bluuebunny/crossref_metadata_2025 huggingface dataset:
Glial regulation of nonsynaptic extracellular glutamate in the substantia nigra
Authors: Rodríguez Díaz Manuel , Alonso Teofilo Jorge , Perdomo Diaz Juan , Gonzalez Hernández Tomás , Castro Fuentes Rafael , Sabate Magdalena , Garcia Dopico Jose
Published: November 2004

GLU is the main neurotransmitter in the brain, where it induces a synaptic excitatory action. There is recent evidence for an extracellular nonsynaptic GLU (EnS‐GLU) pool in different brain nuclei that, released from glial cells, may act on extrasynaptic GLU receptors of cells located far from the position in which it was released. In the present work, the EnS‐GLU pool was studied with microdialysis in the rat substantia nigra (SN). We observed an EnS‐GLU pool that increased in a Ca2+‐dependent manner during cell depolarization. The selective alteration of with methionine sulfoximide (MSO) and fluorocitrate induced marked modifications in EnS‐GLU suggesting that EnS‐GLU is dependent on glial cells. Glutamine administration increased GLU, suggesting that neurons are also involved in EnS‐GLU modulation. GLU administered in the rostral SN showed a long‐distance diffusion to the caudal SN. The ionotropic GLU receptors agonist N‐methyl‐D‐aspartate and kainate and the metabotropic GLU receptors agonist ACPD increased EnS‐GLU and decreased extracellular glutamine. Taken together, these data indicate that nigral glia releases GLU, which probably performs a volume transmitter role. © 2004 Wiley‐Liss, Inc.
Gliosis and brain remodeling after treatment of stroke in rats with marrow stromal cells
Authors: Li Yi , Chen Jieli , Zhang Chun Ling , Wang Lei , Lu Dunyue , Katakowski Mark , Gao Qi , Shen Li Hong , Zhang Jing , Lu Mei , Chopp Michael
Published: November 2004

The long‐term (4‐month) responses to treatment of stroke in the older adult rat, using rat bone marrow stromal cells (MSCs), have not been investigated. Retired breeder rats were subjected to middle cerebral artery occlusion (MCAo) alone, or injected intravenously with 3 × 106 MSCs, at 7 days after MCAo. Functional recovery was measured using an adhesive‐removal patch test and a modified neurological severity score. Bromodeoxyuridine, a cell proliferation marker, was injected daily for 14 before sacrifice. Animals were sacrificed 4 months after stroke. Double immunostaining was used to identify cell proliferation and cell types for axons, astrocytes, microglia, and oligodendrocytes. MSC treatment induced significant improvement in neurological outcome after MCAo compared with control rats. MSC treatment reduced the thickness of the scar wall (P < 0.05) and reduced the numbers of microglia/macrophages within the scar wall (P < 0.01). Double staining showed increased expression of an axonal marker (GAP‐43), among reactive astrocytes in the scar boundary zone and in the subventricular zone in the treated rats. Bromodeoxyuridine in cells preferentially colocalized with markers of astrocytes (GFAP) and oligodendrocytes (RIP) in the ipsilateral hemisphere, and gliogenesis was enhanced in the subventricular zone of the rats treated with MSCs. This is the first report to show that MSCs injected at 7 days after stroke improve long‐term neurological outcome in older animals. Brain tissue repair is an ongoing process with reactive gliosis, which persists for at least 4 months after stroke. Reactive astrocytes responding to MSC treatment of ischemia may also promote axonal regeneration during long‐term recovery. © 2004 Wiley‐Liss, Inc.
The hepatic microcirculation in the isolated perfused human liver
Authors: Villeneuve J , Dagenais M , Huet P , Roy A , Lapointe R , Marleau D
Published: November 2004

In cirrhosis, capillarization of sinusoids could result in impaired exchanges between the hepatocytes and the blood perfusing the liver and contribute to liver failure irrespective of the metabolic capacity of the liver. To characterize anomalies of the hepatic microcirculation, we used the multiple-indicator dilution approach in isolated perfused livers obtained from patients with cirrhosis at the time of transplantation, and from organ donors with normal or near-normal livers or hepatic steatosis. In organ donors, the sinusoidal volume and the permeability of sinusoids to albumin, sucrose, and water were found to be comparable to that of normal dog and rat livers. The sinusoidal volume and the extravascular volume (EVV) accessible to diffusible tracers were larger after hepatic artery than after portal vein injection, probably because of an unshared arterial sinusoidal bed. In cirrhotic livers, two kinds of alterations were found: the appearance of a barrier between the sinusoids and the hepatocytes (capillarization) and intrahepatic shunts. The extravascular space accessible to albumin decreased with increasing severity of cirrhosis, and the diffusion of sucrose in the space of Disse showed a barrier-limited pattern, instead of the normal flow-limited behavior. In cirrhotic livers, a correlation was found between the hepatic extraction of indocyanine green (ICG) and the extravascular space accessible to albumin (r = .84, P < .05), suggesting that the impaired access of this protein-bound dye to the hepatocyte surface contributed to its impaired elimination. Intrahepatic shunts were found between portal and hepatic vein (21% ± 16% of portal flow), but not between hepatic artery and hepatic veins. We conclude that (1) the behavior of diffusible tracers in human livers with normal liver architecture is comparable to that reported in normal animals; (2) the permeability of sinusoids in cirrhotic livers is abnormal, (3) permeability changes are related to changes in liver function in cirrhosis.
The hematologic consequences of transjugular intrahepatic portosystemic shunt
Authors: Sanyal A J , Freedman A M , Purdum P P , Shiffman M L , Luketic V A
Published: November 2004

