bluuebunny/crossref_metadata_2025 huggingface dataset:
Effects of Sho–Saiko–To, A Japanese Herbal Medicine, on Hepatic Fibrosis in Rats
Authors: Shimizu Ichiro , Ma Yue-Rong , Mizobuchi Yoko , Liu Fei , Miura Tetsuo , Nakai Yoichiro , Yasuda Mitugi , Shiba Masako , Horie Takahiro , Amagaya Sakae , Kawada Norifumi , Hori Hitoshi , Ito Susumu
Published: July 2004

It has been shown that lipid peroxidation is associated with hepatic fibrosis and stellate cell activation. Sho–saiko–to (TJ–9) is an herbal medicine, which is commonly used to treat chronic hepatitis in Japan, although the mechanism by which TJ–9 protects against hepatic fibrosis is not known. As a result, we assayed the preventive and therapeutic effects of TJ–9 on experimental hepatic fibrosis, induced in rats by dimethylnitrosamine (DMN) or pig serum (PS), and on rat stellate cells and hepatocytes in primary culture, and assessed the antioxidative activities and the active components of TJ–9. Male Wistar rats were given a single intraperitoneal injection of 40 mg/kg DMN or 0.5 mL PS twice weekly for 10 weeks. In each model, rats were fed a basal diet throughout, or the same diet, which also contained 1.5% TJ–9, for 2 weeks before treatment or for the last 2 weeks of treatment. TJ–9 suppressed the induction of hepatic fibrosis, increased hepatic retinoids, and reduced the hepatic levels of collagen and malondialdehyde (MDA), a production of lipid peroxidation. Immunohistochemical examination showed that TJ–9 reduced the deposition of type I collagen and the number of α–smooth muscle actin (α–SMA) positive–stellate cells in the liver and inhibited, not only lipid peroxidation in cultured rat hepatocytes that were undergoing oxidative stress, but also the production of type I collagen, α–SMA expression, cell proliferation, and oxidative burst in cultured rat stellate cells. In addition, TJ–9 inhibited Fe2+/adenosine 5′–diphosphate–induced lipid peroxidation in rat liver mitochondria in a dose–dependent manner and showed radical scavenging activity. Among the active components of TJ–9, baicalin and baicalein were found to be mainly responsible for the antioxidative activity. These findings suggest that Sho–saiko–to (TJ–9) functions as a potent antifibrosuppressant by inhibition of lipid peroxidation in hepatocytes and stellate cells in vivo.
Organic Osmolyte Transport in Quiescent and Activated Rat Hepatic Stellate Cells (Ito Cells)
Authors: Peters-Regehr Thorsten , Bode Johannes G. , Kubitz Ralf , Häussinger Dieter
Published: July 2004

Activation of hepatic stellate cells (HSCs) results in multiple alterations of cell function, but nothing is known about organic osmolytes in these cells. Organic osmolyte transport and transporter messenger RNA (mRNA) expression was studied in quiescent rat HSCs and after their transformation into α1-smooth muscle actin-positive myofibroblastlike cells. Quiescent stellate cells expressed in an osmosensitive manner the mRNA levels of the transporters for taurine (TAUT) and myoinositol (SMIT), whereas that for betaine was not detectable. However, these cells showed osmosensitive uptake not only of taurine and myoinositol but also of betaine. Osmosensitive betaine uptake was mediated by amino acid transport system A. After transformation into myofibroblasts, taurine and myoinositol uptake increased 5.5–fold and 4.5–fold, respectively, together with the respective transporter mRNA levels. Betaine uptake increased twofold because of osmosensitive induction of BGT1 expression. In both quiescent and activated HSCs, hypoosmotic cell swelling induced a rapid and 4,4′–diisothiocyanatostilbene–2,2′–disulphonic acid-sensitive osmolyte efflux. In quiescent HSCs, hyperosmotic exposure increased the messenger RNA (mRNA) level of cyclooxygenase–2, which was counteracted by taurine but not by betaine or myoinositol. The study identifies taurine, myoinositol, and betaine as osmolytes in HSCs. Transformation of HSCs is accompanied by enhanced osmolyte transport activity and induction of the BGT1 transporter, which may be another activation marker of HSCs.
Growth Hormone Inhibits Rat Liver α–1–Acid Glycoprotein Gene Expression In Vivo And In Vitro
Authors: Mejdoubi Najet , Henriques Cécile , Bui Elisabeth , Durand Geneviève , Lardeux Bernard , Porquet Dominique
Published: July 2004

The gene encoding α–1–acid glycoprotein (AGP), one of the major acute–phase proteins, is positively controlled at the transcriptional level by cytokines (interleukin–1 [IL–1], IL–6, and tumor necrosis factor α) and glucocorticoids. Here, we show that growth hormone (GH) treatment of isolated rat hepatocytes in vitro reduces AGP messenger RNA (mRNA) expression. AGP gene expression remained inducible by IL–1, IL–6, and phenobarbital (PB) in GH–treated hepatocytes. Interestingly, the repressive effect of GH on AGP gene expression was also observed in vivo: liver AGP mRNA content was strongly increased in hypophysectomized rats, and GH treatment of these animals led to a decrease in mRNA to levels lower than those in untreated control animals. Moreover, the inhibitory effect of GH mainly occurs at the transcriptional level and can be observed as little as 0.5 hours after GH adding in vitro to isolated hepatocytes. These results show negative regulation of AGP gene expression and strongly suggest that GH is a major endogenous regulator of constitutive AGP gene expression. Moreover, transfection assays showed that the region of the AGP promoter located at position -147 to -123 is involved in AGP gene regulation by GH. Furthermore, GH deeply modifies the pattern of nuclear protein binding to this region. GH treatment of hypophysectomized rats led to the release of proteins of 42 to 45 and 80 kd and to the binding of proteins of 48 to 50 and 90 kd.
Cost–Effectiveness of Ursodeoxycholic Acid Therapy in Primary Biliary Cirrhosis
Authors: Pasha Tousif , Heathcote Jenny , Gabriel Sherine , Cauch-Dudek Karen , Jorgensen Roberta , Therneau Terry , Dickson Rolland E. , Lindor Keith D.
Published: July 2004

