bluuebunny/crossref_metadata_2025 huggingface dataset:
Sclerotherapy plus octreotide versus sclerotherapy alone in the prevention of early rebleeding from esophageal varices: A randomized, double-blind, placebo-controlled, multicenter trial
Authors: Pkimignani Massimo , Andreoni Bruno , Carpinelli Luca , Capria Alfonso , Rocchi Gabriele , Lorenzini Ivano , Staudacher Carlo , Beretta Luigi , Motta Roberta , DeFranchis Roberto
Published: March 2007

Because of its ability to decrease portal pressure, azygos blood flow, and postprandial splanchnic hyperemia, octreotide administration could be effective in reducing early rebleeding in patients undergoing endoscopic variceal sclerotherapy (EVS). We report the results of a trial comparing EVS + octreotide versus EVS alone. Consecutive patients with cirrhosis and endoscopically proven variceal hemorrhage were considered eligible for the trial if hemodynamically stable for at least 24 hours after bleeding stopped. Patients with advanced liver cancer or having received EVS treatment in the past were not enrolled. After enrollment patients were submitted to EVS (day 1); all patients were randomized to receive octreotide, 100 μg three times a day subcutaneously, or an identical placebo, up to day 29; EVS was repeated at days 8, 15, and 29. Fifty-eight patients were randomized to receive either EVS + octreotide (n = 26) or EVS alone (n = 32). The two groups were evenly balanced for sex, age, Child-Pugh class, history of previous bleeding, endoscopic appearance of varices, or treatment received in emergency. Eight of 26 (31%) patients in the EVS + octreotide group rebled, compared with 11 of 32 (34%) in the EVS group. Four of the eight (50%) patients in the former group and 8 of 11 (73%) in the latter, respectively, bled within day 15. There were 10 (38.5%) deaths in the EVS + octreotide group (seven bleeding-related), compared with seven (21.9%) (five bleeding-related) in the EVS group; these differences did not reach statistical significance. Administration of octreotide, 100 μg three times a day, subcutaneously, to patients undergoing EVS for prevention of recurrent variceal bleeding does not decrease the incidence of early rebleeding.
Which is the best surgery for budd-chiari syndrome: Venous decompression or liver transplantation? a single-center experience with 50 patients
Authors: Ringe Burckhardt , Lang Hauke , Oldhafer Karl-Jürgen , Gebel Michael , Flemming Peer , Georgii Axel , Borst Hans-Georg , Pichlmayr Rudolf
Published: March 2007

The optimal treatment of Budd-Chiari syndrome (BCS) remains an open question. It is still a matter of controversial discussion whether venous decompression or liver transplantation is superior. To elucidate the role and prognosis of both surgical options in our own experience, a consecutive series of 50 patients treated between 1981 and 1993 was retrospectively analyzed. Twelve patients had different types of portosystemic shunts or local decompressive procedures, and transplantation was performed in 43 cases, including five with previous conventional surgery. The overall mortality of 18 of 50 was concentrated within the early postoperative period, with no patient lost after 1 year. In the venous decompression group, the success rate was only 29%, and treatment failure was closely related to the finding of cirrhosis or technical problems like vascular thrombosis. After transplantation, early complications were rejection, primary nonfunction, or graft necrosis, and contributed significantly to the risk of sepsis. Thirty of 43 liver recipients are currently alive, including four rescued after failed decompressive surgery, with 1- and 10-year survival of 69%, and excellent recurrence-free rehabilitation. These results clearly indicate that patient selection plays a dominant prognostic role in the treatment of BCS. Venous decompression and liver transplantation should both be integrated in a common therapeutic concept, and the individual decision for the preferred approach must be based on the leading clinical symptom: portal hypertension or liver failure, together with the assessment of reversibility of hepatic damage, and the potential of cure of the underlying disease.
Regression of bile duct damage and bile duct proliferation in the non-rearterialized transplanted rat liver is associated with spontaneous graft rearterialization
Authors: Zhao Delai , Zimmermann Arthur , Kuznetsova Larisa V. , WheatleyD Anthony M.
Published: March 2007

The aim of this study was to investigate the long-term consequences of non-rearterialization of the graft in rat liver transplantation. Liver transplantation with (AOLT) and without graft rearterialization (NOLT) was performed in anesthetized male Lewis rats. Quantitative morphometry and semiquantitative histopathology of the liver were performed at various times after operation. Volume fractions of tissue components were determined. The number of arteries and bile ducts per portal tract were measured in histological sections from both groups. Hepatic blood flow was measured using the radioactive microsphere technique in rats after NOLT (6 months). AOLT livers had a preserved lobular architecture at all time points and unaltered volume fractions. In addition, AOLT livers maintained approximately one artery and one bile duct per portal tract after transplantation. NOLT livers showed bile duct damage at 3 days, cellular infiltration and ductular proliferation at 1 week, increased ductular proliferation at 4 weeks, and fibrosis at 6 months. The volume fractions for nonhepatocyte parenchyma (3 days, 19.14 ± 1.29; 1 week, 20.44 ± 1.76; 4 weeks, 15.46 ± 3.14), bile ducts/ductules (1 week, 4.88 ± 1.07; 4 weeks, 7.20 ± 2.42), and connective tissue (4 weeks, 4.02 ± 1.66; 6 months, 14.94 ± 0.63) were significantly increased. Hepatocyte volume fraction was significantly decreased at all time points. A total of 1.58 ± 0.08 arteries/portal tract were found in NOLT livers after 4 weeks, rising to 2.44 ± 0.10 arteries/portal tract after 6 months. At 6 months, hepatic arterial blood flow (0.69 mL/min/g) was significantly higher (P < 0.02) than control (0.25 mL/min/g). The findings indicate that NOLT leads to a time-dependent overall derangement of liver structure, possibly due to bile duct damage induced by lack of an arterial blood supply. Spontaneous graft rearterialization may be responsible for the regression of bile duct damage and proliferation noted at the later stages. Nevertheless, biliary fibrosis may be a long-term consequence of NOLT.
Progression of subcellular changes during chemical hypoxia to cultured rat hepatocytes: A laser scanning confocal microscopic study
Authors: Zahrebelski George , Nieminen Anna-Liisa , Al-Ghoul Kristin , Qian Ting , Herman Brian , Lemasters John J.
Published: March 2007

