Background/aims The present study was designed to investigate whether the acute phase protein alpha-1-antitrypsin (α1-AT), which has an inhibitory effect on transferrin (tf) receptor-mediated iron uptake in K562 and THP1 cells, has a similar effect in PLC/PRF/5 cells. This hepatic cell line is of specific interest because it is infected with hepatitis B virus (HBV). Therefore, we addressed the additional question whether α1-AT has any effect on cellular protein synthesis and replication of HBV in PLC/PRF/5 cells. Methods Cells were incubated with various concentrations of α1-AT, dexamethasone, IL-6 and desferrioxamine. HBs-AG, alpha-fetoprotein and albumin concentrations in culture media were measured using commercially available methods. For equilibrium inhibition binding experiments, cells were incubated with 85–182 pmol/l [125l]tf. To study the potential effect of α1-AT on DMA synthesis we measured the incorporation of [3H]thymidine into DNA. Results In equilibrium saturation binding experiments, [125l]tf bound to PLC/PRF/5 cells with KD 17.45 ± 4.57 nM and a maximum density of binding sites of 267 285 ± 39 915 sites/cell. In inhibition studies α1-AT demonstrated an apparently monophasic inhibition of [125l]tf to its receptor. At concentrations > 30 μmo\/\ α1AT inhibited the growth of PLC/PRF/5 cells up to approximately 50%. The inhibitory effect of α1-AT on DNA synthesis was not as potent as that on growth. At the highest concentration of 100 μmol/l, α1-AT produced a 35% maximum inhibition of [3H]thymidine incorporation. Incubating PLC/PRF/5 cells with corticosteroids enhanced HBs-AG release significantly. Interestingly, α1-AT showed the same pattern of effects on cell metabolism and HBs-AG release as the corticosteroids. When we incubated the cells with 50 μmol/1 α1-AT, alpha-fetoprotein production increased significantly and HBs-AG release almost doubled. Conclusion We have to assume that there is a specific mechanism inducing HBs-AG release by α1AT, as has been shown to be the case with steroids.
Background: Biliary strictures and leaks are a major source of morbidity following liver transplantation. Endoscopic retrograde cholangiography (ERC) however, represents an efficient means to successfully treat these types of complications. Aim of this study was to evaluate risk factors for biliary complications in liver transplant recipients and assess the effectiveness of early ERC. Material Methods: We reviewed our institutional database of 820 consecutive liver transplants performed within the last 13 years (01/2000-10/2013) at our institution. Results: The overall biliary complication rate as confirmed by endoscopy was 18.4% (n=151). Incidence of biliary complications was significantly higher in anastomoses with internal draining stents: 23.7% (n=9/38), when compared to anastomoses with T-Tube drainage: 19.9% (n=28/141) and en-end anastomoses: 18.9% (n=104/553) (p<0.01). Mean time from diagnosis to endoscopic treatment was 3.1 days. An average of 4.1 endoscopic re-interventions were necessary to treat 12.3% (n=101) biliary stenoses, 4.3% (n=35) biliary leaks, and 1.9% (n=15) combined complications (stenosis & leak). Early endoscopic treatment (< 3 days after clinical and radiological diagnosis) significantly reduced the need for repeated interventions (<3 days: 3.1±0.3 interventions vs. >3 days: 4.9±0.8 interventions; p<0.01). Sphincterotomy and partial sphincterotomy was performed in 44.6% and 9.6% of cases. The amount of Plastic Stents (62.8%) used was significantly higher when compared to Coated Metal Stents (11.5%) and Pig-Tail Stents (10.3%; p<0.01). Median duration of stent treatment was 43 days (1-265 days). Retransplantation-rate due to persistent complications was 3.8%. Significant risk factors associated with biliary complications were increased cold ischemia times (CIT), recipient-weight, and body mass index, as well as higher donor age, donor- weight and donor-body mass index (all p<0.01). Conclusion: Early endoscopic intervention is key to successful treatment of biliary complications after liver transplantation. Biliary anastomoses with T-Tubes are superior to anastomoses with stents and equal to end-end biliary anastomoses without stents.
The sequence of activation of the components of the cytokine network subsequent to in vivo application of different dosages of IL-2 is still poorly understood. Although side effects of IL-2 therapy are dose dependent, the dose-response relationship for induction of potentially beneficial or harmful cytokine genes still remains to be studied. We examined the patterns of cytokine gene expression after treatment of chronic hepatitis B patients with various doses of IL-2 in a phase 1 trial. Total RNAs were isolated from PBMC harvested at various time points after s.c. injection of natural IL-2 ranging from 30,000 to 1,000,000 U. Dose-dependent effects on mRNA expression of IL-2, GM-CSF, IFN-gamma, TNF-alpha, and IL-6 were assessed using Northern blotting and slot blotting techniques. A single application of 30,000 U nIL-2 induced selective and long-lasting expression of IL-2, IFN-gamma, and GM-CSF genes, which was not accompanied by accumulation of TNF-alpha and IL-6 mRNAs. Larger dosages of IL-2 induced activation of monokine genes and were associated with systemic side effects. mRNA levels of the different cytokines related to biologic activity and correlated with expression of specific proteins and cellular parameters: IL-2 mRNA with soluble IL-2R serum levels and induction of lymphopenia, GM-CSF mRNA with induction of neutrophilia, and IL-6 mRNA with c-reactive protein serum concentrations. Taken together these data indicate unexpected immunoregulatory activities of very low and nontoxic dosages of IL-2 in vivo.
21 patients, who survived acute respiratory insufficiency following shock, are examined for residual pulmonary lesions. Retrospectively various parameters (f.e. blood gaz, radiological alterations) are analysed and correlated with results of a follow-up study (pulmonary function, chest films). Significant impairment of pulmonary functions could not be documented in a single patient. Roentgenologic late sequelae are seen only in two patients, which had a minor impairment of pulmonary function.
