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  • 1
    Electronic Resource
    Electronic Resource
    s.l. : American Chemical Society
    Journal of the American Chemical Society 54 (1932), S. 1644-1647 
    ISSN: 1520-5126
    Source: ACS Legacy Archives
    Topics: Chemistry and Pharmacology
    Type of Medium: Electronic Resource
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  • 2
    Electronic Resource
    Electronic Resource
    Springer
    Inflammation research 41 (1994), S. C56 
    ISSN: 1420-908X
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Modulation of protein kinase C (PKC) activity in basophils (B) can influence IgE-mediated histamine release (HR). The present study investigated the effects of chelerythrine, which inhibits isolated PKC (IC50 0.7 μM), on different activation pathways in B. FcεRI-mediated HR was strongly inhibited by chelerythrine (86.5±5.4%, 5 μM,n=11). TPA-induced mediator release was also suppressed: 77.1±8.5% inhibition (7.5 μM). HR due to non-immunological stimuli (A23187, FMLP) was strongly inhibited by chelerythrine. Previously, other selective PKC-inhibitors have been shown to potentiate IgE-mediated HR from B suggesting a negative modulatory role of PKC, whereas non-specific inhibitors such as staurosporine inhibited HR. Chelerythrine might therefore be less selective for PKC. This may be indicated by the fact that chelerythrine inhibits PKC at its catalytic domain, which is homologous with other protein kinases.
    Type of Medium: Electronic Resource
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  • 3
    ISSN: 1420-908X
    Keywords: Key words: Basophil — Purification — Elutriation — Immunomagnetic bead — Histamine
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract. Objective and Design: We report a method for basophil purification from buffy coats, which avoids positive selection of the cells and gives rise to good purity, yield and functional integrity of the cells.¶Subjects: Buffy coat blood (concentrated leukocyte fraction derived from 450 ml venipuncture donations) obtained from healthy blood donors (n = 51).¶Methods: Basophils were enriched by a three-step process starting with Ficoll density centrifugation (1.6 ± 0.1% basophil purity) followed by counter current centrifugal elutriation (17.7 ± 1.4% basophil purity). The final stage involved negative selection using Dynal immunomagnetic beads directed against CD2, CD14, CD16 and CD19 positive cell contaminants. Functional integrity of which was assessed by comparing the anti-IgE or calcium ionophore A23187 induced histamine release from basophils obtained from each enrichment step. Furthermore, basophil morphology was investigated using light and electron microscopy.¶Results: The final mean basophil purity of 67.3 ± 1.4% with a yield of 3.5 ± 0.5 × 106 basophils and a recovery of 21.8 ± 2.4% was achieved. Net histamine release from basophils stimulated with optimal concentrations of anti-human IgE was 39.1 ± 6.5% after Ficoll centrifugation, 41.6 ± 7.7% following elutriation and 35.7 ± 6.8% from the final purified fraction. Additionally, basophils enriched with our method showed intact morphology by electron microscopy and were functionally active to non-immunological stimulation.¶Conclusions: These results compare favourably with previous studies, which have often required the use of positive selection via the Fc〈epsilon〉RI receptor, which may result in cell degranulation, or cell sorting, which cannot be applied to large cell numbers. Our method provides a reproducible technique for basophil enrichment when large numbers of functionally intact basophils are required.
    Type of Medium: Electronic Resource
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  • 4
    ISSN: 1420-908X
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Arachidonic acid metabolites generated from activated basophils and/or mast cells mediate different types of cutaneous inflammatory reactions. To clarify the mechanisms of leukotriene C4 (LTC4) production from human basophils, cells were purified from the peripheral blood by negative selection with immunobeads. The protein kinase C (PKC) activators 12-O-tetradecanoyl-phorbol-13-acetate, phorbol-12,13-dibutyrate and phorbol-12,13-didecanoate did not induce a significant LTC4 generation from human basophilsin vitro, indicating that phorbol-ester-sensitive PKC isozymes are not involved in the mechanisms of arachidonic acid metabolism in these cells. However, selective PKC inhibitors (Ro 31-7549, ilmofosine, GF109203X, and calphostin C) potentiated the IgE-mediated LTC4 production in a dose-dependent fashion. We therefore suggest that PKC isozymes which are influenced by these inhibitors modulate the degree of LTC4 release after stimulation with anti-IgE antibodies.