Ursodeoxycholic acid (UDCA) is a safe and effective treatment for patients with primary biliary cirrhosis (PBC), but the cost of this drug has raised concerns regarding cost–effectiveness. The aim of our study was to determine the cost–effectiveness of UDCA in PBC. We compared the costs and outcomes of managing PBC patients with and without UDCA. From two previously published trials, the effectiveness of UDCA was determined by comparing the annual reduction in the development of ascites, varices, variceal bleeding, encephalopathy, liver transplantation, and death between the treatment groups. Average annual costs for each of these events were estimated based on literature and institutional data. Approximately twice as many major events occurred in the placebo group compared with the UDCA group. The relative risk (RR) of liver transplantation (1.95; 95% CI: 1.14–3.68) and development of esophageal varices (3.11; 95% CI: 1.57–10.65) were significantly higher in the placebo group compared with the UDCA group. There were no significant increases in the RR of ascites, variceal bleeding, encephalopathy, or death between the two groups. Based on the estimated annual cost of managing these events and the annual costs of UDCA ($2,500), there was an annual cost savings per patient of $1,372. Compared with the placebo group, patients receiving UDCA had a lower incidence of major complications and lower medical care costs.
Differences in Hypervariable Region 1 Quasispecies of Hepatitis C Virus in Human Serum, Peripheral Blood Mononuclear Cells, and Liver
Authors: Okuda Michiari , Hino Keisuke , Korenaga Masaaki , Yamaguchi Yuhki , Katoh Yoshiharu , Okita Kiwamu
Published: July 2004

Hepatitis C virus (HCV) has been reported to potentially replicate in peripheral blood mononuclear cells (PBMCs), but direct information on the pathogenic implication of HCV infection in PBMCs is still limited. To investigate this issue, we compared the complexity of HCV quasispecies in serum, PBMCs, and livers of 13 patients with type C chronic liver disease. Hypervariable region 1 (HVR 1) was amplified by reverse–transcription polymerase chain reaction (RT–PCR), and the PCR products were subcloned and sequenced. Considerable differences in the complexity of HVR 1 quasispecies were found in the serum, PBMCs, and liver in all patients, and the predominant sequences from each source were mutually different in 3 (23%) patients. An amino acid sequence unique to each source existed as well as a sequence common to serum and PBMCs, common to serum and livers, or common to PBMCs and liver. These results suggest infection of PBMCs by HCV, and that HCV in PBMCs may be differently exposed to host immunity from that in liver.
Dynamics of Gb Virus C Viremia Early After Orthotopic Liver Transplantation Indicates Extrahepatic Tissues As the Predominant Site of Gb Virus C Replication
Authors: Berg Thomas , Müller Andrea R. , Platz Klaus P. , Höhne Marina , Bechstein Wolf-Otto , Hopf Uwe , Wiedenmann Bertram , Neuhaus Peter , Schreier Eckart
Published: July 2004

The principal site of GB virus C (GBV–C) replication is unknown. To determine whether hepatic GBV–C replication is important for the maintenance of a measurable viremia level in GBV–C infection, the influence of hepatectomy followed by liver transplantation on GBV–C viremia was investigated. GBV–C RNA levels were determined by a quantitative TaqMan polymerase chain reaction (PCR) in 12 patients with pretransplantation GBV–C infection before and daily after orthotopic liver transplantation (OLT) for 25 to 28 days. Compared with the pretransplantation values (mean, 12.4 ± 3.9 × 107 copies/mL), mean GBV–C RNA levels declined significantly by 1 log by day 1 after OLT (mean, 3.5 ± 1.6 × 107 copies/mL), but subsequently remained relatively stable on this high level for the entire observation period, indicating ongoing high–level virus replication (mean GBV–C RNA levels on days 7 and 28 were: 1.7 ± 0.5 × 107 and 2.8 ± 0.7 × 107 copies/mL; P = ns). Thus, at the end of the follow–up, mean GBV–C RNA levels were not significantly different from that of the 1st and 7th postoperative day and remained significantly lower compared with the pretransplantation values. However, in 2 of the 12 patients, different kinetics were observed. Both already had low–level viremia pre–OLT (0.02 and 0.002 × 107 copies/mL) and became persistently GBV–C RNA-negative 2 days after OLT. In 5 patients, liver tissues were collected 6 to 9 days after OLT and investigated for GBV–C RNA. All but 1 were GBV–C RNA-negative in the liver, although 2 of them had rather high serum GBV–C RNA levels at this time. The kinetics of GBV–C viremia observed in our study were neither influenced by the immunosuppressive therapy nor by the number of blood and blood product transfusions given after OLT. In addition, they were quite different from those observed in patients with chronic hepatitis C in whom early reinfection of the graft could be demonstrated by a steady increase in HCV RNA levels starting 3 days after OLT and exceeding preoperative levels by day 8. From our data, one can conclude that the liver is certainly not the major site of GBV–C replication in most patients. However, one cannot exclude that host or viral factors exist that predispose GBV–C replication predominantly in the liver.
Transjugular Intrahepatic Portosystemic Shunt Versus Endoscopic Sclerotherapy for the Prevention of Variceal Rebleeding After Recent Variceal Hemorrhage
Authors: García-Villarreal Luis , Martínez-Lagares Francisco , Sierra Angel , Guevara Clemencia , Marrero José M. , Jiménez Elena , Monescillo Alberto , Hernández-Cabrero Teresa , Alonso José M. , Fuentes Rafael
Published: July 2004

