bluuebunny/crossref_metadata_2025 huggingface dataset:
Risk of liver and other types of cancer in patients with cirrhosis: A nationwide cohort study in Denmark
Authors: Sørensen Henrik Toft , Friis Søren , Olsen Jørgen H. , Thulstrup Ane Marie , Mellemkjær Lene , Linet Martha , Trichopoulos Dimitrios , Vilstrup Hendrik , Olsen Jørn
Published: July 2004

Cancer risk in patients with cirrhosis could be modified by factors such as changes in hormonal levels, impaired metabolism of carcinogens, or alteration of immunological status. We investigated the risk of liver and various forms of cancer in patients with cirrhosis in a follow-up study. We identified 11,605 1-year survivors of cirrhosis from the files of the Danish National Registry of Patients (NRP) from 1977 to 1989. Occurrence of cancer through 1993 was determined by linkage to the Danish Cancer Registry. For comparison, the expected number of cancer cases was estimated from national age-, sex-, and site-specific incidence rates. Overall, 1,447 cancers were diagnosed among the study subjects, as compared with 708.1 expected, to yield a standardized incidence ratio (SIR) of 2.0 (95% CI: 1.9 to 2.2). In all diagnostic subgroups of cirrhosis, the risk of primary liver cancer, mainly hepatocellular carcinoma, was markedly elevated, with 245 observed cases and an overall 36-fold elevated risk (59.9-fold elevated for hepatocellular carcinoma and 10-fold for cholangiocarcinoma). Substantial and persistent excesses during follow-up were seen for all types of cancer associated with tobacco and alcohol habits (cancer of the lung, larynx, buccal cavity, pharynx, pancreas, urinary bladder, and kidney), while moderate excesses were seen for cancers of the colon and breast. The latter, however, were not complemented by any decrease in the risk of prostate cancer (SIR: 1.0; 95% CI: 0.7 to 1.3). A slightly increased risk was seen for testis cancer, but disappeared after 10 years. We found evidence of an increased risk for liver and several extrahepatic cancers in patients with cirrhosis. Although part of this increase is likely attributable to alcohol and tobacco consumption, our study opens up the possibility that cirrhosis plays a role in the carcinogenesis of types of cancer other than liver cancer.
Endothelial dysfunction and decreased production of nitric oxide in the intrahepatic microcirculation of cirrhotic rats
Authors: Gupta Tarun K. , Toruner Murat , Chung Moon K. , Groszmann Roberto J.
Published: July 2004

Increased intrahepatic resistance in cirrhotic livers is in part caused by increased vascular tone. Several morphological abnormalities have been described in the sinusoidal endothelial cells of cirrhotic livers, but the functional impact of these abnormalities on the intrahepatic vascular tone has not been studied. The aim of this study was to investigate the intrahepatic endothelial function and the role of nitric oxide (NO) with regard to vascular tone in cirrhotic livers. Isolated rat liver perfusions were performed in cirrhotic rats (induced by chronic carbon tetrachloride inhalation) and weight-matched normal controls. After preconstricting the intrahepatic microcirculation with methoxamine (10−4 mol/L), response to cumulative doses of receptor-mediated endothelial agonist, acetylcholine (10−7 mol/L-10−5 mol/L), was obtained. In another series, response to the receptor-independent endothelial agonist, calcium ionophore A23187 (10−7 mol/L and 3 × 10−7 mol/L), was obtained in the absence and presence of N
ω-nitro-l -arginine (NNA) and indomethacin. In a third series of rats, nitrate and nitrite production was measured in the perfusate of perfused normal and cirrhotic livers. There was significantly less vasorelaxation in cirrhotic livers as compared with normal livers in response to acetylcholine and calcium ionophore A23187 (P < .0001). The impaired vasorelaxation was a result of a decrease in both NO-mediated and non-NO-mediated components of vasorelaxation. Cirrhotic livers from ascitic rats had significantly less vasorelaxation as compared with livers from nonascitic rats (P < .005). There was significantly less production of nitrates and nitrites in cirrhotic livers (P < .05). The liver microcirculation of cirrhotic livers is characterized by endothelial dysfunction that results in impaired release of endothelial relaxing factors including NO.
Doppler study of mesenteric, hepatic, and portal circulation in alcoholic cirrhosis: Relationship between quantitative doppler measurements and the severity of portal hypertension and hepatic failure
Authors: Taourel Patrice , Blanc Pierre , Dauzat Michel , Chabre Myriam , Pradel Jean , Gallix Benoit , Larrey Dominique , Bruel Jean-Michel
Published: July 2004

To determine the relationship between quantitative Doppler parameters of portal, hepatic, and splanchnic circulation and hepatic venous pressure gradient (HVPG), variceal size, and Child-Pugh class in patients with alcoholic cirrhosis, we studied forty patients with proved alcoholic cirrhosis who underwent Doppler ultrasonography, hepatic vein catheterization, and esophagoscopy. The following Doppler parameters were recorded: time-averaged mean blood velocity, volume flow of the main portal vein flow, and resistance index (RI) of the hepatic and of the superior mesenteric artery. Doppler findings were compared with HVPG, presence and size of esophageal varices, and Child-Pugh class. There was a significant inverse correlation between portal velocity and HVPG (r = −.69), as well as between portal vein flow and HVPG (r = −.58). No correlation was found between RI in the hepatic artery or superior mesenteric artery and HVPG. No correlation was found between portal vein measurements and presence and size of varices. Severe liver failure was associated with lower portal velocity and flow. In patients with alcoholic cirrhosis, only portal vein blood velocity and flow, but neither hepatic nor mesenteric artery RI, are correlated to the severity of portal hypertension and to the severity of liver failure.
Gene expression of α1-6 fucosyltransferase in human hepatoma tissues: A possible implication for increased fucosylation of α-fetoprotein
Authors: Noda Katsuhisa , Miyoshi Eiji , Uozumi Naofumi , Yanagidani Shusaku , Ikeda Yoshitaka , Gao Cong-xiao , Suzuki Kunio , Yoshihara Harumasa , Yoshikawa Masumi , Kawano Kiyoshi , Hayashi Norio , Hori Masatsugu , Taniguchi Naoyuki
Published: July 2004

