bluuebunny/crossref_metadata_2025 huggingface dataset:
In vivo evaluation of a hollow fiber liver assist device
Authors: Jauregui Hugo O. , Mullon Claudy J-P. , Trenkler Donna , Naik Sharda , Santangini Henry , Press Philip , Muller Thomas E. , Solomon Barry A.
Published: March 2007

Orthotopic liver transplantation is the only effective form of therapy currently available for patients with fulminant hepatic failure (FHF). The use of an extracorporeal (EC) liver assist device (LAD) may result in improved presurgical clinical management. Alternatively, patients treated with LADs could avoid the transplantation procedure if they are able to regenerate a critical mass of hepatocytes that will sustain functional viability. In this study, the efficacy of a prototype hollow fiber LAD seeded with rabbit hepatocytes was assessed in vivo by the use of two different animal models: (1) normal rabbits injected with diazepam or lidocaine, and (2) a galactosamine (Gal)-intoxicated rabbit model of FHF. The EC LAD clearly decreased the blood levels of the two drugs and significantly generated diazepam and lidocaine metabolites indicating the maintenance of active P450 forms in the cellular component of the devices. A 6-hour EC treatment significantly increased the survival time and delayed the onset of hepatic encephalopathy (HE) in the Gal-intoxicated rabbits. Histological evaluations of postmortem livers showed greater hepatocyte regenerative activity in the animals treated with hepatocyte-seeded LADs than in the two control groups, e.g., rabbits not treated or treated with unseeded devices. These findings support the concept that a microporous hollow fiber LAD seeded with rabbit hepatocytes is able to sustain drug detoxification in vivo as well as to modify the course of FHF in a well-characterized animal model. (Hepatology 1995;21:460-469.)
Expression of Von Willebrand factor in normal and diseased rat livers and in cultivated liver cells†
Authors: Knittel Thomas , Neubauer Katrin , Armbrust Thomas , Ramadori Giuliano
Published: March 2007

Von Willebrand factor (vWf) is an adhesive glycoprotein known to play an important role in hemostasis and in tissue injury. Because the latter process resembles hepatic fibrogenesis, we studied the tissue distribution of vWf in diseased livers. In normal rat liver vWf was strongly expressed in the vascular endothelium and as small spots or fiber-like structures in the hepatic parenchyma. During acute liver injury, pronounced staining was observed within the area of necrosis. In fibrotic livers vWf deposits were distributed predominantly at the scar-parenchyma interface but also within the septum and in sinusoidal lining cells. Testing different liver cell populations in vitro demonstrated that vWf gene expression was limited to endothelial cells (ECs) and, therefore, the latter cell population might represent the vWf-positive cells detected in vivo. The distribution of vWf within fibrotic septa suggests that vWf becomes a component of the extracellular matrix (ECM) in fibrotic livers. (Hepatology 1995;21:470-476.)
Stimulated hepatic tissue repair underlies heteroprotection by thioacetamide against acetaminophen-induced lethality
Authors: Chanda Sanjay , Mangipudy Raja S. , Warbritton Alan , Bucci Thomas J. , Mehendale Harihara M.
Published: March 2007

Acetaminophen (APAP) is a widely used analgesic and antipyretic drug that causes massive centrilobular hepatic necrosis at high doses, leading to death. The objectives of this study were to test our working hypothesis that preplaced cell division and hepatic tissue repair by prior thioacetamide (TA) administration provides protection against APAP-induced lethality and to investigate the underlying mechanism. Male Sprague-Dawley rats were treated with a low dose of TA (50 mg/kg, intraperitoneally [i.p.]) before challenge with a 90% lethal dose (1,800 mg/kg, i.p.) of APAP. This protocol resulted in a 100% protection against the lethal effect of APAP. Because TA caused a 23% decrease of hepatic microsomal cytochromes P-450, the possibility that TA protection may be caused by decreased bioactivation of APAP was examined. A 30% decrease in cytochromes P-450 induced by cobalt chloride failed to provide protection against APAP lethality. Time course of serum enzyme elevations (alanine aminotransferase, aspartate aminotransferase, and sorbitol dehydrogenase) indicated that actual infliction of liver injury by APAP peaked between 12 to 24 hours after the administration of APAP, whereas the ultimate outcome of that injury depended on the biological events thereafter. Although liver injury progressed in rats receiving only APAP, it regressed in rats pretreated with TA. Acetaminophen t1/2 was not altered in TA-treated rats, indicating that significant changes in APAP disposition and bioactivation are unlikely. Moreover, hepatic glutathione was decreased to a similar extent regardless of TA pretreatment, suggesting that decreased bioactivation of APAP is unlikely to be the mechanism underlying TA protection. [3H]Thymidine incorporation studies confirmed the expected stimulation of S-phase synthesis, and proliferating cell nuclear antigen studies showed a corresponding stimulation of cell division through accelerated cell cycle progression. Intervention with TA-induced cell division by colchicine antimitosis ended the TA protection in the absence of significant changes in the time course of serum enzyme elevations during the inflictive phase of APAP hepato-toxicity. These studies suggest that hepatocyte division and tissue repair induced by TA facilitate sustained hepatic tissue repair after subsequent APAP-induced liver injury, producing recovery from liver injury and protection against APAP lethality. (Hepatology 1995;21:477-486.)
In vivo microscopy of hepatic metastases: Dynamic observation of tumor cell invasion and interaction with Kupffer cells
Authors: Kan Zuxing , Ivancev Krassnodar , Lunderquist Anders , McCuskey Patricia A. , McCuskey Robert S. , Wallace Sidney
Published: March 2007

