bluuebunny/crossref_metadata_2025 huggingface dataset:
Abnormal Intracellular Distribution of O6–Alkylguanine–Dna–Alkyltransferase in Hepatitis B Cirrhotic Human Liver: A Potential Cofactor in the Development of Hepatocellular Carcinoma
Authors: Lee S M , Portmann B C , Margison G P
Published: November 2004

The expression of O6–alkylguanine–DNA–alkyltransferase (ATase), which is responsible for repair of the promutagenic and cytotoxic DNA lesion O6–alkylguanine, was examined by immunostaining in a series of liver sections from normal and hepatitis–B cirrhosis patients using a polyclonal anti–human ATase antiserum. In 10 normal liver sections, the ATase staining was predominantly nuclear and very intense in the majority of the hepatocytes with a panacinar distribution. In contrast, in 15 hepatitis–B sections, the ATase was located mainly in the cytoplasm of the majority of the hepatocytes and had a perinuclear distribution. Scattered hepatocytes showed a more intense staining involving all or part of their cytoplasm; nuclear staining, however, was reduced in intensity, being inconspicuous in seven and patchy and weak in eight. ATase activity in extracts of 10 of the hepatitis–B livers varied from approximately 90 to 360 fmoles/mg protein and there was no apparent relationship between these levels and the cellular distribution of the enzyme. The sequestration of the ATase protein in the cytoplasm, away from its site of action in the cell nucleus suggests that in hepatitis–B cirrhosis, the repair of O6–alkylguanine lesions by ATase may be less efficient than in normal hepatocytes. The abnormal cellular distribution of ATase may thus be an additional cofactor in the development of hepatitis–B–associated hepatocellular carcinoma.
Inhibition of Hepatitis B Virus Dna Polymerase by Enantiomers of Penciclovir Triphosphate and Metabolic Basis for Selective Inhibition of Hbv Replication by Penciclovir
Authors: Shaw T , Mok S S , Locarnini S A
Published: November 2004

The deoxyguanosine analog penciclovir (PCV; 9–[4–hydroxy–3–hydroxymethyl–but–1–yl]guanine), has shown potent antiviral activity against herpes viruses and hepadnaviruses. Efficacy against chronic hepatitis B virus (HBV) infection has been demonstrated in an animal model and in recent clinical trials of famciclovir, the oral form of PCV. The antiviral activity of PCV is believed to be dependent on the intracellular formation of PCV–triphosphate (PCV–TP) which is presumed to inhibit HBV replication by interfering with viral DNA polymerase activity. The (

S

)–enantiomer is preferentially formed in herpes virus–infected cells, and is the more active against the herpes simplex virus; however, little is known about the biochemical mechanisms of PCV phosphorylation or of interference with viral replication in HBV–infected cells. Here, we report that in contrast with herpes simplex virus, the (R)–enantiomer of PCV–TP is a more potent inhibitor of HBV DNA polymerase–reverse transcriptase (pol–RT) in vitro than the (

S

)–enantiomer. In assays for HBV DNA pol–RT activity, in which purified viral core particles were the enzyme source, the IC50s for (

R

)– and (

S

)–enantiomers of PCV–TP were 2.5 micromol/L and 11 micromol/L, respectively. The estimated Kis for (

R

)– and (

S

)– PCV–TP were approximately 0.03 micromol/L and approximately .04 micromol/L, respectively, about 3–fold lower than the Km for deoxyguanosine triphosphate (dGTP) in the same system. In addition, we report that PCV metabolism is similar in both control (HepG2) and in HBV–transfected (2.2.15) hepatoblastoma cells

in vitro

, indicating that cellular enzyme(s) catalyze PCV phosphorylation. Peak PCV–TP concentrations of about .4 micromol/L were reached in both cell types in less than 12 hours, and intracellular PCV–TP was exceptionally stable with a half–life of about 18 hours. These observations provide a mechanistic basis for the potent activity of PCV against HBV.
The Clinical Significance of Changes in Genetic Heterogeneity of the Hypervariable Region 1 in Chronic Hepatitis C With Interferon Therapy
Authors: Shindo M , Hamada K , Koya S , Arai K , Sokawa Y , Okuno T
Published: November 2004

We examined changes in the hypervariable region 1 of the hepatitis C virus (HCV) RNA that occurred with interferon therapy in 33 patients with chronic hepatitis C to assess the clinical significance of this region. The 33 patients had HCV genotype 1b and were classified into three groups based on serum aminotransferase levels during and after therapy with α interferon; long–term responders (n = 9), short–term responders (n = 11), and nonresponders (n = 13). Changes in the genetic heterogeneity of the hypervariable region 1 were determined by using nonisotopic polymerase chain reaction–single strand conformation polymorphism (PCR–SSCP). HCV RNA levels were evaluated by reverse transcriptase PCR and branched DNA probe assays. Changes in sequences were determined by cloning and sequencing analysis. Before therapy, the long–term responders had significantly lower degrees of heterogeneity and lower viral levels than nonresponders. There were no significant differences between short–term and nonresponders. With interferon therapy, viral levels and degree of heterogeneity decreased to a greater extent among long–term and short–term responders than among nonresponders. Sequencing analysis showed that the three groups had similar clone numbers initially, but long–term responders had rather homogeneous viral populations, whereas short–term and nonresponders had heterogeneous populations, but that there were no nucleotide sequences or amino acid alignments that were specific for any group before, during, and 6 months after therapy. Approximately half of short–term and nonresponders received a second course of interferon 7 to 10 months after the initial therapy; all showed an identical response to the second course of therapy regardless of interim changes in the heterogeneity of hypervariable region 1. These findings suggest that (1) patients who were nonresponders or short–term responders had mixed viral populations that had differing sensitivities to interferon, (2) the changes in the hypervariable region 1 (HVR 1) did not affect responsiveness to interferon, and (3) the lower heterogeneity in the HVR 1 was associated with a long–term response to interferon only when the viral levels were low.
Elevated Plasma Levels of Matrix Metalloproteinase–9 (92–Kd Type Iv Collagenase/Gelatinase B) in Hepatocellular Carcinoma
Authors: Hayasaka A , Suzuki N , Fujimoto N , Iwama S , Fukuyama E , Kanda Y , Saisho H
Published: November 2004

