Journal article
Hydrogen peroxide release from human eosinophils on fibronectin: scopoletin enhances eosinophil activation
Free radical biology & medicine, v 28(11), pp 1652-1660
2000
PMID: 10938462
Featured in Collection : UN Sustainable Development Goals @ Drexel
Abstract
We have examined the release of H
2O
2 from PAF or TNFα-stimulated human eosinophils on fibronectin (FN)-coated polystyrene plates. H
2O
2 release was measured by the standard scopoletin-horseradish peroxidase (SCOP-HRP) method and compared with that measured by a new microplate fluorescent assay for H
2O
2 using a novel HRP substrate A6550. We observed that the SCOP-HRP method gave a 25-fold higher estimate of H
2O
2 release from eosinophils than did the A6550-HRP method. Microscopic examination of PAF or TNFα-stimulated eosinophils in buffer alone or A6550-HRP reaction mixture showed that the cells remained generally round, while eosinophils in SCOP-HRP reaction mixture were spread on the fibronectin-coated surface. Measurement of the cellular ATP content after PAF-stimulation showed that only eosinophils activated in SCOP-HRP had a 50% fall in ATP content. This supported our conclusion that measurement of H
2O
2 release from eosinophils in SCOP-HRP reaction mixture is problematic since the SCOP-HRP system activates eosinophils. However, we also found that A6550-HRP, when present throughout the incubation, resulted in a lower estimate of H
2O
2 release than expected. The method used to detect eosinophil H
2O
2 release greatly influences the absolute amount of H
2O
2 detected.
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Details
- Title
- Hydrogen peroxide release from human eosinophils on fibronectin: scopoletin enhances eosinophil activation
- Creators
- Donald G Raible - MCP Hahnemann School of Medicine, Divisions of Pulmonary and Critical Care Medicine, Philadelphia, PA, USAJoy G Mohanty - MCP Hahnemann School of Medicine, Divisions of Pulmonary and Critical Care Medicine, Philadelphia, PA, USAJonathan S Jaffe - Hahnemann University HospitalHector J Stella - MCP Hahnemann School of Medicine, Divisions of Pulmonary and Critical Care Medicine, Philadelphia, PA, USABoyd E Sprenkle - MCP Hahnemann School of Medicine, Divisions of Pulmonary and Critical Care Medicine, Philadelphia, PA, USAMark C Glaum - MCP Hahnemann School of Medicine, Divisions of Pulmonary and Critical Care Medicine, Philadelphia, PA, USAEdward S Schulman - MCP Hahnemann School of Medicine, Divisions of Pulmonary and Critical Care Medicine, Philadelphia, PA, USA
- Publication Details
- Free radical biology & medicine, v 28(11), pp 1652-1660
- Publisher
- Elsevier
- Resource Type
- Journal article
- Language
- English
- Academic Unit
- Pulmonary, Critical Care, and Sleep (Medicine)
- Web of Science ID
- WOS:000088750400010
- Scopus ID
- 2-s2.0-0034209666
- Other Identifier
- 991019168018904721
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InCites Highlights
Data related to this publication, from InCites Benchmarking & Analytics tool:
- Web of Science research areas
- Biochemistry & Molecular Biology
- Endocrinology & Metabolism