In vivo microscopy was used in the study of the biological behavior of tumor cells and of the activity of Kupffer cells in hepatic tumors in situ. Three tumor models, Friend erythroleukemia inoculated into Dilute Brown Aguti (DBA)/2 mice, murine colon adenocarcinoma (CT)-26 in Bagg Albino inbred albino (BALB)/c mice, and mammary cancer 13762 NF in Fischer rats, were investigated. Tumor cells showed a strong tendency to adhere to the sinusoidal endothelium, most frequently in the sinusoids near the tumors. Mechanical trapping of tumor cells in the narrow portion of hepatic sinusoids, a phenomenon suggested by previous investigators as a predominant pattern for tumor cells to arrest in the liver, was not confirmed. Our study documented that in tumor-bearing livers, as compared with normal control livers, the population size and the phagocytic capacity of Kupffer cells are increased in nontumorous areas but are significantly decreased inside the tumors. In vivo microscopic images showed that Kupffer cells are not only attracted to tumor cells in the hepatic circulation but also have the ability to phagocytose those tumor cells. In vivo microscopy has been shown to be a useful tool for dynamic studies in tumor biology, pathology, and pharmacology. (Hepatology 1995;21:487-494.)
Detection of Gp210 autoantibodies in primary biliary cirrhosis using a recombinant protein containing the predominant autoepitope
Authors: Tartakovsky Flora , Worman Howard J.
Published: March 2007

Autoantibodies against nuclear pore membrane glycoprotein gp210 have been identified in between 10% and 25% of patients with primary biliary cirrhosis (PBC). These antibodies may be useful in diagnosing PBC and in identifying subgroups of patients. Because previous detection procedures relied on the need to purify hydrophobic proteins and perform immunoblotting, the aim of the present study was to develop a simple assay to detect gp210 autoantibodies. A recombinant polypeptide containing glutathione-S-transferase (GST) fused to the region of gp210 that contains its predominant autoepitope(s) was expressed in bacteria. This fusion protein was purified by glutathione-Sepharose chromatography and used in an enzyme-linked immunosorbent assay (ELISA). The ELISA was reproducible in detecting gp210 autoantibodies in serum samples from patients with PBC. Compared with immunoblotting, the ELISA was 93% sensitive and 96% specific for the detection of gp210 autoantibodies. In conclusion, autoantibodies against gp210 can be easily and reliably detected in patients with PBC by an ELISA that uses a purified recombinant polypeptide. (Hepatology 1995;21:495-500.)
Glutamine transport in isolated human hepatocytes and transformed liver cells
Authors: Bode Barrie P. , Kaminski Donald L. , Souba Wiley W. , Li Al P.
Published: March 2007

The transport of L-glutamine was examined in isolated adult and fetal human hepatocytes as well as in the human hepatoma cell lines HepG2 and SK-Hep. In all cells studied, glutamine uptake was at least 85% Na+-dependent. Kinetic analysis of the Na+-dependent component indicated mediation by a single transporter in three human hepatocyte preparations and in SK-Hep cells, whereas two transporters appeared to be responsible for glutamine uptake in HepG2 cells and in hepatocytes from the liver of one male patients. Amino acid inhibition analysis showed primary mediation by System N in fetal and adult hepatocytes, whereas System ASC was principally responsible for glutamine uptake in transformed cells. Similar to the rat transporter, human System N was pH-sensitive, stereospecific, and responsive to treatment with steroid hormones. Although the human carrier was less tolerant of Li+- for Na+ substitution, glutamine transport in primary human hepatocytes was stimulated by treatment with hypotonic buffer (cell swelling), as reported in rat parenchymal cells. In contrast, glutamine transport in hepatoma cells was relatively insensitive to changes in extracellular pH and failed to show enhanced activity in response to hypoosmotic challenge. Collectively, the data suggest that markedly distinct plasma membrane transporters mediate the concentrative uptake of glutamine in normal and transformed human hepatocytes, and that the salient properties of System N have been largely conserved from rat to man. (Hepatology 1995;21:511-520.)
The effects of dietary iron on initiation and promotion in chemical hepatocarcinogenesis
Authors: Stål Per , Hultcrantz Rolf , Möller Lennart , Eriksson Lennart C.
Published: March 2007

The aim of this study was to evaluate the effects of dietary iron on hepatocarcinogenesis in an animal model mimicking noncirrhotic genetic hemochromatosis. Iron overload may lead to liver cirrhosis and an increased risk of developing primary hepatocellular carcinoma. It is unknown if iron is of pathogenic importance for the carcinogenic process, or whether the increased cancer risk results solely from the cirrhotic process. We investigated the initiating, promoting, and mitogenic properties of carbonyl iron in the Solt-Farber model of chemical hepatocarcinogenesis. A diet supplemented with 2.5% to 3.0% carbonyl iron was either added to, or replaced, the initiating and promoting events in the model. None of the animals developed hepatic fibrosis. Hepatic iron was increased 6- to 13-fold in iron-treated animals, and predominantly located in periportal hepatocytes. Iron as an initiator did not increase the number of glutathione-S-transferase-Yp-positive foci. Iron reduced the number of foci when added to low-dose diethylnitrosamine plus partial hepatectomy, which may be explained by a delayed hepatic regeneration in iron-loaded liver. As a promoter, iron did not selectively induce proliferation of initiated cells. Added to a complete promotive regimen, iron decreased the volume density of preneoplastic nodules, possibly because of a mitostimulatory effect of iron on normal hepatocytes surrounding the nodules. Iron increased the hepatocyte labeling index and counteracted the mitoinhibitory effect of 2-acetylaminofluorene on regenerating liver. We conclude that in this animal model, dietary carbonyl iron, mimicking the iron loading pattern of genetic hemochromatosis (1) does not possess genotoxic properties detectable as increased preneoplastic foci formation; (2) does not promote the growth of initiated cells; (3) has mitogenic properties in vivo, but not strong enough to replace partial hepatectomy in promotion; and (4) leads to a decreased number of foci or volume density of nodules when added to a complete initiating or promoting regimen. (Hepatology 1995;21:521-528.)
Increased sinusoidal wall permeability and liver fatty change after two-thirds hepatectomy: An ultrastructural study in the rat
Authors: Morsiani Eugenio , Mazzoni Mary , Aleotti Arrigo , Gorini Paolo , Ricci Daniele
Published: March 2007