Transjugular intrahepatic portosystemic shunts (TIPS) are a recent innovation in the management of portal hypertension. In 1992, we had previously described an instance of severe hemolysis associated with this procedure. This study was undertaken to define and quantify the true incidence of TIPS-associated hemolysis and its clinical spectrum, as well as to test the hypothesis that portal decompression by TIPS would ameliorate hypersplenism in patients with portal hypertension. A total of 60 patients undergoing TIPS for prevention of recurrent variceal hemorrhage (n = 40) or refractory ascites (n = 20) were studied. Forty patients with cirrhosis who were followed concurrently served as controls. At entry, both groups were comparable with the exception of increased ascites in the TIPS group. A total of 7 instances of intravascular hemolysis were identified in 60 TIPS patients, whereas none occurred in controls. Of these, 4 patients were asymptomatic and detected on routine laboratory testing. Hemolysis led to a greater than 4-g/dL decrease in hemoglobin in 2 patients, 2- to 3-g/dL decrease in 2 others and a 3- to 4-gm/dL decrease in 1 patient. Two patients were able to compensate for hemolysis and did not develop anemia. In all but 1 case, the findings of hemolysis subsided by 12 to 15 weeks; in 1 patient, orthotopic liver transplantation was associated with resolution of the hemolysis. Overall, no significant changes in white blood cell or platelet counts were observed in patients undergoing TIPS despite adequate portal decompression. We conclude that TIPS-induced hemolysis occurs in approximately 10% of subjects. However, it is self-limited and rarely requires intervention. Potential mechanisms of such hemolysis are discussed. TIPS is also not recommended as a means of improving platelet counts in patients with severe hypersplenism.
Time course of histological progression in primary biliary cirrhosis
Authors: Locke G R , Therneau T M , Ludwig J , Dickson E R , Lindor K D
Published: November 2004

Histological staging is used for stratification and assessment of treatment efficacy in therapeutic trials for primary biliary cirrhosis (PBC). Knowledge of the rate of progression of the histological changes would be helpful in the design (duration) and conduct of clinical trials. The histological stages were recorded for liver biopsies performed annually on 222 patients during a randomized, placebo- controlled clinical trial in which therapy with D-penicillamine (DPCA) was shown to be ineffective. These data were analyzed using a Markov model to describe the time course of histological progression in PBC. At study entry, 15 patients were stage I, 56 were stage II, 96 were stage III, and 55 were stage IV. Histological progression was observed after 1 year in 41%, 43%, and 35% of the patients, and after 2 years in 62%, 62%, and 50% of the patients who were stage I, stage II, and stage III at entry, respectively. After 4 years biopsies showed cirrhosis in 31% and 50% of the patients in stage I and stage II at entry, respectively. A minority (20%) of the precirrhotic patients showed histological stability; sustained histological regression was rarely observed (2%). Our data suggest that a majority of patients with PBC will progress histologically within 2 years. The distribution of histological stages over time may be helpful in determining the number of patients and length of time necessary to appreciate a treatment effect on histological progression in clinical trials for PBC.
Hyperinsulinemia in preascitic cirrhosis: Effects on systemic and renal hemodynamics, sodium homeostasis, forearm blood flow, and sympathetic nervous activity
Authors: Wong F , Logan A , Blendis L
Published: November 2004

Insulin has been shown to be vasodilatory and antinatriuretic and to stimulate sympathetic nervous activity independent of hypoglycemia in healthy normal subjects. It is hypothesized that hyperinsulinemia, which is commonly observed in cirrhosis, may in part be responsible for the systemic vasodilatation, sympathetic activation, and sodium retention in these patients. The aims of this study, in preascitic cirrhotics, were as follows: (1) to document baseline hyperinsulinemia and its effects on sodium handling, forearm and renal circulations, and sympathetic nervous activity; (2) to determine if pharmacological increases in plasma insulin levels would result in an exaggeration of these physiological effects. Seven male, nonobese, well-compensated, preascitic cirrhotic patients were studied, after being maintained on a 150 mmol sodium per day diet for 7 days, firstly at baseline level, followed by increasing doses of insulin from 10 to 1,200 mU/m2/min using the euglycemic clamp technique. Systemic and renal hemodynamics, urinary sodium excretion, plasma norepinephrine, and forearm blood flow (FBF) were measured at the end of baseline and each hyperinsulinemic period. Baseline measurements in the cirrhotics, when compared with our laboratory standards obtained from a comparable group of male healthy normals, showed significant hyperinsulinemia (P=.01), associated with significantly higher FBF (P=.02), and glomerular filtration rate (GFR) (P=.02), as well as significantly reduced urinary volume (P=.04) and fractional excretion of sodium (P=.04). Insulin infusions in the cirrhotics produced no further sodium retention, but further forearm vasodilatation occurred at doses > or = 10 mU/m2/min. In contrast, there was no further renal vasodilatation except at very high pharmacological levels of insulin together with an unchanged GFR, natriuresis, and diuresis. Hyperinsulinemia produced no significant effects on the sympathetic nervous activity. In conclusion, these results suggest that hyperinsulinemia may be implicated in the glomerular hyperfiltration and sodium retaining tendency of preascitic cirrhotic patients with glucose intolerance. The ability of the kidneys to escape from the sodium retaining effects may serve as an in-built physiological regulatory mechanism on sodium homeostasis.
Fibronectin in human bile fluid for diagnosis of malignant biliary diseases
Authors: Körner T , Kropf J , Hackler R , Brenzel A , Gressner A M
Published: November 2004