The aim of this study was to evaluate changes in the subcellular organelles of cultured hepatocytes by laser scanning confocal microscopy during chemical hypoxia with cyanide and iodoacetate, inhibitors of mitochon-drial respiration and glycolysis, respectively. Parameter-specific fluorophores used were calcein for cell topography and membrane permeability, rhodamine-dextran for lysosomes, rhodamine 123 and tetramethylrhodamine methylester (TMRM) for mitochondrial membrane potential (Δ Ψ) and propidium iodide for loss of cell viability. During the first 30 to 40 minutes of chemical hypoxia to cultured hepatocytes, numerous surface blebs formed and cell volume increased, but Δ Ψ decreased relatively little. Subsequently, the nonspecific permeability of mitochondrial membranes increased, and mitochondria depolarized. These events were followed a few minutes later by disintegration of individual lysosomes. After a few more minutes, viability was lost as indicated by bleb rupture, gross plasma membrane permeability to calcein, and nuclear labeling with propidium iodide. Thus, the following sequence of intracellular events occurred during chemical hypoxia: adenosine triphosphate (ATP) depletion, bleb formation with cellular swelling, onset of a mitochondrial permeability transition, disintegration of lysosomes, plasma membrane failure from bleb rupture, and cell death. Any explanation of the pathophysiology of hypoxic injury must take into account this unique sequence of events.
Is dietary erucic acid hepatotoxic in pregnancy? an experimental study in rats and hamsters
Authors: Reyes Humberto , Ribalta José , Hernández Ismael , Arrese Marco , Pak Nelly , Wells Michael , Kirsch Ralph E.
Published: March 2007

The hypothesis that dietary erucic acid may contribute to the pathogenesis of intrahepatic cholestasis of pregnancy has been examined in pregnant rats and hamsters after prolonged feeding of diets containing 25% rapeseed oil rich in erucic acid (40% of fatty acids) or corn oil, without erucic acid. Both dietary oils were well tolerated, although weight gain was 17% to 20% less in unimals receiving rapeseed oil. Rats and hamsters were studied on the last day of pregnancy and compared with age- and diet-matched nonpregnant animals. Histological examination showed no major morphologic abnormalities in liver, heart, kidneys, and adrenals. Similar microscopic deposits of fat were found in the livers and hearts of pregnant hamsters of both dietary groups. Chromatographic analysis of fatty acids in liver, heart, and kidney homogenates of hamsters and in isolated rat liver cells reflected the fatty acid composition of the dietary oils: oleic (18:1) and linoleic (18:2) acids were among the predominant fatty acids. Erucic acid was found in a higher proportion in the heart (14% by weight of total fatty acids) than in the liver (3%) and kidneys (3%) of animals fed rapeseed oil. Bile flow and biliary lipid composition was similar in rats and hamsters fed rapeseed or corn oil. Bile flow tended to be less in pregnant than in nonpregnant animals. Pregnant hamsters fed rapeseed oil tended to have the lowest bile flow. The lithogenic index of bile was slightly decreased in pregnant rats and increased in pregnant hamsters, although these proportional changes were similar for both diets. In all circumstances the lithogenic index remained below a value of 1. Biliary excretion of sulfobromophthalein (BSP) and maximal transport capacity of BSP into bile (BSP Tm) were similar in rats fed rapeseed or corn oil diets. In conclusion, no morphologic or hepatic functional abnormalities attributable to the diet could be detected in animals fed rapeseed oil with a high content of erucic acid, whereas pregnancy induced changes in bile secretion in rats and hamsters, irrespective of the diet administered. These observations are still insufficient to reject the possibility that rapeseed oil rich in erucic acid could be involved in the pathogenesis of intrahepatic cholestasis of pregnancy because dietary erucic acid could interact with a genetic metabolic predisposition to induce a disease that seems to be specific to humans.
Alterations in bile ducts and peribiliary microcirculation in rats after common bile duct ligation
Authors: Nakano Shu , Haratake Joji , Hashimoto Hiroshi
Published: March 2007

The chronological changes in ductular epithelium and peribiliary capillary plexuses (PBPs) after bile duct ligation are not well understood. Therefore, we examined alterations in both intrahepatic bile ductules and peribiliary microcirculation in rats after ligation of the common bile duct using immunohistochemistry, transmission electron microscopes (TEM), and scanning electron microscopes (SEM). Ductular proliferations appeared first in the peripheral areas of the portal spaces and then gradually advanced along with increasing jaundice. Distorted configuration of the hepatic parenchyma with interconnecting stroma rich in irregular ductules developed 4 weeks after the ligation. Numerous biliary cell types and cells of types intermediate between hepatocytes and biliary cells together with poorly fenestrated capillary-type vessels appeared in the periportal parenchyma, in association with an increased number of canalicular-ductular junctions on TEM. These biliary cells were often found within the lobules and apart from the stroma. SEM examination of hepatic microvascular casts using methacrylated resin showed formation of irregular portal and periportal capillary networks, partly derived from coarsened sinusoids. Direct connections between the newly formed capillary networks and the pre-existing PBPs or sinusoids were numerous, although there were few direct connections between the capillary networks and the hepatic arterial branches. Thus, these proliferated ductules and newly formed complicated capillary networks might play an important role in the effective transport of biliary materials between hepatocytes and native bile ducts or proliferated ductules through the altered microcirculation after bile duct ligation.
Glutathione conjugation of bromosulfophthalein in relation to hepatic glutathione content in the rat in vivo and in the perfused rat liver
Authors: Snel Wilco C. A. , Pang Sandy K. , MulderD Gerard J.
Published: March 2007