Accumulation of lipids in the hepatocyte cytoplasm after partial hepatectomy (PH) has long been recognized, but the mechanism behind this phenomenon is still poorly understood. In this study, rats subjected to a standard two-thirds PH showed early and marked increase in portal venous pressure (P < .01). On scanning electron microscopy, the regenerating liver fixed by portal perfusion under hemodynamic conditions identical to that found in vivo during the first 24 hours showed a significant (P < .01) 10-fold increase in the sinusoidal wall porosity (percentage open area by fenestrations). This was paralleled by the disappearance of the sieveplate arrangement of small fenestrations and by a significant decrease in the number of fenestrations per micrometers squared of sinusoidal surface at 6 (P < .01) and 12 hours (P < .05). In addition, there were major changes in the frequency and distribution of all three classes of fenestrations. At 6 and 12 hours, there was a marked decrease of small class 1 fenestrations and a marked increase of intermediate class 2 fenestrations and large class 3 fenestrations (P < .0001). A concurrent accumulation of lipid droplets in the hepatocyte cytoplasm produced a 20-fold increase in the hepatocyte total lipid volume. A statistically significant linear correlation (r = 0.907; P < .01) was found between the amount of intracellular lipids and the data quantitating the changes in porosity of liver sinusoids at 24 hours. It is concluded that an increased sinusoidal wall permeability to lipids may be the primum movens in the pathogenesis of transient liver fatty change after PH in the rat. (Hepatology 1995;21:539-544.)
Endothelin stimulates platelet-activating factor synthesis by cultured rat Kupffer cells
Authors: Mustafa Shamimunisa B. , Gandhi Chandrashekhar R. , Harvey Stephen A. K. , Olson Merle S.
Published: March 2007

Endothelins are potent peptide mediators that elicit glycogenolytic and vasoconstrictor actions in the liver. Endothelins were found to stimulate the synthesis and release of the lipid mediator platelet-activating factor in cultured rat Kupffer cells. Endothelin-mediated synthesis of platelet-activating factor required extracellular calcium in that the calcium chelator, EGTA and nifedipine, a calcium ion channel blocker, inhibited platelet-activating factor synthesis. The phospholipase A2 inhibitor, 4-bromophenacyl bromide, strongly inhibited endothelin-induced platelet activating factor synthesis. Endothelin-stimulated platelet activating factor synthesis was inhibited after treatment of Kupffer cells with cholera toxin, whereas pertussis toxin inhibited only this response to endothelin-1. Agents that elevate intracellular cyclin AMP levels were found to inhibit endothelin-induced platelet-activating factor synthesis in Kupffer cells. Staurosporine, a protein kinase C inhibitor minimized endothelin-induced platelet-activating factor synthesis but phorbol myristate acetate, an activator of protein kinase C, did not affect endothelin-induced platelet activating factor synthesis. Thus, the current study demonstrates that activation of an endothelin receptor in cultured rat Kupffer cells results in the synthesis and release of platelet-activating factor. The importance of endothelin-mediated platelet-activating factor synthesis relates to the mechanism of intercellular signaling occurring between endothelial cells (i.e., the site of endothelin synthesis) and Kupffer cells (i.e., the site of formation of secondary mediators such as platelet-activating factor and eicosanoids) within the rat liver exposed to various types of pathophysiological episodes. (Hepatology 1995;21:545-553.)
p53 Mutations and hepatitis B virus: Cofactors in hepatocellular carcinoma
Authors: Slagle Betty L.
Published: March 2007

We studied 80 hepatocellular carcinomas from three continents for p53 gene (TP53) mutations and hepatitis B virus (HBV) sequences. p53 mutations were frequent in tumors from Mozambique but not in tumors from South Africa, China, and Germany. Independent of geographic origin, most tumors were positive for HBV sequences. X gene coding sequences of HBV were detected in 78% of tumors, whereas viral sequences in the surface antigen- and core antigen-encoding regions were present in less than 45% of tumors. These observations indicate that hepatocellular carcinomas are genetically heterogeneous. Mozambican-type of hepatocellular carcinomas are characterized by a high incidence of p53 mutations related to aflatoxins. In other tumors, the rarity of p53 mutations combined with the frequent presence of viral X gene coding sequences suggests a possible interference of HBV with the wild-type p53 function.
p53 Mutations in nonmalignant human liver: Fingerprints of aflatoxins?
Authors: Ozturk Mehmet
Published: March 2007

Fifty-eight percent of hepatocellular carcinomas (HCCs) from Qidong, China, contain an AGG to AGT mutation at codon 249 of the p53 tumor suppressor gene, a mutation that is rarely seen in HCCs from Western countries. The population of Qidong is exposed to high levels of aflatoxin B1 (AFB1), a fungal toxin that has been shown to induce the same mutation in cultured human HCC cells. To investigate the role of AFB1 and of these p53 mutations in hepatocarcinogenesis, normal liver samples from the United States, Thailand, and Qidong (where AFB1 exposures are negligible, low, and high, respectively) were examined for p53 mutations. The frequency of the AGG to AGT mutation at codon 249 paralleled the level of AFB1 exposure, which supports the hypothesis that this toxin has a causative—and probably early—role in hepatocarcinogenesis.
Strong, sustained hepatocellular proliferation precedes hepatocarcinogenesis in hepatitis B surface antigen transgenic mice
Authors: Huang Shao-Nan , Chisari Francis V.
Published: March 2007

