In a few patients, the presence of HBeAg within immune complexes may explain HBeAg negativity. In conclusion, our results show that, in patients with anti-HBe—positive chronic hepatitis B: (1) precore mutations creating a stop codon are more frequently found in those without known previous HBeAg positivity; (2) after loss of HBeAg, the patients who have anti-HBe—positive reactivation are infected by wild-type virus, which suggests that reactivation is not related to precore mutations; (3) HBeAg negativity may be caused by immune complexes formation.
Persistent hepatitis C viremia after acute self-limiting posttransfusion hepatitis C
Authors: Barrera Josep M. , Bruguera Miguel , Ercilla Guadalupe M. , Gil Cristina , Celis Raquel , Gil Pilar M. , Del Onorato Valle M. , Rodés Joan , Ordinas Antoni
Published: March 2007

Persistent viremia after clinical or subclinical hepatitis C virus (HCV) infection is believed to occur in patients with chronic hepatitis C, but little is known about the duration of HCV replication in patients with acute hepatitis who have recovered or the relation of HCV viremia with the kinetics of antibodies to HCV (anti-HCV). We tested HCV-RNA and anti-HCV in serial serum samples from 41 patients with posttransfusion non-A, non-B hepatitis, followed for an average of 6 years after transfusion. Serum HCV-RNA was measured by nested polymerase chain reaction, which used primers from the 5′ untranslated region of the HCV genome. Anti-HCV were tested with first- and second-generation enzyme-linked immunosorbent assays (ELISA 1 and ELISA 2), and with a second-generation recombinant immunoblot assay. Of the 41 patients, 10 recovered and 31 progressed to chronic liver disease. HCV-RNA was detected in serum before or simultaneously with the onset of hepatitis in all cases, and lasted between 2 and 6 weeks in 5 of the 10 patients who recovered, whereas it persisted for the entire follow-up period in every case with chronic hepatitis and in the remaining 5 patients with self-limiting hepatitis. Anti-HCV were detected with ELISA 2 in the first serum sample, with raised serum transaminases in 57% of patients, but in only 6% with ELISA 1. In the sample obtained 1 month after the onset of hepatitis, anti-HCV were detected with ELISA 2 in 94% of patients, but in 34% with the ELISA 1. Anti-HCV (anti C-33 and anti-c22) were cleared in the five patients with transient hepatitis C viremia, but remained detectable in those with chronic viremia. In conclusion, serum HCV-RNA is detected at the onset of acute posttransfusion hepatitis C and persists in patients progressing to chronic hepatitis. Some patients with self-limiting hepatitis become HCV-RNA negative soon after the onset of hepatitis, whereas in others it persists throughout follow-up, suggesting the development of a silent carrier state.
The natural history of gallstones: The GREPCO experience
Authors: Attili Adolfo Francesco , de Santis Adriano , Capri Rita , Repice Anna Maria , Maselli Silvana
Published: March 2007

During the cross-sectional studies (February 1981 to July 1984) performed by the Group for Epidemiology and Prevention of Cholelithiasis (GREPCO) in Rome, Italy, 161 subjects were identified as having gallstones. Ten subjects did not participate in the prospective follow-up. At entry, 33 of the 151 remaining subjects were symptomatic, and 118 were asymptomatic. Data on incidence of biliary colics, complications, cholecystectomy, and death were collected at least every 2 years. In the initially asymptomatic group, the cumulative probability (% ± SE) of developing biliary colic was 11.9 ± 3.0 at 2 years, 16.5 ± 3.5 at 4 years, and 25.8 ± 4.6 at 10 years. None of the variables considered as possible modifiers of the natural history were found to be associated with an increased risk of developing biliary colic. The cumulative probability (% ± SE) of developing complications after 10 years was 3.0 ± 1.8 in the initially asymptomatic group and 6.5 ± 4.4 in the symptomatic group (P = NS). Incidence of cholecystectomy was higher in the initially symptomatic than in the asymptomatic group (log-rank test = 2.27; P = .02). Fifteen (53.6%) of the 28 operated in the initially asymptomatic group were submitted to cholecystectomy, although specific symptoms did not occur. Twelve (10.2%) and 2 (6.1%) of the initially asymptomatic and symptomatic subjects died during the follow-up. One patient in the former group died at age 64 of a histologically proven adenocarcinoma of the gallbladder. In conclusion, this study demonstrates that the natural history of gallstones is less benign than is generally considered.
Endoscopic therapy of sclerosing cholangitis†
Authors: Lee John G. , Schutz Stephen M. , England Ruth E. , Leung Joseph W. , Cotton Peter B.
Published: March 2007