    Type of Medium: Electronic Resource
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  • 5
    ISSN: 1420-908X
    Keywords: Key words: Leukocyte — Mast cell — Mediator — Ambroxol — N-acetylcysteine
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract. Objectives and Design: The effects of the mucolytic agents ambroxol and N-acetylcystein (NAC) were studied on the release of histamine, leukotrienes, cytokines and superoxide anions from a variety of cells involved in the pathogenesis of allergic inflammation.¶Subjects: Mast cells were isolated from human adenoids and skin (n = 5–6). Basophils, monocytes and granulocytes were obtained from Buffy-coat blood obtained from healthy blood donors (n = 4–7) and enriched by density centrifugation.¶Treatment and Methods: Ambroxol or NAC were added to the cells for different periods before stimulation with various immunological and non-immunological secretagogues. Histamine release from mast cells, basophils and monocytes was assayed either by radioimmunoassay or spectrofluorometrically. LTC4 (basophils), LTB4 (neutrophil/eosinophil granulocytes or monocytes), IL-4 and IL-13 (basophils) were measured by ELISA.¶Results: Ambroxol inhibited histamine release by more than 50% from human adenoidal mast cells (1000 μM ambroxol) and skin mast cells (100 μM ambroxol) stimulated by Con A and compound 48/80, respectively. Ambroxol (100 μM) strikingly inhibited anti-IgE induced release of both histamine, LTC4, IL-4 and IL-13 from basophils and reduced both histamine and LTB4 release induced by C5a or Zymosan in monocytes. The drug also reduced LTB4 and superoxide anion production in granulocytes stimulated by zymosan or fMLP. In all cell types studied, ambroxol was more efficacious following a short preincubation (5–15 min) of the drug with the cells before stimulation. In contrast, NAC produced no clear effects on any of the different cell types studied, regardless of the preincubation period, the concentration or the stimulus employed.¶Conclusions: Unlike NAC, ambroxol is able to not only inhibit acute mediator release from mast cells and leukocytes but also reduce immunomodulatory cytokine generation from basophils and may have beneficial effects in the treatment of allergic respiratory diseases.
    Type of Medium: Electronic Resource
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  • 6
    ISSN: 1420-908X
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Type of Medium: Electronic Resource
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  • 7
    ISSN: 1420-908X
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Type of Medium: Electronic Resource
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  • 8
    ISSN: 1420-908X
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Type of Medium: Electronic Resource
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  • 9
    Electronic Resource
    Electronic Resource
    Springer
    Inflammation research 49 (2000), S. 17-18 
    ISSN: 1420-908X
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Type of Medium: Electronic Resource
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  • 10
    Electronic Resource
    Electronic Resource
    Springer
    Inflammation research 41 (1994), S. C28 
    ISSN: 1420-908X
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract The present study was performed to investigate the histamine-releasing activity of non-immunological stimuli on cultured mast cell lines in comparison to isolated skin mast cells and basophils as human therapeutic target cells. The ionophore A23187 induced a dose dependent histamine release from all cell populations (enzymatically isolated human skin mast cells, human peripheral basophils and rat basophilic leukemia cells, RBL-1 and RBL-2H3). The lectin concanavalin A and the tripeptide formyl-methionyl-leucyl-phenylalanine activated only basophils, while the neural mediator substance P and compound 48/80 were active only in experiments with skin mast cells. Activators of protein kinase C (different phorbol esters and the non-phorbol mezerein) induced direct histamine release only from basophils. The data provide further evidence for heterogencity of mast cells and indicate different signal transduction mechanisms following non-immunological activation.
    Type of Medium: Electronic Resource
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