The relation between the rate of glutathione (GSH) conjugation and hepatic GSH content was studied in the rat in vivo and the in situ single-pass-perfused rat liver preparation with bromosulfophthalein (BSP) as the model substrate. The biliary excretion of the BSP-GSH conjugate and the hepatic GSH content were monitored simultaneously during intravenous infusions with BSP in the rat in vivo, and during liver perfusions with BSP-containing perfusion medium. Rats were pretreated with single or multiple doses of buthionine sulfoximine, an inhibitor of the de novo synthesis of GSH. Surprisingly, the excretion of the BSP-GSH conjugate was sustained at a high rate, despite a virtually complete depletion of hepatic GSH, both in the rat in vivo as well as in the perfused rat liver. The results indicate that GSH was still available for conjugation with BSP after apparent depletion of the hepatic GSH pool, presumably because of a residual de novo synthesis of GSH in the liver. Despite the multiple pretreatment with buthionine sulfoximine, the de novo GSH synthesis was sufficient to sustain a high rate of GSH conjugation of BSP. The cosubstrate-Km for GSH conjugation of BSP in the liver was estimated to be very small (approximately 0.3μmol/g): the excretion rate of the BSP-GSH conjugate was only impaired at minimal hepatic GSH levels.
Different features of ca2+ oscillations in differentiated and undifferentiated hepatocyte doublets
Authors: Kitamura Tsuneo , Watanabe Sumio , Ikejima Ken-Ichi , Hirose Miyoko , Miyazaki Akihisa , Yumoto Atsuko , Suzuki Satoko , Yamada Toshio , Kitami Noriyuki , Sato Nobuhiro
Published: March 2007

Cytosolic free Ca2+ ([Ca2+]i) oscillations are postulated to play a critical role in cellular proliferation. By using doublets of normal rats (NR) and those 18 hours after two-thirds hepatectomy (PHR), we investigated cytosolic free Ca2+ ([Ca2+]i) responses in liver regeneration. Normal rat hepatocyte doublets that retain their bile canaliculi are polarized and well differentiated. PHR doublets, which also retain their bile canaliculi, were characterized as undifferentiated by (1) decreased canalicular secretion of fluorescein-isothiocyanate-labeled glycocholate; (2) increased labeling index of hepatocytes in BrdU staining (˜30%); and (3) impaired transfer of fluorescent dye injected into one cell of the pair to the other. Addition of phenylephrine to NR and PHR doublets in the presence of extracellular Ca2+ resulted in [Ca2+]i oscillations or a nonoscillatory-sustained increase in [Ca2+]i followed by a gradual return to the baseline. Extracellular Ca2+ was not required for [Ca2+]i oscillations but was necessary for a sustained increase in [Ca2+]i. Simultaneous addition of prazosin, α1-receptor blocker, to doublets immediately abolished these [Ca2+]i responses. The [Ca2+]i level in each of the adjacent cells was synchronous in sustained increase in [Ca2+]i, but asynchronous in [Ca2+]i oscillations. As the phenylephrine concentration was increased (1 to 100 μmol/L), oscillations were replaced by a sustained increase in [Ca2+]i in NR doublets. In contrast, in PHR doublets, oscillations remained, whereas the frequency of oscillations increased in a dose-dependent manner. These results indicate that the mechanisms of phenylephrine-evoked [Ca2+]i responses are different in differentiated and undifferentiated doublets and that the frequency modulation of [Ca2+]i oscillations may be involved in the intracellular signal transduction in the cellular proliferation process during liver regeneration.
Role of cytoskeleton and acidification of endocytic compartment in asialoglycoprotein metabolism in isolated rat hepatocyte couplets
Authors: Harada Masaru , Sakisaka Shotaro , Yoshitake Masao , Ohishi Masahito , Itano Satoshi , Shakado Satoshi , Mimura Yoshihiro , Noguchi Kazunori , Sata Michio , Yoshida Hiroshi , Tanikawa Kyuichi
Published: March 2007

The process of receptor-mediated endocytosis is common to a variety of species and cell types. One of the best characterized receptor-ligand systems is the hepatocyte receptor for asialoglycoproteins. We investigated the morphological features of the uptake and intracellular transport of gold-conjugated asialofetuin in isolated rat hepatocyte couplets. We assessed the effects of colchicine, lumicolchicine, cytochalasin B, and chloroquine on the uptake and intracellular transport of asialoglycoproteins. Isolated rat hepatocyte couplets were incubated with gold-conjugated asialofetuin, and transmission electron micrographs of these cells were analyzed to determine the density and distribution of gold particles in the peripheral and pericanalicular areas. Results were analyzed morphometrically. Colchicine significantly inhibited the uptake and intracellular transport of asialoglycoproteins, but did not affect membrane fusion of endocytic compartments in the peripheral area. Lumicolchicine and cytochalasin B had minimal effects on these processes. Chloroquine inhibited the uptake of asialoglycoproteins, but did not affect the intracellular transport of asialoglycoproteins. Results suggest that the microtubule is essential for intracellular movement of endocytosed asialoglycoproteins and receptor recycling, and that endocytic structures in the peripheral regions can fuse in the absence of intact microtubules. We also found that uptake and intracellular transport of asialoglycoproteins were independent of the microfilaments, and the pH gradient in endocytic compartments was important in receptor-mediated endocytosis of asialoglycoproteins.
Glucagon effect on intracellular proteolysis and pericanalicular location of hepatocyte lysosomes in isolated perfused guinea pig livers
Authors: Lenzen Romana , Stark Peter , Kolb-Bachofen Viktoria , Strohmeyer Georg
Published: March 2007