The efficacy of endoscopic treatment in primary sclerosing cholangitis has not been clearly established. This report presents endoscopic intervention in 53 consecutive patients with this disorder. Pertinent data were abstracted from the GI-TRAC database, medical records, and cholangiograms, and clinical follow-up was obtained by telephone interview of the subjects. Assessed treatment outcomes were clinical symptom, liver function test, and cholangiographic appearance. Between 1986 and 1993, 85 patients with primary sclerosing cholangitis underwent successful ERCP, of which 36 men and 17 women underwent 100 therapeutic endoscopic procedures. Forty-three of 50 dilations, 37 of 38 stentings, 8 of 8 nasobiliary tube placements, and 11 of 17 stone extractions were technically successful. These treatments were complicated by cholangitis or pancreatitis in 15 patients. Clinical follow-up was obtained in 50 of 53 patients who had undergone 85 procedures (median follow-up of 31 months): 28 patients felt better, 21 felt the same, and 1 felt worse. Liver function tests obtained within 3 months of the endoscopic treatment were significantly improved compared with pretreatment values (P < .001). Cholangiograms showed improvement in 36% of the patients, no change in 51%, and the effect of therapy could not be assessed in 13%. Overall, 41 of 53 patients (77%) had improvements of their clinical symptoms, liver function tests, or cholangiograms.
Growth hormone, insulinlike growth factor-1, and insulinlike growth factor binding proteins 1 and 3 in chronic liver disease
Authors: Donaghy Anthony , Ross Richard , Gimson Alexander , Hughes Sian Cwyfan , Holly Jeffrey , Williams Roger
Published: March 2007

The liver is the major source of circulating insulinlike growth factor-I (IGF-I) and has been suggested as a major source of at least two of the major binding proteins that modify its bioavailability. We aimed to assess the direct effects of liver dysfunction on serum levels of IGF-1 and its major binding proteins by measuring fasting levels of growth hormone, IGF-1, IGFBP-1, IGFBP-3, insulin, C peptide, and glucose in 35 patients with cirrhosis and during an oral glucose tolerance test in 16 of those patients. Serum levels of growth hormone (GH) were high in the patients: median, 12.0 mU/L (range, 1 to 87) compared with normals, 0.95 mU/L (0.1 to 20) (P < .0005) and serum IGF-1 levels were low: 81 ng/mL (38 to 153) versus 193 ng/mL (151 to 235) (P < .0001). Serum IGFBP-3 levels were low in the patients: 1.59 mg/L (0.46 to 4.43) compared with normals, 5.41 (4.34 to 6.11) (P < .0001), and there was a significant negative correlation between IGFBP-3 levels and Childs Pugh score (r = .63 P < .0001). Fasting IGFBP-1 levels were significantly higher in the patients 31 ng/mL (11 to 92) than normals, 14 (7 to 20) (P < .0001). There was no correlation between fasting insulin and IGFBP-1 levels despite high fasting insulin levels. A decrease in IGFBP-1 levels was seen during the glucose tolerance test (GTT) in all patients. In conclusion, there are significant changes in the levels of two of the major IGF-1 binding proteins that may further limit the bioavailability of already low circulating IGF-1 levels. Substrate availability appears to be a stronger influence on fasting IGFBP-1 levels than does insulin, and the close correlation of IGFBP-3 with liver function indicates a dominant regulatory role of the hepatocyte.
Extramedullary erythropoiesis in human liver grafts
Authors: Schlitt Hans Jürgen , Schäfers Sabine , Deiwick Andrea , Eckardt Kai-Uwe , Pietsch Torsten , Ebell Wolfram , Nashan Björn , Ringe Burckhardt , Wonigeit Kurt , Pichlmayr Rudolf
Published: March 2007

Extramedullary erythropoiesis in the adult is very rare and is generally confined to situations of severe bone marrow irritation or replacement. In this study, we describe the occurrence of intrahepatic erythropoiesis in patients who have received a liver allograft and who have no evidence of bone marrow dysfunction. By routinely performed transplant aspiration cytology (TAC), marked intrahepatic erythropoiesis could be detected in 39 of 312 patients (12.5%) with liver allograft. In 19 patients, including 5 of 8 (63%) after combined liver and kidney transplantation, intrahepatic erythropoiesis occurred within the first 3 weeks after surgery. Twenty patients showed intrahepatic erythropoiesis between 3 weeks and 4 months after transplantation. Erythropoiesis was usually transient, lasting between 1 and 3 weeks. Cytologically, mature as well as immature erythroblasts of GlyA+ CD36+ CD45− phenotype could be detected in the grafts, whereas they were absent in blood; histologically, the cells could be localized to the sinusoids of the liver. There was no clear correlation of preoperative or postoperative hemoglobin levels, graft function, kidney function, and immunosuppressive medication with the presence or absence of erythropoiesis. Moreover, serum levels of erythropoietin (EPO) and stem cell factor (SCF) in patients with and without intrahepatic erythropoiesis in the early postoperative phase did not show significant differences. These findings show that intrahepatic erythropoiesis can occur transiently in human liver allografts and suggest that systemic stimuli as well as local factors may contribute to it.
Reovirus 3 not detected by reverse transcriptase—mediated polymerase chain reaction analysis of preserved tissue from infants with cholestatic liver disease
Authors: Steele Marilyn I. , Marshall Candaca M. , Lloyd Richard E. , Randolph Vicki E.
Published: March 2007