Variceal hemorrhage continues to be a major cause of morbidity and mortality in cirrhotic patients. Transjugular intrahepatic portosystemic shunt (TIPS) is gaining wide acceptance as a treatment for several complications of portal hypertension. The aim of the current randomized study was to compare the transjugular shunt and endoscopic sclerotherapy (ES) for the prevention of variceal rebleeding (VB) in cirrhotic patients. Forty–six consecutive cirrhotic patients with variceal bleeding were randomly allocated to receive either transjugular shunt (22 patients) or ES (24 patients) 24 hours after control of bleeding. VB (50% vs. 9%) and early (first 6 weeks) VB (33% vs. 5%) were significantly more frequent in sclerotherapy patients; the actuarial probability of being free of VB was higher in the shunt group (P < .002). Eight patients (33%) of the sclerotherapy group and 3 patients (15%) of the shunt group died; the actuarial probability of survival was higher for the shunted patients (P < .05); 6 patients in the sclerotherapy group and none in the shunt group died from VB (P < .05). No difference was found in the proportion of patients with clinically evident hepatic encephalopathy (HE). These results show that the transjugular shunt is more effective than sclerotherapy in the prevention of both early and long–term VB. Moreover, a significant improvement in survival was found in the shunt group.
Platelet Aggregation and Platelet–Derived Growth Factor Inhibition for Prevention of Insufficiency of the Transjugular Intrahepatic Portosystemic Shunt: A Randomized Study Comparing Trapidil Plus Ticlopidine With Heparin Treatment
Authors: Siegerstetter Volker , Huber Michael , Ochs Andreas , Blum Hubert E. , Rössle Martin
Published: July 2004

Intimal proliferation at the interface between prosthetic material and tissue is an intrinsic phenomenon of stenting and the major cause of insufficiency of the transjugular intrahepatic portosystemic shunt (TIPS). For its prevention, a randomized study was performed comparing standard heparin treatment with a combination of trapidil, a drug with anti-platelet–derived growth factor (PDGF) activity, and ticlopidine, a platelet aggregation inhibitor. Ninety patients with cirrhosis who received a transjugular shunt were randomized, and 84 patients completed the trial. Group 1 (n = 42) received a bolus of heparin (12 to 24 U/kg) at shunt placement, followed by 1 week of intravenous and 4 weeks of subcutaneous heparin treatment. Group 2 (n = 42) received the same heparin bolus, followed by a 1–day intravenous heparin treatment and a 6–month treatment with trapidil (400 mg/d) and ticlopidine (250 mg/d). Shunt function was assessed by duplex–sonography and angiography. Stenoses were classified according to their location as type 1 (within the stent) and type 2 (in the draining hepatic vein). The estimated rate of overall stenoses (intention–to–treat analysis) at 1 year showed a significant reduction in patients receiving trapidil and ticlopidine (group 2) as compared with heparin (33 vs. 57%; P = .047). There was no difference in the estimated 1–year rate of type 1 stenoses between the two groups, but there was a significant reduction in type 2 stenoses (group 1: 58%, group 2: 19%; P = .016). The treatment effect continued after withdrawal of the drugs and was accompanied by a decreased incidence of rebleeding. The study demonstrates that the incidence of type 2 stenosis of the transjugular shunt can be reduced by combined inhibition of platelet aggregation and PDGF activity. The findings may be of relevance not only for the transjugular shunt, but also for other stent applications, e.g., vascular and biliary, as well as for bypass and shunt surgery.
Pit Cells (Hepatic Natural Killer Cells) of the Rat Induce Apoptosis in Colon Carcinoma Cells by the Perforin/Granzyme Pathway
Authors: Vermijlen David , Luo Dianzhong , Robaye Bernard , Seynaeve Carine , Baekeland Marijke , Wisse Eddie
Published: July 2004

The high mortality of colon cancer is to a large extent caused by the frequent occurrence of liver metastasis. This is remarkable, because the liver harbors two distinct cell populations that can eliminate invading cancer cells, namely hepatic natural killer (NK) cells and Kupffer cells. These hepatic NK cells, known as pit cells, are the most cytotoxic cells of the naturally occurring NK cells. However, the mechanism by which pit cells eliminate tumor cells is largely unknown. Because we recently found an indication that apoptosis is involved, we tried to assess the role of this mode of cell death using an in vitro system with isolated pure pit cells (>90%) and CC531s cells, a rat colon carcinoma (CC) cell line. Pit cells induced apoptosis in CC531s cells as shown by quantitative DNA fragmentation, agarose gel electrophoresis, and different modes of microscopy. When extracellular Ca2+ was chelated by ethylene glycol–bis(β–aminoethyl ether)–N,N,–tetraacetic acid (EGTA) during coincubation or when the pit cells were preincubated with the granzyme inhibitor 3,4–dichloroisocoumarin (DCI), the induction of apoptosis was abolished. These results show that pit cells use the Ca2+–dependent perforin/granzyme pathway to induce apoptosis in the CC531s cells, and not the alternative Ca2+–independent Fas pathway. To further exclude the possibility of the involvement of the Fas pathway, we treated CC531s cells with recombinant Fas ligand. This treatment did not result in the induction of apoptosis, indicating that CC531s cells are resistant to Fas–mediated apoptosis. We conclude therefore that pit cells induce apoptosis in CC cells in vitro by the perforin/granzyme pathway.
Propensity of Ectopic Liver to Hepatocarcinogenesis: Case Reports and A Review of the Literature
Authors: Arakawa Masahiro , Kimura Yoshio , Sakata Kenji , Kubo Yasuhiko , Fukushima Takashi , Okuda Kunio
Published: July 2004