Background

Matrix metalloproteinase 9 (MMP–9), a 92–kd gelatinase/type IV collagenase, has been implicated as playing an important role in cancer invasion and metastasis. A previous study showed that serum type IV collagenase activity correlated with metastasis by hepatocellular carcinoma (HCC). The aims of this study were to determine the plasma levels of immunoreactive MMP–9 in patients with HCC and to compare the levels with the clinical features including vascular invasion.

Patients and methods:

This study included 100 patients with HCC, 21 patients with chronic hepatitis (CH), 24 patients with liver cirrhosis (LC), and 138 healthy control subjects. Plasma MMP–9 levels were measured with a specific one–step sandwich enzyme immunoassay.

Results:

Plasma MMP–9 levels in HCC (62 [33 to 130 ng/mL] median [25%, 75%], 13 to 660 ng/mL, minimum, maximum) were significantly elevated compared with those in normal controls (36 [25 to 45], range, 2.8–70 ng/mL), in CH (28 [18 to 30], 13 to 66 ng/mL) and in LC (35 [26 to 58], 16 to 86 ng/mL) (

P

< .0000001;

P

= .0000003; and

P

= .00205, respectively). When the cut–off level was defined as 60 ng/mL from a receiver operating characteristic curve, plasma MMP–9 concentrations had a sensitivity of 53% and a specificity of 89% for the detection of HCC from CH and LC. The levels were significantly higher in HCC patients with macroscopic portal venous invasion (79 [36 to 160], 15–660 ng/mL) than those without the invasion (44 [27 to 80], 13 to 210 ng/mL) (

P

= .00726). Plasma MMP–9 levels in patients with HCC were not correlated with tumor number, size, volume, or serum α–fetoprotein levels.

Conclusions:

The present data suggest that plasma MMP–9 levels can be a candidate for a novel marker for HCC. The levels appear to reflect its potential and ongoing activity of vascular invasion. A long–term follow–up of the patients will be necessary to determine whether increased plasma MMP–9 levels are predictive of more invasive and metastatic HCC.
Azygos Blood Flow: Phase Contrast Quantitation in Volunteers and Patients With Portal Hypertension Pre– and Postintrahepatic Shunt Placement
Authors: Debatin J F , Zahner B , Meyenberger C , Romanowski B , Schopke W , Marincek B , Fuchs W A
Published: November 2004

Flow volume in the azygos venous system was quantitated with Cine–phase contrast (PC) velocity mapping in volunteers and compared with patients with known portal hypertension, who were referred for transjugular intrahepatic portosystemic shunt (TIPS) placement. Subsequently, the TIPS–induced hemodynamic effects on portal and azygos flow were analyzed. To assess intra– and intersubject variability, flow in the azygos veins was measured in each of 10 normal subjects at three different times. Subsequently, portal and azygos flow was quantitated in 20 patients with portal hypertension, before and after TIPS placement. All imaging was performed on a 1.5–tesla magnetic resonance imaging (MRI) system. Azygos flow was measured transaxially at the midthoracic level. Cine–PC flow measurements of the main portal vein followed morphological evaluation of the portal venous system with axial and coronal breath–held magnet resonance angiogram. Azygos flow in normal subjects was characterized by high inter– and low intrasubject variability. Azygos flow in patients with portal hypertension was significantly higher (

P

< .001). Following successful TIPS placement in 18 patients, Cine–PC revealed a mean decrease in azygos flow of 46.3% (

P

< .001) and a 134% (

P

< .001) increase of portal flow. Reflecting the complexity of portal hemodynamics, there was no correlation between the changes in the portal and azygos systems. In 2 patients with early occlusion of TIPS, documented invasively, portal and azygos flow values remained largely unchanged. Cine–PC enables the noninvasive, quantitative assessment of flow within the azygos venous system. Azygos flow was found to be markedly elevated in patients with portal hypertension. Combined portal and azygos PC flow measurements quantitate the therapeutic effect of TIPS placement.
Fibrolamellar Carcinoma of the Liver: Composition of the Extracellular Matrix and Expression of Cell–Matrix and Cell–Cell Adhesion Molecules
Authors: Scoazec J , Flejou J , D'Errico A , Fiorentino M , Zamparelli A , Bringuier A , Feldmann G , Grigioni W F
Published: November 2004

We have analyzed the composition of the tumor stroma and the expression of cell–matrix and cell–cell adhesion molecules in 11 cases of fibrolamellar carcinoma of the liver (FLC), in comparison with 34 cases of hepatocellular carcinoma and 8 cases of focal nodular hyperplasia. Fibrolamellar carcinoma was characterized by the presence of large amounts of tenascin in tumor stroma and by the scarce expression of basement membrane components at the contact of neoplastic clusters. Like normal hepatocytes, neoplastic cells constantly expressed the α1 integrin chain, lacked the β4 integrin chain, and coexpressed E–cadherin and the hepatocyte N–related cadherin. Abnormalities in the expression of cell adhesion molecules, including altered cadherin expression, αV integrin chain induction, and CD44 expression, were detected in the majority of cases. The composition of the tumor stroma and the pattern of expression of cell adhesion molecules in fibrolamellar carcinoma were reminiscent of those observed in grade III and grade IV hepatocellular carcinomas. Our results therefore show that, despite its slow local growth and good prognosis, fibrolamellar carcinoma expresses many characteristics usually associated with clinically aggressive malignancies. Further studies are needed to identify the factors responsible for the apparent dissociation between clinical behavior and biological characteristics in this tumor.
T–Cell Receptor Vβ Gene Utilization in Primary Biliary Cirrhosis
Authors: Mayo M J , Combes B , Jenkins R N
Published: November 2004