Reovirus type 3 has been implicated in the origin and pathogenesis of extrahepatic biliary atresia and idiopathic neonatal hepatitis, but routine detection of this virus in hepatobiliary tissues from affected infants by culture and histological techniques has been unsuccessful. In this study, oligonucleotide primers specific to the M3 genome segment of reovirus 3 (Dearing) were used in a reverse transcriptase-mediated polymerase chain reaction technique to develop a sensitive and specific assay for the detection of reovirus 3 RNA in formalin-fixed, paraffin-embedded patient samples. Optimal reaction conditions were determined by testing infected murine tissues and preserved human liver tissue supplemented with reovirus 3. Archival specimens from 50 infants, including 14 with extrahepatic biliary atresia, 20 with idiopathic neonatal hepatitis, and 16 age-matched controls, were evaluated. Successful amplification of human albumin complementary DNA from the preserved tissues confirmed the presence of intact RNA in every patient specimen tested. Analysis of the amplification reactions by agarose gel electrophoresis and Southern blot hybridization detected the presence of reoviral RNA only once in a single patient sample. These results do not support a strong role for reovirus 3 in the development of neonatal cholestatic liver disease. The recent association of other RNA viruses of the Reoviridae family with murine liver disease and human extrahepatic biliary atresia indicates that continued investigation into a viral cause for idiopathic neonatal hepatobiliary disease is warranted.
Macroregenerative nodules in a series of adult cirrhotic liver explants: Issues of classification and nomenclature†
Authors: Hytiroglou Prodromos , Theise Neil D. , Schwartz Myron , Mor Eytan , Miller Charles , Thung Swan N.
Published: March 2007

Macroregenerative nodules (MRNs), probably representing a pathway for human hepatocarcinogenesis, are generally classified into type I MRNs (or ordinary adenomatous hyperplasia) and type II MRNs (or atypical adenomatous hyperplasia), on the basis of imprecise definitions of cytological and architectural atypia. It is currently believed that type II MRNs are probably true precursors of hepatocellular carcinoma (HCC), whereas type I lesions may simply represent large regenerative nodules. A series of 155 consecutive adult cirrhotic liver explants were examined for evidence of MRNs, HCC, and liver cell dysplasia (LCD) of large and small cell types, and their appearance, in terms of proposed classification schemes, was reviewed. There was evidence indicating that the presence of either type of MRN was associated with an increased incidence of HCC (all MRNs, P < .00019; type I MRNs, P < .067; type II MRNs, P < .012) compared with cirrhotic livers without MRNs. A subset of younger patients with a large (uncountable) number of MRNs in their livers, who did not show any increased incidence of carcinoma, was identified. Excluding these cases from statistical analysis, all associations were strengthened, implying either that malignant progression had not had time to occur in this younger population or that these nodules were simply large regenerative nodules without malignant potential. MRNs from these livers were histologically indistinguishable from MRNs occurring in more limited numbers, although atypical changes other than large cell type LCD were less frequent. No independent association between LCD of large cell type and HCC was found in the entire series. Deleting this feature from the criteria for cytological atypia resulted in a stronger association of both types of MRNs with HCC (redefined type II MRNs/HCC, P < .0001; redefined type I MRNs/HCC, P < .0306). Some of the type II MRNs remaining after exclusion of large cell type LCD showed “borderline” changes insufficient for a diagnosis of HCC, but most type II MRNs (82%) contained expansile “nodule-in-nodule” growth patterns. The conclusions of this report are that (1) histological examination of type I MRNs is insufficient in many cases to distinguish large regenerative nodules from neoplastic ones; (2) LCD of large cell type should not be used as a criterion for terming an MRN atypical; and (3) expansile “nodule-in-nodule” formation in MRNs should be considered to represent evidence of architectural atypia.
Systemic hemodynamic, forearm vascular, renal, and humoral responses to sustained cardiopulmonary baroreceptor deactivation in well-compensated cirrhosis
Authors: Wong Florence , Logan Alexander , Blendis Laurence
Published: March 2007

The aim of this study was to assess baroreceptor function in well-compensated cirrhosis by determining the forearm vascular, renal, and humoral responses to sustained baroreceptor deactivation. The effect of sodium status on baroreceptor function was also assessed. Eight cirrhotic patients and 10 age- and sex-matched controls were studied twice after a 20 mmol and 200 mmol of sodium/d diet for 7 days. Systemic and renal hemodynamics, renal sodium handling, forearm blood flow, and neurohumoral factors were assessed before, during, and after the application of lower body negative pressure (LBNP) for 1 hour. Controls and cirrhotic patients had similar baseline mean arterial pressure, heart rate, forearm and renal hemodynamics. High-sodium intake resulted in suppression of sympathetic nervous activity in the controls (plasma norepinephrine, 1.06 ± 0.11 nmol/L on low vs. 0.76 ± 0.08 nmol/L on high sodium; P = 0.01) but not in the cirrhotic patients (1.35 ± 0.22 nmol/L on low vs. 1.26 ± 0.11 nmol/L on high sodium; P > 0.05). Both groups responded to LBNP with significant further increases in plasma norepinephrine, resulting in significant decreases in forearm blood flow on both sodium diets. Controls also responded with a significant worsening of renal hemodynamics on low-sodium diet only, but this was not observed in the cirrhotic patients on either diet. Therefore, in well-compensated cirrhotic patients: (1) sympathetic activation occurs despite an adequate, effective arterial filling, and this may contribute to sodium retention; and (2) baroreceptor function is normal. Apparent end organ unresponsiveness within the renal circulation may account for the lack of renal hemodynamic changes to reflex sympathetic stimulation.
Expression of the hepatitis C virus genome in rat liver after cationic liposome-mediated in vivo gene transfer
Authors: Takehara Tetsuo , Hayashi Norio , Miyamoto Yasuhide , Yamamoto Masato , Mita Eiji , Fusamoto Hideyuki , Kamada Takenobu
Published: March 2007