Hepatocyte turnover rates were studied in two lineages of transgenic mice that overproduce the hepatitis B virus (HBV) large envelope protein and retain filamentous hepatitis B surface antigen (HBsAg) particles in the endoplasmic reticulum, resulting in the formation of ground glass hepatocytes. The high producer lineage (50-4) develops a necroinflammatory liver disease that progresses to hepatocellular carcinoma (HCC), whereas the low producer lineage (107-5) displays no histopathologic changes other than ground glass hepatocytes. Bromodeoxyuridine (BrdU)-labeling studies of S-phase hepatocytes provide quantitative evidence for a strong, sustained proliferative response in the hepatocytes in lineage 50-4 that occurs after the onset of hepatocellular injury but long before the development of liver cell tumors. In contrast, the level of hepatocellular proliferation in lineage 107-5 is the same as nontransgenic controls. The findings support the concept that sustained hepatocellular proliferation plays an important role in the development of hepatocellular carcinoma (HCC).
Mapping of immunodominant CD4+ T lymphocyte epitopes of hepatitis C virus antigens and their relevance during the course of chronic infection
Authors: Hoffmann Robert M. , Diepolder Helmut M. , Zachoval Reinhart , Zwiebel Franz-Maximilian , Jung Maria-Christina , Scholz Siegfried , Nitschko Hans , Riethmüller Gert , Pape Gerd R.
Published: March 2007

In acute and chronic viral disease the specific response of CD4+ T lymphocytes to certain viral proteins is an essential part of antiviral effector mechanisms. In hepatitis C virus infection, the contribution of the immune system and particularly of CD4+ T lymphocytes to the pathogenesis of disease is unknown. We serially determined the peripheral blood CD4+ T lymphocyte response to several recombinant hepatitis C virus proteins (core, NS3, NS4, NS5) and 17 overlapping synthetic peptides derived from the core sequence over up to 48 months in 43 patients with chronic hepatitis C; of these, 16 had been treated with interferon alfa (IFN). Twelve of 27 untreated patients, 4 of 4 sustained responders to IFN, 7 of 8 patients with a transient response, and 1 of 4 nonresponders showed a proliferative response to hepatitis C virus proteins. The hepatitis C virus core protein was the most immunogenic protein, and fine analysis with peptides indicated amino acids 23 to 42, 66 to 85, and 131 to 150 as immunodominant regions. In a subgroup of nine patients, proliferation assays were performed before or during IFN. In this subgroup, sustained responders but not those with a transient or no response to IFN showed a specific CD4+ immune reaction to hepatitis C viral antigens (P < .05). Infection with hepatitis C virus genotype 3a was significantly associated with a sustained response to IFN (P < .05). In general, a CD4+ T lymphocyte response was more common in patients with chronic hepatitis C who responded to interferon-alpha as compared with nonresponders. Thus a strong CD4+ reaction before and during IFN therapy may be a predictor of sustained response.
Breakthrough during recombinant interferon alfa therapy in patients with chronic hepatitis C virus infection: Prevalence, etiology, and management
Authors: Roffi Luigi , Mels Guido Colloredo , Antonelli Guido , Bellati Giorgio , Panizzuti Fabio , Piperno Alberto , Pozzi Massimo , Ravizza Davide , Angeli Giovanni , Dianzani Ferdinando , Mancia Giuseppe
Published: March 2007

Recombinant interferon alfa (r-IFNα2) has been shown to normalize the aminotransferase levels in approximately 50% of patients with chronic hepatitis C virus (HCV). Few patients experience a relapse during the treatment, in spite of a complete initial response (breakthrough). We studied 191 HCV Ab-positive patients with histologically proven chronic hepatitis. All of them were treated with r-IFNα2 (3 MU three times a week). A complete response was seen in 54.4%. However, 12 of 104 responders experienced a breakthrough. At the time of breakthrough, neutralizing IFN antibodies were positive in 6 of 12 patients. Binding IFN antibodies were positive in all of these 12 patients. Continued treatment with r-IFNα2, even at higher doses, did not restore the previous response in any patient. All of them were then switched to natural lymphoblastoid IFN, and this rapidly restored a complete response in all of the patients.
Incidence of hepatocellular carcinoma in chronic hepatitis B and C: A prospective study of 251 patients
Authors: Takano Susumu , Yokosuka Osamu , Imazeki Fumio , Tagawa Masami , Omata Masao
Published: March 2007

The incidence of hepatocellular carcinoma (HCC) was prospectively studied in 251 chronic hepatitis patients, and was compared between the 127 cases of hepatitis B and 124 cases of hepatitis C. All patients were diagnosed by needle biopsy on entering the study, and the cases consisted of chronic persistent hepatitis (CPH), chronic active hepatitis (CAH)2a, and CAH2b (cirrhosis was not included). Of the cases of chronic hepatitis B, 5 cases of HCC (3.9%) were detected; among the chronic hepatitis C cases, 13 cases (10.4%) were detected. Thus, although the mean follow-up periods were in the same range, the incidence of hepatocellular carcinoma was 2.7 times higher in hepatitis C than in hepatitis B (χ2 = 3.116, P < .05). Using the Kaplan-Meier method, the incidence of HCC was significantly higher in chronic hepatitis C (P = .0194, generalized Wilcoxon test). In hepatitis C, the incubation period until HCC was detected was shorter when the liver disease was more advanced. Such a tendency was not observed in hepatitis B. In the 13 cases of HCC occurring in chronic hepatitis C, noncirrhotic liver was seen in only 1 case (7.7%), whereas 2 of the 5 cases of HCC (40%) in chronic hepatitis B were noncirrhotic. The prevalence of hepatitis C virus (HCV) genotypes II and III was the same in the total followed cases and HCC cases.
Escherichia coli capsular polysaccharide and spontaneous bacterial peritonitis in cirrhosis
Authors: Soriano Germán , Coll Pere , Guarner Carlos , Such José , Sánchez Ferran , Prats Guillem , Vilardell Francisco
Published: March 2007

