Hepatotoxic substance(s) removed by high-flux membranes enhances the positive acute phase response

Hepatotoxic substance(s) removed by high-flux membranes enhances the positive acute phase response.BackgroundAcute phase proteins (APPs) are enhanced in end-stage renal disease patients (ESRD) requiring dialysis treatment. They are involved in a variety of pathologic processes like muscle proteolysis, cachexia, regulation of appetite, and atherosclerosis. They are predictive for mortality. APPs are not only makers but also active substances. They are mainly produced in liver cells and are primarily, but not exclusively, regulated by proinflammatory cytokines. To what extent hepatic APPs are influenced by uremic toxins is still unclear. Therefore, we investigated the effects of different ultrafiltrates (UFs) on the synthesis of α1-acid glycoprotein (AGP) in HepG2 cells.MethodsA cross-sectional as well as a crossover study with high-/low-flux membranes was conducted to investigate the impact of UFs on bioactivity of liver cell cultures. Metabolic activity (MTT test), cytotoxicity (lactate dehydrogenase release), and the positive APP AGP were measured in HepG2 cells.ResultsCultured hepatocytes treated with UFs from high-flux membranes exhibited a higher cytotoxicity (18.6 ± 0.3% high-flux vs. 13.9 ± 0.2% low-flux, P < 0.001) and a lower metabolic activity (29.3% high-flux vs. 50.3% low-flux, P < 0.001) in comparison with low-flux UFs. In addition, enhanced APP secretion could be observed under costimulatory conditions (high-flux 5.0 ± 0.7 vs. low-flux 3.1 ± 0.6 ng/μg protein, P < 0.05). The effects of high- and low-flux UFs were strongly expressed at the beginning and were still significantly different after 120 minutes of hemodialysis (HD) treatment. The cross-over experiments confirmed that UFs collected during high-flux HD had a higher capacity to stimulate AGP synthesis in liver cells.ConclusionThe effects of UFs from dialysis patients demonstrate that hepatotoxic substances can be removed by dialysis. Stimulating the acute phase response UF collected during high-flux HD had a higher impact on liver cells in comparison with low-flux UF. These substances are putative cofactors involved in cytokine regulation

Hemofiltration in human sepsis: Evidence for elimination of immunomodulatory substances

Hemofiltration in human sepsis: Evidence for elimination of immunomodulatory substances. Continuous hemofiltration is widely used for renal replacement therapy in patients with acute renal failure. It has been suggested that hemofiltration may also eliminate toxic mediators thought to be important in the pathophysiology of sepsis. The present study examined whether hemofiltration can activate or eliminate inflammatory mediators in patients with sepsis, and whether ultrafiltrate can alter specific functions of peripheral blood mononuclear leukocytes (PBMC) in vitro. Veno-venous hemofiltration was performed in 16 patients and in 5 healthy volunteers. Pre-filter (afferent), post-filter (efferent) and ultrafiltrate concentrations of cytokines (IL-1β, IL-6, IL-8, TNFα) and of complement components (C3, C3adesArg, C5adesArg, terminal complement complex) were measured after the beginning of hemofiltration (t0), and 60 minutes later (t60). PBMC, and monocyte and lymphocyte subfractions were incubated with ultrafiltrate, and cytokines were determined in the supernatants. Hemofiltration did not induce significant mediator activation. There was no evidence for significant cytokine elimination. However, pre-filter C3adesArg concentration showed a significant decline during hemofiltration (patients: t0 = 676.9 ± 99.7 ng/ml, t60 = 545.4 ± 83.2, P < 0.001; volunteers: t0 = 54.8 ± 13.3 ng/ml, t60 = 33.9 ± 10.7, P < 0.001). Ultrafiltrate from septic patients significantly stimulated PBMC and monocyte TNFα release, but suppressed lymphocyte IL-2 and IL-6 production. Ultrafiltrate from volunteers was without effect. Hemofiltration effectively eliminates certain mediators such as C3adesArg. Ultrafiltrate contains compounds with significant immunomodulatory qualities. Therefore, hemofiltration may represent a new modality for removal of immunomodulatory mediators