Semiquantitative reverse–transcriptase polymerase chain reaction (RT– PCR) was used to study the T–cell receptor (TCR) β–chain variable (Vβ) region gene families expressed by T cells in 28 patients with primary biliary cirrhosis (PBC), 20 normal controls, and 9 patients with other chronic inflammatory hepatic diseases. We hypothesized that activation and clonal proliferation of T cells would lead to biases in the T–cell repertoire of patients with PBC. Freshly harvested T cells from both peripheral blood and liver tissue were examined for evidence of biased Vβ utilization. Individuals varied considerably in their pattern of Vβ expression, but several significant differences were noted in the PBC group. In peripheral blood, the mean level of Vβ6.1,3 expression was greater in PBC patients than in normal controls. In the liver of PBC patients, the mean level of Vβ6.1,3 expression was even higher than in the peripheral blood, indicating intrahepatic accumulation of these T cells. The mean level of Vβ6.1,3 expression was not significantly different in the blood compared with liver in patients with other liver diseases. Expression of Vβ7 and Vβ13.1 was also significantly greater in the liver than in the blood of PBC patients, but similar trends were also seen in the control liver group. These data show that specific alterations in the TCR repertoire are present in the blood and liver of patients with PBC.
Chemokine Levels in Human Liver Homogenates: Associations Between Gro α and Histopathological Evidence of Alcoholic Hepatitis
Authors: Maltby J , Wright S , Bird G , Sheron N
Published: November 2004

Alcoholic hepatitis is characterized by parenchymal neutrophil infiltration. Hepatic synthesis of the neutrophil chemokine interleukin–8 (IL–8) is highly elevated in alcoholic hepatitis and levels correlate with the degree of neutrophil infiltration. The aim of this study was to further determine the spectrum of synthesis of chemokines in liver tissue from patients with alcoholic liver disease and a range of disease control subjects. Subjects were composed of 24 patients with alcoholic liver disease of whom 15 had histopathological evidence of alcoholic hepatitis (10 cirrhotic) and 9 no evidence of alcoholic hepatitis (5 cirrhotic); other controls included; normal liver (n = 6), viral hepatitis (n = 16), primary biliary cirrhosis (n = 5), acute liver failure (n = 4), and miscellaneous liver disease (n = 13). Levels of the C–X–C neutrophil chemokine GRO α and the mononuclear cell C–C chemokines: macrophage inflammatory protein 1 α, macrophage chemotactic protein 1 and RANTES, were determined by ELISA in liver homogenates. Levels of the neutrophil chemokine GRO α were specifically elevated (mean 46 pg/mg, compared with normal liver 11 pg/mg) in patients with alcoholic hepatitis. GRO α levels correlated with IL–8 levels and were higher in patients with alcoholic liver disease and parenchymal neutrophil infiltration. Hepatic RANTES was elevated in diseased liver, with the highest levels found in viral hepatitis (mean 117 pg/mg, compared with 24 pg/mg in normal liver). No significant changes in hepatic levels of macrophage inflammatory protein 1 α (MIP–1 α) or macrophage chemotactic protein 1 (MCP–1) were found. These data provide further supportive evidence that parenchymal neutrophil infiltration in alcoholic hepatitis may be determined by selective upregulation of C–X–C chemokine synthesis.
No Association Between Genetic Hemochromatosis and α1–Antitrypsin Deficienc
Authors: Fargion S , Bissoli F , Fracanzani A L , Suigo E , Sergi C , Taioli E , Ceriani R , Dimasi V , Piperno A , Sampietro M , Fiorelli G
Published: November 2004

Genetic hemochromatosis and α1–antitrypsin (AAT) deficiency are frequent in white populations. Conflicting data on the association of the two conditions and on the severity of the disease in those in whom these disorders coexist have emerged from analyses of small numbers of patients. To determine if the frequency of AAT deficiency is increased in genetic hemochromatosis, we characterized this protein by isoelectric focusing and DNA analysis in 115 Italian patients with the disease and 290 controls. The frequency of AAT deficiency in patients with genetic hemochromatosis was similar to that in controls (10% and 9%, respectively). The prevalence of cirrhosis in patients with genetic hemochromatosis with MM phenotype was 53%, compared with 58% in those with non–MM phenotype; that of hepatocellular carcinoma, occurring only in cirrhotic patients, was 22% and 28%, respectively. In conclusion, the frequency of AAT deficiency was not increased in our large series of Italian patients with genetic hemochromatosis. Patients in whom the two defects coexisted did not appear to have a more severe disease, but the limited number of subjects with non–MM phenotype does not allow a conclusive evaluation of clinical differences between them and patients with genetic hemochromatosis with MM phenotype.
Kupffer Cell Production of Cytokine–Induced Neutrophil Chemoattractant Following Ischemia/Reperfusion Injury in Rats
Authors: Hisama N , Yamaguchi Y , Ishiko T , Miyanari N , Ichiguchi O , Goto M , Mori K , Watanabe K , Kawamura K , Tsurufuji S , Ogawa M
Published: November 2004