In guinea pigs, glucagon choleresis is accompanied by a significant, but transient, stimulation of biliary protein secretion, which can be accounted for mainly by biliary discharge of lysosomal enzymes. To clarify whether intracellular proteolysis—a process regulated by glucagon and taking place predominantly in the lysosomes—may interact with biliary protein secretion, we determined hepatic proteolytic activity and bile secretory function during substrate deprivation, amino acid supplementation, and glucagon administration in isolated perfused guinea pig livers. To further elucidate the nature of transient lysosomal enzyme release into bile during glucagon infusion, we analyzed pericanalicular distribution of lysosomes by quantitative electron microscopy. The results demonstrate that intracellular proteolysis is accompanied by biliary excretion of lysosomal enzymes. Glucagon-induced secretion of these enzymes as well as labeled proteins into bile occurs independent of protein breakdown and cannot be modulated by addition of amino acids as potent inhibitors of intracellular proteolysis. During glucagon administration, bile canalicular area and pericanalicular distribution of secondary lysosomes show a rapid increase, which persists during the entire infusion period and thus does not explain the transient biliary release of lysosomal enzymes. We therefore postulate that regulation of this process must be located beyond the lysosomal compartment, either involving transport processes or intracellular kinetics of lysosome formation or altered fusion kinetics at the bile canalicular membrane compartment. Metabolic and biliary effects of glucagon seem to occur independent of each other and to underly different regulatory mechanisms.
Insulinlike growth factor—ii/mannose 6-phosphate receptor is expressed on ccl4-exposed rat fat-storing cells and facilitates activation of latent transforming growth factor-β in cocultures with sinusoidal endothelial cells
Authors: De Bleserc Pieter J. , Jannes Peggy , Van Buul-Offers Sylvia C. , Hoogerbrugge Cok M. , Van Schravendijk Christiaan F. H. , Niki Toshiro , Rogiers Vera , Van Den Brande Jan L. , Wisse Eddie , Geerts Albert
Published: March 2007

Transforming growth factor β (TGF-β), a potent fibrogenic cytokine, is secreted in latent form. We examined which cell type in both normal and carbon tetrachloride (CCl4)-induced fibrotic rat liver bears surface type II IGF/mannose 6-phosphate (IGF-II/M6P) receptor, known to facilitate activation of TGF-β. In addition, the role of the IGF-II/M6P receptor in activation of latent TGF-β was investigated in a coculture system with sinusoidal endothelial cells. Northern hybridization analysis for IGF-II/M6P receptor messenger RNA (mRNA) was performed on total RNA of different isolated and purified liver cell types. In normal liver, cells expressed little IGF-II/M6P receptor mRNA. In fibrotic liver, we found significant expression only in fat-storing cells. The presence of IGF-II/M6P receptors was established by [125I]IGF-II binding assays on freshly isolated fat-storing cells from normal and CCl4-exposed rat livers. We found specific binding of [125I]IGF-II only on CCl4 exposed fat-storing cells. As determined by polyacrylamide gel electrophoresis after affinity labeling, the specific binding involved 220 kD type II IGF receptors. Scatchard analysis revealed the presence of 3,043 ± 1,378 IGF-II/M6P high-affinity receptors/fat-storing cell, with a Kd of 387 ± 165 pmol/L. With a mink lung epithelial cell (Mv1Lu) proliferation inhibition assay, inhibition of proliferation (a measure of active TGF-β function) was determined using conditioned media of activated fat-storing cells, cocultures of fat-storing cells, and endothelial cells and pure endothelial cell cultures. We found that conditioned media of cocultures of fat-storing cells and endothelial cells inhibits the growth of Mv1Lu cells more strongly than conditioned media of homotypic cultures. Addition of neutralizing anti-TGF-β antibodies neutralizes this effect. The contribution of IGF-II/M6P receptors on the cell membrane of activated fat-storing cells to the activation of latent TGF-β was demonstrated by incubating the cells with M6P, or antibodies directed to the IGF-II/M6P receptor, both of which diminishes activation of TGF-β. We conclude that IGF-II/M6P receptors are present on activated fat-storing cells and that the presence of this receptor at the cell surface is necessary, but not sufficient, for activation of latent TGF-β. Other factors, derived from endothelial cells, are involved.
Rapid activation of the stat3 transcription complex in liver regeneration
Authors: Cressman Drew E. , Diamond Robert H. , Taub Rebecca
Published: March 2007

