Abstract
Background: Oxidative stress is an imbalance between reactive species and antioxidants production, leading to protein, DNA and lipid damage. The aim of this study was to evaluate the oxidative stress levels in buccal cells of ten non-shisha and heavy shisha smokers using MAWI iSWAB protein tubes. Materials and findings: The presence of S-glutathionylation, NOX2 and iNOS was analyzed in buccal cells of ten non-smokers and ten heavy shisha smokers using MAWI i-SWAB protein tubes, knowing that they were able to detect previously several protein oxidation biomarkers (S-nitrosylation and nitrotyrosine). Proteins concentrations were assessed by Bradford assay then western blot method was used to analyze the presence of the different proteins of interest. Conclusion: Results confirmed the findings about the ability of these tubes to detect S-nitrosylation, nitrotyrosine, as well as S-glutathionylation. Results showed also the presence of immature NOX2 in human buccal cells equally. Levels of iNOS were the same in non-shisha and heavy shisha smokerss.
Highlights
Oxidative stress is the disruption of the redox signaling due to an overproduction of reactive species, or an impairment of the antioxidant defense system resulting in the pathogenesis of many diseases such as cancer, cardiovascular, pulmonary and neurodegenerative diseases as well as aging processes [1,2]
The presence of the different protein oxidation biomarkers in nonsmokers and smokers samples were detected by western blots
GAPDH is one of the many housekeeping proteins. It was used as an internal control for Western blot analysis to normalize different proteins expressions [19] in order to compare the levels of protein oxidation biomarkers in non- smokers and heavy shisha smokers
Summary
Oxidative stress is the disruption of the redox signaling due to an overproduction of reactive species, or an impairment of the antioxidant defense system resulting in the pathogenesis of many diseases such as cancer, cardiovascular, pulmonary and neurodegenerative diseases as well as aging processes [1,2]. It’s a membrane - embedded glycoprotein that needs p22phox protein for its stabilization These proteins are inactive until they interact with other cytosolic components: p47phox, p67phox, p40phox and Rac. The activation consists of the phosphorylation of p47phox that enables the binding to p22phox and membrane lipids. Rac is maintained in an inactive cytoplasmic complex with guanine nucleotide dissociation inhibitor (GDI) Upon activation, they dissociate and Rac is translocated to the membrane, following a GDP to GTP exchange promoted by a guanine nucleotide exchange factors (GEFs) [4]. Materials and findings: The presence of S-glutathionylation, NOX2 and iNOS was analyzed in buccal cells of ten non-smokers and ten heavy shisha smokers using MAWI i-SWAB protein tubes, knowing that they were able to detect previously several protein oxidation biomarkers (S-nitrosylation and nitrotyrosine). Proteins concentrations were assessed by Bradford assay western blot method was used to analyze the presence of the different proteins of interest
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