The α1-6 fucosylated α-fetoprotein (AFP) present in serum of patients with hepatocellular carcinoma (HCC) has been employed for the differential clinical diagnosis of HCC from chronic liver diseases. The molecular mechanism by which this alteration occurs, however, remains largely unknown. To address this issue, we purified GDP-l -Fuc:N-acetyl-β-d-glucosaminide α1-6 fucosyltransferase (α1-6 FucT), an enzyme involved in the α1-6 fucosylation of N-glycans from porcine brain, as well as from a human gastric cancer cell line, and cloned their genes. In this study, levels of α1-6 FucT mRNA expression and the activity of this enzyme for 12 human HCC tissues were examined and compared with that in surrounding tissues and normal livers. The mean ± SD for α1-6 FucT activity was 78 ± 41 pmol/h/mg in normal control liver, 202 ± 127 pmol/h/mg in adjacent uninvolved liver tissues (chronic hepatitis: 181 ± 106 pmol/h/mg; liver cirrhosis: 233 ± 164 pmol/h/mg), and 195 ± 72 pmol/h/mg in HCC tissues. The mRNA expression of α1-6 FucT was also enhanced in proportion to enzymatic activity except for a few cases, suggesting that α1-6 FucT expression is increased in chronic liver diseases, especially liver cirrhosis. Transfection of α1-6 FucT gene into cultured rat hepatocytes markedly increased α1-6 FucT activity and led to an increase in lens culinaris agglutinin (LCA) binding proteins in both cell lysates and condition media. When the α1-6 FucT gene was transfected into a human HCC cell line, Hep3B, which originally showed low levels of α1-6 FucT expression, α1-6-fucosylated AFP was dramatically increased in the condition media. Collectively, these results suggest that the enhancement of α1-6 FucT expression increased the fucosylation of several proteins, including AFP, and that the level of α1-6-fucosylated AFP in patients with HCC was in part caused by up-regulation of the α1-6 FucT gene expression.
Dietary juniper berry oil minimizes hepatic reperfusion injury in the rat
Authors: Jones Shannon M. , Zhong Zhi , Enomoto Nobuyuki , Schemmer Peter , Thurman Ronald G.
Published: July 2004

Juniper berry oil is rich in 5,11,14-eicosatrienoic acid, a polyunsaturated fatty acid similar to one found in fish oil, yet less prone to peroxidation. Dietary fish oil treatment has been shown to effectively reduce reperfusion injury; therefore, the effects of a diet containing juniper berry oil on hepatic reperfusion injury in a low-flow, reflow reperfusion model were investigated in the rat. Rats were fed semisynthetic diets containing either juniper berry oil, fish oil, or corn oil for 14 to 16 days. Daily food consumption averaged around 20 g/d in both the control and treatment groups; average daily weight gain was around 4 g per 100 g rat weight in all three groups studied, and there were no significant differences in these parameters. Livers were initially perfused at low-flow rates to induce pericentral hypoxia followed by a 40-minute reperfusion period. Peak lactate dehydrogenase (LDH) release during reflow averaged 44 U/g/h in the corn oil group and 32 U/g/h in the fish oil group, but was only 21 U/g/h as a result of juniper berry oil treatment. Malondialdehyde (MDA), an end-product of lipid peroxidation, reached a maximum value of 62 nmol/g/h in the corn oil group, but only reached 43 nmol/g/h and 34 nmol/g/h in the fish oil and juniper berry oil groups, respectively. Both juniper berry oil and fish oil treatment improved rates of bile flow from 25 μL/g/h (corn oil) to 36 and 38 μL/g/h, respectively. Importantly, juniper berry oil reduced cell death in pericentral regions of the liver lobule by 75%. Trypan blue distribution time, an indicator of the hepatic microcirculation, was reduced by approximately 25% with fish oil and over 50% by juniper berry oil diets compared with corn oil controls. The rates of entry of fluorescein-dextran, a dye confined to the vascular space, were increased 1.8- and 2.6-fold, and rates of outflow were increased 4.4- and 4.3-fold by fish oil and juniper berry oil, respectively, also reflecting improved microcirculation. Juniper berry oil also blunted increases in intracellular calcium and release of prostaglandin E2 (PGE2) by cultured Kupffer cells stimulated by endotoxin. These results are consistent with the hypothesis that feeding a diet containing juniper berry oil reduces reperfusion injury by inhibiting activation of Kupffer cells, thus reducing vasoactive eicosanoid release and improving the hepatic microcirculation in livers undergoing oxidant stress.
Increased risk of chronic liver failure in adults with heterozygous α-antitrypsin deficiency
Authors: Graziadei Ivo W. , Joseph Jacob J. , Wiesner Russell H. , Therneau Terry M. , Batts Kenneth P. , Porayko Michael K.
Published: July 2004