Liver regeneration in response to partial hepatectomy is a physiological growth response observed in the intact animal. Understanding the early signals that trigger liver regeneration is of vital importance to understand the liver's response to injury. It has been observed that several growth factors and cytokines, including epidermal growth factor (EGF) and interleukin-6 (IL-6), can activate members of the signal transducers and activators of transcription (Stat) family of transcription factors resulting in tyrosine phosphorylation of these factors, nuclear translocation, and an active DNA binding transcriptional complex. Because Stat3 participates in the regulation of primary growth response genes, we wondered if it is induced in the early phase of liver regeneration. We found that Stat3 DNA-binding activity is increased in the remnant liver within 30 minutes of partial hepatectomy and peaks at more than 30-fold at 3 hours. This induction is not observed after sham surgery. The induction of Stat3 appears to be part of the initial response of the remnant liver to partial hepatectomy, because it occurs in the presence of cycloheximide-mediated protein synthesis blockade. Activation of Stat3 is unusual, because it extends beyond the immediate-early time period and remains near peak level at 5 hours posthepatectomy. Although insulin-treated H35 cells activate many of the same immediate-early genes as regenerating liver, Stat3 is not induced in these cells. Because Stat factors are known to be inactivated by protein tyrosine phosphatases (PTPase), we showed that a PTPase is able to eliminate the DNA binding of hepatic Stat3. It is likely that Stat3 contributes to the transcriptional activation of a subset of immediate-early genes that are induced over a prolonged time in the G1 phase of hepatic cells following partial hepatectomy. The identification of Stat3 as an early factor in liver regeneration provides clues to the activation of signal transduction pathways in the remnant liver within the first minutes to several hours after partial hepatectomy.
The enigma of anti-neutrophil antibodies in ulcerative colitis primary sclerosing cholangitis: Important genetic marker or epiphenomenon?
Authors: Chapman Roger W.
Published: March 2007

Background/Aims:

Perinuclear antineutrophil cytoplasmic antibodies (pANCA) were found at high frequency in patients with primary sclerosing cholangitis and ulcerative colitis. In this study, to accumulate further evidence for the importance of genetic factors in pathogenesis of inflammatory bowel disease, sera of patients with inflammatory bowel disease and primary sclerosing cholangitis and their unaffected family members were tested for pANCA.

Methods:

Three hundred twenty-seven sera from 11 families of patients with primary sclerosing cholangitis, 43 families of patients with ulcerative colitis, 11 families of patients with Crohn's disease, and 11 healthy families were tested for pANCA in immunofluorescence on cytospin slides with isolated neutrophils.

Results:

pANCA were found in 82% of the patients with primary sclerosing cholangitis and in 25% of their relatives. In ulcerative colitis, 70% of the patients and 30% of their relatives had pANCA. pANCA were found only in low titers in 27% of patients with Crohn's disease and in 6% of their relatives. pANCA were not detected in members of healthy families. Only 16% of the patients with ulcerative colitis and their families and none of the patients with primary sclerosing cholangitis and their families were completely negative for pANCA.

Conclusions:

These data show that pANCA may be a genetic marker in families of patients with ulcerative colitis and primary sclerosing cholangitis.
Spectrum of liver disease and duck hepatitis B virus infection in a large series of chinese ducks with hepatocellular carcinoma
Authors: Duflot Agnes , Mehrotra Raj , Yu Shun-Zhang , Barraud Luc , Trepo Christian , Cova Lucyna
Published: March 2007

The striking difference in the geographical distribution of liver cancer in ducks raised the question of whether duck hepatitis B virus (DHBV), like mammalian hepadnaviruses, could be an oncogenic agent. Hepatocellular carcinomas (HCCs) have been found only in domestic ducks in Qidong, China, where hepatitis B virus infection and aflatoxin B1 (AFB1) are both risk factors and where a high frequency of human HCCs has been reported. To date, the study of liver pathology occurring in Chinese ducks has been hampered by the small number of samples available. We describe here a series of 59 Chinese brown duck livers that were collected in Qidong more than 20 years ago and formalin fixed. Thirty-six HCCs, which ranged from well-differentiated trabecular to highly anaplastic type, were identified in relatively young ducks (average age, 3.3 years). Several unique features not previously reported, such as tumor giant cells, tumor necrosis, tumor thrombi in blood vessels, and inactive cirrhosis, were observed. Bile ductule proliferation, known to be a prominent feature of AFB1 exposure in ducks, was present in 86% of livers. Using polymerase chain reaction (PCR) and two primer pairs, located within conserved portions of the DHBV S and C genes, we demonstrated the presence of DHBV DNA in 23 of 34 HCCs analyzed (68%). The spectrum of liver pathology that we report in brown ducks from Qidong was never observed in Pekin ducks of the same age chronically infected with DHBV and followed under controlled conditions outside China, suggesting that causative factors other than virus infection may be involved in duck liver carcinogenesis observed in this area. However, because DHBV DNA was present in most HCCs, and given the lower sensitivity of PCR in formalin-fixed tissue, we cannot rule out the importance of DHBV infection in the liver disease of ducks from this study. The specific role of Chinese DHBV isolates and brown duck breed in liver pathology and oncogenesis observed in Qidong ducks should be further investigated in controlled experimental transmission studies with long-term follow-up in the absence of external carcinogens.
Low prevalence of hepatitis C virus infection in porphyria cutanea tarda in germany
Authors: Stölzel Ulrich , Köstler Erich , Koszka Christiane , Stöffler-Meilicke Maricka , Schuppan Detlef , Somasundaram Rajan , Doss Manfred O. , Habermehl Karl O. , Riecken Ernst O.
Published: March 2007

