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A cytotoxic protein isolated from Pseudomonas aeruginosa damages the plasma membranes of many mammalian cells by forming pores. We studied binding of the 125I-cytotoxin and the resulting increase of cation permeability in erythrocytes of various mammalian species. The sensitivity of red blood cells was inversely related to the relative sphingomyelin content(More)
Effects of Staphylococcus aureus alpha-toxin and Pseudomonas aeruginosa cytotoxin on the permeability of an endothelial monolayer were studied. Porcine pulmonary artery endothelial cells were grown on a polycarbonate membrane, mounted in a chamber, and exposed to a continuous hydrostatic pressure of 10 cmH2O. On application of this trans-endothelial(More)
Rabbit erythrocyte membrane glycosylated 28-kDa protein was investigated in the membrane-bound as well as in the soluble state on an example of a Pseudomonas aeruginosa cytotoxin-binding component. When membranes were treated with trypsin/N-glycosidase F, a 13.5-kDa-binding active peptide residue is obtained as revealed by a ligand-blot technique after(More)
The mode of phospholipase C activation initiated with platelet-derived growth factor (PDGF) has been studied in comparison with that initiated with vasopressin and bombesin in a rat fibroblast line, WFB. Stimulation of WFB cells by PDGF, vasopressin, and bombesin elicites rapid hydrolysis of polyphosphoinositides and an increase in cytoplasmic free Ca2+(More)
The effects of highly purified Pseudomonas aeruginosa cytotoxin were investigated on cultured pulmonary artery endothelial cells. This toxin dose-dependently (7.5-60 micrograms/ml) and time-dependently (20-75 minutes) stimulated the release of radiolabeled arachidonic acid and metabolites and the synthesis of prostacyclin in the absence of overt cell damage(More)
The site-specific integration of the phage phi CTX genome, which carries the gene for a pore-forming cytotoxin, into the Pseudomonas aeruginosa chromosome was analysed. The 1,167 bp integrase gene, int, located immediately upstream of the attachment site, attP, was characterized using plasmid constructs, harbouring the integration functions, and serving as(More)
Presence of cytotoxin was studied in extracts of 57 strains of Pseudomonas aeruginosa (46 bacteremia, 4 environmental, and 7 Fisher immunotype), 10 Pseudomonas species, and 7 nonpseudomonas isolates. Cytotoxin was identified by Western immunoblot in extracts of all P. aeruginosa isolates. None of the Pseudomonas species or nonpseudomonas isolates were shown(More)
Increased plasma membrane permeability induced by a purified cytotoxic protein from Pseudomonas aeruginosa was studied using mouse Ehrlich ascites tumor cells in incubation medium containing an osmotic stabilizer. In the presence of serum albumin, 40 nM of the cytotoxin was required for cationic imbalance of 2.7 X 10(7) cells per ml at pH 7.4. The rate of(More)
Quin2 loaded human leukemic, JURKAT and K562 cells, were exposed to various doses of Pseudomonas aeruginosa cytotoxin. This cytotoxin induced an increase in quin2 fluorescence indicating an increase in the cytoplasmic free Ca2+ concentration. The rate of the fluorescence increase and the lag time before the response were dependent on the doses of the(More)