Two patients with ectopic liver are described. In one patient, a small ectopic liver attached to the gastric serosa developed hepatocellular carcinoma (HCC). The preoperative diagnosis was an α–fetoprotein (AFP)–producing carcinoma and a malignant ulcer of the stomach. Total gastrectomy and esophago–jejunostomy were performed. The tumor that measured 4 × 2 × 2 cm contained an AFP–producing HCC and normal liver tissue. In another patient who had alcoholic cirrhosis, ectopic liver on the serosa of the gallbladder was found to have the same histological changes as the mother liver. A survey of the literature disclosed more than 20 cases in which HCC developed outside the liver; the liver did not have HCC. By contrast, there was only one report on HCC occurring in the liver in the presence of a noncancerous, relatively large accessory liver lobe. Because ectopic liver does not have a complete vascular and ductal system as a normal liver, it is perhaps functionally handicapped and more prone to hepatocarcinogenesis.
Natural History of Untreated Nonsurgical Hepatocellular Carcinoma: Rationale for the Design and Evaluation of Therapeutic Trials
Authors: Llovet Josep M. , Bustamante Javier , Castells Antoni , Vilana Ramon , Del Carmen Ayuso Maria , Sala Margarita , Brú Concepció , Rodés Joan , Bruix Jordi
Published: July 2004

This study analyzed the natural history and prognostic factors of patients with nonsurgical hepatocellular carcinoma (HCC). Twenty variables from 102 cirrhotic patients with HCC who were not treated within prospective randomized controlled trials (RCT) were investigated through uni– and multivariate analyses. None of them was suitable for radical therapies (surgical resection, liver transplantation, or ethanol injection) or presented end–stage disease as reflected by an Okuda stage 3 or a Performance Status ≥3. Sixty–five patients were Child–Pugh A, 34 were B, and 3 were C. Most of them exhibited a preserved Performance Status Test (PST) (0 = 56; 1 = 38; 2 = 8). Tumor was solitary in 26 (≤5 cm in 16) and multinodular/massive in 76. After a median follow–up of 17 months, 79 patients died, the 1–, 2–, and 3–year survival being 54%, 40%, and 28%. The multivariate study identified PST (P = .01), constitutional syndrome (P = .04), vascular invasion (P = .001), and extrahepatic spread (P = .04) as independent predictors for mortality. The 1–, 2–, and 3–year survival for the 48 patients without adverse factors (Stage 0) was 80%, 65%, and 50%, respectively, and 29%, 16%, and 8% in the 54 patients with at least one adverse parameter (Stage I). Therefore, Stage 0 would correspond to an intermediate stage, while Stage I would represent an advanced status, before reaching an end–stage phase. In conclusion, the outcome of nonsurgical HCC is not homogeneously grim and may be predicted by assessing the presence of symptoms and of an invasive tumoral pattern. Therapeutic trials should be designed and evaluated considering these characteristics.
Restoration of Serum Albumin Levels in Nagase Analbuminemic Rats by Hepatocyte Transplantation
Authors: Oren Ran , Dabeva Mariana D. , Petkov Petko M. , Hurston Ethel , Laconi Ezio , Shafritz David A.
Published: July 2004

Recently, we described a new strategy for hepatocyte transplantation, using retrorsine/partial hepatectomy (PH) in a DPPIV- mutant Fischer rat model. Treatment of rats with retrorsine, a pyrrolizidine alkaloid, blocks endogenous hepatocytes from proliferating, so that after exposure to this agent coupled with PH and hepatocyte transplantation, transplanted hepatocytes selectively repopulate the liver. In the present study, we determined whether this method of cell transplantation can restore biosynthetic and physiological function in the liver by transplanting normal hepatocytes into rats genetically deficient in albumin synthesis, the Nagase analbuminic rat (NAR). After hepatocyte transplantation, albumin mRNA and protein were identified in the liver by in situ hybridization and immunohistochemistry, respectively, and serum albumin levels were determined using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS–PAGE), Western blot, and enzyme–linked immunosorbent assay (ELISA) methods. At 1 month posttransplantation, large clusters of cells expressing albumin mRNA and protein were identified in the liver, representing ≈50% of hepatocytes for albumin mRNA and ≈61% for protein. At 2 months' posttransplantation, cells expressing albumin mRNA represented ≈77% of hepatocyte mass, and cells expressing albumin protein represented ≈81% of total hepatocyte mass. Hepatocyte–transplanted NAR also exhibited normal or near–normal serum albumin levels (3.0 ± 0.2 g/dL). High levels of serum albumin were sustained for the 2–month duration of experiments. These results demonstrate the ability of this protocol for hepatocyte transplantation to restore a major biosynthetic and physiological function of the liver, and suggest its potential use as a method to treat genetic–based or acquired liver diseases.
Altered Gq/G11 Guanine Nucleotide Regulatory Protein Expression in A Rat Model of Hepatocellular Carcinoma: Role in Mitogenesis
Authors: McKillop Iain H. , Schmidt Max C. , Cahill Paul A. , Sitzmann James V.
Published: July 2004

