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Protocols to Measure Oxidative Stress and DNA Damage in Asthma

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Asthma

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2506))

Abstract

Asthma is associated with oxidative stress and oxidative damage of biomolecules, including DNA. Here, we describe the protocols to quantify reactive oxygen species (ROS) and oxidative stress markers in a mouse model of allergic airway inflammation. We also provide detailed methods to measure DNA damage by long-run real-time PCR for DNA-damage quantification (LORD-Q) assay and gene-specific DNA damage analyses by long amplicon (LA)-qPCR. Additionally, we describe methods to quantify oxidized DNA base lesions in lung genomic DNA by mass spectrometry, and to measure enzymatic activity of 8-oxoguanine DNA glycosylase (OGG1). Using these methods, the levels of oxidative stress and DNA damage in allergic inflammation and asthma can be elucidated.

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Acknowledgments

We thank Dr. Pawel Jaruga, Dr. Erdem Coskun, and Dr. Miral Dizdaroglu (Biomolecular Measurement Division National Institute of Standards and Technology, Gaithersburg, Maryland, USA) for their scientific input and for critical writing and editing of the manuscript. We also thank Dr. Nisha Tapryal (Department of Internal Medicine, Division of Pulmonary, Critical Care and Sleep Medicine, University of Texas Medical Branch, Galveston, Texas, USA) for her scientific input and for critical writing and editing of the manuscript.

This research was supported in parts by the National Heart, Lung, and Blood Institute (grant nos. 5R01HL145477-02 and 3R01HL145477-01S1) and the Department of Defense (grant no. PR171425 W81XWH-18-1-0743).

Disclosure of Potential Conflict of Interest

The authors declare that they have no relevant conflicts of interest.

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Authors and Affiliations

  1. Department of Medicine, Immunology Allergy and Rheumatology, Baylor College of Medicine, Houston, TX, USA

    Koa Hosoki & Sanjiv Sur

  2. Department of Internal Medicine, Division of Pulmonary, Critical Care and Sleep Medicine, University of Texas Medical Branch, Galveston, TX, USA

    Anirban Chakraborty & Tapas K. Hazra

Authors
  1. Koa Hosoki
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  2. Anirban Chakraborty
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  3. Tapas K. Hazra
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  4. Sanjiv Sur
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Corresponding author

Correspondence to Sanjiv Sur .

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Editors and Affiliations

  1. Division of Allergy and Clinical Immunology, Department of Medicine, National Jewish Health, Denver, CO, USA

    Magdalena M. Gorska

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Hosoki, K., Chakraborty, A., Hazra, T.K., Sur, S. (2022). Protocols to Measure Oxidative Stress and DNA Damage in Asthma. In: Gorska, M.M. (eds) Asthma. Methods in Molecular Biology, vol 2506. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2364-0_22

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  • DOI: https://doi.org/10.1007/978-1-0716-2364-0_22

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-2363-3

  • Online ISBN: 978-1-0716-2364-0

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