Controversy exists whether patients who are genetically heterozygous for α1-antitrypsin deficiency (α1 ATD), carrying a single PI*Z allele, are at increased risk of developing chronic liver disease. In these investigations, we determined the prevalence of heterozygous α1 AT phenotypes (PI MZ, PI SZ) in a well-characterized cohort of patients presenting with chronic liver failure before orthotopic liver transplantation (OLT). We analyzed data collected from all adult patients (n = 641) who underwent OLT at our tertiary referral center between March 1985 and December 1996. Study patients entered a prospective protocol designed to test for all known etiologies of liver disease. Complete testing including α1 AT phenotyping was successfully performed in 599 adults. We compared the overall number of heterozygous PI*Z carriers in our OLT cohort with established prevalence figures for general and regional American populations, and examined their distribution among various liver disease subgroups. Fifty-one patients were found to be heterozygous carriers of a single PI*Z allele for α1 AT. The predominant phenotype in our transplantation cohort was PI MZ, identified in 49 patients (8.2%), which is a significantly higher prevalence than that reported from previous American population studies (2%-4%). Additionally, a significantly greater number of PI MZ carriers existed in patients with cryptogenic cirrhosis compared with other liver disease categories (26.9%; P < .001). These data suggest that individuals carrying a single PI*Z allele for α1 AT may be at increased risk of developing cirrhosis and liver failure, even in the absence of an identifiable coexisting liver disease.
Cholesterol 7α-hydroxylase (CYP7A): Patterns of messenger RNA expression during rat liver development
Authors: Massimi Mara , Lear Steven R. , Huling Sandra L. , Jones Albert L. , Erickson Sandra K.
Published: July 2004

Cholesterol 7α-hydroxylase is a rate-limiting enzyme in bile acid synthesis, a major pathway for cholesterol catabolism. It plays a crucial role in postnatal development and survival. In an adult liver, its activity and messenger RNA (mRNA) are heterogeneously distributed with concentration in the pericentral area. We defined how the pattern of cholesterol 7α-hydroxylase mRNA evolves during rat liver development, correlated this with its total liver mRNA levels, and determined when its heterogeneous pattern of expression is established. Cholesterol 7α-hydroxylase mRNA was undetectable in 18-day-old fetal livers by Northern blot. It was increased markedly in newborns with a homogeneous liver lobular distribution as determined by in situ hybridization. At postnatal day four, mRNA levels were markedly decreased with concomitant appearance of a lobular gradient: mRNA was detected only in a few hepatocytes located around efferent venules. At 22 days, the time of highest mRNA expression, a marked extension of the gradient towards the periportal area was observed, indicating that the increase in total liver cholesterol 7α-hydroxylase mRNA level was a result of recruitment of hepatocytes upstream from the central vein area. By 28 days, the adult pattern was observed. Thus, expression of cholesterol 7α-hydroxylase mRNA is tightly regulated during rat liver development, both temporally and spatially supporting its critical role in normal postnatal development.
Activation of protein kinase Cα couples cell volume to membrane Cl− permeability in HTC hepatoma and Mz-ChA-1 cholangiocarcinoma cells
Authors: Roman Richard M. , Bodily Kurt O. , Wang Yu , Raymond John R. , Fitz Gregory J.
Published: July 2004

Physiological increases in liver cell volume lead to an adaptive response that includes opening of membrane Cl− channels, which is critical for volume recovery. The purpose of these studies was to assess the potential role for protein kinase C (PKC) as a signal involved in cell volume homeostasis. Studies were performed in HTC rat hepatoma and Mz-ChA-1 human cholangiocarcinoma cells, which were used as model hepatocytes and cholangiocytes, respectively. In each cell type, cell volume increases were followed by: 1) translocation of PKCα from cytosolic to particulate (membrane) fractions; 2) a 10- to 40-fold increase in whole-cell membrane Cl− current density; and 3) partial recovery of cell volume. In HTC cells, the volume-dependent Cl− current response (−46 ± 5 pA/pF) was inhibited by down-regulation of PKC (100 nmol/L phorbol 12-myristate 13-acetate for 18 hours [PMA]; −1.97 ± 1.5 pA/pF), chelation of cytosolic Ca2+ (2 mmol/L EGTA; −5.3 ± 4.0 pA/pF), depletion of cytosolic adenosine triphosphate (ATP) (3 U/mL apyrase; −12.58 ± 1.45 pA/pF), and by the putative PKC inhibitor, chelerythrine (25 μmol/L; −7 ± 3 pA/pF). In addition, PKC inhibition by chelerythrine and calphostin C (500 nmol/L) prevented cell volume recovery from swelling. Similar results were obtained in Mz-ChA-1 biliary cells. These findings indicate that swelling-induced activation of PKC represents an important signal coupling cell volume to membrane Cl− permeability in both hepatic and biliary cell models.
Effect of interferon therapy on hepatitis C virus RNA in whole blood, plasma, and peripheral blood mononuclear cells
Authors: Schmidt Warren N. , Wu Ping , Brashear Donna , Klinzman Donna , Phillips Mary Jeanne , LaBrecque Douglas R. , Stapleton Jack T.
Published: July 2004

Fifty-two patients with chronic hepatitis C virus (HCV) infection were treated with standard doses of interferon alfa-2b. During treatment, HCV RNA detection was studied in samples of whole blood (WB), plasma (Pl), and peripheral blood mononuclear cells (PBMCs). Individuals were classified as sustained responders (SRs), complete responders with relapse (CRs), partial responders (PRs), or nonresponders (NRs) according to normalization of serum alanine transaminase (ALT) during treatment and follow-up. Before treatment, 100% of WB samples and more than 95% of Pl and PBMC samples were positive for HCV RNA. During treatment, there was progressive clearance of HCV RNA from Pl and PBMCs in SRs and CRs, but CRs had significantly more positive WB samples during and following treatment (P <.0001). At 6 months, only 10% of CR patients were positive by Pl assay, but 50% were positive by WB assay (P <.01). In the PR group, all WB samples remained positive throughout treatment, although 25% to 40% of PBMC and Pl samples became negative for HCV RNA during the first 2 months of therapy (WB > Pl or PBMC; P < .001). However, at later times during treatment most Pl and PBMC samples in the PR group were positive. Samples from the NR group showed no clearance of HCV RNA from WB, Pl, or PBMC fractions. These data document the increased sensitivity of WB assays for detecting HCV RNA in the peripheral blood of patients during interferon therapy. Furthermore, our findings suggest that WB analysis of HCV RNA may be a useful parameter to monitor in determining the end point of interferon therapy.
10-year follow-up after interferon-α therapy for chronic hepatitis C
Authors: Lau Daryl T. , Kleiner David E. , Ghany Marc G. , Park Yoon , Schmid Peter , Hoofnagle Jay H.
Published: July 2004