Spontaneous bacterial peritonitis (SBP) is a frequent and severe complication of cirrhosis. Escherichia coli is the most frequent bacterium isolated in this condition. The presence of capsular antigens, mainly the K1 capsular polysaccharide, has been associated with invasiveness in E coli infections. Capsular serotypes of E coli causing SBP were determined in 37 cirrhotic patients. Twenty-seven strains were encapsulated (72.9%), 9 of them (24.3%) with K1 capsular polysaccharide, and 10 were nonencapsulated. Patients with encapsulated E coli showed a significantly higher incidence (92.5% vs. 50%; P < .01) and number of complications per patient (1.9 ± 1.1 vs. 0.8 ± 1.0; P < .01) than patients with nonencapsulated strains. Although mortality was higher in patients with encapsulated strains (44.4% vs. 20%), the difference did not reach statistical significance. Considering patients infected by encapsulated strains, the incidence of complications and mortality were similar in patients with or without K1 strains. These data suggest that the presence of encapsulated strains could have a prognostic significance in SBP caused by E coli in cirrhotic patients.
Two different dosages of cefotaxime in the treatment of spontaneous bacterial peritonitis in cirrhosis: Results of a prospective, randomized, multicenter study
Authors: Rimola Antoni , Salmerón Joan M. , Clemente Gerardo , Rodrigo Luis , Obrador Antoni , Miranda Luisa M. , Guarner Carlos , Planas Ramon , Solá Ricard , Vargas Victor , Casafont Fernando , Marco Francesc , Navasa Miquel , Bañares Rafael , Arroyo Vicente , Rodés Joan
Published: March 2007

Cefotaxime (CTX) is considered one of the first-choice antibiotics in the therapy of spontaneous bacterial peritonitis (SBP) in cirrhosis. Because CTX is largely metabolized in the liver, this drug may also be effective in SBP by administering lower doses than those habitually used. To investigate this possibility, a prospective, randomized, multicenter study was performed to compare the therapeutic efficacy of two different dosages of CTX in 143 patients with SBP: 71 (group I) were allocated to receive a high dose (2 g every 6 hours, which is one of the most frequently recommended doses in this infection), and 72 (group II) were allocated to receive a low dose (2 g every 12 hours). At inclusion, both groups were similar in relation to clinical and laboratory data, with the exception of a higher incidence of positive ascitic fluid culture in group I than in group H (59% vs. 40%; P = .029). The rate of infection resolution was similar for both groups (77% vs. 79%). Hospital survival was also similar in both groups (69% vs. 79%). No difference was observed between patients with positive or negative ascitic fluid cultures with regard to infection resolution and patient survival. The duration of antibiotic therapy was similar in both groups (9.0 ± 3.3 days in group I vs. 8.8 ± 3.1 days in group II). In a subset of 13 patients from group I and 11 patients from group II CTX levels were determined in serum (peak and trough) and ascitic fluid (concomitantly with trough serum). Peak serum levels were similar in patients from both groups. In contrast, trough serum and/or ascitic fluid levels were significantly lower or more frequently undetectable in group II patients than in group I patients. Nevertheless, this feature did not correlate with infection resolution or patient survival. These results indicate that the High efficacy of CTX in SBP can be maintained by using doses lower than those habitually recommended.
Transjugular intrahepatic portosystemic shunts for refractory ascites: Assessment of clinical and hormonal response and renal function†
Authors: Somberg Kenneth A. , Lake John R. , Tomlanovich Stephen J. , Laberge Jeanne M. , Feldstein Vickie , Bass Nathan M.
Published: March 2007

Cirrhosis is frequently complicated by ascites that may become resistant to diuretic therapy. Transjugular intrahepatic portosystemic shunts (TIPS) represent a new treatment for this debilitating condition. The aim of this study was to ascertain the clinical efficacy of TIPS, as well as its impact on renal function and on hormonal parameters. Five inpatients with refractory ascites were studied prospectively before TIPS, and 3 and 14 days after TIPS. After TIPS, ascites completely resolved or was minimal in all patients. Diuretics were discontinued in three subjects and decreased by at least 50% in two. One patient developed liver failure after TIPS and required liver transplantation; the others remained stable after a mean follow-up of 14 months. Mean urinary sodium excretion increased from 2.1 ± 0.6 mEq/24 hr before TIPS to 13.0 ± 4.3 mEq/24 hr 14 days after TIPS. Mean serum creatinine and glomerular filtration rate also tended to improve during the study period. With the exception of the patient who developed liver failure, plasma aldosterone concentration decreased from a mean of 126.0 ± 29.9 ng/dL to 22.8 ± 6.8 ng/dL (P = .04), and plasma renin activity decreased from a mean of 9.0 ± 3.0 μg/L/h to 0.9 ± 0.1 μg/L/h (P = .08). Additionally, 19 patients who underwent TIPS for refractory ascites outside of this protocol were followed prospectively for a mean of 282 days. Clinical improvement in ascites control was noted in 74%, and the mean dose of diuretics was decreased by more than 50%. Nonresponders more often had underlying renal disease. In conclusion, TIPS is an effective therapy for refractory ascites in most patients. TIPS improves renal function and in most patients reverses underlying hormonal derangements. TIPS may have an adverse effect on hepatic function, necessitating careful patient selection.
Prevalence, severity, and risk factors of liver disease in blood donors positive in a second-generation anti—hepatitis C virus screening test†
Authors: Serfaty Lawrence , Nousbaum Jean Baptiste , Elghouzzi Marie Helene , Giral Phillipe , Legendre Claire , Poupon Raoul
Published: March 2007