Guanine nucleotide regulatory proteins (G–proteins) represent an important transmembrane pathway whereby extra–cellular signals are transduced to intracellular signaling pathways. The mitogen–activated protein kinase (MAPK) cascade has been identified as a key factor in transducing numerous mitogenic stimuli. MAPK activity is regulated via numerous receptor types, including those linked to Gq/G11–proteins, which regulate phospholipase–C activity. We hypothesized that alterations in a Gq/G11–PLC pathway may contribute to the enhanced cellular mitogenesis characteristic of hepatocellular carcinoma (HCC), possibly via a MAPK–dependent pathway. By using an in vivo model of HCC we investigated changes in Gq/G11–protein expression in tumorigenic tissue versus adjacent, non–neoplastic liver. In addition we addressed the role of Gq/G11–proteins in the regulation of MAPK–linked mitogenesis by using rat hepatic tumorigenic cells (H4IIE) and isolated hepatocytes in culture. Western blot analysis showed significant increases in Gqα and G11α expression in tumorigenic liver versus normal liver specimens, an effect that was augmented in cultured H4IIE cells versus isolated cultured hepatocytes. Furthermore, phosphoinositol specific phospholipase–C (PLC) activity was significantly increased in HCC versus normal liver. A specific PLC inhibitor (Et–18–OCH3) caused a dose–dependent decrease in serum stimulated DNA synthesis in both cultured H4IIE cells and isolated rat hepatocytes, the H4IIE cell line showing greater sensitivity to Et–18–OCH3. In addition, serum–stimulated MAPK activity was significantly enhanced in H4IIE versus cultured hepatocytes. Moreover, treatment with Et–18–OCH3significantly attenuated serum stimulated MAPK activity in both cultured hepatocytes and H4IIE cells. Furthermore, U73122 (Gqα–PLC specific uncoupler) and GP2A (Gqα specific inhibitor) mirrored the effects of those observed for Et–18–OCH3 whereas PD98059 (specific MEK inhibitor) completely abolished serum–stimulated DNA synthesis in tumorigenic H4IIE cells. We conclude that HCC is associated with enhanced Gq/G11–PLC expression/activity as compared with normal liver. Furthermore, a PLC–linked MAPK cascade plays a significant role in the progression of the enhanced mitogenesis characteristic of HCC.
Effect of Losartan, An Angiotensin Ii Receptor Antagonist, on Portal Pressure in Cirrhosis
Authors: Schneider Arved Winfried , Kalk Johann Friedrich , Klein Christian Peter
Published: July 2004

Administration of angiotensin II causes an increase in portal pressure, and plasma concentration of angiotensin II is elevated in patients with cirrhosis, suggesting that angiotensin II may be involved in the pathogenesis of portal hypertension in cirrhosis. We evaluated the effect of the orally active angiotensin II receptor antagonist, losartan, on portal pressure in patients with cirrhosis and portal hypertension. Thirty patients with severe (hepatic venous pressure gradient [HVPG] ≥ 20 mm Hg) and 15 patients with moderate (HVPG < 20 mm Hg) portal hypertension at baseline measurement were treated with an oral dose of 25 mg losartan once daily for 1 week and compared with 15 (HVPG ≥ 20 mm Hg) and 10 (HVPG < 20 mm Hg), respectively, cirrhotic controls. On the seventh day, HVPG was determined again, and blood pressure, heart rate, body weight, and parameters of liver and kidney function were recorded. Losartan induced a significant (P < .001) decrease of HVPG in the patients with severe (-46.8% ± 15.5%) and moderate (-44.1% ± 14.7%) portal hypertension, while no significant change was seen in the controls. Losartan caused a slight but significant (P < .01) fall in mean arterial blood pressure (-3.1 ± 5.0 and -3.5 ± 4.3 mm Hg, respectively). One patient treated with losartan had a short symptomatic hypotensive reaction after the first dose of losartan that did not recur despite continued treatment. No deterioration of liver or kidney function was observed. The present study indicates that angiotensin II blockade with orally administered losartan is safe and highly effective in the treatment of portal hypertension.
Nitric Oxide Reduces Nontransferrin–Bound Iron Transport in Hepg2 Cells
Authors: Barisani Donatella , Cairo Gaetano , Ginelli Enrico , Marozzi Anna , Conte Dario
Published: July 2004

Nitric oxide (NO) donors S–nitroso–N–acetylpenicillamine (SNAP) and sodium nitroprusside (SNP) modulate iron regulatory protein (IRP) activity and may, therefore, affect iron uptake through transferrin receptor expression. However, iron also enters the cell as nontransferrin–bound iron (NTBI), and the aim of this study was to evaluate the effects of NO donors on NTBI transport in HepG2 cells, a model of liver physiology. Incubation with SNP and SNAP led to a time– and concentration–dependent reduction in Fe3+ and Fe2+ uptake, thus indicating an effect on the transporter rather than on the reductase. In terms of Fe2+ uptake, no variations in the Michaelis–Menten constant (K
m ) and a reduction in maximum uptake (V
max ) (50, 33, and 16.6 fmol/μg protein/min in control, SNP–, and SNAP–treated cells, respectively) were detected, which suggested a decrease in the number of putative NTBI transport protein(s). Gel shift assays showed that IRP activity was reduced by SNP and slightly increased by SNAP. Northern blot analysis of transferrin receptor messenger RNA (mRNA) levels showed variations similar to those observed for IRPs, but both NO donors increased L–ferritin mRNA levels and had no effect on the stimulator of Fe transport (SFT) mRNA. In conclusion, NO donors significantly reduce NTBI transport in HepG2 cells, an effect that seems to be IRP and SFT independent. Moreover, the reduction in NTBI uptake after NO treatment suggests that this form of iron may play a minor role in the increased hepatic iron stores observed in inflammation or that other liver cells are more involved in this pathological condition.
Recurrent Familial Intrahepatic Cholestasis in the Faeroe Islands. Phenotypic Heterogeneity But Genetic Homogeneity
Authors: Tygstrup Niels , Steig Bjarni Á. , Juijn Jenneke A. , Bull Laura N. , Houwen Roderick H.
Published: July 2004