Sustained responses to interferon-α occur in 10% to 25% of patients with chronic hepatitis C, but the long-term outcome is not well defined. We evaluated the long-term clinical, histological, and virological outcomes of 10 patients with chronic hepatitis C who were treated between 1984 and 1987 with interferon-α-2b for 52 ± 6 weeks (total doses of 492 ± 116 MU). Before therapy, all 10 had hepatitis C virus (HCV) RNA, elevations of serum aminotransferases, and chronic hepatitis with fibrosis on liver biopsy. Clinical follow up was 6 to 13 years, and liver biopsies were done 5 to 11 years after initiation of therapy. HCV RNA was assayed by qualitative and quantitative reverse transcriptase-polymerase chain reaction assays. Among 5 patients who had a 6-month sustained response after therapy, all remained HCV RNA negative, and at last follow-up, 4 had normal and 1 minimally elevated serum aminotransferase levels. Liver biopsy specimens were nonreactive for HCV RNA, and all the patients showed improvements in both inflammation and fibrosis and were either normal or had mild, nonspecific inflammatory changes. Among 5 patients without a sustained response, all continued to have HCV RNA in serum and persistent or intermittent aminotransferase elevations. Liver biopsy specimens showed little or no change in necrosis and inflammation; all except 1 patient had progression of fibrosis scores or cirrhosis. All 5 patients had symptoms of chronic hepatitis, 1 underwent liver transplantation, and another had progressive hepatic decompensation. In conclusion, patients with a 6-month posttreatment virological response have a favorable long-term clinical and histological outcome.
Mutations in the interferon-sensitivity determining region of hepatitis C virus and transcriptional activity of the nonstructural region 5A protein
Authors: Fukuma Toshiko , Enomoto Nobuyuki , Marumo Fumiaki , Sato Chifumi
Published: July 2004

Amino acid (aa) mutations in the interferon-sensitivity determining region (ISDR) (aa position 237-276 of the nonstructural region 5A [NS5A] protein consisting of 447 amino acids) of hepatitis C virus (HCV) are related to increased interferon sensitivity and low viral load, but its mechanism has not been clarified. Recently, the NS5A protein has been reported to have a transcriptional activation function, like other viral transactivator proteins known to repress interferon-induced gene expression, and the ISDR overlaps one of the acidic amino acid regions, putative domains conferring this activity. In the present study, we investigated the transcriptional activation function of the ISDR itself and the effect of amino acid mutations in the ISDR on this activity. The full-length or truncated NS5A cDNA with different ISDR sequences was cloned into a yeast or mammalian expression vector to form a fusion protein consisting of the GAL4 DNA-binding domain (GAL4-DBD) and NS5A protein. Following transfection, the transcriptional activities of these constructs were determined using β-galactosidase (yeast) or chloramphenicol acetyltransferase (CAT) (mammalian cell) reporter gene expression under the control of GAL4 binding sites. In yeast, both the full-length sequence of NS5A-R (a clone with one aa mutation in the ISDR) and NS5A-S (a derivative of NS5A-R with six aa mutations in the ISDR) had no distinguishable transcriptional activity, whereas an amino-terminal deletion construct of NS5A-R (aa position 228-447) lacking 227 aa, showed remarkable activity with the relative value of 117.0 over that of the backbone vector. The same deletion mutant of NS5A-S produced five times higher activity with the relative value of 575.0, indicating that aa mutations in the ISDR profoundly affect this transcriptional activity. In a hepatoma cell line, HuH-7, the transcriptional activity was more prominent with a construct consisting of only the ISDR and short flanking sequences (aa 228-284) than larger deletion constructs of NS5A-R. Analysis using six different ISDR clones revealed that different mutations enhanced this activity to various extent compared with the wild-type ISDR. In particular, site-directed mutagenesis targeted to the aa position 252 showed that this aa residue had profound influence on the activity. These results suggest that the ISDR has a transcriptional activity, and it is enhanced by aa mutations that are also related to decreased viral load and increased interferon sensitivity. The possible association between transcriptional activation and interferon sensitivity or viral replication should be studied further.
Hepatic Artery Resistance in Alcoholic Liver Disease
Authors: Colli Agostino , Cocciolo Massimo , Mumoli Nicola , Cattalini Nadia , Fraquelli Mirella , Conte Dario
Published: July 2004