In a preliminary study, we demonstrated a strong association between the concentration of the glycoprotein fibronectin (FN) in human bile fluid and the presence of malignant biliary diseases. We now present the results of measurements of total FN (tFN) and cellular FN (cFN) within a larger group of 71 patients. Bile fluid was collected during routine endoscopic retrograde cholangiography or by transhepatic puncture, respectively, from patients admitted for examination/treatment of biliary obstruction. Determination of tFN in bile was performed using a previously described time-resolved fluorescence immunoassay (TRFIA). For cFN, a newly developed TRFIA, using a specific monoclonal antibody for the EDA epitope of cFN, was applied. Within the noncarcinoma group of patients (n=50), consistently low concentrations of tFN (median = 5 ng/mL) were found. In most of these cases, the corresponding concentrations of cFN were below the detection limit (2.6 ng/mL) of this assay. Highly significantly elevated concentrations were found for both tFN (median = 1,220 ng/mL) and cFN (median = 243 ng/mL) in the carcinoma group (n = 21) in comparison with the noncarcinoma group (P ≤ .01). By adopting cutoff values of 60 ng/mL for tFN and > 0 ng/mL for cFN, diagnostic sensitivities for carcinoma of the biliary tract of 0.89 and 0.92, and specificities of 0.96 and 0.98, respectively, were computed. FN in bile fluid is suggested as a sensitive, specific, and easily determined marker for differential diagnosis of malignant and benign diseases of the biliary tract.
Interphase cytogenetic studies of human hepatocellular carcinomas by fluorescent in situ hybridization
Authors: Hamon-Benais C , Ingster O , Terris B , Couturier-Turpin M , Bernheim A , Feldmann G
Published: November 2004

Although numerous allelic chromosome losses have been reported in hepatocellular carcinomas (HCC), chromosome analysis by cytogenetic methods has rarely been performed in these tumors, unlike other solid malignant tumors. The purpose of the current study was to analyze primary liver tumors by conventional cytogenetic methods and by a new molecular cytogenetic technique, called fluorescent in situ hybridization (FISH), a technique that has been recently proposed to count the number of chromosome copies in interphase nuclei with chromosome centromeric probes. Primary cultures of tumoral cells were prepared to obtain metaphases. Specific chromosomes probes 7, 17, and 20 were used to perform in situ hybridization on isolated intact tumoral cells. Seven cases of primary liver tumors (six cases of HCC and one case of benign focal hepatic nodular hyperplasia) were investigated. A few metaphases were obtained in five of the seven tumors, and in most cases numerical abnormalities were difficult to interpret. In contrast with in situ hybridization, all cases of HCC showed losses and/or gains of chromosomes. Loss of one to three chromosomes occurred in five tumors. A gain of two chromosomes was observed in two of these five tumors. In only one case, a gain of only three chromosomes occurred. In addition, a loss of chromosome 17 was recorded for the benign tumor. These results demonstrate that FISH with specific probes can provide information on chromosome number in the tumoral cells of primary liver tumors even in the absence of analyzable metaphases. This technique opens new possibilities for the investigation of chromosome abnormalities in HCC.
Regenerative changes in hepatic morphology and enhanced expression of CYP 2B10 and CYP 3A during daily administration of cocaine
Authors: Pellinen P , Stenback F , Kojo A , Honkakoski P , Gelboin H V , Pasanen M
Published: November 2004

The effects of daily cocaine administration for up to 14 days were studied in terms of hepatic morphology and the expression of cytochrome P450 (CYP) enzymes in the mouse liver. Daily intraperitoneal doses of 60 mg/kg of cocaine for 3 days induced severe hepatocellular necrosis in the pericentral zone and decreased activities of CYP1A2, CYP2A4/5, and CYP2Cx. The microsomal CYP2B10 protein content was increased by about 2.5-fold, but 2B10-dependent 7-pentoxyresorufin O-dealkylase (PROD) activity was barely detectable. Five or seven daily cocaine doses caused prominent pericentral inflammation and a significant (up to 14-fold) increase in the microsomal protein content and PROD activity. An increase in microsomal CYP3A was also evident, but CYP2A5 and CYP1A2 still remained at a low level. Immunohistochemical examination showed that the relative induction of CYP2B10 and CYP3A after treatment with cocaine was strongest in perivenous hepatocytes. Immunoinhibition experiments showed that CYP2B10 accounted for catalysis of only 15% to 20% of the enhanced microsomal cocaine N-demethylase (CNDM) activity, which correlated well with immunoreactive 3A protein, and could be blocked 70% to 90% by triacetyloleandomycin. After 10 or 14 daily doses of cocaine, regenerative changes with hepatocyte ballooning were observed, coinciding with increases in CYP1A2, CYP2A4/5, and CYP3A. These results suggest the following: (1) cocaine enhances its own cytochrome P450-dependent metabolism; (2) increased production of norcocaine in microsomes is catalyzed mainly by CYP3A enzyme(s), whereas 2B10, although markedly increased by cocaine treatment, has only a minor role in cocaine hepatotoxicity; and (3) despite increased microsomal CNDM activity, cocaine-induced liver injury is reversible in mice.
Release of soluble intercellular adhesion molecule 1 into bile and serum in murine endotoxin shock
Authors: Jaeschke H , Essani N A , Fisher M A , Vonderfecht S L , Farhood A , Smith C W
Published: November 2004

