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Biomedical subjects

C Pusineri

Publications and source records attributed to C Pusineri.

8 recordsLinked to original sources

Minimodule dialyser for quantitative ex vivo evaluation of membrane haemocompatibility in humans: comparison of acrylonitrile copolymer, cuprophan and polysulphone hollow fibres.

Minimodule hollow fibre dialysers, representing clinical dialysis modules on a scale of 1/25, enable quantitative evaluation of the haemocompatibility of hollow fibre membranes in an ex vivo flow system in humans. On line heparinization, adjusted for donor sensitivity, is maintained at a minimal level (approximately 0.14 units/ml). Blood samples collected at the minimodule exit over 30 min are analysed for heparin (anti-Xa activity), activated partial thromboplastin time, fibrinopeptide A, platelet count and beta-thromboglobulin, complement fragment C3a, leucocyte count and polymorphonuclear neutrophil elastase. Initial experiments were performed using well-characterized reference materials: acrylonitrile copolymer (AN 69 HF), polysulphone and cuprophan (CUP). Activation of coagulation and platelets was low for AN 69 HF, intermediate for CUP and greatest for polysulphone, while complement activation was negligible in the presence of AN 69 HF, moderate for polysulphone and most important for CUP. Future applications will be directed towards haemocompatibility screening of prototype membranes with the aim of developing clinical dialysers with improved biocompatibility.

Acrylic Resins

Influence of a preadsorbed terpolymer on human platelet accumulation, fibrinogen adsorption, and ex vivo blood activation in hemodialysis hollow fibers.

Results are presented on kinetics of platelet accumulation in charged polyacrylonitrile (AN69) hollow fibers by continuous data recording under flow conditions (wall shear rate 108-1050 s-1), using suspensions of washed 111In-labeled human platelets in Tyrode's-albumin buffer, containing washed red blood cells (0-40%). Preadsorption of a terpolymer of acrylonitrile, poly(ethyleneoxide) methacrylate and trimethylaminoethyl chloride methacrylate leads to very efficient passivation with respect to platelet accumulation and fibrinogen adsorption. In human ex vivo tests, evaluation of complement peptide C3a, platelet beta-thromboglobulin, leucocyte-polymorphonuclear neutrophile elastase and fibrinopeptide A shows no detectable activation. Furthermore, preadsorption appears to result in simultaneous improvement in hemocompatibility of the blood lines leading to and from the dialysis module. This single pretreatment of dialysis membranes should allow injection of lower doses of anticoagulant to patients submitted to hemodialysis.

Acrylic Resins

High in vitro spontaneous histamine release in long-term hemodialyzed patients.

Adverse reactions, such as skin flush and pruritus, have been observed among hemodialyzed (HD) patients. A potential relationship between hemodialysis and allergic phenomena prompted us to investigate some allergologic parameters. Total IgE, specific IgE antiethylene oxide and anti-isocyanate toluene diisocyanate, and in vitro spontaneous histamine release (SHR) were measured in 105 long-term stable HD patients. Thirty-two of the patients had occasionally developed skin flush reactions and/or pruritus during dialysis sessions. Blood samples were taken at the start of dialysis, sera were kept frozen until assay, and in vitro SHR was performed on washed leukocytes at 37 degrees C for 40 minutes. A high (range, 20% to 80%) and reproducible in vitro SHR was observed among 24% of patients. No correlation was found between SHR and the type of dialysis membrane used (copolymer of acrylonitrile-methallyl-sodium sulfonate, cuprophan, and polysulfone). High SHR (i.e., greater than 20%) was observed mainly among patients with detectable antiethylene oxide IgE antibodies (p less than 0.001). High levels of IgE (greater than 300 ng/ml), specific IgE antibodies, and a high in vitro SHR were all observed concomitantly in 26% of the patients with skin flush reactions and/or pruritus, whereas all three of these parameters together were present in only 3% of the symptom-free patients. To our knowledge, this is the first time that a high in vitro SHR has been described among HD patients, and putative correlations with in vivo phenomena are discussed.