We investigated the role of hepatic macrophages in the inflammatory response following reperfusion injury by blocking Kupffer cell phagocytosis with gadolinium chloride (GdCl3). Liver ischemia was induced in rats by occluding the portal vein for 30 minutes. A bolus of GdCl3 (7 mg/kg) was injected intravenously 1 and 2 days before surgery. The serum levels of cytokine–induced neutrophil chemoattractant (CINC) in untreated rats increased following reperfusion, peaked after 6 hours, and then gradually decreased. GdCl3 or heparin alone significantly decreased the serum levels of CINC (

P

< .05). In addition, pretreatment with GdCl3/heparin further inhibited the rise in the serum levels of CINC following reperfusion compared with those in untreated animals (

P

< .01). The in vitro production of CINC by Kupffer cells, obtained from animals pretreated with heparin or GdCl3, was significantly lower than that of cells isolated from untreated animals. Pretreatment with GdCl3/heparin further decreased CINC production by Kupffer cells compared with that of cells from animals that were pretreated with heparin or GdCl3 alone. The expression of CINC transcripts in Kupffer cells or in liver tissue peaked 3 hours after reperfusion in untreated animals. Pretreatment with heparin, GdCl3, or both significantly decreased the levels of CINC messenger RNA (mRNA) transcripts. Pretreatment with heparin, GdCl3, or GdCl3/heparin significantly decreased the number of neutrophils that accumulated in the liver 24 hours following reperfusion, compared with those in untreated animals. These results suggest that Kupffer cells release CINC and may play an important role in early neutrophil infiltration into the liver following ischemia/reperfusion.
Uncoupling of Biliary Lipid From Bile Acid Secretion by Formyl– Methionyl–Leucyl–Phenylalanine in the Rat
Authors: Mizuno K , Hoshino M , Hayakawa T , Yamada H , Nakazawa T , Inagaki T , Takeuchi T , Kunimatsu M
Published: November 2004

A neutrophil chemotactic factor N–formyl–methionyl–leucyl–phenylalanine (fMLP), produced by

Escherichia coli

under conditions of intestinal inflammation, is reported to circulate enterohepatically in the presence of experimental colitis, but its effect on bile secretion is unclear. Therefore, we investigated the effect of fMLP on bile secretion in a single–pass isolated perfused rat liver system. Infusion of fMLP at different concentrations (2 μmol/L, 10 μmol/L, and 20 μmol/L) into the portal vein resulted in excretion into bile in the native form, independent of sodium taurocholate (1 μmol/min) infusion. Excretion of fMLP increased dose dependently, and approximately 12% of the infused dose was detected at each concentration. With constant infusion of sodium taurocholate (1 μmol/min), fMLP (20 μmol/L) increased bile flow but decreased phospholipid and cholesterol secretion. Bile acid secretion was not affected. Phospholipid/bile acid molar ratios decreased from 0.069 ± 0.002 to 0.038 ± 0.002, and cholesterol/bile acid molar ratios decreased from 0.0074 ± 0.0009 to 0.0029 ± 0.0008. Thus, administration of fMLP resulted in the uncoupling of biliary excretion of phospholipid and cholesterol from that of bile acids; this effect proved reversible. The increase in bile flow caused by fMLP infusion appeared to result from osmotic choleresis. When 25 mg of horseradish peroxidase, a conventional marker of transcytotic vesicle transport pathway, was infused for 1 minute as a pulse load into the portal vein after continuous infusion of taurocholate, its late peak excretion was reduced by fMLP (10 μmol/L) from 9.59 ± 1.09 to 6.05 ± 0.66 (ng/g liver). Gel–permeation chromatography of bile showed a specific association of fMLP with bile acids. These results suggest an uncoupling of biliary lipids from bile acids by fMLP because of inhibition of transcellular vesicle transport and interaction between fMLP and bile acid micelles in the bile canaliculus.
Hla Class Ii Genes in Chronic Hepatitis C Virus–Infection and Associated Immunological Disorders
Authors: Congia M , Clemente M G , Dessi C , Cucca F , Mazzoleni A P , Frau F , Lampis R , Cao A , Lai M E , De Virgiliis S
Published: November 2004

To investigate the factors that may confer susceptibility or protection to hepatitis C virus (HCV) infection and to HCV–associated immunological disorders, we designed two studies on 420 Sardinian transfusion–dependent thalassemia patients followed in our department in Cagliari since 1974. The first one was an epidemiological survey aimed to evaluate the prevalence of HCV infection and HCV–associated immunological disorders. In the second study, the distribution of different HLA class II genes was examined by DNA analysis in 116 HCV positive patients, 30 HCV negative patients, and 606 healthy controls. Three hundred fourteen patients became infected with HCV (74.7%) after 5.6 ± 2.8 years of regular transfusion program. Mixed cryoglobulinemia, purpura, arthritis, proteinuria, decreased complement levels, rheumatoid factor and anti–GOR, smooth muscle antibody (SMA), anti–nuclear antibody (ANA), and liver, kidney microsome (LKM) autoantibodies were significantly more represented in HCV positive patients than in negative ones (

P

< .05). A significant increase of HLA class II DR2 subtype (DRB1*1601,DQB1*0502) was observed in a group of 30 HCV negative patients who despite 10.3 ± 2.2 years in a regular blood transfusion program did not show any evidence of HCV infection (

P

c < .0092). Our results represent clear evidence for a relationship between HCV infection and immune extrahepatic abnormalities. A gene(s) located in the human major histocompatibility complex (MHC) region may play an important role in conferring protection against HCV infection.
Evidence That the Hla Dqa1*03 Allele Confers Protection From Chronic Hcv–Infection in Northern European Caucasoids
Authors: Tibbs C , Donaldson P , Underhill J , Thomson L , Manabe K , Williams R
Published: November 2004