In a cohort of 483 blood donors positive for antibody to hepatitis C virus on second-generation enzyme-linked immunosorbent, the confirmatory second-generation recombinant immunoblot assay (Ortho Diagnostic Systems) was positive in 172 cases (36%), indeterminate in 113 (23%), and negative in 198 (41%). We further studied 94 of the donors (recombinant immunoblot assay positive in 85, indeterminate in 6, and negative in 3). Alanine transaminase (ALT) activity, assayed on three occasions, was elevated in at least one assay in 85% of the 85 recombinant immunoblot assay-positive donors. Liver disease was present in 95% of these patients (chronic persistent hepatitis, 35%; chronic active hepatitis, 53%; cirrhosis, 7%). Ten of the 13 recombinant immunoblot assay-positive donors with normal ALT activity had liver disease; polymerase chain reaction testing for viral RNA was predictive of liver disease in most cases. Donors with cirrhosis differed significantly from cirrhosis-free donors in terms of age, sex ratio, ALT activity, and excessive alcohol consumption. Three of the 6 recombinant immunoblot assay-indeterminate donors (isolated C 22) who underwent histological examination had elevated ALT activity and liver disease. The 3 recombinant immunoblot assay-negative donors evaluated were free of liver disease. This study shows that anti-HCV second-generation enzyme-linked immunosorbent positivity is confirmed in fewer than 40% of blood donors by the second-generation recombinant immunoblot assay, and that liver disease is present in 95% of recombinant immunoblot assay-positive donors. Recombinant immunoblot assay positivity combined with viremia is frequently associated with the existence of liver disease, regardless of transaminase activity. Excessive alcohol consumption may be an important factor in the onset of cirrhosis in anti-HCV-positive blood donors.
Hepatocellular codistribution of c100, c33, c22, and NS5 hepatitis C virus antigens detected by using immunopurified polyclonal spontaneous human antibodies
Authors: Ballardini Giorgio , Groff Paolo , Giostra Fabrizio , Francesconi Raffaella , Miniero Rita , Ghetti Sabrina , Zauli Daniela , Bianchi Francesco B.
Published: March 2007

Hepatitis C virus (HCV) antigens in liver biopsy have been detected by immunohistochemistry using both spontaneous human IgG and murine monoclonal or rabbit polyclonal monospecific reagents. Conflicting results have been obtained in different studies. This was probably because of the incapacity of single experimental antibodies, raised against synthetic or recombinant peptides, to recognize native tissue antigens. To overcome this possibility, we immunopurified monospecific spontaneous polyclonal human Ig, therefore induced by native antigens, from the single antigen-containing bands of RIBA 3 strips. Antibodies to c100, c33, c22, and NS5 antigens were obtained from the serum of a patient affected by chronic hepatitis C. The IgG fraction of this serum had proved to stain tissue HCV antigens. Eight biopsies were selected on the basis of strong hepatocellular reactivity with the whole IgG fraction in a variable number (from 5% to 75%) of cells. The four antigens were detected in all biopsies; a clear cellular codistribution was observed on serial sections. These data demonstrate that the possibility to identify HCV antigens in liver biopsies is higher when using human antibodies induced by native antigens rather then experimental antibodies. The approach of immunopurification of human antibodies can be extended to other HCV-related epitopes to obtain reagents useful for the selection and optimization of monoclonal or polyclonal antibodies.
Endothelin-1 and -3 plasma concentrations in patients with cirrhosis: Role of splanchnic and renal passage and liver function†
Authors: Gerbes Alexander L. , Møller Søren , Gülberg Veit , Henriksen Jens H.
Published: March 2007

Increased as well as decreased plasma concentrations of the endothelins, endogenous vasoactive peptides, have been reported in cirrhosis. This might be caused by alterations of hepatic or renal clearance or release. Therefore, the aim of the current study was to investigate the influence of splanchnic and renal passage and of liver function on plasma concentrations of endothelin-1 (ET-1) and endothelin-3 (ET-3) in patients with cirrhosis compared with controls. Eighteen patients with cirrhosis and 8 normotensive controls of similar age were investigated. Arterial and venous plasma samples were obtained simultaneously, and ET-1 and ET-3 concentrations were determined in extracted plasma by two separate radioimmunoassays. Arterial as well as hepatic and renal venous concentrations of ET-1 in cirrhosis (17.8 ± 0.8 pg/mL, 19.1 ± 0.9 pg/mL, and 16.8 ± 0.8 pg/mL) were significantly (P < .001) higher than in controls (9.2 ± 1.7 pg/mL, 9.0 ± 2.0 pg/mL, and 8.4 ± 1.9 pg/mL, respectively). The same held true for the corresponding ET-3 plasma concentrations in cirrhosis (19.3 ± 1.6 pg/mL, 20.5 ± 1.5 pg/mL, and 18.4 ± 1.5 pg/mL, respectively) compared with controls (11.1 ± 1.8 pg/mL, 11.3 ± 1.5 pg/mL, and 10.1 ± 1.7 pg/mL, respectively; P < .01). There was a significant (P < .05) renal net extraction of ET-1 and ET-3 in cirrhosis. In contrast, a significant (P < .05) net release of ET-1 and ET-3 (2.40 ± 0.80 ng/min and 1.75 ± 1.16 ng/min) during splanchnic passage was observed in cirrhosis, but not in controls (−0.24 ± 0.51 ng/min, and −0.46 ± 0.64 ng/min). Plasma concentrations of ET-3 in cirrhosis were correlated to the Child-Turcotte score (r = .66, P < .01) and inversely to the functional liver cell mass, determined by the galactose elimination capacity (r = −.72,P < .01). Hepatic venous ET-1 concentrations correlated to the hepatic blood flow assessed by the indocyanine green clearance (r = .48; P < .05). Net splanchnic release may contribute to elevated ET-1 and ET-3 plasma concentrations in patients with cirrhosis. Splanchnic ET-1 and ET-3 handling in cirrhosis may be influenced by different mechanisms.
Isolation and purification of large quantities of fresh human Kupffer cells, which are cytotoxic against colon carcinoma
Authors: Heuff Gijsbert , De Van Loosdrecht Arjan A. , Betjes Michiel G. H. , Beelen Robert H. J. , Meijer Sybren
Published: March 2007