Recurrent familial intrahepatic cholestasis is an autosomal recessive disorder characterized by episodes of severe pruritus and jaundice lasting for weeks to months without extrahepatic bile duct obstruction. Symptom–free intervals may last for months to years, and chronic liver damage does not develop. We recently studied four of the five patients from the Faeroe Islands described by us 30 years ago (one had recently died) and an additional five patients that were identified after the initial report. The episodes of cholestasis were more frequent and severe in patients with early onset, but tended to reduce in frequency with age. The youngest patient, aged 25 years, who had had 16 episodes each lasting about 6 months, had a liver transplant after which no further episodes were recorded (1 year after surgery). Signs of chronic liver disease were absent in all patients. The FIC1 gene was investigated for mutations in the surviving patients. A single mutation (I661T) was found on both chromosomes in all nine patients, indicating that they are genetically identical for the disease–causing defect. Nevertheless, considerable differences among patients were observed clinically
Tauroursodeoxycholate and S–Adenosyl–L–Methionine Exert An Additive Ameliorating Effect on Taurolithocholate–Induced Cholestasis: A Study in Isolated Rat Hepatocyte Couplets
Authors: Milkiewicz Piotr , Mills Charles O. , Roma Marcelo G. , Ahmed-Choudhury Jalal , Elias Elwyn , Coleman Roger
Published: July 2004

The monohydroxy bile acid, taurolithocholate (TLC), causes cholestasis in vivo and in isolated perfused livers. It is also cholestatic in vitro and, in this study using isolated rat hepatocyte couplets, causes a reduction of the accumulation of (fluorescent) bile acid in the canalicular vacuoles (cVA) of this polarized cell preparation. The hepatoprotective bile acid, tauroursodeoxycholate (TUDCA), partially protects against the action of TLC when added at the same time. It also partially reverses the cholestatic effect if added after the cells have been exposed to TLC. A second hepatoprotective compound, S–adenosyl–L–methionine (SAMe) also not only partially protects against the action of TLC when added at the same time, but it too is able to partially reverse the cholestatic effect. Neither hepatoprotective agent is fully effective alone, but their effects are additive. In combination, a full restoration of cVA is observed in moderate cholestasis, but not in severe cholestasis. We discuss briefly some possible mechanisms involved in the additive mode of action of both hepatoprotective compounds. In summary, we show for the first time that SAMe and TUDCA can exert an additive effect in the amelioration of TLC–induced cholestasis in isolated rat hepatocyte couplets. This finding may be of possible clinical relevance
Differences in the Metabolism and Disposition of Ursodeoxycholic Acid and of Its Taurine–Conjugated Species in Patients With Primary Biliary Cirrhosis
Authors: Invernizzi Pietro , Setchell Kenneth D. , Crosignani Andrea , Battezzati Pier Maria , Larghi Alberto , O'Connell Nancy C. , Podda Mauro
Published: July 2004

The clinical effectiveness of ursodeoxycholate in the treatment of liver disease may be limited by its poor absorption and extensive biotransformation. Because in vitro and in vivo studies suggest that the more hydrophilic bile acid tauroursodeoxycholate has greater beneficial effects than ursodeoxycholate, we have compared for the first time the absorption, metabolism, and clinical responses to these bile acids in patients with primary biliary cirrhosis (PBC). Twelve female patients with PBC were sequentially administered tauroursodeoxycholate and ursodeoxycholate (750 mg/d for 2 months) in a randomized, cross–over study. Bile acids were measured in serum, duodenal bile, urine, and feces by gas chromatography-mass spectrometry (GC–MS). Biliary ursodeoxycholate enrichment was higher during tauroursodeoxycholate administration (32.6% vs. 29.2% during ursodeoxycholate; P < .05). Lithocholic acid concentration was consistently higher in all biological fluids during ursodeoxycholate administration. Fecal bile acid excretion was the major route of elimination of both bile acids; ursodeoxycholate accounted for 8% and 23% of the total fecal bile acids during tauroursodeoxycholate and ursodeoxycholate administration, respectively (P < .05). Tauroursodeoxycholate was better absorbed than ursodeoxycholate, and, although it was partially deconjugated and reconjugated with glycine, it underwent reduced biotransformation to more hydrophobic metabolites. This comparative study suggests that tauroursodeoxycholate has significant advantages over ursodeoxycholate that may be of benefit for long–term therapy in PBC.
A Model to Predict Survival Following Liver Retransplantation
Authors: Rosen Hugo R. , Madden Joseph P. , Martin Paul
Published: July 2004

In the current era of critical–organ shortage, one of the most controversial questions facing transplantation teams is whether hepatic retransplantation, which has historically been associated with increased resource utilization and diminished survival, should be offered to a patient whose first allograft is failing. Retransplantation effectively denies access to orthotopic liver transplantation (OLT) to another candidate and further depletes an already–limited organ supply. The study group was comprised of 1,356 adults undergoing hepatic retransplantation in the United States between 1990 and 1996 as reported to the United Network for Organ Sharing (UNOS). We analyzed numerous donor and recipient variables and created Cox proportional–hazards models on 900 randomly chosen patients, validating the results on the remaining cohort. Five variables consistently provided significant predictive power and made up the final model: age, bilirubin, creatinine, UNOS status, and cause of graft failure. Although both hepatitis C seropositivity and donor age were significant by univariate and multivariate analyses, neither contributed independently to the estimation of prognosis when added to the final model. The final model was highly predictive of survival (whole model χ2= 139.63). The risk scores for individual patients were calculated, and patients were assigned into low–, medium–, and high–risk groups (P< .00001). The low degree of uncertainty in the probability estimates as reflected by confidence intervals, even in our high–risk patients, underscores the applicability of our model as an adjunct to clinical judgment. We have developed and validated a model that uses five readily accessible “bedside” variables to accurately predict survival in patients undergoing liver retransplantation.
An Economic Analysis of Different Strategies of Immunization Against Hepatitis A Virus in Developed Countries
Authors: Das Ananya
Published: July 2004

