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  • 1
    ISSN: 1550-7408
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: The production of free radicals in human neutrophils was studied in both Pneumocystis carinii derived from cultures of L2 rat lung epithelial-like cells and Pneumocystis carinii purified from human lung. Using the cytochrome C technique, which selectively measured extracellular superoxide generation, hardly any free radical production was observed after stimulation with cultured rat-derived P. carinii. A chemiluminescence technique, which separately measured intra- and extracellular free radical production, was subsequently employed to differentiate the free radical generation. It was established that 1) P. carinii stimulated intra- but not extracellular free radical production in human neutrophils. 2) opsonized cultured rat-derived P. carinii stimulated human neutrophils to a strong intra-cellular response of superoxide production, and 3) opsonized P. carinii. purified from human lung also stimulated human neutrophils to produce intracellular free radicals.
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  • 2
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 44 (1987), S. 0 
    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: The phagocytic capacity of Acanthamoeba castellanii was compared with that of human polymorphonuclear leukocytes (PMNL) using non-fimbriated and fimbriated Escherichia coli with different physicochemical properties as phagocytic preys. The E. coli strain PN7 with mannose-sensitive (MS-)fimbriae adhered both to Acanthamoeba and to PMNL. The adherence to leukocytes but not to the amoebas, was completely inhibited by mannose. E. coli strain ABU2 with MS-fimbriae adhered in a mannose-sensitive manner to the PMNL. To Acanthamoeba the adhesion was only partly inhibited by mannose. The ability to ingest the adhered bacteria differed between the two phagocytes. Thus the PMNL ingested hydrophobic, weakly negative charged rough-like bacteria while hydrophilic smooth-like bacteria resisted ingestion. In contrast, the amoeba ingested both strains irrespective of the physicochemical surface properties.
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  • 3
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 22 (1984), S. 0 
    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Abstract The association of Chlamydia trachomatis serotypes E and L1 with human polymorphonuclear leukocytes (PMNL) and the ability of these chlamydiae to induce a chemiluminescence response in PMNL were recorded. C. trachomatis unequivocally activated a respiratory burst in PMNL. Despite similar association with PMNL, serotype E activated the PMNL significantly more than serotype L1. Type-specific, but not group-specific antisera increased the chemiluminescence but not association of chlamydiae with PMNL. These results indicate that recording of chemiluminescence may be of value to estimate potential antichlamydial activity of PMNL and that type-specific antibodies have opsonizing effect exerted subsequent to the attachment phase of chlamydiae to PMNL.
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  • 4
    ISSN: 0884-3996
    Keywords: Chemiluminescence ; leukocytes ; N-ethylmaleimide ; NADPH-oxidase inactivation ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Sustained generation of reactive oxygen metabolites following respiratory burst activation in neutrophils is a result of continued replenishment of a pool of active NADPH-oxidase. The sulphydryl-modifying reagent N-ethylmaleimide (NEM) has been shown to be without effect on the turnover of activated NADPH-oxidase but to inhibit the replenishment of active oxidase molecules (Akard et al., 1988). NEM was thus used to determine the rate of deactivation of extracellularly and intracellularly generated chemiluminescence in human neutrophils. We have shown that deactivation is more rapid when activation leads to a release of oxygen metabolites (extracellular chemiluminescence) than when the metabolites are generated intracellularly. The results indicate that the rate of deactivation of NADPH-oxidase is higher when the oxidase system is localized on the plasma membrane than when it is localized on the phagosomal membrane.
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  • 5
    Electronic Resource
    Electronic Resource
    New York : Wiley-Blackwell
    Journal of Bioluminescence and Chemiluminescence 10 (1995), S. 353-359 
    ISSN: 0884-3996
    Keywords: chemiluminescence ; neutrophil ; NADPH-oxidase ; luminol ; isoluminol ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: In search for a luminol with very high output of light, 20 different luminol samples were tested for their ability to enhance the chemiluminescence reaction in phorbol myristate acetate activated human neutrophils. We found that the majority of luminols tested (17 samples) gave almost the same light output from neutrophils, and that the major part of the activity was from an intracellular origin. Owing to the fact that three isoluminol samples were unable to monitor respiratory burst activity taking place intracellularly, a very low level of chemiluminescence was obtained with these samples. Their light output was, however, greatly increased when horseradish peroxidase or myeloperoxidase was added, showing that the light-generating reaction with isoluminol as well as with luminol is peroxidase-dependent.The fact that isoluminol could also use myeloperoxidase as amplifying peroxidase, suggests that the lack of measurable intracellular activity in the presence of isoluminol is somehow related to a limited or restricted diffusion of the molecule to intracellular sites. The isoluminol system constitutes a sensitive system for measuring release of oxygen metabolites from phagocytic cells.
    Additional Material: 4 Ill.