Patency and direction of flow in portal veins and their branches are generally assessed by duplex Doppler ultrasonography (DDUS), whereas few data are available on hepatic arterial hemodynamics. In this study, resistive (RI) and pulsatility indexes (PI) were calculated at DDUS in 21 controls, 22 chronic alcoholic patients without evidence of liver damage, 19 patients with acute alcoholic hepatitis (AAH), 30 patients with chronic viral hepatitis (CVH), 23 patients with alcoholic cirrhosis, and 22 patients with viral–related cirrhosis. Diagnosis was based on clinical and histological findings. Mean ± SD RI was similar in controls and CVH patients (0.64 ± 0.02 and 0.66 ± 0.04, respectively), significantly decreased in alcoholic patients without liver damage and AAH patients (0.61 ± 0.07 and 0.60 ± 0.07) (P < .05), and significantly increased in patients with alcoholic (0.72 ± 0.04) and viral–related cirrhosis (0.74 ± 0.04) (P < .05). It was <0.60 in 9 of the 19 AAH patients (47%) and 11 of the 22 alcoholic patients without liver damage (50%), and >0.70 in 39 of the 45 cirrhotic patients (87%) and 12 of the 71 noncirrhotic patients pooled together (17%). A significant correlation was observed between RI and PI (r = .83; P < .05). The coefficients of variation for intraobserver variability were 6.3% ± 5.1% for RI and 10.1% ± 6.2% for PI, and the corresponding figures for interobserver variability were 5.2% ± 3.5% and 9.3% ± 4.6%. These findings support the existence of ethanol–related hepatic arterial vasodilation in AAH and alcoholic patients without liver damage. Progression of liver damage from AAH to cirrhosis profoundly impairs the hepatic responsiveness as a consequence of fibrosis with vascular distortion.
Small Intestine Dysmotility and Bacterial Overgrowth in Cirrhotic Patients With Spontaneous Bacterial Peritonitis
Authors: Chang Chi-Sen , Chen Gran-Hum , Lien Han-Chung , Yeh Hong-Zen
Published: July 2004

Patients with bacterial overgrowth of the small intestine developed spontaneous bacterial peritonitis (SBP) more frequently than patients without bacterial overgrowth of the small intestine. The objective of this study was to determine whether the incidences of small intestine dysmotility and bacterial overgrowth are higher in cirrhotic patients with a history of SBP than in cirrhotic patients without SBP. Forty cirrhotic patients were enrolled in this study. There were 20 patients with a history of SBP and 20 patients without a history of SBP. Small intestine bacterial overgrowth was diagnosed by breath hydrogen test. Small intestine motility was recorded, by a 3–channel solid–state manometric catheter, for 24 hours. There were no statistical differences in age or sex between the two groups. The Child–Pugh scores in the SBP group were higher than in the non–SBP group (10.5 ± 2.1 vs. 8.1 ± 1.9, P< .01). The incidence of bacterial overgrowth of the small intestine was higher in the SBP group than in the non–SBP group (70% vs. 20%, P < .01). The amplitude and duration of migrating motor complex (MMC) activity fronts, as well as the number of clusters per hour, were similar in both groups. However, the frequency of MMC activity fronts was higher in the non–SBP group than in the SBP group (4.8 ± 2.3/24 hours vs. 3.5 ± 1.2/24 hours, P < .05). In addition, the MMC velocity was significantly higher in the non–SBP group (8.3 ± 2.6 vs. 5.3 ± 2.1 cm/min, P < .01). The incidence of bacterial overgrowth of the small intestine was higher in cirrhotic patients with history of SBP than in those without SBP. Small intestine motility dysfunction was more severe in cirrhotic patients with history of SBP. Impaired motility of the small intestine, causing bacterial overgrowth of the small intestine, may be one of the explanations for recurrent SBP in cirrhotic patients.
Lipoprotein–X in Patients With Cirrhosis: Its Relationship to Cholestasis and Hypercholesterolemia
Authors: Sörös Peter , Böttcher Joachim , Maschek Hansjörg , Selberg Oliver , Müller Manfred James
Published: July 2004

Lipoprotein–X (Lp–X) is an abnormal low–density lipoprotein frequently found in liver disease. It is regarded as the most sensitive and specific biochemical parameter for the diagnosis of intra– and extrahepatic cholestasis. Moreover, Lp–X is supposed to contribute to the development of hypercholesterolemia in cholestatic liver disease, because it fails to inhibit de novo cholesterol synthesis. This investigation will focus on the relationship between the presence of Lp–X and serum lipid concentrations in cirrhosis. The significance of Lp–X in the diagnosis of cholestasis, compared with alkaline phosphatase (AP), γ–glutamyl transferase (GGT), and bilirubin levels, will be assessed as well. The present cross–sectional study includes 212 patients with histopathologically proven cirrhosis. The detection of Lp–X and the quantification of α–, β–, and pre-β–cholesterol was based on agar gel electrophoresis and polyanion precipitation. For the characterization of liver function, the concentrations of albumin and bilirubin, the activities of liver enzymes, and coagulation times were assessed. In a subgroup of 40 individuals, liver biopsies were re–evaluated to confirm or exclude intrahepatic cholestasis. As a result, there was no association between the appearance of Lp–X and total cholesterol concentrations. While all patients with Lp–X showed intrahepatic cholestasis (predictive value of the positive test = 1), only 16 of 28 patients with cholestasis formed Lp–X (sensitivity = 0.57). The activities of AP and of GGT, as well as the concentrations of bilirubin, were strongly elevated in most patients, with and without cholestasis. The predictive values of AP, GGT, and bilirubin were 0.77, 0.69, and 0.74 for the positive test and 0.5, 0, and 0.6 for the negative test, respectively. We conclude that Lp–X is not related to hypercholesterolemia in cirrhosis. The positive, but not the negative, Lp–X test has high predictive value for the diagnosis of cholestasis in cirrhosis. The biochemical parameters traditionally used for the assessment of extrahepatic cholestasis, AP, GGT, and bilirubin, do not support the diagnosis of intrahepatic cholestasis caused by cirrhosis.
Octreotide Compared With Placebo in A Treatment Strategy for Early Rebleeding in Cirrhosis. A Double Blind, Randomized Pragmatic Trial
Authors: D'Amico Gennaro , Politi Flavia , Morabito Alberto , D'Antoni Adele , Guerrera Daniela , Giannuoli Gandolfo , Traina Mario , Vizzini Giovanni , Pasta Linda , Pagliaro Luigi
Published: July 2004