Acute hepatitis A is a major public health problem in developed countries, and because a large proportion of patients with acute hepatitis A do not have any identifiable risk factors, current practice of targeting the high–risk groups for vaccination against hepatitis A virus (HAV) is unlikely to have a significant impact on the overall incidence of acute hepatitis A. No economic analysis of strategies of mass immunization against HAV is available. Three different strategies of immunization against HAV using commercially available inactivated vaccine were compared in a Markov model analysis of a cohort of 2–year–old healthy children in a developed country. In strategy I, universal vaccination was pursued. In strategy II, children were initially screened for antibody and, if susceptible, they were vaccinated. In strategy III, no vaccination was offered. Cost per person and quality adjusted life–years (QALY) gained in each strategy were the outcome measures compared. The baseline analysis showed that strategy II is more cost–effective than strategy I, with marginal cost–effectiveness ratios of $7,267.67 and $12,833.34, respectively, compared with a strategy of no intervention. Sensitivity analysis showed that if the cost of the two–dose vaccine could be reduced to less than $57, the strategy of universal vaccination would be the preferred immunization strategy. Different strategies of mass immunization against HAV in the developed countries are cost–effective by current standards of health care interventions and should be considered for incorporation into current childhood immunization programs.
Activation of Cdk4 and Cdk2 During Rat Liver Regeneration Is Associated With Intranuclear Rearrangements of Cyclin–Cdk Complexes
Authors: Jaumot Montserrat , Estanyol Josep-Maria , Serratosa Joan , Agell Neus , Bachs Oriol
Published: July 2004

Partial hepatectomy (PH) triggers the entry of rat liver cells into the cell cycle. The signals leading to cell–cycle activation converge into a family of kinases named cyclin–dependent kinases (cdks). Specific cyclin–cdk complexes are sequentially activated during the cell cycle. Cyclin D–cdk4 and cyclin E–cdk2 are activated during the G1 phase, cyclin A–cdk2 is activated during the S phase, and cyclin B–cdk1 during mitosis. In the present study, we have examined the timing of the activation of cdk4 and cdk2, the intracellular location of G1/S cyclins and cdks, and the relationship between location and cdk4 and cdk2 activities during rat liver regeneration after a PH. Results showed that the activity of both kinases started at 13 hours and showed maximal levels at 24 hours after hepatectomy. In quiescent cells, cyclin D3 and cdk4 were cytoplasmatic, whereas cyclin D1 was nuclear. At 5 hours after hepatectomy, cyclin D3 and cdk4 began to move into the nucleus, and at 13 hours, they were mostly nuclear. During the first 13 hours after hepatectomy, significant amounts of cyclin D1–cdk4 and cyclin D3–cdk4 complexes were formed, but they were mostly inactive. At 24 hours, these complexes were maximally activated. This activation was associated with the accumulation of cyclin D1, cyclin D3, and cdk4 in a nuclear subfraction extractable with nucleases. At 28 hours, the activity of cdk4 in this nuclear subfraction decreased when cyclin D1 moved from this fraction to the nuclear matrix (NM) and the levels of cyclin D3 diminished. The maximal activation of cdk2 at 24 hours was also associated with the accumulation of cyclin E, cyclin A, and cdk2 in this nuclease–sensitive fraction. The inactivation of cdk2 at 28 hours was associated with a strong decrease in cdk2 in this nuclear subfraction. Thus, results reported here indicate that the activation of cdk4 and cdk2 observed in rat liver cells after a PH is associated with a specific intranuclear location of these cdks and their associated cyclins
The Hepatitis B Virus-Trimera Mouse: A Model for Human Hbv Infection and Evaluation of Anti–Hbv Therapeutic Agents
Authors: Ilan Ehud , Burakova Tatjana , Dagan Shlomo , Nussbaum Ofer , Lubin Ido , Eren Rachel , Ben-Moshe Ofer , Arazi Joseph , Berr Shoshana , Neville Lewis , Yuen Leonard , Mansour Tarek S. , Gillard John , Eid Ahamed , Jurim Oded , Shouval Daniel , Reisner Yair , Galun Eithan
Published: July 2004