In the search for factors which may influence susceptibility to and outcome from chronic hepatitis C virus (HCV) infection, few studies have considered the influence of host genes. In the present investigation we have performed HLA DRB1, DQA1, DQB1, and DPB1 genotyping on 104 northern European patients with chronic HCV infection and 177 racially and geographically matched controls. Three HLA class II alleles, DRB1*0403, DQA1*03, and DQB1*0302 were present at a significantly lower frequency in patients compared with controls (4.9% vs. 13%, 20.7% vs. 41.2%, and 11.4% vs. 30.5%, respectively) though only two DQB1*0302 and DQA1*03 were significant after correction for multiple testing (pc = 0.038, and pc = 0.046, respectively). No further HLA associations with chronic HCV infection were observed and there was no correlation between stage of disease and HLA genotype. These data provide the first suggestion that susceptibility to chronic HCV infection may be influenced by the hosts' HLA DQ alleles.
Humoral and Cellular Immune Responses to Hepatitis B Vaccination in Hepatitis B Surface Antigen–Carrier Children Who Cleared Serum– Hepatitis B Surface Antigen
Authors: Hsu H , Chang M , Hsieh R , Ni Y , Chi W
Published: November 2004

The immune responses to hepatitis B vaccine were studied in 11 hepatitis B surface antigen (HBsAg) carrier children who had cleared HBsAg but failed to develop hepatitis B surface antigen antibodies (anti–HBs) in sera (group 1), 5 HBsAg carrier children who had cleared HBsAg and developed detectable anti–HBs in sera (group 2), and 5 healthy subjects seronegative for all hepatitis B virus (HBV) markers (group 3). After receiving three doses of HB vaccine, group 1 subjects failed to develop detectable anti–HBs. Subsequently, each subject of the three groups was given one dose of the same vaccine for a cellular immunity study, and a measurable proliferation of peripheral blood mononuclear cells (PBMC) to HBsAg was detected in 1 of 8 (12.5%), 0 of 5, and 4 of 5 (80%) of the cases in each group, respectively, after vaccination. The removal of CD8+ cells enhanced the HBsAg blastogenic response in group 3 but did not reverse the unresponsiveness in group 1 and group 2 subjects. The addition of interleukin (IL)–2 in culture reversed unresponsiveness in all cases except one case in group 1. Compared with before vaccination, PBMC from group 2 subjects produced significantly less interferon gamma (IFN–gamma) and more IL–4 in response to HBsAg after vaccination, a cytokine response not observed in group 1 subjects. HLA typing indicated that 3 of 10 patients in group 1 (30%) and 1 of 5 patients in group 2 (20%) had HLA–DRw14–DRw52, a marker previously linked to low anti–HBs response to hepatitis B vaccine in Taiwan. We conclude that the underlying causes of poor anti–HBs response in group 1 subjects are multifactorial, including specific failure of antigen presentation or T–cell activation, or the lack of T helper (Th)2 cell–like response to HBsAg. HLA–DRw14–DRw52 does not confer absolute nonresponsiveness to HBsAg. These patients are not benefited by hepatitis B immunization.
Preoperative Portal Vein Embolization for Extension of Hepatectomy Indications
Authors: de Baere T , Roche A , Elias D , Lasser P , Lagrange C , Bousson V
Published: November 2004

To render hepatectomy feasible in patients with an initially deficient volume of the future remnant liver (FRL), we redistributed portal blood flow rich in hepatotrophic substances toward the FRL. Redistribution was achieved with preoperative portal vein embolization (POPE) feeding the future resected liver. POPE was performed in 31 patients, under fluoroscopic guidance, via a percutaneous access. POPE was well tolerated and surgery was practicable in 24 patients without severe postoperative liver failure. Seven operations were cancelled, but only one due to insufficient hypertrophy of the FRL. FRL volume values were 90 to 560 mL (mean 260 mL) before POPE and 160–783 mL (mean 443 mL) after POPE, which represents a median increase of 79% ± 50%. Hypertrophy of the FRL was 90% ± 52% after 30 days with cyanoacrylate, 53% ± 6% after 43 days with Gelfoam, and 44% ± 30% after 35 days with coils. Slight shrinkage was obtained in the volume of the embolized liver, for which resection was planned. Overall survival was 2–62 months (mean 26 months), disease–free survival was 0–60 months (mean 19 months), and 7 patients are disease–free and alive 14 to 60 months (mean 43 months) after surgery. Although exclusively applicable in a limited subset of patients, POPE widens the possibilities of curative hepatectomies, because it induces sound hypertrophy of unembolized liver segments. Cyanoacrylate seems to ensure better and faster hypertrophy.
Increased Nitric Oxide Synthase Expression in Arterial Vessels of Cirrhotic Rats With Ascites
Authors: Morales-Ruiz M , Jimenez W , Perez-Sala D , Ros J , Leivas A , Lamas S , Rivera F , Arroyo V
Published: November 2004

Arterial vasodilatation is thought to play a major role in the pathogenesis of systemic hemodynamics and renal disturbances occurring in cirrhotic patients. Recent investigations suggest that an increased vascular nitric oxide (NO) production could be implicated in this abnormality. The current study assessed whether increased expression of inducible and/or endothelial nitric oxide synthase (iNOS and eNOS, respectively) occurs in arterial vessels of cirrhotic rats. The investigation was performed in thoracic and abdominal aortas and mesenteric arteries of 10 control rats and 16 cirrhotic rats with ascites. iNOS and eNOS messenger RNA (mRNA) expression were evaluated by polymerase chain reaction and ribonuclease protection assay, respectively. Endothelial NOS protein expression was assessed by Western blot. No iNOS mRNA was detected in arterial vessels of control rats. In contrast iNOS mRNA was consistently detected in all arteries of cirrhotic rats with ascites, the weakest signal being observed in the thoracic aorta and the strongest in the mesenteric artery. Enhanced eNOS mRNA abundance was found in the aorta of cirrhotic animals as compared with controls. Higher eNOS protein expression was noted in the thoracic aorta of cirrhotic rats. These results indicate the existence of increased eNOS and iNOS expression in arterial vessels of cirrhotic rats, suggesting that transcriptional activation of vascular NOSs and the associated nitric oxide hyperproduction may be of major importance in the pathogenesis of arterial vasodilation in cirrhosis.
Transverse Movement of Spin–Labeled Phospholipids in the Plasma Membrane of A Hepatocytic Cell Line (Hepg2): Implications for Biliary Lipid Secretion
Authors: Muller P , Pomorski T , Porwoli S , Tauber R , Herrmann A
Published: November 2004