Neutrophil-induced liver injury during endotoxemia is dependent on the adhesion molecules Mac-1 (CD11b/CD18) on neutrophils and its counterreceptor on endothelial cells and hepatocytes, intercellular adhesion molecule 1 (ICAM-1). To investigate a potential release of a soluble form of ICAM-1 (sICAM-1), animals received 100 μg/kg Salmonella abortus equi endotoxin alone or in combination with 700 mg/kg galactosamine. In endotoxin-sensitive mice (C3Heb/FeJ), injection of endotoxin did not cause liver injury but induced a time-dependent increase of sICAM-1 in serum (300%) and in bile (615%) without affecting bile flow. In galactosamine/endotoxin-treated animals, which developed liver injury, the increase in both compartments was only 97% and 104%, respectively. In either case, the increase in sICAM-1 concentrations paralleled the enhanced ICAM-1 expression in the liver. The endotoxin-resistant strain (C3H/HeJ) did not show elevated sICAM-1 levels in serum or bile after endotoxin administration. In contrast, the intravenous injection of murine tumor necrosis factor α (TNF-α), interleukin-1α (IL-1α) or IL-1β (13-23 μg/kg) into endotoxin-resistant mice induced a 225% to 364% increase in serum sICAM-1 and a 370% elevation of the biliary efflux of sICAM-1, again independent of changes in bile flow. These data indicate that cytokines are major inducers of sICAM-1 formation during endotoxemia in vivo. The described experimental model can be used to investigate the role of sICAM-1 in the pathophysiology of inflammatory liver disease.
Changes in liver membrane potentials after partial hepatectomy in rats
Authors: Zhang X K , Gauthier T , Burczynski F J , Wang G Q , Gong Y , Minuk G Y
Published: November 2004

Changes in potential differences (PD) across hepatocyte membranes after partial hepatectomy may play an important role in hepatic homeostatic mechanisms and regenerative activity. To date, few studies have attempted to describe the extent and duration of such changes. In the present study, we documented PD values immediately before and for a 24-hour period after a partial hepatectomy (PHx) of 70% in healthy adult rats. We also documented PD changes after 30% and 90% PHx and PD values in different lobes of the same liver before and after PHx. Our findings showed that the liver depolarizes promptly (within 5 minutes of PHx) and that changes in PD are sustained for approximately 18 hours before returning to baseline values. We also observed that the magnitude of hepatic depolarization correlates with the extent of PHx. Finally, we did not observe regional differences in PD recordings from various lobes before and after PHx. These findings enhance our understanding of the physiological changes that occur in regenerating livers after PHx in rats.
The herbal medicine Inchin-ko-to inhibits liver cell apoptosis induced by transforming growth factor β1
Authors: Yamamoto M , Ogawa K , Morita M , Fukuda K , Komatsu Y
Published: November 2004

Transforming growth factor β1 (TGF-β1) is reported to play an important role in the induction of liver cell apoptosis. In this report, we demonstrate that TGF-β1 induces apoptosis in a rat Morris hepatoma McA-RH8994 cell line and rat primary cultured hepatocytes at similar doses and in a similar manner. Using McA-RH8994 cells, we screened a number of chemical reagents, aqueous extracts of crude drugs, and herbal medicines for their inhibitory activities on TGF-β1-induced apoptosis. The results indicate that Artemisiae capillaris spica (ACS) and the ACS-containing herbal medicine Inchin-ko-to, which are used for treatment of various liver disorders, exhibited the most potent anti-apoptotic activity. Various chemicals that were reported as inhibitors of apoptosis in other experimental systems showed no evident activity. By contrast, two of nine ACS ingredients we tested, capillin and capillene, showed activity at concentrations of submicrogram per milliliter. The inhibitory effects of Inchin-ko-to, capillin and capillene were also confirmed on TGF-β1-induced apoptosis of rat primary cultured hepatocytes. Inhibition of undesired apoptosis induced by TGF-β1 is expected to be beneficial for the treatment of various inflammatory liver diseases. Our findings therefore suggest the possibility that therapeutic effects of Inchin-ko-to on liver diseases might be associated with its inhibitory activity on TGF-β1-induced liver cell apoptosis.
Toxicity of azathioprine and monocrotaline in murine sinusoidal endothelial cells and hepatocytes: The role of glutathione and relevance to hepatic venoocclusive disease
Authors: DeLeve L D , Wang X , Kuhlenkamp J F , Kaplowitz N
Published: November 2004

The mechanisms leading to hepatic venoocclusive disease (HVOD) remain largely unknown. Azathioprine and monocrotaline were studied as part of a series of studies looking at a variety of toxins that induce HVOD to find common features that might be of pathogenic significance. In a previous study, dacarbazine showed selective in vitro toxicity to sinusoidal endothelial cells (SEC) compared with hepatocytes and a key role for SEC glutathione (GSH) was demonstrated. Murine SEC and hepatocytes were isolated and studied in culture. Azathioprine and monocrotaline were found to be selectively more toxic to SEC than to hepatocytes. The relative resistance of hepatocytes to azathioprine was due to enhanced GSH defense: hepatocytes exposed to azathioprine maintained intracellular GSH levels better than SEC, particularly when supplemental GSH precursors were added, and hepatocyte resistance was completely overcome by depletion of intracellular GSH. In contrast, monocrotaline toxicity in hepatocytes was largely unaffected by depletion of GSH, which suggests that selectivity of monocrotaline for SEC may be attributable to differences in metabolic activation. Both compounds are detoxified by GSH in SEC, as demonstrated by enhanced toxicity in the presence of buthionine sulfoximine (BSO) and attenuation of toxicity with exogenous GSH. SEC GSH levels were more than 70% to 80% depleted by monocrotaline and azathioprine, respectively, before cell death. Azathioprine and monocrotaline are selectively toxic to SEC; the mechanism of toxicity in the SEC may be caused by profound GSH depletion.
Adaptive response of the enterohepatic circulation of bile acids to extrahepatic cholestasis
Authors: Dumaswala R , Berkowitz D , Heubi J E
Published: November 2004