The lack of a small animal model of hepatitis C virus (HCV) infection has impeded elucidation of the pathogenesis of HCV. The aim of this study was to develop an HCV-expressing animal model by means of cationic liposome-mediated in vivo gene transfer. To examine the feasibility of this strategy, pActLacZ, an expression vector composed of the LacZ gene driven by the β-actin promoter, complexed with lipofectin, was injected retrogradely into the common bile ducts of adult rats. X-Gal histochemical staining clearly showed that the LacZ gene was expressed in hepatocytes, but not in biliary epithelial cells. Maximal expression was observed at a DNA to lipofectin ratio of 1:4. Based on this observation, pAGS3M091, an expression vector containing the full length of HCV complementary DNA (cDNA) preceded by the β-actin promoter, was evaluated. Two days after in vivo intrabiliary administration of pAGS3M091 complexed with lipofectin, polymerase chain reaction (PCR) amplification of reverse-transcribed liver RNA demonstrated the 5′ and 3′ portions of HCV transcripts derived from pAGS3M091. Immunohistochemical analysis showed the HCV core protein in a small number of hepatocytes scattered in the hepatic lobules. We conclude that the full-length HCV genome was successfully expressed in adult rat liver by means of cationic liposome-mediated in vivo gene transfer. This model will be useful for detemining the immunopathological role of HCV in vivo.
Extracorporeal high-intensity focused ultrasound for VX2 liver tumors in the rabbit
Authors: Prat Frédéric , Centarti Maria , Sibille Alain , El Fadil Fatima Abou , Henry Luc , Chapelon Jean-Yves , Cathignol Dominique
Published: March 2007

High-Intensity Focused Ultrasound (HIFU) can produce radical tissue necrosis. We wanted to assess tumor destruction, proliferation, and tumorigenesis after HIFU, in an animal model of hepatic tumor. New Zealand rabbits bearing VX-2 solitary liver tumors were treated with extracorporeal HIFU under ultrasound (US) guidance and standardized conditions. Groups differed only for the administration of either one or two consecutive HIFU procedures. Tissue destruction was assessed by steremicroscopy and planimetry, cell proliferation was estimated by in vivo intra-arterial injection of 1200 μCi [3H]thymidine, and tumorigenesis was tested by reimplantation of treated or untreated pieces of liver tumors into the thighs of nontumor-bearing animals. Mortality was 0. Tumor destruction rates were 76.3% ± 16% after one procedure and 94.2% ± 7.3% after two procedures. Nuclear staining was heavy in control tumors and was absent in treated tumors. Untreated hepatic tumors induced measurable tumors at 3 weeks in thighs of all recipients, 7.8 ± 2.4 cm3 in volume. Hepatic tumors treated with on HIFU procedure induced tumors in the thigh of recipients in 31.3% of cases (0.47 ± 0.06 cm3), and those treated with one HIFU procedure induced tumors in 0% even after 8 weeks of follow-up. In conclusion, HIFU allows a noninvasive approach to the destruction of liver tumors in this model, with little toxicity but significant effects on proliferation and tumorigenesis. The repetition of HIFU procedures may improve results.
Conditional immortalization of gunn rat hepatocytes: An ex vivo model for evaluating methods for bilirubin-UDP-glucuronosyltransferase gene transfer
Authors: Fox Ira J. , Chowdhury Namita Roy , Gupta Sanjeev , Kondapalli Ravi , Schilsky Michael L. , Stockert Richard J. , Chowdhury Jayanta Roy
Published: March 2007

Viral vectors and protein carriers utilizing asialoglycoprotein receptor (ASGR)-mediated endocytosis are being developed to transfer genes for the correction of bilirubin-UDP-glucuronosyltransferase (bilirubin-UGT) deficiency. Ex vivo evaluation of these gene transfer vectors would be facilitated by a cell system that lacks bilirubin-UGT, but expresses differentiated liver functions, including ASGR. We immortalized primary Gunn rat hepatocytes by transduction with a recombinant Moloney murine leukemia virus expressing a thermolabile mutant SV40 large T antigen (tsA58). At 33° C, the immortalized hepatocyte clones expressed SV40 large T antigen, synthesized DNA, and doubled in number every 2 to 3 days. At this temperature, differentiated hepatocyte markers, e.g., albumin, ASGR, and androsterone-UGT, were expressed at 5% to 10% of the levels found in primary hepatocytes maintained in culture for 24 hours. Glutathione-S-transferase Yp (GST-Yp), an oncofetal protein, was expressed in these cells at 33° C, but was undetectable in primary hepatocytes. In contrast, when the cells were cultured at 39° C or 37° C, the large T antigen was degraded, DNA synthesis and cell growth stopped, and morphologic characteristics of differentiated hepatocytes were observed. The expression of albumin, ASGR, and androsterone-UGT, and their corresponding mRNAs, increased to 25% to 40% of the level in primary hepatocytes, whereas GST-Yp expression decreased. Functionality of ASGR was demonstrated by internalization of Texas red-labeled asialoorosomucoid, and binding and degradation of 125I-asialoorosomucoid. After liposome-mediated transfer of a plasmid containing the coding region of human bilirubin-UGT1, driven by the SV40 large T promoter, active human bilirubin-UGT1 was expressed in these cells. The immortalized cells were not tumorigenic after transplantation into severe combined immunodeficiency mice. These conditionally immortalized cells will be useful ex vivo evaluation of builirubin-UGT gene transfer vectors.
Lipoproteins and the liver sieve: The role of the fenestrated sinusoidal endothelium in lipoprotein metabolism, atherosclerosis, and cirrhosis
Authors: Fraser Robin , Dobbs Bruce R. , Rogers George W. T.
Published: March 2007