The redistribution of spin–labeled phospholipid analogs across the plasma membrane of HepG2 cells, either in suspension or grown as monolayers, was investigated. After incorporation into the outer membrane leaflet spin–labeled aminophospholipids phosphatidylserine (PS) and phosphatidylethanolamine (PE) moved rapidly to the inner monolayer, whereas the analog of phosphatidylcholine (PC) disappeared more slowly from the outer leaflet. The fast, inward movement of the aminophospholipids was abolished after adenosine triphosphate (ATP)– depletion of cells, suggesting the presence of an aminophospholipid translocase in the plasma membrane of these cells. Compared with human red blood cells, the activity of the aminophospholipid translocase is two orders of magnitude higher in HepG2 cells. From these data, a transverse phospholipid asymmetry can be inferred with the aminophospholipids mainly concentrated on the inner monolayer and the choline–containing phospholipids on the outer leaflet. The relevance of the enrichment of PC in the outer membrane leaflet for the formation and composition of the bile is discussed.
Does hippocampus associate discontiguous events? Evidence from event‐related fMRI
Authors: Luo Jing , Niki Kazuhisa
Published: November 2004

To examine the hypothesis that the hippocampus is necessary to overcome temporal or spatial “discontiguity” (Wallenstein et al., Trends Neurosci 1998; 21:317–323), subjects were imaged by functional magnetic resonance imaging (fMRI) when they were making judgments as to whether two words were semantically related. Two words were presented, either at the same time (the Simultaneous Presentation Condition) or one after the other with a short unfilled rest period (the Delayed Presentation Condition). The latter condition, relative to the former, was proposed to involve the process of “discontiguity association.” Event‐related fMRI results of eight subjects showed that, relative to the binding of simultaneously presented words, the binding of delay presented words was associated with left hippocampus activity. This result provided direct neuroimaging evidence for the role of the hippocampus in “discontiguity association.” © 2004 Wiley‐Liss, Inc.
Serenoa repens (Permixon®) inhibits the 5α‐reductase activity of human prostate cancer cell lines without interfering with PSA expression
Authors: Habib Fouad K. , Ross Margaret , K.H. Ho Clement , Lyons Valerie , Chapman Karen
Published: November 2004

The phytotherapeutic agent Serenoa repens is an effective dual inhibitor of 5α‐reductase isoenzyme activity in the prostate. Unlike other 5α‐reductase inhibitors, Serenoa repens induces its effects without interfering with the cellular capacity to secrete PSA. Here, we focussed on the possible pathways that might differentiate the action of Permixon from that of synthetic 5α‐reductase inhibitors. We demonstrate that Serenoa repens, unlike other 5α‐reductase inhibitors, does not inhibit binding between activated AR and the steroid receptor‐binding consensus in the promoter region of the PSA gene. This was shown by a combination of techniques: assessment of the effect of Permixon on androgen action in the LNCaP prostate cancer cell line revealed no suppression of AR and maintenance of PSA protein expression at control levels. This was consistent with reporter gene experiments showing that Permixon failed to interfere with AR‐mediated transcriptional activation of PSA and that both testosterone and DHT were equally effective at maintaining this activity. Our results demonstrate that despite Serenoa repens effective inhibition of 5α‐reductase activity in the prostate, it did not suppress PSA secretion. Therefore, we confirm the therapeutic advantage of Serenoa repens over other 5α‐reductase inhibitors as treatment with the phytotherapeutic agent will permit the continuous use of PSA measurements as a useful biomarker for prostate cancer screening and for evaluating tumour progression. © 2004 Wiley‐Liss, Inc.
Retracted: Frequent genetic and biochemical alterations of the PI 3‐K/AKT/PTEN pathway in head and neck squamous cell carcinoma
Authors: Pedrero Juana Maria Garcia , Carracedo Dario Garcia , Pinto Cristina Muñoz , Zapatero Agustín Herrero , Rodrigo Juan Pablo , Nieto Carlos Suarez , Gonzalez Maria Victoria
Published: November 2004

We investigated the status of the PI 3‐kinase/AKT/PTEN signaling pathway in a series of 117 head and neck squamous cell carcinomas (HNSCC) in a search for molecular alterations in genes/proteins with potential prognostic value. For this purpose, PIK3CA and AKT2 gene amplification was assessed by multiplex and Quantitative Real‐Time PCR. Protein expression of AKT, p‐AKT, p110α and PTEN was determined by Western blot. PTEN allelic loss was evaluated by microsatellite analysis. PTEN‐exon 5 was screened for point mutations by PCR‐SSCP. Homozygous deletions were determined by multiplex PCR. PIK3CA gene was amplified in 43/117 (37%) fresh tumor samples, a frequency that did not differ from that found in archival premalignant tissues: 15/38 (39%); 12/40 (30%) fresh tumors harbored AKT2 gene amplification. AKT was found activated in 6/36 (17%) fresh tumor samples, when compared to their normal tissue counterparts. Of these 6 cases, 1 showed p110α overexpression and 5 displayed PTEN protein downregulation. Neither allelic loss (found in 11/77 informative cases) nor point mutations or homozygous deletions accounted for the reduced PTEN protein expression observed in our tumor series. The histologically normal mucosa of 4 patients displayed some of the molecular alterations analyzed. Dysregulation of the PI 3‐K/AKT/PTEN pathway might contribute to early HNSCC tumorigenesis and might constitute a potential clinical target. Overall, 17/36 (47%) cases showed at least 1 of the molecular alterations studied here, which makes the PI 3‐kinase‐initiated signaling pathway one of the most frequently altered in HNSCC. © 2004 Wiley‐Liss, Inc.
Self‐reported health and use of health care services in long‐term cancer survivors
Authors: Nord Carina , Mykletun Arnstein , Thorsen Lene , Bjøro Trine , Fosså Sophie D.
Published: November 2004