Experimental cholestasis induced by ligation of the common bile duct results in morphological and functional changes in the rat hepatocyte. The aim of this study was to evaluate the adaptive response of the transport process involved in the enterohepatic circulation of bile salts to obstructive cholestasis. Male Sprague-dawley rats with common bile duct ligation were killed after 48 and 72 hours. Portal and systemic blood and duodenal aspirates were collected. Taurocholate transport was measured in isolated ileal brush border (BBM) and liver basolateral membranes (BLM). Drastic reduction in intraluminal bile salt concentrations in ligated rats accompanied decreases in saturable taurocholate uptake by ileal BBMs. Kinetic analysis indicated that the decrease was attributable to reduction in transporter density and alteration in affinity for the substrate, both of which accentuated with increase in postligation time. In contrast, despite 20-fold higher portal venous bile salt concentrations, taurocholate uptake by hepatic sinusoidal membrane was lower in rats. Kinetic analysis and immunoblots developed using polyclonal antisera to the liver BLM bile acid transporter demonstrated a gradual decrease in transporter density with increase in postligation time accompanied by reduced taurocholate uptake by basolateral membrane. These results further support the concept that the ileal brush border membrane transporter is regulated by the availability of the substrate, whereas regulation of the bile salt transport across the liver BLM is independent of portal venous bile acid concentration.
Quantitative image analysis of cytokeratin filament distribution during fetal rat liver development
Authors: Vassy J , Beil M , Irinopoulou T , Rigaut J P
Published: November 2004

Recent reports suggest that not only chemical but also mechanical influences from the cellular environment could have profound effects on gene activity and could act on cell differentiation and proliferation. These mechanisms involve a tissue matrix system, which includes extracellular matrix, nuclear matrix, and cytoskeleton. Supposing that the cytoskeleton mediates mechanical transduction from the cell environment to the nucleus, significant differences in the spatial distribution of the cytoskeleton network could reflect variations in environmental signals. Hepatocytes during fetal development provide a useful model to study such variations, because a progressive establishment of intercellular contact is reported. The aim of this work is to discriminate steps in hepatocyte differentiation from fetal to adult livers, using computerized quantitative image analysis of cytokeratin (C8) immunofluorescent localization, visualized by confocal scanning laser microscopy. The filament structure is represented by the gray-scale skeleton of the digital images obtained by specially designed segmentation methods. A set of line features was investigated, including number and length of lines, orientation of lines, and the fractal dimension of the filament network. The features studied showed highly significant differences throughout liver development, with an increase of the total amount of cytokeratin filaments. We could also demonstrate a modification in the structure of the network, being more and more dense, with an increase of connecting points. Moreover, it has been shown that filaments have some orientation in fetal hepatocytes, and that directionality. Thus, significant differences in the pattern of the cytokeratin filament network according to the stages of hepatocyte differentiation have been demonstrated with objective and quantitative methods.
A pilot study of corticosteroid priming for lymphoblastoid interferon alfa in patients with chronic hepatitis C
Authors: Chayama K , Tsubota A , Kobayashi M , Hashimoto M , Miyano Y , Koike H , Koida I , Arase Y , Saitoh S , Murashima N , Ikeda K , Kumada H
Published: November 2004

Interferon treatment reduces the serum level of hepatitis C virus (HCV) and improves inflammatory activity, but relapse is frequently observed. In an attempt to develop a new therapeutic strategy that may reduce relapse and cure the disease, we evaluated the effect of corticosteroid priming on lymphoblastoid interferon alfa in an open randomized clinical trial. The level of HCV RNA increased significantly during corticosteroid priming (from 5.60 [median] to 21.0 x 105 Eq/mL; P = .0004) but decreased to the pretreatment level 4 weeks after cessation of corticosteroid (7.0 x 105 Eq/mL; P = .07). Sustained normalization of alanine transaminase (ALT) level and virus clearance, confirmed by negative results for HCV RNA using reverse-transcription nested polymerase chain reaction (PCR), were observed over a period of 6 months in 8 of 19 (42.1%) corticosteroid-primed patients, compared with 6 of 19 patients (31.6%) treated with interferon only. A “rebound” of ALT after the withdrawal of corticosteroid was observed in only 2 of 19 patients primed with corticosteroid, but both showed sustained responses. Multivariate analysis for factors predictive of the sustained response indicated that HCV titers measured immediately before interferon therapy and HCV genotype were statistically significant (P = .006 and P = .025, respectively). Our results indicated that corticosteroid priming has a marginal benefit over treatment with interferon alone and that large-scale clinical trials are necessary to determine whether interferon with corticosteroid priming is more effective than interferon alone.
Productive infection of primary cultures of endothelial cells from the cat liver sinusoid with the feline immunodeficiency virus
Authors: Steffan A , Lafon M , Gendrault J , Smedsrod B , Nonnenmacher H , Koehren F , Gut J , de Monte M , Martin J , Royer C , Kirn A
Published: November 2004