A new rapid method is described for the isolation and purification of functional active human Kupffer cells without the need of in situ perfusion techniques. Liver wedge biopsies (3 to 5 g), obtained after laparotomy, were incubated with pronase under continuous pH registration. Human Kupffer cells were subsequently separated from other nonparenchymal cells by Nycodenz gradient centrifugation and purified by counterflow centrifugal elutriation. Kupffer cells, 1.7 ± 0.4 × 106 per gram liver, were isolated with a purity of 95% ± 3%. Cell-mediated cytotoxicity of Kupffer cells was assayed against a human colon carcinoma cell line (SW948). Kupffer cell cytotoxicity was 42% ± 9% (mean ± SD) at an effector-to-target cell ratio of 10 and significantly increased to 73 ± 17% (P < .05) after activation of Kupffer cells with interferon-gamma. In conclusion, a reliable and relatively simple method is provided to isolate and purify fresh human Kupffer cells in large yields, which show spontaneous as well as gamma-interferon-induced cytotoxicity against a human colon carcinoma cell line.
Ex vivo hepatic gene transfer in mouse using a defective herpes simplex virus-1 vector
Authors: Lu Bing , Gupta Sanjeev , Federoff Howard
Published: March 2007

A defective amplicon herpes simplex virus-1 (HSV-1) vector, HSVlac, was used to transfer an E. coli lacZ reporter gene into primary hepatocytes. The lacZ gene was driven by the HSV immediate early (IE) 4/5 promoter. Use of the HSVlac vector resulted in highly efficient gene transfer. Because difficulties in culturing primary hepatocytes impose limitations in ex vivo gene therapy, we sought to determine whether use of the HSVlac vector could simplify gene transfer. Therefore, we incubated HSVlac with primary hepatocytes in suspension and found that the lacZ gene was still transferred with great rapidity and efficiency. To examine lacZ expression in transduced hepatocytes in vivo, we used a mouse hepatocyte transplantation system. In congeneic recipients of primary hepatocytes transduced with HSVlac in suspension, the lacZ gene was expressed in liver and spleen up to 2 weeks. However, survival of transplanted hepatocytes, as well as persistence of HSVlac genome in recipient organs, was demonstrated for up to an 11-week duration of the experiment. These findings suggest that in vivo regulation of the HSV IE4/5 promoter was responsible for the short-term expression of lacZ, which should be overcome by the use of liver-specific promoters. Therefore, our results indicate the feasibility of hepatic gene transfer with a defective HSV-1 vector.
Transforming growth factor—beta 1 (TGF-β1) and TGF-β1 receptors in normal, cirrhotic, and neoplastic human livers
Authors: Bedossa Pierre , Peltier Eric , Terris Benoit , Franco Dominique , Poynard Thierry
Published: March 2007

Transforming growth factor-Beta 1 (TGF-β1) is an important mediator of control of liver cell proliferation and replication. The aim of the current study was to compare TGF-β1 gene expression, protein synthesis, and cell membrane receptors in normal liver, cirrhotic nodules, and neoplastic human livers. Five surgical resections for metastasis in an otherwise normal liver and 25 resections for hepatocellular carcinoma with cirrhosis were included in this study. Messenger RNA (mRNA) and TGF-β1 protein were detected on serial tissue sections of normal, cirrhotic and tumoral livers using in situ hybridization and immunohistochemistry. TGF-β1 type II receptors were detected on tissue sections using immunohistochemistry. In normal livers, TGF-β1 mRNA and protein were not significantly expressed. In cirrhotic nodules, a few sinusoidal cells and mesenchymal cells of fibrous septa displayed TGF-β1 mRNA. By immunohisto-chemistry, protein was detected in the extracellular matrix along the fibrous septa. Hepatocytes from normal and cirrhotic livers did not express TGF-β1. In contrast, the cytoplasm of hepatocytes in neoplastic nodules showed intense staining for TGF-β1 mRNA and protein. Although TGF-β1 receptor II was expressed on the plasma membrane of normal liver cells, tumoral hepatocytes no longer displayed membrane labeling but rather diffuse intracytoplasmic staining with perinuclear accumulation. This study suggests that the escape of tumoral hepatocytes from control of cell proliferation by TGF-β1, despite its overexpression by these cells, might be related to a defect in TGF-β1 receptor II processing on the liver cell membrane.
Spontaneous acceptance of rat liver allografts is associated with an early downregulation of intragraft interleukin-4 messenger RNA expression
Authors: Farges Oliver , Morris Peter J. , Dallman Margaret J.
Published: March 2007