The liver sieve, formed by the fenestrated hepatic sinusoidal endothelium, is a dynamic biofilter separating the hepatic blood from the plasma within the space of Disse. It filters macromolecules of differing sizes, especially lipoproteins. More specifically, it acts as a barrier to the large triglyceride-rich parent chylomicrons, while permitting the smaller triglyceride-depleted but cholesterol- and retinol-rich remnants to enter the space of Disse. There the remnants contact specific receptor sites on the hepatocyte microvilli. Thus, the liver sieve is the first site of hepatic selection and consequent metabolism of dietary cholesterol and fat-soluble vitamins, as well as rejection of dietary triglycerides. Therefore, perturbations of the porosity of the sieve, whether from changes in size, number of fenestrae, or composition of the underlying extracellular matrix within the space of Disse, will have a profound influence on the metabolism of lipoproteins. This disturbance of the homeostasis of lipids, including fat-soluble vitamins and cholesterol, as well as other macromolecules, may tilt the balance between health and disease in a variety of organs and tissues, such as the liver, kidney and arteries.
Is hepatitis B virus smarter than the immune system?
Authors: Peters Marion
Published: March 2007

It has been suggested that mutations within immunodominant cytotoxic T-lymphocyte (CTL) epitopes may be exploited by viruses to evade protective immune responses critical for clearance.1-4 Viral escape could originate from passive mechanisms, such as mutations within curcial CTL epitopes, either affecting major histocompatibility complex binding or T-cell antigen receptor (TCR) recognition. Additionally, it has recently been shown that substitutions of TCR contact sites can yield analogue peptides that can still interact with the T-cell receptor but be unable to deliver a full stimulatory signal, thus inducing anergy5 or acting as an antagonist for the TCR.6-8 We report here that hepatitis B virus isolates derived from two chronically infected patients display variant epitopes that act as natural TCR antagonists with the capacity to inhibit the CTL response to the wild-type epitope. During natural infection, TCR antagonist mutations of CTL epitopes could contribute to the development of viral persistence, especially if the antiviral CTL response is monospecific or the epitope is strongly immunodominant.
Hepatitis B virus escape mutants: “pushing the envelope” of chronic hepatitis B virus infection
Authors: Mimms Larry
Published: March 2007

Hepatitis B virus (HBV) DNA was extracted from sera of six carriers with hepatitis B e antigen as well as antibody to hepatitis B surface antigen and sequenced within the pre-S regions and the S gene. HBV DNA clones from five of these carriers had point mutations in the S gene, resulting in conversion from IIe-126 or Thr-126 of the wild-type virus to Ser-126 or Asn-126 in three carriers and conversion from Gly-145 to Arg-145 in three of them; clones with Asn-126 or Arg-145 were found in one carrier. All 12 clones from the other carrier had an insertion of 24 bp encoding an additional eight amino acids between Thr-123 and Cys-124. In addition, all or at least some of the HBV DNA clones from these carriers had inphase deletions in the 5′ terminus of the pre-S2 region. These results indicate that HBV escape mutants with mutations in the S gene affecting the expression of group-specific determinants would survive in some carriers after they seroconvert to antibody against surface antigen. Carriers with HBV escape mutants may transmit HBV either by donation of blood units without detectable surface antigen or through community-acquired infection, which would hardly be prevented by current hepatitis B immunoglobulin or vaccines.
Simultaneous detection of both hepatitis B virus DNA and hepatitis C virus RNA using a combined one-step polymerase chain reaction technique
Authors: Hu Ke-Qin , Yu Chang-Hong , Lee Sunny , Villamil Federico G. , Vierling John M.
Published: March 2007

Hepatitis C virus (HCV) RNA polymerase chain reaction (PCR) is widely used for diagnosis of HCV infection and evaluation of therapy. The sensitive hepatitis B virus (HBV) DNA PCR is often reserved for detection of quantities of HBV DNA that are insufficient for hybridization. Application of both PCR techniques is limited by their labor-intensity, potential for contamination, and substantial time required for analysis. To study HCV and HBV infections, occurring alone or in combination, we developed a combined one-step PCR method to detect HCV RNA and HBV DNA in a single serum specimen using oligoprimers from the HCV 5′ untranslated region and the HBV preS/S region. Specificity of the HBV and HCV PCR products was confirmed on the basis of their molecular sizes in positive samples, Southern blot hybridization, and negative controls. The sensitivities of the combined PCR were assessed using samples containing a wide range of defined amounts of HBV DNA and HCV RNA and were comparable with those obtained with conventional HBV DNA or HCV RNA PCR methods. The sensitivity of the combined method was further validated by the 100% concordance between results of its HBV and HCV components and those of conventional PCR methods in patients with HBV and/or HCV infections. The combined one-step HBV/HCV PCR is a sensitive, specific, rapid, and cost-effective method, especially suited for epidemiological screening and clinical diagnosis of HBV and HCV infections occurring alone or in combination. (Hepatology 1995; 21:901-907.)
Two-year biochemical, virological, and histological follow-up in patients with chronic hepatitis C responding in a sustained fashion to interferon alfa-2b treatment
Authors: Reichard Olle , Glaumann Hans , Frydén Aril , Norkrans Gunnar , Schvarcz Robert , Sönnerborg Anders , Yun Zhi-Bing , Weiland Ola
Published: March 2007