β–Blockers and sclerotherapy prevent long–term upper digestive rebleeding in cirrhosis but they seem ineffective for early rebleeding. We compared octreotide with a placebo for the prevention of early rebleeding in cirrhotic patients. After control of acute upper digestive bleeding, 262 consecutive cirrhotic patients were randomized to octreotide 100 μg subcutaneously three times a day for 15 days (n = 131) or to the placebo (n = 131), in a double blind pragmatic trial in which β–blockers and/or sclerotherapy were allowed together with the experimental treatment. Separate randomization and analysis were performed according to whether patients were eligible for β–blockers and/or sclerotherapy (101 placebo, 97 octreotide) or not (30 placebo, 34 octreotide). Rebleeding within 15 days was the primary measure of treatment efficacy; 6–week rebleeding rate was also assessed as a secondary measure. Fifteen–day cumulative proportions of patients rebleeding were 28% in the placebo group and 24% in the octreotide group (P = .40); corresponding figures among the 198 patients eligible to β–blockers and/or sclerotherapy were 26% and 16% (P = .05) and among the 64 not eligible for these treatments 33% and 49% (P = .29). Among patients eligible to β–blockers and/or sclerotherapy, a significant reduction of rebleeding episodes (35 vs. 18, P = .03), blood transfusions (75 vs. 50, P = .04), and days of stay in hospital (1,544 vs. 1,190, P = .0001) was also found in the octreotide group: this beneficial effect was confirmed 6 weeks after randomization. Mortality was not affected by octreotide in either group of patients. It is suggested that octreotide may reduce the risk of early rebleeding in cirrhotic patients treated with β–blockers and/or sclerotherapy after control of acute upper digestive bleeding. Further studies are needed to confirm this result.
Portosystemic Hepatic Encephalopathy After Transjugular Intrahepatic Portosystemic Shunt in Patients With Cirrhosis: Clinical, Laboratory, Psychometric, and Electroencephalographic Investigations
Authors: Nolte Wilhelm , Wiltfang Jens , Schindler Christian , Münke Hans , Unterberg Knut , Zumhasch Uta , Figulla Hans R. , Werner Gerald , Hartmann Heinz , Ramadori Giuliano
Published: July 2004

A prospective study of hepatic encephalopathy (HE) including neuropsychiatric and psychometric evaluation, electroencephalography, and determination of arterial ammonia levels was performed in 55 cirrhotic patients treated consecutively by transjugular intrahepatic portosystemic shunt (TIPS). The cumulative HE rate increased from 23.6% within the 3–month interval before TIPS to 50.9% within the first 3–month interval post–TIPS (P = .003). Significant and independent predictors of HE post–TIPS were the presence of HE pre–TIPS and reduced liver function. The cumulative HE rate declined in the second 3–month interval post–TIPS and reached the pre–TIPS level. Chronic forms of HE exceeding grade I were not observed. In a subgroup of 22 nonencephalopathic TIPS patients, the prevalence of subclinical HE did not change after TIPS. Among individual psychometric tests, the block design test gave the highest proportion of pathological results (about 50%), whereas selective reminding gave the lowest (10%–25%). Electroencephalography (EEG) showed a temporary increase of pathological results at 1 month after TIPS, when patients with overt HE (grade I) were included (proportion of 21.1% before vs. 57.1%, P = .005). Arterial ammonia concentration increased from a mean of 94 ± 26 μg/dL to 140 ± 28 μg/dL at 3 months after TIPS (P < .001). Elevated ammonia levels persisted. TIPS led to a temporary increase of HE incidence within 3 months. The decline of the HE rate beyond 3 months despite a sustained increase of arterial ammonia levels could not entirely be explained by reduction of shunt flow, nor by alteration of liver function. Instead, cerebral adaptation to gut–derived neurotoxins might be anticipated.
Tumor Necrosis Factor Primes Hepatocytes for Dna Replication in the Rat
Authors: Webber Eric M. , Bruix Jordi , Pierce Robert H. , Fausto Nelson
Published: July 2004

Signaling through tumor necrosis factor receptor type 1 (TNFR–1) using a pathway that involves nuclear factor κB (NF–κB), interleukin–6 (IL–6), and STAT3 is required for the initiation of liver regeneration. We have proposed that TNF primes hepatocytes to respond to the mitogenic effect of growth factors, but so far, there has been no experimental demonstration that TNF enhances growth factor responses of hepatocytes. To test this hypothesis, we infused hepatocyte growth factor (HGF) and transforming growth factor α (TGF–α) (40 μg/24 h) directly into the portal vein of rats for 24 hours using osmotic pumps and determined whether TNF injection (5 μg per rat) would significantly increase hepatocyte DNA labeling in these animals. All rats received 5–bromo–2′–deoxyuridine (BrdU) by intraperitoneal delivery during a 48–hour period (i.e., BrdU infusion continued for 24 hours after the end of growth factor administration). BrdU labeling in the liver was measured by both immunohistochemistry and flow cytometry, and the results obtained by these methods showed excellent concordance. The results demonstrate that TNF transiently activates NF–κB and STAT3 and increases the proliferative response of hepatocytes to HGF or TGF–α by fourfold. Priming effects on hepatocyte DNA replication were also obtained with injection of lipopolysaccharide (LPS) and gadolinium chloride (GdCl3), agents that release TNF in the liver. Similarly to TNF, GdCl3 injection caused the activation of NF–κB and STAT3, reaching a maximum 8 to 12 hours after the injection. The results show that TNF acts as a primer to sensitize hepatocytes to the proliferative effects of growth factors and offers a mechanism to explain the initiation and progression phases of liver regeneration after partial hepatectomy (PH).
Early Hepatocellular Carcinoma As An Entity With A High Rate of Surgical Cure
Authors: Takayama Tadatoshi , Makuuchi Masatoshi , Hirohashi Setsuo , Sakamoto Michiie , Yamamoto Junji , Shimada Kazuaki , Kosuge Tomoo , Okada Shuichi , Takayasu Kenichi , Yamasaki Susumu
Published: July 2004