Adolescent

Dissociation between the interleukin 1-inducing capacity and Limulus reactivity of lipopolysaccharides from gram-negative bacteria.

In this study we compared the interleukin 1 (IL 1)-inducing capacity and the reactivity in the Limulus amoebocyte assay (LAL) of purified lipopolysaccharides (LPSs) from various bacterial strains. LPSs differed greatly in their capacities (on a weight basis) to induce IL 1 release from serum-free cultured human monocytes. LPS species that induced high levels of IL 1 release from human monocytes exhibited a high thiobarbiturate-reactive 2-keto-3-deoxy-octonic acid (KDO) content. No relationship was found between the IL 1-inducing activity and the LAL reactivity of purified LPSs. Filtration experiments in which membranes of decreasing size-exclusion limits were used demonstrated that molecular species of LPS with an apparent Mr below 3,000 may induce IL 1, whereas only species with an apparent Mr above 8,000 are recognized in the LAL assay. The latter observation suggests that the reaction with LAL requires an aggregated form of LPS. These results indicate that biologically active LPS species can cross dialysis membranes in vivo although no LAL reactive material is detected in the blood compartment. The Limulus assay is an insufficient criterion for the absence of LPS in biological fluids.

Gram-Negative Bacteria

Induction of IL-1 during hemodialysis: transmembrane passage of intact endotoxins (LPS).

Circulating monocytes of patients undergoing chronic hemodialysis are triggered to produce interleukin-1 (IL-1) in vivo. Intradialytic induction of IL-1 is associated with complement activation in patients dialyzed with first-use cellulose membranes. Chronic stimulation of IL-1 production occurs because of an yet unidentified mechanism in patients dialyzed with high permeability membranes. The present study demonstrates that intact bacterial lipopolysaccharide (LPS) molecules may cross cuprophan, AN69 and polysulfone membranes under in vitro conditions simulating in vivo hemodialysis. The experiments used purified LPS from Neisseria meningitidis and LPS from Pseudomonas testosteroni, a bacterial strain grown out from a clinically used dialysate. LPS were purified to homogeneity and radiolabeled. Transmembrane passage of 3H-labeled LPS was observed within the first five minutes of dialysis. A total of 0.1 to 1% of 3H-labeled LPS were recovered in the dialysate compartment after one hour of dialysis. High amounts of LPS, representing 40 to 70% of the amount originally present in the dialysate, were absorbed onto high permeability membranes. Low amounts of LPS were absorbed onto cuprophan membranes. The amount of LPS absorbed decreased with the concentration of LPS in the dialysate. LPS recovered from the blood compartment exhibited the same molecular weight as that used to contaminate the dialysate. Biochemically detectable transmembrane passage of LPS was not associated with that of material detectable using the limulus amebocyte lysate (LAL) assay. An IL-1-inducing activity was, however, detected in the blood compartment upon dialysis with high permeability membranes, as previously found by others with cuprophan membranes.(ABSTRACT TRUNCATED AT 250 WORDS)

Acrylic Resins

Plasma contact activation and decrease of factor V activity on negatively-charged polyelectrolytes.

The effect of negatively-charged polymers, used in some artificial devices, on plasma clotting and kinin systems was studied in vitro using polyelectrolyte complexes. Contact activation was observed as an immediate, transient and surface-dependent phenomenon. After incubation of the plasma with the polymer a small decrease of factor XII activity was noticed, which corresponded to a greater reduction of prekallikrein activity and to a marked kinin release. No significant decrease of factor XII, prekallikrein, HMW kininogen could be detected immunologically. Only the initial contact of the plasma with the polyelectrolyte lead to activation, subsequently the surface became inert. Beside contact activation, factor V activity also decreased in the plasma. The decrease was surface and time-dependent. It was independent of contact factor activation, and appeared to be related to the sulfonated groups of the polymer. If purified factor V was used instead of plasma factor V, inactivation was immediate and not time-dependent suggesting a direct adsorption on the surface. A second incubation of the plasma-contacted polymer with fresh plasma resulted in a further loss of Factor V activity.

Electrolytes