Given the similarities between the two viruses, the feline immunodeficiency virus (FIV) is becoming an interesting animal model for human immunodeficiency virus (HIV) studies. To explore the still controversial role of the liver in the development of HIV infection, sinusoidal endothelial cells (SEC) were isolated, and primary cultures were infected with the FIV Villefranche IFFA strain. The isolated cells were characterized by their typical fenestrations, the presence of von Willebrand factor (vWf), and their ability to take up acetylated low-density lipoproteins and denatured collagen. Two weeks after infection, significant amounts of FIV p24 antigen were detected by immunofluorescence in both multinucleated giant and single cells and by enzyme-linked immunosorbent assay in the culture medium. High amounts of viral particles were observed together with different steps of budding at the plasma membrane or at the membrane of intracytoplasmic vacuoles. The released viral particles were shown to be infectious for a permissive cell line. During the first 3 weeks of infection, the only cytopathic effect of FIV was syncytia formation. No noticeable impairment of the pattern of fenestrations and the modulation of their number by a cytoskeleton-mediated process occurred. The productive infection of SEC may contribute to the progression of the infection.
Severe recurrent cholestatic hepatitis C following orthotopic liver transplantation
Authors: Schluger L K , Sheiner P A , Thung S N , Lau J Y , Min A , Wolf D C , Fiel I , Zhang D , Gerber M A , Miller C M , Bodenheimer H C
Published: November 2004

Recurrent infection with hepatitis C virus (HCV) is almost universal following orthotopic liver transplantation although clinical severity varies. Data on 135 patients who underwent transplantation for hepatitis C cirrhosis were reviewed. We describe a progressive, severe cholestatic form of hepatitis occurring in a subgroup of patients with recurrent hepatitis C. Ten patients with severe recurrent hepatitis C were identified; 1 has died, 1 awaits retransplantation, and 8 have undergone retransplantation. All 10 developed severe progressive cholestatic hepatitis, with a mean rise in bilirubin to 24.7 mg/dL at the time of retransplantation. Histology at initial recurrence was of mild hepatitis without evidence of rejection. The failed grafts showed either cirrhosis or confluent hepatic necrosis. The onset of cholestasis preceded retransplantation by less than 5 months. Our study suggests that a minority of patients with recurrent hepatitis C after undergoing liver transplantation develop a severe progressive cholestatic hepatitis and liver failure.
Pulmonary hypertension after transjugular intrahepatic portosystemic shunt: Effects on right ventricular function
Authors: Van der Linden P , Le Moine O , Ghysels M , Ortinez M , Deviere J
Published: November 2004

The short- and mid-term hemodynamic effects of transjugular intrahepatic portosystemic shunt (TIPS) were studied in 16 sedated cirrhotic patients. Indications included relapsing variceal bleeding (n = 10) and refractory ascites (n = 6). The decrease of porto-atrial pressure gradient (from 20.4 ± 4.2 mm Hg to 10.1 ± 2.4 mmHg; P < .05) was associated with an increase of mean pulmonary artery pressure (MPAP) (from 12.3 ± 3.0 mm Hg to 20.3 ± 5.3 mm Hg; P < .05) and of right atrial pressure (RAP) from 3.4 ± 2.6 mm Hg to 8.3 ± 3.7 mm Hg; P < .05), whereas right ventricular end-diastolic volume (RVEDVI) remained unchanged. The significant increase of cardiac index (CI) (from 4.5 ± 1.2 L/min/m2 to 5.0 ± 1.1 L/min/m2; P < .05) was essentially attributable to an increase of heart rate (HR) (from 81 ± 11 to 88 ± 10 beats/min; P < .05). Systemic vascular resistance (SVR) decreased (from 812 ± 281 to 666 ± 191 dynes/sec/cm5; P < .05), whereas pulmonary vascular resistance (PVR) increased (from 60.6 ± 29.6 to 82.0 ± 34.6 dynes/sec/cm5; P < .05). After transient shunt occlusion with a balloon catheter, all of the hemodynamic parameters returned to baseline values, except pulmonary artery pressure, which also decreased but remained significantly increased. One month after TIPS, pulmonary pressure remained elevated, and CI further increased. It is concluded that increased PVR is the major hemodynamic alteration occurring after TIPS placement. It correlates with the decrease of porto-atrial gradient and is probably mediated by both mechanical and neurohumoral factors.
Prognosis of small hepatocellular carcinoma (less than 3 cm) after percutaneous acetic acid injection: Study of 91 cases
Authors: Ohnishi K , Nomura F , Ito S , Fujiwara K
Published: November 2004

To assess the efficacy of ultrasound (US)-guided percutaneous acetic acid (in concentrations of 15%, 20%, 30%, 40%, and 50%) injection for small hepatocellular carcinomas (HCCs) for long-term prognosis, percutaneous acetic acid injection using 15% to 50% acetic acid was performed in 91 patients with one to four HCCs smaller than 3 cm during the past 6.5 years. During the series of treatment sessions for each patient, the same concentration of acetic acid was used. All tumors could be treated successfully with percutaneous acetic acid injection despite the differences in acetic acid concentration used. The number of treatment sessions to treat similar size of tumor was less when the higher concentration of acetic acid was used. No serious complications occurred as a direct sequela to percutaneous acetic acid injection. None of the tumor treated regrew. The 1-, 2-, 3-, 4-, and 5-year survival rates for 91 patients were 95%, 87%, 80%, 63%, and 49%, respectively. The 1-, 2-, 3-, 4-, and 5-year cancer-free survival rates of these patients were 83%, 54%, 50%, 37%, and 29%, respectively. Both liver function and size of tumor affected both survival rate and cancer-free survival rate significantly, but the number of tumors did not. The concentration of acetic acid did not affect the survival rate. Percutaneous acetic acid using 15% to 50% acetic acid will be effective therapy for small HCCs for long-term prognosis.
Relationship between hepatocyte proliferative activity and liver functional reserve in human cirrhosis
Authors: Delhaye M , Louis H , Degraef C , Le Moine O , Deviere J , Gulbis B , Jacobovitz D , Adler M , Galand P
Published: November 2004