Previous studies from Spain, Italy, and France have demonstrated a high prevalence (71% to 91%) of antibodies against hepatitis C virus in patients with porphyria cutanea tarda (PCT). To determine the role of hepatitis C virus (HCV) in PCT in Germany, we have assessed the prevalence of antibodies against HCV and hepatitis B virus (HBV) in 106 patients (mean age, 60 ± 14 years) with the disease. Eight of 106 patients (8%) were positive for HCV antibodies and HCV RNA using second-generation enzyme-linked immunosorbent assay (ELISA), recombinant immunoblot assay, and polymerase chain reaction. Antibodies against HBV core antigen were found in 14 patients (13%). Of the patients with antibodies against HCV alanine transaminase (ALT) (aspartate transaminase [AST]) levels above normal occurred in 71% (86%). Because elevated ALT (AST) levels were also found in 51% (64%) of 88 patients without markers of HCV or HBV, we suggest that liver damage in PCT may exist in absence of these viruses. This is supported by the finding that in patients without HCV or HBV markers, higher serum ALT and AST activities were found in patients with overt disease or relapse (ALT, 59 ± 44 U/L; AST, 37 ± 21 U/L), whereas patients in remission displayed significantly lower serum enzyme activities (ALT, 16 ± 8 U/L; AST, 16 ± 7 U/L), (P < 0.001). These results indicate that HCV infection does not play a major role in the pathogenesis of PCT in Germany.
Comparison of endoscopic variceal injection sclerotherapy and ligation for the treatment of esophageal variceal hemorrhage: A prospective randomized trial
Authors: Hou Ming-Chih , Lin Han-Chieh , Kuo Benjamin Ing-Tiau , Chen Chen-Hsiang , Lee Fa-Yauh , Lee Shou-Dong
Published: March 2007

To determine the efficacy of endoscopic variceal sclerotherapy (EVS) and ligation (EVL) in the management of esophageal variceal bleeding, 134 cirrhotic patients were randomized to receive either treatment. The clinical and endoscopic characteristics were similar in both groups. Active bleeding was controlled with ligation (20 of 20) as efficiently as with sclerotherapy (14 of 16). Elective sclerotherapy consumed less time than ligation (7.9 ± 1.8 minutes vs 11.5 ± 2.7 minutes, P < 0.001), but there was no difference between emergent sclerotherapy (14.5 ± 5.8 minutes) and ligation (14.9 ± 4.1 minutes). Ligation reduced one grade of variceal size more quickly than sclerotherapy (1.1 ± 0.4 vs 2.0 ± 1.7 session, P < 0.001). The rebleeding rate was lower with h'gation (13 of 67 vs. 28 of 67, P < 0.01). Esophageal ulcer was the most common source of rebleeding. Recurrence of varices appears more probable with ligation (P = 0.079). The complication rate was higher with sclerotherapy (15 of 67 vs. 3 of 67, P < 0.01), with esophageal stricture being the most common cause. Survival rate was the same in both groups even after stratifying patients into good and poor hepatic reserve groups. Hepatic failure was the major cause of death, followed by exsanguination. In summary, EVL was superior to EVS regarding rebleeding and complications but not in other aspects such as tune consumption in elective treatment and recurrence of varices. Substantial results for long-term follow-up are required before conclusion of the treatment of choice.
The effects of the 3-hydroxy, 3-methylglutaryl coenzyme a reductase inhibitor pravastatin on bile composition and nucleation of cholesterol crystals in cholesterol gallstone disease
Authors: Smit Jan W. A. , Van Erpecum Karel J. , Renooij Willem , Stolk Mark F. J. , Edgar Patrick , Doornewaard Heleen , Vanberge-Henegouwen Gerard P.
Published: March 2007

3-hydroxy, 3-methylglutaryl coenzyme A (HMG-CoA) reductase inhibitors reduce biliary cholesterol saturation index (CSI) in duodenal bile in hypercholesterolemic patients and might be useful for gallstone dissolution. However, preliminary data suggest that these drugs are not effective in this respect. We therefore studied 33 patients with radiolucent gallstones in an opacifying gallbladder who were scheduled for elective cholecystectomy. Patients were treated with 40 mg pravastatin day−1 or placebo during the 3 weeks before surgery. Six patients could not be evaluated. Baseline characteristics (age, sex, body mass index, serum cholesterol, and the solitary/multiple gallstone ratio) were similar in both groups. Serum cholesterol fell by 39% in the pravastatin group (P < 0.001) and remained unchanged in the placebo group. Biliary cholesterol (9.5 ± 1.3 vs. 14.3 ± 1.5 mmol/L, P = 0.026), and phospholipid concentrations (24.8 ± 3.9 vs. 36.7 ± 3.9 mmol/L, P = 0.043) were lower in the pravastatin group. Although bile salt concentrations were lower in the pravastatin group (114 ± 21 vs. 152 ± 15 mmol/L), this difference was not significant. CSI was not different between both groups (142 ± 27% [pravastatin] vs. 113 ± 6% [placebo], P = NS). Cholesterol crystals were present in fresh bile in 7 of 13 patients in the pravastatin group and in 11 of 14 controls (P = NS). Nucleation time was comparable between the 2 groups (13 ± 3 vs. 9 ± 3 days, P = NS). Bile salt species and molecular species of phospholipids determined with high-performance liquid chromatography did not differ either between both groups. Not only cholesterol but also phospholipid concentrations decrease in gallbladder bile during pravastatin treatment in cholesterol gallstone patients, with comparable CSI and nucleation time. This study does not support the use of HMG-CoA reductase imhibitors for dissolution of cholesterol gallstones.
Serial hemodynamic measurements in well-differentiated hepatocellular carcinomas
Authors: Saitoh Satoshi , Ikeda Kenji , Koida Isao , Tsubota Akihito , Arase Yasuji , Chayama Kazuaki , Kumada Hiromitsu
Published: March 2007

