In hepatorenal syndrome (HRS), renal insufficiency is often progressive, and the prognosis is extremely poor under standard medical therapy. The molecular adsorbent recirculating system (MARS) is a modified dialysis method using an albumin-containing dialysate that is recirculated and perfused online through charcoal and anion-exchanger columns. MARS enables the selective removal of albumin-bound substances. A prospective controlled trial was performed to determine the effect of MARS treatment on 30-day survival in patients with type I HRS at high risk (bilirubin level, > or =15 mg/dL) compared with standard treatment. Thirteen patients with cirrhosis with type I HRS were included from 1997 to 1999. All were Child's class C, with Child-Turcotte-Pugh scores of 12.4 +/- 1. 0, United Network for Organ Sharing status 2A, and total bilirubin values of 25.7 +/- 14.0 mg/dL. Eight patients were treated with the MARS method in addition to hemodiafiltration (HDF) and standard medical therapy, and 5 patients were in the control group (HDF and standard medical treatment alone). None of these patients underwent liver transplantation or received a transjugular intrahepatic portosystemic shunt or vasopressin analogues during the observation period. In the MARS group, 5.2 +/- 3.6 treatments (range, 1 to 10 treatments) were performed for 6 to 8 hours daily per patient. A significant decrease in bilirubin and creatinine levels (P <.01) and increase in serum sodium level and prothrombin activity (P <.01) were observed in the MARS group. Mortality rates were 100% in the control group at day 7 and 62.5% in the MARS group at day 7 and 75% at day 30, respectively (P <.01). We conclude that the removal of albumin-bound substances with the MARS method can contribute to the treatment of type I HRS.

Background: Pancreatic stellate cells (PSCs) have been implicated in pancreatic fibrosis as they synthesise increased amounts of extracellular matrix proteins in response to activation by profibrogenic mediators such as cytokines. Aims: The purpose of this study was to analyse cytokine receptor stimulated signalling pathways involved in PSC activation. Using a rat culture model of PSCs, we have also tested the potential of the platelet derived growth factor (PDGF) antagonist trapidil and PD98059, a specific inhibitor of extracellular signal regulated kinase (ERK) activation, to suppress PSC growth. Methods: Cultured PSCs were stimulated with PDGF, and the signal transduction pathways activated in response to the mitogen were analysed by immunoblotting, kinase assays, and electrophoretic mobility shift assays. Furthermore, comparison of signalling cascades activated in PSCs before and after completing transdifferentiation to α-smooth muscle actin expressing myofibroblasts was performed. Biological effects of PDGF, trapidil, and PD98059 were analysed by proliferation assays and correlated with molecular effects of the substances. Results: PDGF induced rapid activation of Raf-1, ERKs 1 and 2, as well as AP-1 proteins. The transforming growth factor β activated transcription factor Smad2 was found to be constitutively phosphorylated in PSCs of different transdifferentiation grades. Furthermore, the results indicate a correlation between ERK activities and induction of PSC activation. Trapidil efficiently inhibited both PDGF induced ERK activation and, in common with PD98059, PSC proliferation. Conclusions: Our data suggest that ERKs play a key role in the regulation of PSC growth and that inhibition of the ERK signalling pathway may become a strategy to prevent activation of these cells.

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Background/Aims: Chronic pancreatitis is histologically characterized by an extended fibrosis and infiltration of leukocytes. We intended to differentiate the infiltration to evaluate the inflammatory process. Methods: Samples of tissues of normal pancreas (NP, n = 12), of chronic pancreatitis (CP, n = 7), and pancreatic tissues surrounding pancreatic carcinoma (CA, n = 7) were investigated by immunohistochemical staining using the APAAP technique. Results: In normal pancreas, mononuclear cells (47.1 ± 26.0 cells/mm²) were observed with a predominance of macrophages (56.3%) and T lymphocytes (31.3%) which were differentiated in CD8+ lymphocytes (9.3 ± 7.2 cells/mm²) and CD4+ lymphocytes (6.7 ± 3.2 cells/mm²). Rarely, plasma cells (5.3%) and B lymphocytes (7.1%) could be detected. In pancreatic tissue of patients with CP and in CA there was a significant increase of mononuclear cells to 264.4 ± 120.3 cells/mm² and 284.3 ± 67.8 cells/mm², respectively. In both diseases percentages of T lymphocytes (CP: 50.5%; CA: 48.1%) were higher than in normal controls. CD4+/CD8+ ratio of 0.77 in CP and 0.82 in CA demonstrated a predominance of CD8+ cells compared to the peripheral blood. In NP and CA, nearly all T lymphocytes expressed CD45R0 identifying memory cells, while only 58% of T lymphocytes were CD45R0 positive in CP. Conclusion: Our data suggest that the investigated cases of CP were of a common inflammatory type rather than due to an autoimmunological reaction. CD8+ T lymphocytes were the predominant T cell subset in the inflammatory infiltrates in both CP and CA.

Pancreatic stellate cells (PSCs) are involved in, among other things, the pathogenesis of pancreatic fibrosis. Here, we present the generation of immortalized PSCs 7 and 14 days after isolation by retroviral gene transfer of the SV40 large T antigen encoding region. Propagated cell lines [large T immortalized cells (LTC)-7, LTC-14] retained characteristics of primary cells in terms of morphology, responsiveness to mediators regulating cellular functions such as proliferation, and expression profile of a number of investigated genes. Whereas LTC-14 kept the morphological features of the differentiation status of the primary cells they were made of, LTC-7 appeared similar to an earlier stage. Thus the established cell lines represent a versatile tool to investigate various aspects of PSC biology.