Liver allografts are not rejected in the fully incompatible Lewis-RT11 (LEW) to blood group D Agouti-RT1a (DA) rat strain combination despite an early infiltration by recipient mononucleated cells that initially display a phenotype, an ability to respond to interleukin-2 (IL-2) and donor-specific cytotoxicity indistinguishable from that observed in the rejected, DA to LEW combination. To further analyze the mechanism of this tolerance, we have compared in these two combinations, as well as in syngeneic grafts and in normal livers, the presence of intrahepatic cytokine transcripts (IL-1α, IL-2, IL-4, IL-6, IL-10, tumor necrosis factor [TNF]-α, TNF-β, and transforming growth factor [TGF]-β) by a semiquantitative polymerase chain reaction (PCR) or by northern-blotting. In normal livers or syngeneic grafts, IL-1α, TNF-β, and TGF-β were the only cytokines detected by these methods. The levels of all cytokine transcripts were increased in allogeneic grafts. Expression of cytokine transcripts was very similar in the two allogeneic strain combinations except IL-4, which was expressed at a much lower level in the nonrejected strain than in the rejected strain from day 2 onward. We conclude that selective downregulation of IL-4 gene expression is associated with, and a potential mediator of, the induction of tolernce in this model.
Nitric oxide is released in regenerating liver after partial hepatectomy
Authors: Hortelano Sonsoles , Dewez Beatrice , Genaro Ana M. , Díaz-Guerra María J. M. , Boscá Lisardo
Published: March 2007

The induction of hepatic nitric oxide synthase (NOS) and the biosynthesis of nitric oxide (NO) were studied in liver after partial hepatectomy (PH). NOS activity in the liver remnant was observed 4 to 6 hours after PH, and no differences were evidenced between the proximal and distal surgical areas. The form of NOS expressed in liver was independent of calcium and calmodulin, and the messenger RNA levels were first detected 2 hours after hepatectomy using a probe corresponding to the cytokine-induced macrophase NOS. The seric concentration of nitrites remained unchanged after hepatectomy, whereas the content in nitrates and in S-nitrosylated proteins progressively increased in parallel with the NOS activity. The spectra of hemoglobin in the 400- to 460-nm region failed to exhibit the characteristic shift caused by the formation of the nitrosyl-hemoglobin complex, suggesting that NO was rapidly metabolized in liver. Treatment of the animals with substrate analogue NOS inhibitors blocked the pattern of DNA ploidy elicited after hepatectomy, suggesting a role for NO in the regenerative process. Peritoneal resident macrophages were used as an alternative reporter cell system for the assessment of NOS expression. Incubation ex vivo of peritoneal macrophages from animals that underwent hepatectomy induced the expression of NOS in a cyto-kine-modulated fashion, suggesting that macrophages were primed as a result of the hepatectomy. When peritoneal macrophages from control rats were incubated with the sera of animals that underwent hepatectomy, a time-dependent induction of NOS was observed, with a maximal induction corresponding to sera collected 2 hours after PH. These results indicate that NO might be involved in the control of early responses after PH.
Increased expression of matrix metalloproteinase-II in experimental liver fibrosis in rats
Authors: Takahara Terumi , Furui Kei , Funaki Jun , Nakayama Yoshihide , Itoh Hiroyuki , Miyabayashi Chiharu , Sato Hiroshi , Seiki Motoharu , Ooshima Akira , Watanabe Akiharu
Published: March 2007

Matrix metalloproteinase-II (MMP-II, 72-kd type IV collagenase, or gelatinase) is one of the gene families of zinc enzymes capable of degrading extracellular matrix molecules, and specifically of degrading type IV and V collagens, gelatin, fibronectin, and elastin. In this study, we used both the liver fibrosis model and the reversibility model of experimental cirrhosis to clarify how MMP-II participates in liver fibrosis of rats. To produce fibrosis model, rats received subcutaneous injections of CCI4 twice weekly for 7, 9, or 14 weeks. For the reversibility model, rats were treated with CCl4 three a week for 8 weeks and killed at 3, 7, 14, 28, or 42 days after discontinuation of treatment. MMP-II gene expression was studied by Northern hybridization technique, and gelatinase activity of MMP-II was examined by zymography using gelatin substrate. At the same time, an immunohistochemical study using anti-type IV collagen antibody was carried out. In liver fibrosis model, nodule formation was established at 14 weeks. Immunodeposit of type IV collagen was increased in wide fibrous septa and was clearly observed along sinusoidal wall. Gene expression of MMP-II increased up to 7 to 12 times compared with that of controls, with the expression rate being maximum at an intermediate stage of fibrosis. Zymography showed the expressions of both 65-kd latent MMP-II, which is confirmed to be activated by adding p-aminophenylmercuric acetate, and 62-kd active MMP-II during fibrosis. The expression of both forms increased 13 to 28 times as the fibrosis progressed. By contrast, little latent MMP-II was detected in control livers. The percent active form to total MMP-II at each stage was elevated most at an intermediate stage of fibrosis up to 30% and decreased to 16% in the cirrhotic stage. As cirrhosis reversed, fibrous septa became thin but still persisted at 42 days in the reversibility model. Immunostaining of type IV collagen was increased in thin septa and faintly observed along sinusoid. Gene expression was elevated 18-fold and recovered gradually to remain elevated 18-fold and recovered gradually to remain elevated at 42 days after the discontinuation of intoxication. Expressions of both active and latent forms detected by zymography were elevated 15-fold during the early reversible stage and decreased gradually after the discontinuation of intoxication. These results indicated MMP-II may participate in pathogenesis of liver fibrosis and cirrhosis.