The role of polymer surface degradation and barium sulphate release in the pathogenesis of catheter-related infection

Background. Susceptibility to infection and thrombosis of intravascular catheters is increased by surface irregularities, which might be prevented by coating. Methods. BaSO4 release from conventional haemodialysis catheters (CC) and modified catheters (MC) which had been coated with a surface-modifying additive (SMA) was assessed in vivo and in vitro. For the in vivo part, patients were randomized to receive a temporary CC or MC, with crossover after 1 week. After retrieval, catheters were examined using scanning electron microscopy to assess surface integrity, and an in vitro model of catheter exposure to the bloodstream was used to evaluate surface morphology and susceptibility to bacterial adhesion and proliferation. Results. BaSO4 moieties covered 14.7 +/- 3.7% of the surface of unused CC. After in vivo use in 16 patients, 62.7 +/- 32.9 x 10(3) holes/mm-were detected, indicating BaSO4 detachment from 3.3 +/- 1.7% of the catheter surface. No defects were observed in unused CC and in MC, whether used or unused. After incubation of four catheters (two of each type) with Staphylococcus epidermidis, the two degraded CC showed an immediate and strong bacterial growth as indicated by an increase in medium impedance of 0.512%/10 min compared to -0.021%/10 min in MC (P < 0.001). Conclusions. Short-term exposure of CC to the bloodstream causes BaSO4 particle release, resulting in surface irregularities predisposing to bacterial proliferation. BaSO4 release can be prevented by SMA coating

Review on uraemic toxins III: recommendations for handling uraemic retention solutes in vitro - towards a standardized approach for research on uraemia

International audienceNo abstract availabl

Review on uremic toxins : classification, concentration, and interindividual variability

Background. The choice of the correct concentration of potential uremic toxins for in vitro, ex vivo, and in vivo experiments remains a major area of concern; errors at this level might result in incorrect decisions regarding therpeutic correction of uremia and related clinical complications. Methods. An encyclopedic list of uremic retention solutes was composed, containing their mean normal concentration (C-N), their highest mean/median uremic concentration (C-U), their highest concentration ever reported in uremia (C-MAX), and their molecular weight. A literature search of 857 publications on uremic toxicity resulted in the selection of data reported in 55 publications on 90 compounds, published between 1968 and 2002. Results. For all compounds, C-U and/or C-MAX exceeded C-N. Molecular weight was lower than 500 D for 68 compounds; of the remaining 22 middle molecules, 12 exceeded 12,000 D. C-U ranged from 32.0 ng/L (methionine-enkephalin) up to 2.3 g/L (urea). C-U in the ng/L range was found especially for the middle molecules (10/22; 45.5%), compared with 2/68 (2.9%) for a molecular weight <500 D (P < 0.002). Twenty-five solutes (27.8%) were protein bound. Most of them had a molecular weight <500 D except for leptin and retinol-binding protein. The ratio C-U/C-N, an index of the concentration range over which toxicity is exerted, exceeded 15 in the case of 20 compounds. The highest values were registered for several guanidines, protein-bound compounds, and middle molecules, to a large extent compounds with known toxicity. A ratio of C-MAX /C-U <4, pointing to a Gaussian distribution, was found for the majority of the compounds (74/90; 82%). For some compounds, however, this ratio largely exceeded 4 [e.g., for leptin (6.81) or indole-3-acetic acid (10.37)], pointing to other influencing factors than renal function, such as gender, genetic predisposition, proteolytic breakdown, posttranslation modification, general condition, or nutritional status. Conclusion. Concentrations of retention solutes in uremia vary over a broad range, from nanograms per liter to grams per liter. Low concentrations are found especially for the middle molecules. A substantial number of molecules are protein bound and/or middle molecules, and many of these exert toxicity and are characterized by a high range of toxic over normal concentration (C-U/C-N ratio). Hence, uremic retention is a complex problem that concerns many more solutes than the current markers of urea and creatinine alone. This list provides a basis for systematic analytic approaches to map the relative importance of the enlisted families of toxins