We performed serial hemodynamics in 15 patients with 21 well-differentiated hepatocellular carcinomas. The total length of the observation period ranged from 129 to 678 days (median, 368). We investigated both arterial and portal blood flow at intervals of at least 4 months. Arterial blood flow was measured with carbon dioxide—enhanced ultrasonography (US), and portal blood flow was measured with computed tomographic arterial portography (CTAP). None of the tumors were hypervascular on the initial study; however, by the final study nine nodules (42.9%) had become homogeneously hypervascular, two (9.5%) had become partially hypervascular, and the others did not change. Of 10 nodules without portal blood supply on any study, 8 became hypervascular by the final study. Of 11 nodules with portal blood supply on the initial study, a follow-up study showed no change in portal flow in 6 and the loss of portal flow in 5. Four of these five became hypervascular, with a documented loss of portal flow before the increase in arterial flow. The doubling time of tumors with a homogeneous increase in arterial blood flow ranged from 89 to 333 days (median, 172), whereas the doubling time of other tumors ranged from 227 to 607 days (median, 392). Thus, growth rate and vascularity in well-differentiated hepatocellular carcinoma are closely correlated. When the well-differentiated hepatocellular carcinoma has portal blood flow and is not hypervascular, it grows slowly. On the other hand, when it loses portal blood flow and becomes hypervascular, it grows rapidly.
Treatment of hepatocellular carcinoma with combined suppression and inhibition of sex hormones: A randomized, controlled trial
Authors: Manesis Emanuel K. , Giannoulis Gregory , Zoumboulis Pavlos , Vafiadou Irini , Hadziyannis Stephanos J.
Published: March 2007

We tested the hypothesis that a combination of sex hormone suppression and inhibition of their target receptors might improve survival for patients with hepatocellular carcinoma (HCC). Eighty-five consequent, previously untreated HCC patients with inoperable disease, were randomized to receive the luteinizing hormone-releasing hormone (LR-RH)-analogue triptorelin and the antiestrogen tamoxifen (33 patients) or triptorelin plus the antiandrogen flutamide (23 patients), or only placebo (29 patients) in a double blind fashion. All groups were comparable as to age, sex, tumor extension, underlying cirrhosis and biochemical parameters. The tamoxifen (TMX) group had a significantly longer survival (282 days) compared with flutamide (112 days) and with placebo (127 days) groups (P = 0.0238, log rank test). The upper quartile of patients in the TMX group lived 384 days or longer, and most of them (57.1%) were women (P < 0.0005), in contrast to the upper quartile of the placebo (170 days, 16.7% women) and the flutamide group (134 days, 33.3% women). The calculated tumor volume doubling time (TVDT) was significantly higher in the TMX group (296 days) than in the other two groups (99 and 101 days for placebo and flutamide groups, respectively, P = 0.023). In a Cox proportional hazards model, the TMX treatment, along with the baseline Okuda's HCC stage, the hepatitis B surface antigen, the portal vein diameter, the carcino embryonic antigen (CEA) and a self-assessment score of quality of life, were covariates predicting survival. Although the degree of serum sex hormone suppression was not a significant predictor of survival, the interaction of female sex and TMX treatment, it was (P = 0.0052). We conclude that TMX treatment significantly prolongs survival and the TVDT in unresectable HCC. The effect is most pronounced in female patients and is unrelated to sex hormone suppression.
Overexpression of c- met protooncogene product and raised Ki67 index in hepatocellular carcinomas with respect to benign liver conditions
Authors: Grigioni Walter F. , Fiorentino Michelangelo , d'Errico Antonia , Ponzetto Antonio , Crepaldi Tiziana , Prat Maria , Comoglio Paolo M.
Published: March 2007

The c-met protooncogene encodes a 190-kd transmembrane tyrosine kinase. This molecule is the receptor for the hepatocyte growth factor (HGF), which is an important mitogen for hepatocytes both in vitro and in vivo. In experimental models, the c-met transcripts appeared strongly expressed by actively proliferating oval cells (OCs). We evaluated the phenotypic modulation of the c-met protooncogene product (c-met pp), in 10 hepatocellular carcinomas (HCCs), 5 focal nodular hyperplasias (FNHs), 4 cases of fulminant hepatitis (FH), and 1 regenerated liver, selected to include the different biological states of hepatocyte (mature normal hepatocytes, transformed ones, and OCs). The supposed mitogenic effect of HGF was analyzed by comparing c-met pp overexpression with the Ki67 index, whereas anti—OV-6 antibody was used for comparison with the presence of OCs. The anti—c-met pp showed a typical plasma membrane-specific staining in all cases. The signal was much stronger in the HCCs than in the benign conditions. The anti—OV-6 monoclonal antibody showed positive immunostaining in many of the cells expressing c-met pp. The percentage of Ki67+ nuclei in high-grade HCCs paralleled c-met protooncogene overexpression. The c-met pp in OV-6+ cells suggests that the paracrine mechanism postulated in experimental models could also apply to human liver.
The frequency of 4977 base pair deletion of mitochondrial DNA in various types of liver disease and in normal liver
Authors: Fukushima Shu , Honda Kazuo , Awane Masaaki , Yamamoto Eiji , Takeda Ryouji , Kaneko Ichiro , Tanaka Akira , Morimoto Taisuke , Tanaka Koichi , Yamaoka Yoshio
Published: March 2007