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  • 6
    ISSN: 0884-3996
    Keywords: Chemiluminescence ; luminol ; scavengers ; human neutrophils ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: When polymorphonuclear leukocytes (PMNL) interact with the soluble stimulus formylmethionyl-leucyl-phenylalanine (FMLP), the cells increase their production of oxidative metabolites. This increased production can be measured as lumino-amplified light emission or chemiluminescence (CL). In the present report, experimental systems which allow a quantitation of extracellularly and intracellularly generated metabolites have been used, and the effect of mannitol, benzoate, taurine, indomethacin and nordihydroguaiaretic acid has been investigated. The presence of the hypochlorous acid scavenger taurine had no effect on the intracellular response, whereas the extracellular response was reduced with around 50%. The hydroxyl radical scavenger mannitol had only minor effects on the response, whereas benzoate, another hydroxyl radical scavenger, reduced the extracellular response with around 50% and the intracellular response with more than 90%. Indomethacin, an inhibitor of arachidonic acid metabolism, did not influence the response, whereas NDGA, also an inhibitor of the arachidonic acid metabolism, totally abolished both the extracellular and the intracellular response. The use of scavengers/inhibitors as a means of determining the mechanisms of light emission, and the origin of chemiluminescence produced by neutrophils stimulated by FMLP is discussed.
    Additional Material: 1 Ill.
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  • 7
    Electronic Resource
    Electronic Resource
    New York : Wiley-Blackwell
    Journal of Bioluminescence and Chemiluminescence 2 (1988), S. 25-33 
    ISSN: 0884-3996
    Keywords: Chemiluminescence ; leukocytes ; luminol ; peroxidase ; cytoplasts ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: When neutrophil cytoplasts (granule-free vesicles of cytoplasm enclosed by plasmalemma) were exposed to the chemotactic peptide formylmethionyl-leucyl-phenylalanine, no luminol-dependent chemiluminescence was detected, despite a pronounced production of superoxide anions and hydrogen peroxide. Addition of purified myeloperoxidase (MPO) or human serum albumin (HSA) to the cytoplasts before the stimulus resulted in a chemiluminescence response. In contrast to the bimodal response obtained from normal PMNL, only a single peak of chemiluminescence was obtained from the cytoplasts responding to the peptide. The time-course of the response obtained in the presence of albumin was more prolonged than the response obtained in the presence of MPO. Furthermore, the involvement of different oxidative metabolites in the MPO and the HSA systems, respectively, was demonstrated by the accumulated chemiluminescence effect obtained when MPO, but not HSA, was introduced in the measuring system after FMLP addition.From these results it can be concluded that there are at least two different light-generating mechanisms in FMLP-induced luminol-dependent chemiluminescence of neutrophil cytoplasts. One of these is dependent on myeloperoxidase and possibly related to the myeloperoxidase-hydrogen peroxide reaction, whereas the other one is hydrogen peroxide independent.
    Additional Material: 7 Ill.
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  • 8
    Electronic Resource
    Electronic Resource
    New York : Wiley-Blackwell
    Journal of Bioluminescence and Chemiluminescence 4 (1989), S. 263-266 
    ISSN: 0884-3996
    Keywords: Leukocytes ; respiratory burst ; granule ; calcium ionophore ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: The granulocyte luminol-dependent chemiluminescence (CL) reaction is linked to the enzyme myeloperoxidase reacting with products of the respiratory burst activation. The results presented in this paper, show that the light generated in granulocytes originate both from intracellular and extracellular reactions; however, depending on the stimulus used the one or the other will dominate the activity measured. Furthermore, lysosomal fusion is proposed to be required for the intracellular CL reaction.
    Additional Material: 2 Ill.
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  • 9
    Electronic Resource
    Electronic Resource
    New York : Wiley-Blackwell
    Journal of Bioluminescence and Chemiluminescence 5 (1990), S. 37-41 
    ISSN: 0884-3996
    Keywords: Monocytes ; chemiluminescence ; oxidative metabolites ; mye operoxidase ; luminol ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: The characteristics of the luminol-amplified chemiluminescence (CL) induced in mononuclear phagocytes interacting with PMA, FMLP or ionomycin were determined. Azide reduced the CL activity by more than 80%, while superoxide dismutase and catalase had minor effect on the monocyte CL response. The sensitivity of the monocyte CL response, to the addition of extra peroxidase differed depending on the stimulus used. Furthermore, no direct correlation was obtained between the CL response and superoxide anion or hydrogen peroxide production. In comparison with the response in granulocytes, minor quantitative differences were observed. The mechanism for the light-generating reaction, seems to be the same in both cell types.
    Additional Material: 3 Ill.
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  • 10
    ISSN: 0263-6484
    Keywords: Ligand binding ; respiratory burst ; neutrophils ; cytoplasts ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: When polymorphonuclear leukocytes (neutrophils) and soluble or particulate matter interact, the cells produce superoxide anions (O2-) and hydrogen peroxide (H2O2). The chemotactic peptide formylmethionyl-leucylphenylalanine (FMLP) induced a very weak response in normal neutrophils. The cellular response was changed, however, as a result of in vitro aging of the cells, i.e. the magnitude of the response was increased following storage of the cells at 22°C for up to 120 min, in the absence of any stimulus, and before the addition of the peptide. When phorbol myristate acetate was used as a stimulus, there was a pronounced production of O2- and H2O2, but no change in magnitude as a result of in vitro aging. When neutrophil cytoplasts (granule-free vesicles of cytoplasm enclosed by plasmalemma) were exposed to the peptide FMLP of PMA, the vesicles produced both O2- and H2O2. There was, however, no increase in oxidative metabolite production in cytoplasts as a result of in vitro aging when either FMLP or PMA was used as a stimulus. The results thus indidate that mere incubation at room temperature primed the cells to increase their production of oxidative metabolites as a result of spontaneous exposure of hidden receptors. The fact that no such effects were observed with cytoplasts indicates that spontaneous receptor recruitment is a granule-dependent process.
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