Fourteen patients with chronic hepatitis C who had a sustained response to a 60-week interferon alfa-2b treatment course were followed, biochemically and virologically, 2 years after treatment cessation. Biopsies were repeated in 12 of 14 for histological and virological evaluation at 2-year follow-up. All 14 patients had normal serum alanine transaminase (s-ALT) levels and were negative for hepatitis C virus (HCV) RNA in serum during treatment and at short-term follow-up 6 months posttreatment. At 2-year follow-up, 13 patients still had normal ALT levels (<0.6 μkat/L for women; <0.8 μkat/L for men), 1 a near normal level (0.76 μkat/L); all were HCV RNA negative in serum, and 11 of 12 also in the liver. Liver histology improved during treatment and remained stable during the 2-year follow-up. The authors conclude that most sustained responders, who have normal ALT levels and are nonviremic at short-term follow-up 6 months after interferon treatment, will continue to have a durable long-term response without relapse of the viremia. (Hepatology 1995; 21:918-922.)
Utilization of tyrosine-containing dipeptides and N-acetyl-tyrosine in hepatic failure
Authors: Druml Wilfred , Hübl Wolfgang , Roth Erich , Lochs Herbert
Published: March 2007

The impact of hepatic dysfunction on the elimination and hydrolysis of three potential tyrosine sources for total parenteral nutrition, the dipeptides L-alanyl-L-tyrosine (Ala-Tyr) and glycyl-L-tyrosine (Gly-Tyr), and N-acetyl-L-tyrosine (Nac-Tyr) were evaluated in six patients with hepatic failure (five chronic, one acute) and seven healthy subjects. In controls, whole-body clearance (Cltot) of Ala-Tyr was higher than of Gly-Tyr (3,169 ± 214 vs. 1,780 ± 199 mL/kg/min, P < .01), and both exceeded clearance of Nac-Tyr (309 ± 29 mL/kg/min, P > .01). Both dipeptides were hydrolyzed and released tyrosine immediately. In hepatic failure, elimination and hydrolysis of Ala-Tyr and Gly-Tyr were comparable to controls, but Cltot of Nac-Tyr was reduced (236 ± 26 mL/kg/min). Neither in controls nor in patients an increase in plasma tyrosine concentration was seen after Nac-Tyr, and the major part of Nac-Tyr infused was lost in urine. The Cltot of tyrosine as evaluated after Ala-Tyr infusion (with the immediate release of tyrosine) was severely reduced in hepatic failure (152.7 ± 38.4 vs. 484.4 ± 41.4 mL/kg/min, P < .001) and half-life (kle) was retarded from 14.4 ± 1.4 to 90.2 ± 32.2 minutes (P > .03). The authors conclude that acute and chronic hepatic dysfunction does not affect elimination and hydrolysis of the dipeptides Ala-Tyr and Gly-Tyr and the constituent amino acids are released immediately. Nac-Tyr elimination was not grossly affected by hepatic failure, but neither in healthy subjects nor in hepatic failure patients was an increase of tyrosine seen. Both dipeptides but not Nac-Tyr may serve as a tyrosine source in parenteral nutrition. Moreover, by its rapid hydrolysis, the use of Ala-Tyr, for the first time, enables a simple rapid nonisotope evalution of tyrosine kinetics for assessement of liver function. (Hepatology 1995; 21:923-928.)
Insulin secretion and plasma levels of glucose-dependent insulinotropic peptide and glucagon-like peptide 1 [7-36 amide] after oral glucose in cirrhosis
Authors: Kruszynska Yolanta T. , Ghatei Mohammad A. , Bloom Stephen R. , McIntyre Neil
Published: March 2007

A blunted initial insulin secretory response may contribute to oral glucose intolerance in cirrhosis. Oral glucose is a better stimulant to insulin secretion than intravenous (IV) glucose in part because of release of gut peptides, notably glucose-dependent insulinotropic peptide (GIP) and glucagon-like peptide 1 [7-36 amide] (GLP-1 [7-36 amide]). Because impaired release of or resistance to these gut peptides could explain impaired insulin secretion after oral glucose, we measured insulin secretion, plasma GIP, and GLP-1 [7-36 amide] levels, basally and after 75 g oral glucose, in 10 cirrhotics and 10 controls. Insulin secretion was calculated from a two-compartment analysis of serum C-peptide levels using kinetic parameters derived from IV injection of recombinant human C-peptide. C-peptide metabolic clearance rate, and the fractional rate constants for C-peptide (using the two-compartment model) were not significantly different, but the volume of the central compartment was 15% greater in cirrhotics (P < .01). Fasting blood glucose levels were similar (cirrhotics, 4.9 ± 0.2; controls, 4.6 ± 0.1 mmol/L) but serum insulin was six times higher in cirrhotics (P < .001). Cirrhotics had higher fasting GIP (215 ± 72 vs. 42 ± 18 pmol/L) and GLP-1 [7-36 amide] levels (25 ± 3 vs. 16 ± 1 pmol/L) (both P < .05). After oral glucose, blood glucose levels were significantly higher in cirrhotics. The timing of the gut peptide response to oral glucose was similar in the two groups, but peak levels of both peptides were ˜ × 2 higher in the cirrhotics. For the 1st 30 minutes, C-peptide levels and insulin secretion were similar in the two groups. In controls, insulin secretion peaked at 20 to 25 minutes and coincided with the GLP-1 [7-36 amide] peak; insulin secretion then decreased despite a persistent elevation in plasma GIP and glucose levels. In cirrhotics, peak insulin secretion was delayed until 50 to 55 minutes (coincident with the glucose peak). Total insulin secretion during the 4 hours after glucose ingestion was 55% higher in the cirrhotics, but peak insulin secretion rates were similar despite higher gut peptide and glucose levels in cirrhotics. The study shows that, after oral glucose, cirrhotics have a blunted serum C-peptide response and insulin secretion rate. This is not attributable to reduced levels of GIP or GLP-1 [7-36 amide] but could be attributable to resistance to their action.(Hepatology 1995; 21:933-941.)
HLA DPB polymorphism in primary sclerosing cholangitis and primary biliary cirrhosis
Authors: Underhill James A. , Donaldson Peter T. , Doherty Derek G. , Manabe Koji , Williams Roger
Published: March 2007