Owing to an increasing number of long‐term cancer survivors, the use of health care services and somatic health problems were compared between cancer survivors and a noncancer population. Data from the Nord‐Trøndelag Health Survey 2 (HUNT 2, 1995–1997) was merged with the Cancer Registry of Norway. Six cancer subgroups were constructed with diagnosis 5 years prior HUNT 2: testicular cancer (n= 59), colorectal cancer (n= 175), prostate cancer (n= 87), breast cancer (n= 258), gynaecological cancer (n= 153) and lymphoma/leukaemia (n= 83). For each cancer survivor 3 matched noncancer controls were selected from the HUNT 2 survey. The prevalence of common health problems, use of health care services and unfavourably life style parameters were compared between the 2 groups. Cancer survivors used health care services and received social welfare benefits more often than the controls. There was an increased risk of perceiving poor health after a history of cancer. Common health problems and/or unfavourable life style parameters could not explain poor health or the increased use of health care services among cancer survivors. Further studies are needed to investigate the reasons for increased use of health care services and perceived poor health in cancer survivors. © 2004 Wiley‐Liss, Inc.
Accelerated bone repair with the use of a synthetic BMP‐2‐derived peptide and bone‐marrow stromal cells
Authors: Saito Atsuhiro , Suzuki Yoshihisa , Ogata Shin‐Ichi , Ohtsuki Chikara , Tanihara Masao
Published: November 2004

A novel synthetic peptide corresponding to BMP‐2 residues 73–92 that can induce bone formation and can form a conjugate with a carrier to localize its effect has been reported previously. The synthetic peptide was bound to a BMP‐2–specific receptor, and it elevated both the alkaline phosphatase activity and the osteocalcin mRNA in the murine multipotent mesenchymal cell line, C3H10T1/2. The 73–92 peptide also induced ectopic bone formation when conjugated to a covalently crosslinked alginate gel and implanted into a rat's calf muscle. Here, it is reported that the 73–92 peptide‐conjugated alginate gel particles significantly promoted the repair of rat tibial bone defects, whereas the alginate gel sponge that the peptide was conjugated with was less effective. Further acceleration and denser bone regeneration was achieved when the 73–92 peptide‐conjugated alginate gel particles were coimplanted with syngeneic rat bone‐marrow stromal cells. Therefore, the 73–92 peptide can induce differentiation of osteoblast precursor cells into osteoblasts, and can activate osteoblasts to promote the repair of bone defects. © 2004 Wiley Periodicals, Inc. J Biomed Mater Res 72A: 77–82, 2005
Adenylyl cyclase‐cAMP system inhibits thrombin‐induced HSP27 in vascular smooth muscle cells
Authors: Hirade Kouseki , Tanabe Kumiko , Niwa Masayuki , Ishisaki Akira , Nakajima Keiichi , Nakamura Mitsuhiro , Sugiyama Tadashi , Katagiri Yoshihiro , Kato Kanefusa , Kozawa Osamu
Published: November 2004

We previously reported that thrombin stimulates the induction of heat shock protein (HSP) 27 via p38 mitogen‐activated protein (MAP) kinase activation in aortic smooth muscle A10 cells. In the present study, we investigated the effect of the adenylyl cyclase‐cAMP system on the thrombin‐stimulated induction of HSP27 in A10 cells. Forskolin, a direct activator of adenylyl cyclase, reduced the thrombin‐induced p38 MAP kinase phosphorylation, and significantly suppressed the thrombin‐stimulated accumulation of HSP27. However, dideoxyforskolin, a forskolin derivative that does not activate cAMP, failed to suppress the HSP27 accumulation. Furthermore, dibutyryl‐cAMP (DBcAMP), a permeable analog of cAMP, significantly suppressed the accumulation of HSP27. On the other hand, calphostin C, an inhibitor of protein kinase C (PKC), reduced the thrombin‐induced p38 MAP kinase phosphorylation, and significantly suppressed the thrombin‐stimulated accumulation of HSP27. Moreover, forskolin reduced the p38 MAP kinase phosphorylation induced by the 12‐O‐tetradecanoylphorbol‐13‐acetate (TPA), a PKC‐activating phorbol ester, and significantly suppressed the TPA‐stimulated accumulation of HSP27. These results indicate that adenylyl cyclase‐cAMP system has an inhibitory role in thrombin‐stimulated HSP27 induction in aortic smooth muscle cells, and the effect seems to be exerted on the thrombin‐induced PKC‐ p38 MAP kinase signaling pathway. © 2004 Wiley‐Liss, Inc.
Do bcl‐2 and survivin help distinguish benign from malignant B‐cell lymphoid aggregates in bone marrow biopsies?
Authors: Gandhi Amish M. , Ben‐Ezra Jonathan M.
Published: November 2004