Hepatocyte proliferative activity is elevated in cirrhotic patients who develop hepatocellular carcinoma (HCC) and decreased in alcohol-induced hepatitis patients with poor outcome. Hepatocyte proliferative activity has not been evaluated in an unselected population of cirrhotic patients regarding the severity of the disease. Forty-six cirrhotic patients (21 alcoholic, 20 viral, and 5 other) were prospectively analyzed by proliferating cell nuclear antigen (PCNA) immunostaining on methanol-fixed, paraffin-embedded liver biopsy specimens. In these conditions, the PCNA-labeling index (PCNA-LI) measures the number of cells in the S-phase and assesses tissue proliferation. The median value of the PCNA-LI for all samples was 4.3% (range, 0%-20.2%). It declined with worsening Child-Pugh score: 9.15% (range, 3.3%-20.2%), 5.3% (range, 1.2%-18%), and 2.4% (range, 0%-4.4%) in Child classes A, B, and C, respectively (P < .05). Using the best cutoff PCNA-LI value to divide cirrhosis into slowly and rapidly proliferating tissue subsets, the PCNA index was independently associated with serum albumin. The probability of survival in patients with a high PCNA-LI (> 4.4%) was significantly higher than in those with a lower PCNA-LI (0.93 vs. 0.53, at a median follow-up of 153 days; P = .01). In all 6 patients undergoing placement of a transjugular intrahepatic portosystemic shunt (TIPS), the PCNA-LI decreased after the procedure. This early impairment of hepatocyte proliferative activity after TIPS placement might reflect the functional alterations induced by this treatment. In conclusion, hepatocyte proliferative activity assessed by PCNA-LI is increased in cirrhotic patients and decreases with worsening of the disease.
Cell cycle progression proteins (cyclins), oncogene expression, and signal transduction during the proliferative response of human hepatocytes to hepatocyte growth factor
Authors: Gomez-Lechon M J , Guillen I , Ponsoda X , Fabra R , Trullenque R , Nakamura T , Castell J V
Published: November 2004

Human hepatocytes stimulated with human recombinant hepatocyte growth factor (h-rHGF) (10 ng/mL) displayed a characteristic lag period before entering into the S phase. The duration of this delay was dependent on the timing of h-rHGF addition to cultures. The highest peak of DNA synthesis was observed at 120 hours of culture when hepatocytes were stimulated with h-rHGF at 72 hours of culture. This was accompanied by an early peak of c-jun and c-fos synthesis (3 hours after addition of h- rHGF) followed by c-myc (6 hours) and increased expression of cyclins A, B, D, and E (12 hours after h-rHGF). A significant dose-dependent increase in inositol 1,4,5-P3 was observed within 45 seconds after stimulation with the factor. This was followed by an immediate increase in the cytosolic-free calcium. Cyclic adenosine monophosphate (cAMP) levels did not change after stimulation with the factor. Tyrosine phosphorylation seems to be an early event in the course of the stimulatory effect of h-rHGF on DNA synthesis of hepatocytes; genistein, a tyrosine kinase inhibitor, impaired the stimulatory effect of h-rHGF on DNA synthesis dose dependently. On the other hand, the action of the factor was negatively regulated by protein kinase C activation, as shown by the increased stimulatory effect of h-rHGF on DNA synthesis upon inhibition of protein kinase C by H7.
Prediction of risk of liver disease by alcohol intake, sex, and age: A prospective population study
Authors: Becker U , Deis A , Sorensen T I , Gronbaek M , Borch-Johnsen K , Muller C F , Schnohr P , Jensen G
Published: November 2004

The association between self-reported alcohol intake and the risk of future liver disease was studied in a large population-based prospective cohort with 12-year follow-up. Alcohol intake was assessed in 13,285 men and women aged 30 to 79 years by a self-administered questionnaire. Diagnoses indicating alcohol-induced liver disease (n − 261) or alcohol-induced cirrhosis (n − 124) were obtained from death certificates and the hospital discharge register, and data were analyzed by multiplicative Poisson regression models. The total cumulated observation time was 130,558 person-years. The overall incidence rates of alcohol-induced cirrhosis were 0.2% per year in men and 0.03% per year in women. The nadir of the estimated relative risk of developing liver disease was observed at an alcohol intake of 1 to 6 beverages per week, and above this level a steep increase in relative risk was observed. The risk function was independent of age and stable over time. The level of alcohol intake above which the relative risk was significantly greater than 1 was observed at 7 to 13 beverages per week for women and 14 to 27 beverages per week for men. Women had a significantly higher relative risk of developing alcohol-related liver disease than men for any given level of alcohol intake. We observed a dose-dependent increase in relative risk of developing alcohol-induced liver disease for both men and women, with the steepest increase among women. In the general population, self-reported current alcohol intake is a good predictor of the future risk of alcohol-induced liver disease.
Liver function tests in normal pregnancy: A prospective study of 103 pregnant women and 103 matched controls
Authors: Bacq Y , Zarka O , Brechot J , Mariotte N , Vol S , Tichet J , Weill J
Published: November 2004