In recent years there has been an increasing interest in the link between susceptibility to autoimmune liver disease and genes of the HLA system, although the role of the DPB1 locus in British patients has only been investigated in autoimmune hepatitis. The aim of the current study was to determine the distribution of DPB1 alleles in a large series of British patients with the two other autoimmune liver diseases, primary biliary cirrhosis and primary sclerosing cholangitis, and compare the allele frequencies obtained with those of a geographically matched control group. Polymerase chain reaction sequence-specific oligonucleotide probing was used to assign 18 DPB1 alleles in 82 patients with primary biliary cirrhosis (PBC), 71 patients with primary sclerosing cholangitis (PSC), and 103 controls. The frequencies of the DPB1 alleles were not significantly different comparing patients and controls. However, two important observations were made. Firstly, in primary sclerosing cholangitis, the previously reported association with the haplotype A1-B8-DR3-DQ2 does not extend to the DPB1 locus, suggesting that the genetic determinants of susceptibility for this disease lie closer to the DRB loci. Secondly, in primary biliary cirrhosis there is evidence that the reported association with DR8-DQB1*0402 includes the DPB1*0301 allele. The weak HLA association reported here is in contrast with recent data from Japan, where susceptibility is strongly linked to a particular amino acid residue encoded by the DPB1*0501 allele. These data clearly demonstrate that the alleles of the DPB1 locus are not associated with susceptibility to or protection from either primary biliary cirrhosis or primary sclerosing cholangitis in British patients. (Hepatology 1995; 21:959-962.)
Plasma calprotectin: A new prognostic marker of survival in alcohol-induced cirrhosis
Authors: Homann Christian , Garred Peter , Graudal Niels , Hasselqvist Philip , Christiansen Michael , Fagerhol Magne K. , Thomsen Åge Christian
Published: March 2007

Plasma levels of calprotectin were determined in 84 patients with alcohol-induced cirrhosis. Calprotectin is released from disintegrating neutrophils, and plasma levels seem to reflect activation and turnover of such cells. The purpose of the study was to investigate the degree of activation of neutrophils, which has been indicated to be increased and a cause of neutrophil exhaustion in these patients. Additionally, on follow-up after a median observation period of 559 days, we investigated the prognostic value of calprotectin for survival. No difference was found in calprotectin levels when comparing healthy controls with patients with compensated cirrhosis and those with decompensated cirrhosis. However, high calprotectin concentrations (>median) were a significant prognostic marker of poor survival (P = .001, log-rank test). Using a multivariate Cox proportional hazard model, the prognostic value of calprotectin seemed independent of severity of liver disease evaluated on eight clinical and biochemical variables of liver disease. Divided into groups by the median calprotectin concentration, analysis of survival was performed in the whole series of patients (n = 84) as well as in patients who were completely without signs of recent or actual infection (n = 54). In both groups, calprotectin levels (>median) showed a much higher prognostic value than albumin, prothrombin complex, bilirubin, and ascites. During follow-up, calprotectin levels (>median) were also a predictor of recurrent infection (P = .009). Thus, in patients with alcoholic cirrhosis, plasma calprotectin seems to be a new prognostic marker of survival, which seems independent of the severity of liver disease. Furthermore, high plasma calprotectin levels may characterize a group of patients with cirrhosis with recurring bacterial infections. (Hepatology 1995; 21:979-985.)
Treatment of bleeding from portal hypertensive gastropathy by portacaval shunt
Authors: Orloff Marshall J. , Orloff Mark S. , Orloff Susan L. , Haynes Kevin S.
Published: March 2007

