A chemiluminescence (CL) study of diphenyleneiodonium-inhibited NADPH oxidase was performed on a cellular system containing neutrophils stimulated by phorbol myristate acetate, indicating a complex bimodal structure of CL processes corresponding to different stages of the inhibition. The complex structure of these processes was described by a superposition of two logistic-exponential (LE) models, characterizing these processes as bimodal ones. To determine the mechanistic foundation of the LE model-described processes, a generalized form of the second-order dynamic system of CL reactions, the solution to which corresponds to the LE model, was constructed.
View Article and Find Full Text PDFA human leukaemia cell line--HL-60--can be differentiated into neutrophils or macrophages and both differentiation processes are accompanied by changes of the lipid composition. Various methods were described for the extraction of lipids from cellular systems, but only two of them were applied to the HL-60 cell line so far. In this study we compared five selected extraction methods for the lipid extraction from HL-60 cells with regard to their qualitative analysis by matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry (MALDI-TOF MS): chloroform/methanol at volume ratios 2:1 and 1:2, isopropanol/ chloroform, isopropanol/hexane and butanol.
View Article and Find Full Text PDFA small library of peptide analogues of the chemotactic tripeptide For-Met-Leu-Phe-NH2 modified by substitution of Leu at position 2 by three different fluorinated amino acids varying in content of fluorine, length of the fluorinated side chain, and alkylation degree at the alpha-carbon atom was synthesized. The influence of the fluorine substitution on the biological activity was investigated by measuring the oxidative activity of neutrophils using a luminol-dependent chemiluminescence assay.
View Article and Find Full Text PDFBackground/aim: Polymorphonuclear leukocytes (neutrophils) release a variety of toxic agents--proteins and reactive oxygen species (ROS)--that are used to inactivate foreign microorganisms in the non-specific immune response. This study was undertaken to compare intracellular signalling pathways that lead to the ROS production as well as degranulation of azurophilic granules of human fMet-Leu-Phe/cytochalasin B stimulated neutrophils.
Methods: Luminol-amplified chemiluminescence was used for monitoring the oxidative activity of human neutrophils in the presence of various inhibitors.