Using a polymerase chain reaction (PCR) method, we tested for the hepatic mitochondrial DNA (mtDNA) deletion in 40 hepatic tumors (28 hepatocellular carcinomas [HCCs], 9 other malignant tumors, and 3 benign tumors) and in the livers of 71 patients, including 16 pediatric patients with end-stage liver disease who underwent living related donor liver transplantation and 16 liver donors. A 4977 base pair (bp) deletion of mtDNA was detected in 36 of 55 specimens of non—tumor portions of adult liver (65.5%). However, none of the specimens obtained from cirrhotic livers of the 16 pediatric patients younger than 13 years of age had the 4977 bp deletion. The frequency of mtDNA deletion was significantly decreased compared with normal liver in HCCs (7 of 28) and other malignant liver tumors (2 of 9). The frequency of this deletion was unrelated to the presence of liver cirrhosis, patient's gender, hepatitis B virus surface antigen status, and hepatitis C virus antibody status.
Further characterization and comparison of inducible nitric oxide synthase in mouse, rat, and human hepatocytes
Authors: Nussler Andreas K. , Di Silvio Mauricio , Liu Zhi-Ze , Geller David A. , Freeswick Paul , Dorko Kenneth , Bartoli Fabio , Billiar Timothy R.
Published: March 2007

Marked differences in induced nitric oxide (NO) synthesis occur between species. We have previously shown that both human and rat hepatocytes express an inducible NO synthase in response to cytokines and lipopolysaccharide. In this study, we compare the expression and regulation of cytokine-induced NO synthase in hepatocytes isolated from three species, human, rat, and mouse. On stimulation with tumor necrosis factor alpha (TNFα), interleukin-1β (IL-1β), interferon gamma (IFNγ), and lipopolysaccharide (LPS), it was found that hepatocytes from all three species produce high levels of NO with levels of production exhibiting the following hierarchy: rat hepatocytes > mouse hepatocytes > human hepatocytes. Whereas rat and mouse hepatocytes express inducible NO synthase messenger RNA (mRNA) in response to TNFα, IL-1β, or IFNγ as a single stimulus, human hepatocytes respond to LPS alone. Inhibition of NO generation through transforming growth factor (TGF-β1) was seen in mouse (77% ± 5.9) and rat hepatocytes (17% ± 2.6) whereas only about 10% was seen in human hepatocytes. Epidermal growth factor (EGF) was shown to inhibit NO synthesis in human and mouse hepatocytes but not rat. A marked NO-dependent inhibition of total protein synthesis was seen in rat and human hepatocytes, whereas mouse hepatocytes showed almost no inhibition in protein synthesis when stimulated. NO-dependent cyclic guanosine monophosphate (cGMP) release was found in all three species. Comparative studies on cytosol for inducible NO synthase enzyme activity showed that mouse and rat hepatocyte cytosol needed only L-arginine and reduced form of nicotinamide-adenine dinucleotide phosphate (NADPH) to exhibit NO formation, whereas cytosol from human hepatocytes required the addition of 5,6,7,8-tetrahydrobiopterin, flavin mononucleotide (FMN), and flavin adenine dinucleotide (FAD) to exhibit maximal NO synthase activity. Our results show even though hepatocytes from all three species can express considerable inducible NO synthase activity, important differences exist in the characteristics and effects of the NO synthesis.
Hypocalcemia decreases the early and late responses to epidermal growth factor in rat hepatocytes
Authors: Bilodeau Marc , Provencher Sébastien J. , Néron Stéphane , Haddad Pierre , Vallières Sylvie , Gascon-Barré Marielle
Published: March 2007

Extreme variations in extracellular Ca2+ concentrations ([Ca2+]e) modify the signaling generated by many hormones and growth factors. However, the influence of physiological changes in [Ca2+]e on the response to hepatic mitogens remains largely unknown. To study the influence of [Ca2+]e on the response to epidermal growth factor (EGF), hepatocytes from normal rat livers were equilibrated in vitro at [Ca2+]e similar to those observed in normocalcemia or hypocalcemia. To further investigate the effect of hypocalcemia in vivo, hepatocytes were obtained from chronically hypocalcemic rats and kept in vitro at the [Ca2+]e prevailing in vivo. Intracellular Ca2+ concentrations ([Ca2+]i) and DNA synthesis were evaluated after increasing doses of EGF. [Ca2+]e strongly influenced the [Ca2+]i response to EGF with significantly smaller [Ca2+]i increases in hepatocytes of normal rats kept in low [Ca2+]e compared with those kept in normal [Ca2+]e. In hypocalcemic rat hepatocytes, the response was further decreased and found to be significantly lower than that obtained in control cells kept in vitro at either 1.25 mmol/L or 0.8 mmol/L [Ca2+]e. In normal [Ca2+]e, the EGF-induced increases in [Ca2+]i were abolished by inhibiting EGF receptor autophosphorylation and by blocking calcium channels. Low in vitro [Ca2+]e significantly dampened the EGF-mediated DNA synthesis in normal rat hepatocytes but hypocalcemia in vivo further reduced the proliferative response compared with that obtained in control rat hepatocytes maintained in normal, or low [Ca2+]e. Furthermore, the blunted responses in [Ca2+]i mobilization and DNA synthesis associated with hypocalcemia could not be overcome by increasing concentrations of EGF nor by normalization of [Ca2+]e
in vitro. These data demonstrate that [Ca2+]e within the physiological concentration range can strongly influence the hepatocyte response to EGF. At [Ca2+]e comparable to that of hypocalcemia in vivo, our data point to the appearance of a phenomenon of hepatocellular resistance to the early (increases in [Ca2+]i) and late (DNA synthesis) cellular responses to EGF.