Early hepatocellular carcinoma (HCC) has been defined as a well–differentiated cancer containing Glisson's triad, but it remains unknown whether this lesion is curable. We prospectively studied 70 patients (enrolled from 1,172 referrals between 1982 and 1991) who had a diagnosis of a single HCC 2 cm or less in diameter (Stage T1) and who underwent curative hepatectomy and long–term follow–up (range, 0.2 to 14.3 years). Patients were eligible for surgery if they had a tumor that met the diagnostic criteria for HCC and were in Child–Pugh class A (n = 59) or B (n = 11) status. Among the 70 patients, there was 1 operative death. Based on our typing system, the tumors were assigned as early HCC (n = 15), overt HCC (n = 52), and non–HCC tumor (n = 3). The rate of microscopic regional spread was lower in early HCCs than in overt HCCs (7% vs. 42%; P = .01). The early HCC group had a longer time to recurrence than did the overt HCC group (3.9 vs. 1.7 years; P < .001) and had no local recurrence. After a median follow–up of 6.3 years, both overall survival and recurrence–free survival in the early HCC group were significantly better than those in the overt HCC group (P = .01; P = .001). In these two groups, the 5–year rates of overall survival were 93% and 54% (P = .01), and those of recurrence–free survival were 47% and 16% (P = .05), respectively; a significant survival benefit persisted over a decade (57% vs. 21%; P = .05). The early HCC group was at a lower risk of recurrence (relative risk, 0.31; 95% CI, 0.15 to 0.65; P = .002) and death (relative risk, 0.26; 95% CI, 0.09 to 0.73; P = .01) than was the overt HCC group. Early HCC is a distinct clinical entity with a high rate of surgical cure, thereby justifying its definition. It can be a lesion that corresponds to “Stage 0” cancer in other organs.
Establishment of A Functional Hgf/C– Met Autocrine Loop in Spontaneous Transformants of Wb–F344 Rat Liver Stem–Like Cells
Authors: Presnell Sharon C. , Hooth Michelle J. , Borchert Kristen M. , Coleman William B. , Grisham Joe W. , Smith Gary J.
Published: July 2004

A model of spontaneous malignant transformation was used to evaluate the molecular changes that take place in WB–F344 rat liver epithelial cells during neoplastic transformation and tumorigenesis. A comparison of wild–type low–passage WB–F344 cells to spontaneously transformed tumor cell lines revealed that the majority of the tumor cell lines have an increased capacity for autonomous proliferation and motility when maintained in serum–free media. In the current study, we show that c–met is expressed at some level in wild–type WB–F344 cells and in all of the spontaneously transformed tumor cell lines, and that 9/16 of the tumor cell lines have acquired hepatocyte growth factor (HGF) expression. In vitro growth of HGF–expressing tumor cell lines is inhibited as much as 68% by the addition of neutralizing antibodies to HGF or antisense HGF oligonucleotides, indicating that the production of HGF by the tumor cells is partially responsible for driving autonomous proliferation in a subset of tumor cell lines. Furthermore, conditioned media collected from HGF–expressing tumor cell lines stimulates DNA synthesis in wild–type WB–F344 cells, and this effect can be abrogated by pre–incubation of the conditioned media with neutralizing antibodies to HGF. Because HGF is a motility–promoting growth factor, all cell lines were evaluated to determine if expression of HGF stimulated motogenesis. All tumor cell lines (regardless of HGF expression) were highly motile in comparison with wild–type WB–F344 cells, with a 3.5–fold to 20–fold greater number of motile cells. The high basal rate of motility characteristic of the tumor cell lines is not a result of the production of HGF, because it is also a property of the cell lines that do not express HGF messenger RNA. Furthermore, tumor cell motility is not inhibited by antisense oligonucleotides or neutralizing antibodies. Establishment of an autocrine HGF/c–met loop in a subset of spontaneously transformed WB–F344 cell lines may influence development and/or expression of the tumorigenic phenotype by driving cellular proliferation.
Interleukin–6, Hepatocyte Growth Factor, and Their Receptors in Biliary Epithelial Cells During A Type I Ductular Reaction in Mice: Interactions Between the Periductal Inflammatory and Stromal Cells and the Biliary Epithelium
Authors: Liu Zejin , Sakamoto Toshiki , Ezure Tsukasa , Yokomuro Shigeki , Murase Noriko , Michalopoulos George , Demetris Anthony J.
Published: July 2004