Bcl‐2 and survivin are cellular proteins that are known to be inhibitors of apoptosis and are commonly found in malignant tissues, including lymphomas. In previous studies, it has been shown that staining for bcl‐2 can help distinguish between benign and malignant lymphoid aggregates in bone marrow biopsies. To determine whether staining for survivin expression in lymphoid aggregates can aid investigators in making this clinically important distinction, we stained bone marrow biopsies from 10 patients with benign lymphoid aggregates, and 15 malignant ones derived from B cells (six mantle cell, four follicular cells, two diffuse large cell, two small lymphocytic cell, and one marginal zone lymphoma) with antibodies to CD3, CD20, bcl‐2, and survivin by an indirect immunoperoxidase technique. Whereas staining for bcl‐2 was significantly stronger in the malignant lymphoid aggregates (P=0.001), both the control and malignant cases were almost uniformly negative for survivin expression. Only three cases (two mantle cell and one small lymphocytic lymphoma) showed very faint expression of survivin. Although bcl‐2 and survivin both act to inhibit apoptosis, their expressions do not parallel each other. Survivin is not significantly expressed in either benign or malignant bone marrow aggregates, and therefore measuring its expression does not help distinguish benign from malignant B‐cell bone marrow lymphoid aggregates. J. Clin. Lab. Anal. 18:285–288, 2004. © 2004 Wiley‐Liss, Inc.
Insulin‐like growth factors in patients with liver cysts
Authors: Nedić Olgica , Nikolić J. Anna , Baričević Ivona , Jovanović Biljana , Ilić Nataša
Published: November 2004

Insulin‐like growth factors (IGFs) play an important role in cell growth and differentiation, and the liver is the main source of IGFs and IGF‐binding proteins (IGFBPs) that appear in the circulation. The effect of liver cysts on the circulating IGF system was studied in this work. Serum concentrations of IGF‐I and ‐II were measured by radioimmunoassay, IGFBP patterns were characterised by ligand‐affinity and immunoblotting, and a lectin‐binding assay was used to investigate the glyco component of IGFBP‐3 complexes. IGF‐I and ‐II concentrations in patients with cysts were significantly lower compared to those in healthy individuals (P<0.0001 and P<0.01, respectively), and the decrease was related to age but not sex. The overall mean concentrations of IGF‐I and ‐II were not significantly different whether the cysts were caused by Echinococcus granulosus, cross‐reactive pathologies, or some other factor. IGFBP profiles correlated with the amount of IGF present: patients with lower IGF‐I concentrations expressed decreased IGFBP‐3 and elevated IGFBP‐2 levels. Increased IGFBP‐3 proteolytic activity in the patients' blood was not detected by immunoblotting. In the lectin‐binding assay, IGFBP‐3 complexes in the circulation of patients demonstrated reactivity similar to that in healthy persons, suggesting that the overall structure of the saccharide moieties of the IGFBP‐3 complexes was not significantly altered due to liver cyst formation. J. Clin. Lab. Anal. 18:299–304, 2004. © 2004 Wiley‐Liss, Inc.
Five polymorphisms in gene candidates for cardiovascular disease in Afro‐Brazilian individuals
Authors: Sakuma Tatsuya , Hirata Rosario D.C. , Hirata Mario H.
Published: November 2004

Associations of polymorphisms in the angiotensin I‐converting enzyme (ACE), apolipoprotein B (APOB) and apolipoprotein E (APOE) genes with hypertension and variations in lipid serum levels were evaluated in 184 Afro‐Brazilians with a familial history of coronary artery disease (CAD). ACE (Ins/Del) and APOB (Ins/Del, XbaI, and EcoRI) and APOE (HhaI) polymorphisms were determined by polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP) analyses on agarose, and polyacrylamide gel electrophoresis. Serum lipids were measured by means of routine enzymatic assays. The results showed a high frequency of hypertension (44%) in Afro‐Brazilians that was increased in subjects >40 years old and those with a blood mass index (BMI) higher than 25 kg/m2 (P<0.001). The ACE Del allele was associated with hypertension in men >40 years old (P<0.05). APOE (HhaI) and APOB (XbaI and Ins/Del) polymorphisms were not associated with hypertension or variations in serum concentrations of lipids, while subjects with the APOB E− allele had higher low‐density lipoprotein cholesterol (LDL‐C) levels than E+ carriers (P<0.05). These results suggest that ACE Ins/Del polymorphism is associated with hypertension, and APOB EcoRI polymorphism is associated with LDL‐C variation in Afro‐Brazilians. J. Clin. Lab. Anal. 18:309–316, 2004. © 2004 Wiley‐Liss, Inc.
Lignans in resin of Araucaria angustifolia by gas chromatography/mass spectrometry
Authors: Yamamoto Shuichi , Otto Angelika , Simoneit Bernd R. T.
Published: November 2004

Total extract of resin from Araucaria angustifolia was analyzed by gas chromatography/mass spectrometry and 32 lignans were identified. Lignan acetates are present in the resin and consist of four secoisolariciresinol acetates, six lariciresinol acetates, two 7′‐hydroxylariciresinol acetates and an isolariciresinol acetate, which have hitherto not been reported in the plant kingdom. Shonanin and 7′‐hydroxylariciresinol type lignans are also present in A. angustifolia resin. Lignans containing syringyl moieties, characteristic for angiosperms, occur in the resin and consist of 5‐methoxylariciresinol‐9‐acetate, 5′‐methoxylariciresinol‐9‐acetate, 5‐methoxypinoresinol dimethyl ether and 5‐methoxypinoresinol. This is noteworthy because syringyl moieties have only been reported for Thuja species (Cupressaceae) among the gymnosperms. The mass spectra of the various lignan trimethylsilyl derivatives are discussed with the interpretations of the fragmentation patterns. Copyright © 2004 John Wiley & Sons, Ltd.