Except for increased serum alkaline phosphatase (AP) levels, the changes in liver function test (LFT) values during normal pregnancy have not been clearly established, mainly because most studies do not include matched controls. We therefore measured the serum values of routine liver tests including 5′-nucleotidase and total bile acids in 103 healthy pregnant women (first trimester, n = 34; second trimester, n = 36; third trimester, n = 33) and in 103 age-matched controls not receiving oral contraception. Fasting blood samples were taken. Because of hemodilution, serum albumin levels were significantly lower during all trimesters. As expected, AP activity was significantly higher in the third trimester. Serum aspartate transaminase (AST) activity and total bile acid (TBA) concentrations did not differ between pregnant and nonpregnant women. Serum alanine transaminase (ALT) activity was slightly higher in the second-trimester pregnant women than in controls (6.8 ± 4.5 vs. 8.2 ± 5.8, P = .04), although all values remained within normal limits. In pregnant women, total and free bilirubin concentrations were significantly lower during all three trimesters, as was conjugated bilirubin during the second and third trimesters. Serum γ-glutamyl transpeptidase (GGT) activity was significantly lower in the second and third trimesters. Serum 5′-nucleotidase activity was slightly but significantly higher in the second and third trimesters. The knowledge of these results is useful for the interpretation of LFT values and the management of liver diseases during pregnancy.
Does malnutrition affect survival in cirrhosis?
Authors: Merli M , Riggio O , Dally L
Published: November 2004

A total of 1,053 cirrhotic patients were included in a prospective study to determine whether malnutrition is a risk factor for mortality in cirrhotic patients. Child-Pugh classification as well as clinical and biochemical variables were used to assess the severity of cirrhosis. Nutritional status was evaluated both by anthropometric and clinical measurements. Patients were defined as malnourished when midarm muscle area (MAMA) and/or midarm fat area (MAFA) were below the 5th percentile of an age- and sex-matched population. During follow-up, 419 patients died. The estimated survival rate was 82.7% at 1 year, 65.1% at 3 years, and 50.7% at 5 years. The presence of muscle depletion and/or of a steep reduction in fat deposits was associated with a higher risk of mortality (midarm muscle area, < 5th percentile, relative risk = 1.79; midarm fat area, < 5th percentile, relative risk = 1.35). When patients were stratified according to the Child-Pugh classification, cumulative survival was lower in patients with a reduction in muscle mass in Child-Pugh classes A and B (log rank: P = .027; P = .022, respectively) but not in class C. Conversely, a significant reduction in adipose tissue deposits appeared to have no independent impact on survival in any Child-Pugh class. When examined using a multivariate Cox proportional hazard analysis, age, sex, bilirubin, cholinesterase, ascites, and esophageal varices were selected, whereas the parameters of nutritional status were not. This suggests that malnutrition, while strongly associated with the deterioration of liver function, cannot be considered an independent risk factor for mortality in a general population of cirrhotic patients.
Predictive score for the development of hepatocellular carcinoma and additional value of liver large cell dysplasia in Western patients with cirrhosis
Authors: Ganne-Carrie N , Chastang C , Chapel F , Munz C , Pateron D , Sibony M , Deny P , Trinchet J , Callard P , Guettier C , Beaugrand M
Published: November 2004

The aim of this study was to identify high-risk patients for hepatocellular carcinoma (HCC). Among 151 patients with histologically proven cirrhosis hospitalized from 1987 to 1990 and prospectively followed-up until June 1994, 31 developed HCC. We assessed the predictive value of 22 variables recorded at enrollment for HCC occurrence by the log rank test and the Cox proportional hazards model. Six clinical and biological variables summarized predictive information of HCC: age > or = 50 years (P = .01), male (P = .01), large esophageal varices (EV) (P = .03), prothrombin activity < 70% (P = .04), serum α-fetoprotein (AFP) > or = 15 ng/L (P = .06), and anti-hepatitis C virus antibodies (P = .08). A clinicobiological predictive score identified two groups of patients at low (n = 67; 3-year cumulative incidence, 0%) and high risk for HCC (n = 84; 3-year cumulative incidence, 24%). The predictive value of this score was confirmed using an independent population of 49 patients with cirrhosis. Furthermore, liver large-cell dysplasia (LCD) had an additional predictive value in high-risk patients (P = 10-4, which thus helped to define a subgroup at very high risk for HCC (n = 12; 3-year cumulative incidence, 72%). In Western patients with cirrhosis, a limited number of usual variables can identify a group of patients at high risk for HCC. Among these patients, liver biopsy allows for the determination a subgroup of patients at very high risk for HCC requiring intensive screening or preventive measures.
Effect of liver transplantation on QT interval prolongation and autonomic dysfunction in end-stage liver disease
Authors: Mohamed R , Forsey P R , Davies M K , Neuberger J M
Published: November 2004

Both a prolonged QT interval and disturbance of autonomic nervous system function are markers of poor prognosis in patients with diabetes mellitus and alcoholic liver disease (ALD). We studied the prevalence of abnormal QT interval and autonomic nervous system dysfunction in 53 consecutive patients with end-stage liver disease before and after orthotopic liver transplantation (OLT). The maximum QT interval in any lead (QTmax) was assessed by two independent observers. The QTmax, corrected for heart rate (QTcmax) was prolonged in 44 patients (83%), although increased QT dispersion was not found. There was a significant correlation between the QTcmax and Child-Pugh score but not with etiology. Evidence of parasympathetic dysfunction was present in 41 patients (77%), and sympathetic dysfunction was present in 20 patients before OLT. Fifty-two patients underwent transplantation. There was significant improvement in the QTcmax interval after OLT (P < .001); 32 of the 44 patients with prolonged QTcmax (> 440 milliseconds) improved. Repeat testing was not performed in 7 patients, because they had died or had not undergone transplantation. Indices of parasympathetic function improved in 27 patients after OLT, but no improvement was observed in 8. Improvement in sympathetic dysfunction was observed in 13 of the 19 patients tested. There was no association between QTcmax, autonomic dysfunction, and survival. These results suggest that both prolonged QTcmax and some tests of autonomic function are temporary and arise as a consequence of liver dysfunction.