Portal hypertensive gastropathy is a vascular disorder of the gastric mucosa distinguished by ectasia of the mucosal capillaries and submucosal veins without inflammation. During 1988 to 1993, 12 patients with biopsyproven cirrhosis (10 alcoholic, 2 posthepatitic) were evaluated and treated prospectively by portacaval shunt for active bleeding from severe portal hypertensive gastropathy. Eleven patients had been hospitalized for bleeding three to nine times previously, and one was bleeding uncontrollably for the first time. Requirement for blood transfusions ranged from 11 to 39 units cumulatively, of which 8 to 30 units were required specifically to replace blood lost from portal hypertensive gastropathy. Admission findings were ascites in 9 patients, jaundice in 8, severe muscle wasting in 10, hyperdynamic state in 9. Child's risk class was C in 7, B in 4, A in 1. Ten of the 12 patients had previously received repetitive endoscopic sclerotherapy for esophageal varices, which has been reported to precipitate portal hypertensive gastropathy. Eight patients had failed propranolol therapy for bleeding. Portacaval shunt was performed emergently in 11 patients and electively in 1, and permanently stopped bleeding in all by reducing the mean portal vein-inferior vena cava pressure gradient from 251 to 16 mm saline. There were no operative deaths, and two unrelated late deaths after 13 and 24 months. During 1 to 6.75 years of follow-up, all shunts remained patent by ultrasonography, the gastric mucosa reverted to normal on serial endoscopy, and there was no gastrointestinal bleeding. Recurrent portal-systemic encephalopathy developed in only 8% of patients. Quality of life was generally good. It is concluded that portacaval shunt provides definitive treatment of bleeding portal hypertensive gastropathy by eliminating the underlying cause, and makes possible prolonged survival with an acceptable quality of life. (Hepatology 1995; 21:1011-1017.)
Translocation of different forms of transferrin from blood to bile in the rat
Authors: Regoeczi Erwin , Chindemi Paul A.
Published: March 2007

Five different forms of transferrin (rat apo [iron-free], rat diferric, diferric rat asialo, human diferric, and diferric human asialotransferrin type 3) were used to monitor the passage of this protein and its metal to the bile. Cumulative biliary excretion of the dose over 3 hours was determined. In addition, an excretion profile was constructed from the concentration of tracer in bile samples collected over 10-minute intervals. The profile obtained with apotransferrin was very similar to that found in an earlier study with albumin, the implication being that the apo form is transferred passively (e.g., by diffusion). Behavior of rat diferric transferrin, however, was consistent with the assumption that this form is transferred both passively and actively (i.e., in vesicles). The three other transferrins were investigated with the intent of broadening the spectrum of ligand affinities for the plasmalemma of hepatocyte. The higher this attraction was, the larger fraction of the dose appeared in bile. When transferrin was targeted to lysosomes, the bile contained several intermediate proteolytic fragments. Double-labeled (125I, 59Fe) transferrin was used to measure recovery of iron (Fe) relative to the protein (P) in bile. With rat diferric transferrin, the Fe/P ratio was 0.72. Lower values were recorded with transferrins (human or asialo) that had higher affinities for the plasmalemma and therefore were expected to be transported to a larger extent in vesicles. Of the biliary 59Fe, 85% to 92% was protein bound. The proportion of the protein-bound fraction was essentially independent of the magnitude of Fe/P ratios. (Hepatology 1995; 21:1063-1069.)
Cell proliferation and oncogene expression after bile duct ligation in the rat: Evidence of a specific growth effect on bile duct cells
Authors: Polimeno Lorenzo , Azzarone Alessandro , Zeng Qui Hua , Panella Carmine , Subbotin Vladimir , Carr Brian , Bouzahzah Boumediene , Francavilla Antonio , Starzl Thomas E.
Published: March 2007

The proliferative response of the rat liver was measured after temporary or permanent total biliary obstruction (BDO) and in different regions after selective ligation of the lobar ducts draining the right 60% of the hepatic mass. The results were compared with those after 70% partial hepatectomy (PH). Cell proliferation was assessed globally by measuring DNA synthesis and stratified to the separate cell populations with cytostaining techniques that allowed distinction of hepatocytes, duct cells, and nonparenchymal cells (NPCs). In selected experimental groups, gene expression was determined of transforming growth factor-β1 (TGFβ-1), prothrombin, c-erb-B2, transforming growth factor alpha (TGFα), human Cyclophilin (CyP), and 28S ribosomal RNA. The stimulation of a proliferative response to total BDO required obstruction for longer than 24 hours, but after this deligation did not switch off regeneration. In the first week after permanent BDO, there was progressive infiltration of NPCs, fibrous linkage of some portal areas, and a crescendo of DNA synthesis that was obvious at 24 hours, maximal at 48 hours, and back nearly to baseline at 6 days. At the 2-day mark, the bile duct cells had a 17-fold increase in proliferation, accompanied by a threefold to fourfold increase in hepatocyte renewal. Litle or no increase in expression of TGFα or the hepatocyte-specific prothrombin gene was detectable in the first 48 hours, whereas levels of the oncogene c-erb-B2 that is associated with cholangiocarcinoma were expressed from 48 to 96 hours. Livers subjected to regional BDO with or without immunosuppressive treatment with FK 506 and cyclosporine had an inflammatory reaction only on the side with ligated ducts. DNA synthesis increased in both the obstructed and freely draining lobes to approximately half the level that occurred after total BDO. The proliferation of the obstructed side was similar to the mixed duct cell/hepatocyte response after total BDO, but this almost exclusively involved duct cells on the freely draining side. In contrast to the findings after BDO, livers after PH regenerated maximally at 24 hours rather than 48 hours, had a predominantly noninflammatory hepatocyte as opposed to duct cell response, and had marked expression of the prothrombin and TGFα genes but only weakly and late of c-erb-B2 messenger RNA. The results show that the liver responds as a whole and in a biologically intelligent way to the nature of the injury inflicted on any part of it. It further implies the presence of humoral communications and control networks that assure organ homeostasis and relate this to total body homeostasis. (Hepatology 1995; 21:1070-1078.)