Previous studies have demonstrated the feasibility of implantation of human blood cells or tissues in lethally irradiated mice or rats, radioprotected with SCID mouse bone marrow cells: The Trimera system. In the present study, we describe the development of a mouse Trimera model for human hepatitis B virus (HBV) infection. In this model, viremia is induced by transplantation ofex vivoHBV–infected human liver fragments. Engraftment of the human liver fragments, evaluated by hematoxylin–eosin staining and human serum albumin mRNA expression, was observed in 85% of the transplanted animals 1 month postimplantation. Viremia levels were determined in these mice by measuring serum HBV DNA using polymerase chain reaction (PCR), followed by dot–blot hybridization. HBV DNA is first detected 8 days after liver transplantation. Viremia attains a peak between days 18 and 25 when HBV infection is observed in 85% of the transplanted animals. The HBV–Trimera model was used to evaluate the therapeutic effects of human polyclonal anti–HBs antibodies (Hepatect) and of two reverse–transcriptase inhibitors, lamivudine (3TC) and β–l–5–fluoro–2′,3′–dideoxycytidine (β–l–5FddC). Treatment of HBV–Trimera mice with these drugs effectively reduced both the percentage of infected animals and the viral load in their sera. Treatment cessation resulted in rebound of viral load, indicating HBV replication upon drug withdrawal. These results show that the HBV–Trimera model represents a novel experimental tool for simulating human HBV infection and evaluating potential anti–HBV therapeutic agents.
Quality of Life Before and After Liver Transplantation for Cholestatic Liver Disease
Authors: Gross Cynthia R. , Malinchoc Michael , Kim Ray W. , Evans Roger W. , Wiesner Russell H. , Petz Janice L. , Crippin Jeffrey S. , Klintmalm Goran B. , Levy Marlon F. , Ricci Paola , Therneau Terry M. , Dickson Rolland E.
Published: July 2004

Liver transplantation (LT) is an established therapy for patients with end–stage primary biliary cirrhosis (PBC) or primary sclerosing cholangitis (PSC). In this report, we describe the health status and quality of life (QOL) in patients with these cholestatic liver diseases before and after LT. A QOL questionnaire was completed by 157 adult patients with PBC or PSC before and 1 year after liver transplantation at the Mayo Clinic or Baylor University Medical Center. This questionnaire measured four aspects of QOL, including symptoms; physical, social, and emotional functioning; health perceptions; and overall QOL. Changes in these QOL parameters before and after LT were described, and regression analysis was used to assess the relationships between clinical and QOL factors. There were no differences in QOL parameters between patients with PBC and PSC. QOL following transplantation was substantially better than before transplantation. This was observed in all four aspects of QOL. The degree of improvement as measured by effect size (difference in mean scores divided by the pretransplantation standard deviation) was 0.53 for symptoms (P<.01), 1.16 for function (P < .01), 2.37 for health satisfaction (P < .01), and 1.16 for overall QOL (P< .01). Patients' overall QOL before transplantation was significantly related to subjective and objective health status indicators and clinical factors such as ascites and renal dysfunction. QOL at 1–year follow–up, however, could not be adequately predicted by the pretransplantation subjective health status and clinical factors. Patients with end–stage cholestatic disease undergoing LT experience substantial improvement in all aspects of QOL addressed in this study. The patients' QOL 1 year after LT could not be predicted by pretransplantation variables used in this study
Clinical and Histological Outcome After Hepatitis B E Antigen to Antibody Seroconversion in Children With Chronic Hepatitis B
Authors: Ruiz-Moreno Mercedes , Otero Manuela , Millán Antonio , Castillo Inmaculada , Cabrerizo María , Jiménez Javier F. , Oliva Horacio , y Cajal Santiago Ramón , Carreño Vicente
Published: July 2004

Data regarding the outcome of children with chronic hepatitis B after seroconversion are scarce. We describe the long–term evolution of these patients. One hundred and three children with antibody against hepatitis B e antigen and normal alanine aminotransferase (ALT) levels were followed for 0.6 to 12.5 years (mean, 6.3 years). Paired liver biopsies (before and after seroconversion) were available in 83 cases. Final biopsies were obtained 0.5 to 12.5 years (mean, 4.5 years) after seroconversion. ALT levels remained normal in most of the children (79%) throughout the follow–up. All children, except five who lost hepatitis B surface antigen, had serum viral DNA detected by polymerase chain reaction. When comparing baseline and final liver biopsies, a significant improvement (P < .001) was found in the histological activity index and in the necrosis, cytolysis, inflammation, and fibrosis scores. The histological diagnosis improvement in the final biopsy was significantly related (P< .001) to the time from seroconversion to the biopsy performance. All children had viral DNA on their final liver biopsy. In summary, seroconversion and ALT normalization are quite stable findings in children, and no differences in the long–term outcome between treated and untreated children were found. In light of the histological outcome, it seems unnecessary to perform a follow–up liver biopsy in these cases.
Hepatitis C Virus Rna Profiles in Chronically Infected Individuals: Do They Relate to Disease Activity?
Authors: Pontisso Patrizia , Bellati Giorgio , Brunetto Maurizia , Chemello Liliana , Colloredo Guido , Di Stefano Rosellina , Nicoletti Massimo , Rumi Maria Grazia , Ruvoletto Maria Grazia , Soffredini Roberta , Valenza Lilli Mario , Colucci Giuseppe
Published: July 2004

Fluctuations of hepatitis C virus (HCV)–RNA serum levels were monitored in a multicenter study in 76 chronic HCV carriers who had been followed longitudinally without receiving antiviral therapy to assess their relation with the course of liver disease activity. Forty–four patients had normal transaminases over more than 2 years, while 32 additional patients had fluctuating levels. Viral load was measured in serial serum samples prospectively collected for 10 to 12 months in 54 patients and in sera stored yearly up to 8 years in an additional 22 patients. In patients tested monthly, a lesser extent of fluctuations was detected in cases with constantly normal transaminases as compared with those with fluctuating transaminases. In the former group, the mean difference between maximum and minimum values observed in each individual patient was 0.7 Log, while in the latter group, it was 1.3 Log (P = .0004). Most of these patients experienced, on average, three peaks of viremia over 1 year. The range of variation observed upon yearly testing was between 0.2 and 2.2 Log and did not reach statistical significance between the two groups. In conclusion, a careful viral replication profile can be achieved only by monthly testing, because longer time intervals could miss viremia fluctuations. HCV–RNA levels are more stable in asymptomatic HCV carriers than in patients with biochemical activity of liver disease.