The interleukin–6 (IL–6)/gp–80 and hepatocyte growth factor (HGF)/met ligand/receptor systems have been shown to stimulate biliary epithelial cell (BEC) DNA synthesis in vitro. The mRNA and protein production of these two in vitro mitogens were mapped in vivo during the first week after bile duct ligation (BDL) when peak BEC DNA synthesis is seen. Changes around the biliary tree were compared with those seen in the peripheral liver using a combination of Northern blotting and a unique biliary tree isolation technique, in which the bile ducts and the surrounding portal stroma and inflammatory cells are separated from the hepatocytes by perfusion digestion. Further localization was performed with in situ hybridization and immunohistochemistry. In the normal liver, there is low–level expression of HGF mRNA by periportal stellate cells, and HGF protein localizes to these cells and to neutrophils; extracellular HGF protein is present in the bile. There is no detectable IL–6 mRNA by Northern analysis or IL–6 protein expression in the normal liver, but both met and IL–6 receptor (IL–6R) mRNA are detectable; met mRNA is expressed strongly in the biliary tree, and met protein is expressed weakly on hepatocytes and strongly on BEC. IL–6R mRNA is weakly expressed in the biliary tree, and IL–6R protein is detectable on hepatocytes, with a periportal–to–perivenular gradient, but not on BEC. During the first 3 days after BDL, HGF mRNA expression is increased in both the biliary tree and in the peripheral liver, and production is localized to stellate cells, periductal neutrophils, and stromal cells, which typically accompany the proliferating ductules. IL–6 mRNA and protein were detected only near the biliary tree after BDL, and not in the peripheral liver, and the production was localized to periductal hematolymphoid cells, which had the morphological appearance of macrophages and/or dendritic cells. There is also a distinct up–regulation of met and gp–80 mRNA and protein in the biliary tree, which is stronger than that seen in the peripheral liver. Met protein expression is increased, and IL–6R(gp–80) protein is induced on the proliferating BEC, consistent with the participation of both the HGF/met and IL–6/gp–80 systems in the early phases of type I ductular reactions. These observations show that periductal hematolymphoid and stromal cells are the source of BEC growth factors, and receptors for these factors are up–regulated on BEC during active ductular proliferation. Complex interactions between the inflammatory, stromal, and BEC results in a dysmorphogenic repair response that eventually leads to cirrhosis.
Splenorenal Shunt Blood Flow by Transit–Time Ultrasound As An Index of Collateral Circulation in Portal Hypertensive Rats
Authors: Calès Paul , Oberti Frédéric , Veal Nary , Fort Joël , Kaassis Mehdi , Moal Frédéric , Aubé Christophe , Vuillenin Eric , Pilette Christophe , Rifflet Hervé , Trouvé Renaud
Published: July 2004

The aim of this study was to develop a technique that could serve as an index of portosystemic shunt (PSS) blood flow in portal hypertensive rats whose main shunt is the splenorenal shunt (SRS). The main hemodynamic measurements performed were: SRS blood flow by the transit–time ultrasound (TTU) method, percentage of PSS, and regional blood flows by the microsphere method. We determined the accuracy and reproducibility of SRS blood flow measurements under baseline and pharmacological (octreotide) conditions. SRS blood flow was compared with other hemodynamic characteristics. Two models of portal hypertension were used: secondary biliary and dimethylnitrosamine cirrhosis. The SRS blood flow was correlated with mesenteric (r = .76; P < .001) and splenic (r = .67; P < .01) PSS percentages. The intra– and interobserver agreements for SRS blood flow were excellent: ric = .99 and ric = .98, respectively. SRS blood flow was six times higher in portal hypertensive rats (0.6 ± 0.7 mL · min-1 · 100 g-1) than in sham rats (0.1 ± 0.1 mL · min-1 · 100 g-1 [P < .01]). Octreotide significantly decreased SRS blood flow but not mesenteric or splenic PSS percentages. SRS is the main PSS in rats. The measurement of SRS blood flow by TTU is accurate and reproducible. This method can be used to identify new mechanisms in hemodynamic studies that differ from those identified by the measurement of the percentage of PSS by the microsphere method, especially in pharmacological studies.
Macrophage Pacification Reduces Rodent Pancreatitis–Induced Hepatocellular Injury Through Down–Regulation of Hepatic Tumor Necrosis Factor α and Interleukin–1β
Authors: Yang Jun , Denham Woody , Carter Gay , Tracey Kevin J. , Norman James
Published: July 2004

Overproduction of tumor necrosis factor α (TNF–α), interleukin–1β (IL–1β), and nitric oxide (NO) is believed to be detrimental during the progression of acute pancreatitis, yet little is known about the hepatic production of these mediators and their role in mediating pancreatitis–induced hepatic dysfunction. Rats were randomized to receive a single intraperitoneal injection of the macrophage–pacifying compound, CNI–1493 (1.0 mg/kg), or vehicle 1 hour before the induction of retrograde bile salt pancreatitis. Sham–operated animals served as controls. Animals were killed 18 hours later, with serum and livers harvested to determine the degree of hepatocellular injury and the induction of TNF–α, IL–1β, and inducible nitric oxide synthase (iNOS). In addition, serum TNF–α and nitrites (end–product of NO breakdown) were determined in each group to assess the mechanism of action of CNI–1493. TNF–α, IL–1β, and iNOS gene expression (by reverse–transcription polymerase chain reaction) as well as aspartate transaminase (AST), alanine transaminase (ALT), and lactic dehydrogenase (LDH) (but not alkaline phosphatase [ALP]) increased following the development of pancreatitis (all P < .05). Macrophage pacification significantly prevented the induction of TNF–α and IL–1β mRNA (but not iNOS), resulting in lessened serum AST, ALT, and LDH (all P < .05). Serum TNF–α protein and nitrites correlated with gene induction in that both were increased following the onset of pancreatitis, and TNF–α protein production was significantly attenuated in animals receiving CNI–1493. Hepatocellular, but not bile duct, injury occurs during experimental pancreatitis that is associated with hepatic TNF–α, IL–1β, and iNOS mRNA gene induction, as well as TNF–α protein and nitrite production. Preventing the production of TNF–α and IL–1β by macrophage pacification attenuates the hepatocellular damage, suggesting that these mediators play a role in pancreatitis–induced hepatic injury.